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  • Cartilage
  • Springer  (110)
  • American Institute of Physics
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  • 1
    ISSN: 1435-604X
    Keywords: Cartilage ; Laser-induced reshaping ; Nd:YAG laser ; Plastic surgery ; Reconstructive surgery ; Stress relaxation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Physics , Technology
    Notes: Abstract Cartilage undergoes characteristic mechanical stress relaxation following laser irradiation below the ablation threshold. Porcine auricular cartilage (1–2 mm thickness) was irradiated with a Nd:YAG laser (λ=1.32 μm) at two power levels (W/cm2). Surface temperature (S c (t) (°C)) (monitored using a single element HgCdTe infrared detector, 10-14 μm spectral range), and integrated back scattered light intensityI(t) were measured during laser irradiation. A HeNe laser beam (λ=632.8 nm) was incident on the back surface of the cartilage specimen and fractional integrated backscattered light intensity was measured using an integrating sphere anda silicon photodiode. Laser irradiation (5.83 W/cm2, 50 Hz pulse repetition rate (PRR)) continued until surface temperature reached approximately 70°C, during which cartilage mechanical stress relaxation was observed. Integrated back scattered light intensity reached a plateau at about 70°C). At higher laser power (39.45 W/cm2, 50 Hz PRR), a feedback-controlled cryogen spray was used to maintain surface temperature below 50°C. A similar plateau response was noted in integrated backscattered light intensity. This signal may be used to optimise the process of stress relaxation in laser cartilage reshaping. Several clinical applications involving reconstructive surgery are proposed.
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  • 2
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    Springer
    Cellular and molecular life sciences 49 (1993), S. 393-402 
    ISSN: 1420-9071
    Keywords: Cartilage ; proteoglycan ; link protein ; hyaluronic acid ; extracellular matrix
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Aggregates of chondroitin-keratan sulfate proteoglycan (aggrecan) and hyaluronic acid (hyaluronan) are the major space-filling components of cartilage. A glycoprotein, link protein (LP; 40–48 kDa) stabilizes the aggregate by binding to both hyaluronic acid and aggrecan. In the absence of LP, aggregates are smaller (as estimated by rotary shadowing of electron micrographs) and less stable (they dissociate at pH 5) than they are in the presence of LP. The proteoglycan aggregate, including LP, is dissociated in the presence of chaotropes such as 4 M guanidine hydrochloride. On removal of the chaotrope, the complex will reassociate. This forms the basis of the isolation of LP from cartilage and has been described in detail elsewhere. Tryptic digestion of the proteoglycan aggregates results in a high molecular weight product that consists of hyaluronic acid to which is bound LP and the N-terminal globular domain of aggrecan (hyaluronic acid binding region; HABR) in a 1∶1 stoichiometry. The amino acid sequences of LP and HABR are surprisingly similar. The amino acid sequence can be divided into three domains; an N-terminal domain that falls into the immunoglobulin super-family and two C-terminal domains that are similar to each other. The DNA structure echoes this similarity, in that the major domains are reflected in three separate exons in both LP and HABR. The two C-terminal domains are largely responsible for the association with HA and are related to two recently described hyaluronate-binding proteins, CD44 and TSG-6. A variety of approaches, including analysis of the forms of LP found in vivo, rotary shadowing and analysis of the sequence in the immunoglobulin-like domain, have shed considerable light on the structure-function relationships of LP. This review describes the structure and function of LP in detail, focusing on what can be inferred from the similarity of LP, HABR and related molecules such as immunoglobulins and lymphocyte HA-receptors.
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  • 3
    ISSN: 1432-0827
    Keywords: Vitamin D ; Vitamin D deficiency ; Cartilage ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary To test the importance of 24-hydroxylation of vitamin D3 on bone mineralization, rat pups born to vitamin D-deficient females were given either 25-hydroxyvitamin D3 or 24,24-difluoro-25-hydroxyvitamin D3 for 16 days beginning at the time of weaning. Following such treatment analysis of blood samples revealed no detectable 24R,25-(OH)2D3 and 1,25-(OH)2D3 in the rats given the difluoro compound while revealing the expected 24,24-difluoro-25-hydroxyvitamin D3 and 24,24-difluoro-1,25-dihydroxyvitamin D3. The rats given 25-hydroxyvitamin D3 had the expected levels of 25-hydroxyvitamin D3, 24,25-dihydroxyvitamin D3, and 1,25-dihydroxyvitamin D3. Following sacrifice at day 17, postweaning bone mineralization and modeling were studied in long bones using histological methods. Bones taken from vitamin D-deficient rats at the beginning and end of the experimental period had lesions typical of rickets. These included wide growth plates, excessive amounts of osteoid, and metaphyseal fibrosis. Following treatment with either 25-hydroxyvitamin D3 or 24,24-difluoro-25-hydroxyvitamin D3, bone mineralization returned to normal. Growth plate widths and the amount of osteoid on bone surfaces were both substantially reduced and to a similar degree in both treatment groups. Normal cartilage core formation and trabecularization of the metaphyseal primary spongiosa were also restored to a similar degree in both groups. In effect, no difference was observed in any bone parameter studied between the 25-hydroxyvitamin D3- and the 24,24-difluoro-25-hydroxyvitamin D3-treated animals. These results provide strong evidence that 24-hydroxylation of the vitamin D molecule plays little or no role in the modeling and mineralization of bone.
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  • 4
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    Calcified tissue international 10 (1972), S. 31-37 
    ISSN: 1432-0827
    Keywords: Molecular weight ; Chondroitin sulphate ; Cartilage ; Aging ; Osteoarthritis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Le poids moléculaire du sulfate de chondroitine de cartilage articulaire normal et de l'épiphyse inférieure fémorale de cas d'autopsie, d'âges variés, et de cartilage ostéo-arthritique a été déterminé par chromatographie de gel sur Sephadex G-200. Les échantillons de polysaccharide se situent dans les mêmes variations de poids moléculaire, mais diffèrent par la distribution de ce dernier. Le poids moléculaire moyen de sulfate de chondroitine du cartilage articulaire normal est plus élevé chez le nouveauné et les sujets jeunes que chez l'adulte ou le vieillard. Le sulfate de chondroitine de cartilage ostéo-arthritique a un poids moléculaire inférieur que celui du cartilage macroscopiquement normal. La signification des variations dans la longueur des chaines polysaccharidiques est discutée.
    Abstract: Zusammenfassung Die Molekulargewichts-Verteilung von Chondroitinsulfat im normalen Gelenkknorpel der unteren Femora-Epiphyse (menschliche Autopsien von verschiedenen Altersgruppen) und im osteo-arthritischen Knorpel wurde mittels Gel-Chromatographie auf Sephadex G-200 bestimmt. Die Polysaccharidproben fielen in denselben Molekulargewichts-Bereich, aber die Molekulargewichts-Verteilung war unterschiedlich. Das durchschnittliche Molekulargewicht des Chondroitinsulfates von normalem Gelenkknorpel war bei Neugeborenen und jungen Individuen höher als bei erwachsenen oder älteren Personen. Chondroitinsulfat von osteoarthritischem Knorpel hatte das niedrigere Molekulargewicht als dasjenige von makroskopisch normalem Knorpel. Die beobachteten Variationen in den Längen der Polysaccharidketten werden diskutiert.
    Notes: Abstract The molecular weight distributions of chondroitin sulphate from normal articular cartilage of the lower femoral epiphysis from human autopsy cases of different ages and from osteoarthritic cartilage have been determined by gel chromatography on Sephadex G-200. The polysaccharide samples fell within the same molecular weight range but differed with respect to molecular weight distribution. The average molecular weight of chondroitin sulphate from normal articular cartilage was higher in newborn and young individuals than in adults or the aged. Chondroitin sulphate from osteoarthritic cartilage had a lower molecular weight than that from macroscopically normal cartilage. The observed variations in polysaccharide chain length are discussed.
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  • 5
    ISSN: 1432-0827
    Keywords: Pyrophosphate ; Bone ; Cartilage ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les concentrations de pyrophosphate inorganiques (PPi), d'orthophosphate et de calcium ont été déterminées dans le cartilage au repos, en prolifération, hypertrophique et calcifié au niveau d'épiphyses foetales de veaux et aussi dans l'os spongieux, périosté et compact. Dans les échantillons de cartilage, le contenu en PPi augmente progressivement dans l'ordre énoncé ci-dessus, de 8.59 μg P/g en poids sec de cartilage au repos à 236 μg P/g en poids sec dans le cartilage calcifié. Cependant, le rapport PPi sur orthophosphate suit une relation inverse: il est plus élevé dans la zone de repos et diminue considérablement lorsque le tissu se calcifie. Le rôle possible du PPi, en inhibant la précipitation de phosphate de calcium amorphe (ACP) et en ralentissant la transformation d'ACP en hydroxyleapatite (HA) dans les tissus calcifiés, est envisagé par rapport à d'autres facteurs, tels que le collagène, le magnésium, les phospholipides et les protéines-polysaccharides, qui peuvent aussi intervenirin vivo. Pour l'instant, aucun facteur isolé ne peut être considéré comme un régulateur physiologique certain.
    Abstract: Zusammenfassung Der Gehalt an anorganischem Pyrophosphat (PPi), Orthophosphat und Calcium wurde in ruhendem, proliferierendem, hypertrophischem und verkalktem Knorpel von foetalen Kalbsepiphysen und auch in spongiösem, periostalem und kompaktem Knochen gemessen. Bei den Knorpelproben nahm der Gehalt an PPi progressiv in der oben angegebenen Reihenfolge zu: von 8,59 μg P/g Trockengewicht in ruhendem Knorpel bis zu 236 μg P/g Trockengewicht in verkalktem Knorpel. Das PPi/Orthophosphat-Verhältnis hingegen verlief in umgekehrter Richtung; es war am höchsten in der ruhenden Zone und nahm dramatisch ab, wenn das Gewebe verkalkte. Die mögliche Rolle von PPi bei der Hemmung der Ausfällung von amorphem Calciumphosphat (ACP) und bei der Verlangsamung der Umwandlung von ACP in Hydroxyapatit (HA) in verkalkenden Geweben wird in bezug auf folgende anderen Faktoren, welche den Vorgang in vivo ebenfalls beieinflussen könnten, diskutiert: Collagen, Magnesium, Phospholipide und Proteinpolysaccharide. Bis jetzt war es nicht möglich, einen einzelnen Faktor als sicheren physiologischen Regulator zu identifizieren.
    Notes: Abstract The amounts of inorganic pyrophosphate (PPi), orthophosphate and calcium have been measured in resting, proliferating, hypertrophic and calcified cartilage from foetal calf epiphyses and also in cancellous, periosteal and compact bone. In the cartilage samples, the content of PPi increased progressively in the order named above, from values of 8.59 μg P/g dry weight in resting cartilage to 236 μg P/g dry weight in calcified cartilage. However, the ratio of PPi to orthophosphate followed the reverse relationship and was highest in the resting zone and fell dramatically as the tissue calcified. The possible role of PPi in inhibiting the precipitation of amorphous calcium phosphate (ACP) and in the slowing the transformation of ACP to hydroxyapatite (HA) in calcifying tissues is discussed in relation to other factors, such as collagen, magnesium, phospholipids and proteinpolysaccharides, which might also influence the processin vivo. At present, no single factor can be identified as a proven physiological regulator.
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  • 6
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    Calcified tissue international 13 (1973), S. 271-285 
    ISSN: 1432-0827
    Keywords: Glycosaminoglycans ; Extraction ; Epiphysis ; Cartilage ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des glycosaminoglycanes de zones au repos, sériées, hypertrophiques et calcifiées de la métaphyse épiphysaire et du cartilage du septum nasal ont été extraits à l'aide d'un solvant, 3 M GuCl, selon Sajdera et Hascall (1969), pour séparer les glycosaminoglycanes en une partie que l'on peut extraire et une partie qui reste liée au tissu. Le cartilage épiphysaire nécessite des temps d'extraction plus prolongés que le cartilage du septum nasal pour extraire des quantités similaires de glycosaminoglycanes acides suggérant une liaison plus étroite de ces derniers dans le tissu. Vers le front de calcification, les glycosaminoglycanes sont extraits plus facilement alors que dans la zone calcifiée pas plus de 30% ne peut être extrait. Des résultats obtenus par le procédé de microfractionnement CPC d'Antonopoulos et coll. (1964) indiquent une distribution similaire selon le poids moléculaire et/ou la densité de charge pour le chondroitine sulfate qui se laisse extraire ou non du cartilage du septum nasal et le chondroitine sulfate que l'on ne peut extraire du cartilage du septum nasal et des zones au repos et sériées de la métaphyse. Les glycosaminoglycanes des zones hypertrophiques et calcifiées que l'on ne peut extraire sont surtout de faible poids moléculaire et/ou leur densité de charge se compare au pool que l'on peut extraire. L'acide hyaluronique ne peut être extrait du septum nasal et des zones au repos, sériées et hypertrophiques avec des concentrations croissantes vers le front de calcification. Dans la zone calcifiée, un changement avec augmentation de l'acide hyaluronique que l'on peut extraire est noté. Ces résultats sont discutés.
    Abstract: Zusammenfassung Die sauren Glycosaminoglycane von ruhenden, säulenförmigen, hypertrophischen und verkalkten Zonen der Epiphysen-Wachstumsplatte und des Nasenseptum-Knorpels vom Kalb wurden mittels eines dissoziierenden Lösungsmittels, 3 M Guanidinchlorid, nach Sajdera und Hascall (1969) extrahiert, um die Glycosaminoglycane in einen extrahierbaren und einen unextrahierbaren Anteil — dieser bleibt ans Gewebe gebunden — zu trennen. Der Epiphysenknorpel beanspruchte längere Extraktionszeiten als der Nasenseptumknorpel, um vergleichbare Mengen saurer Glycosaminoglycane abzugeben. Dies deutet auf eine stärkere Bindung der Proteoglycane im Gewebe. Die Glycosaminoglycane ließen sich leichter aus dem Bereich der Verkalkungsfront extrahieren, während aus der verkalkten Zone nicht mehr als 30% extrahiert werden konnten. Die Resultate, die mit dem Cetylpyridinchlorid-Mikrofraktionierungsverfahren von Antonopouloset al. (1964) erhalten wurden, deuteten auf eine ähnliche Verteilung entsprechend dem Molekulargewicht und/oder der Ladungsdichte für extrahierbares und unextrahierbares Chondroitinsulfat im Nasenseptumknorpel und in ruhenden und säulenförmigen Zonen der Epiphysen-Wachstumsplatte. Die unextrahierbaren Glycosaminoglycane in hypertrophischen und verkalkten Zonen zeigten vorwiegend niederes Molekulargewicht und/oder niedere Ladungsdichte im Vergleich zum extrahierbaren Anteil. Die Hyaluronsäure ließ sich aus Nasenseptum und ruhenden, säulenförmigen und hypertrophischen Zonen nicht extrahieren und zeigte höhere Konzentrationen als in der Verkalkungsfront. In der verkalkten Zone erfolgte eine Verschiebung zu meist extrahierbarer Hyaluronsäure. Die Bedeutung dieser Befunde wird diskutiert.
    Notes: Abstract The acid glycosaminoglycans of resting, columnar, hypertrophic and calcified zones of calf epiphyseal growth plate and of nasal septum cartilage were extracted with a dissociative solvent, 3M GuCl, according to Sajdera and Hascall (1969), to separate the glycosaminoglycans into an extractable pool and an unextractable pool which remains bound within the tissue. Epiphyseal cartilage required longer extraction times than did nasal septum cartilage to extract comparable amounts of acid glycosaminoglycans suggesting a stronger binding of proteoglycans within the tissue. Towards the calcification front the glycosaminoglycans were extracted more easily while in calcified zone not more than 30% could be extracted. Data obtained by the CPC microfractionation procedure of Antonopouloset al. (1964) indicated similar distribution according to molecular weight and/or charge density for extractable and unextractable chondroitin sulphate in nasal septum cartilage and in resting and columnar zones of epiphyseal growth plate. Unextractable glycosaminoglycans in hypertrophic and calcified zones were of predominantly low molecular weight and/or charge density compared to the extractable pool. Hyaluronic acid was unextractable in nasal septum and in resting, columnar and hypertrophic zones with increasing concentrations towards the calcification front. In calcified zone a shift to mainly extractable hyaluronic acid occurred. The significance of these findings is discussed.
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  • 7
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    Calcified tissue international 13 (1973), S. 131-142 
    ISSN: 1432-0827
    Keywords: Mucopolysaccharide ; Synthesis ; Vitamin A ; Cartilage ; Tissue culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé L'effet de la vitamine A sur la synthèse des mucopolysaccharides acides a été étudiéin vitro dans des cultures cellulaires de chondrocytes sternaux d'embryons de poulets. La vitamine A provoque une inhibition, liée à la concentration, de la synthèse des mucopolysaccharides acides. L'effet d'inhibition de la vitamine A sur la synthèse des mucopolysaccharides acides n'est pas la conséquence d'une cytotoxicité générale, étant donné que dans les conditions utilisées, provoquant une inhibition de 50% de la synthèse mucopolysaccharidique, l'incorporation de leucine en matériel insoluble acide ou la synthèse de collagène n'est pas altérée. L'activation des lysosomes ne semblent pas intervenir dans l'inhibition de l'incorporation de sulfate, étant donné que l'acide E-amino caproique (un inhibiteur protéique) et la chloroquine (un stabilisateur de membrane des lysosomes) n'inversent pas cette inhibition. En plus, l'augmentation de la dégénérescence polysaccharidique est trop faible pour rendre compte de l'incorporation de sulfate inhibé par la vitamine A. L'action de la vitamine A n'est pas facilement modifiée et les chondrocytes deviennent fibroblastiques après 24 heures de traitement avec 3 μg/ml de vitamine A. Etant donné que l'incorporation de glucose en polysaccharides est plus rapidement inhibée que la glucosamine ou sulfate, il semble que la vitamine A agit par un mécanisme inconnu pour inhiber une étape préliminaire de la biosynthèse des mucopolysaccharides.
    Abstract: Zusammenfassung Die Wirkung von Vitamin A auf die Synthese von sauren Mucopolysacchariden wurde in vitro in Zellkulturen von Chondrocyten aus dem Sternum von Kükenembryonen studiert. Vitamin A verursacht eine Konzentrations-abhängige Hemmung der Synthese saurer Mucopolysaccharide. Die Hemmwirkung von Vitamin A auf die Synthese saurer Mucopolysaccharide ist nicht die Folge einer allgemeinen Cytotoxicität, denn unter den hier untersuchten Bedingungen besteht wohl eine über 50%ige Hemmung der Mucopolysaccharid-Synthese, aber weder der Einbau von Leucin in Säure-unlösliches Material noch die Collagensynthese werden verändert. Eine Lysosomen-Aktivierung scheint an der Hemmung des Sulfateinbaus nicht beteiligt zu sein, da E-Aminocapron-Säure (ein Hemmstoff der Protease) und Chloroquin (ein Stabilisator der Lysosomen-Membran) diese Hemmung nicht aufheben. Außerdem ist der erhöhte Polysaccharid-Abbau zu gering, um als Ursache für einen durch Vitamin A gehemmten Sulfateinbau in Betracht zu kommen. Die Wirkung von Vitamin A läßt sich nicht leicht rückgängig machen und die Chondrocyten werden nach einer 24 stündigen Behandlung mit 3 μg Vitamin A/ml fibroblastisch. Da der Glucoseeinbau durch die Polysaccharide schneller gehemmt wird als derjenige von Glucosamin oder Sulfat, wird vermutet, daß Vitamin A durch einen unbekannten Mechanismus eine frühe Stufe im Verlauf der Mucopolysaccharid-Biosynthese hemmt.
    Notes: Abstract The effect of vitamin A on the synthesis of acid mucopolysaccharides has been studiedin vitro in cell cultures of chick embryo sternal chondrocytes. Vitamin A produces a concentration-dependent inhibition of acid mucopolysaccharide synthesis. the inhibitory effect of vitamin A on acid mucopolysaccharide synthesis is not the result of a general cytotoxicity, since, under the conditions studied here, while there is over a 50% inhibition of mucopolysaccharide synthesis, the incorporation of leucine into acid insoluble material or the synthesis of collagen is unaltered. Lysosomal activation does not appear to be involved in the inhibition of sulfate incorporation, since E-amino caproic acid (a protease inhibitor) and chloroquine (a lysosomal membrane stabilizer) do not reverse this inhibition. In addition, enhanced polysaccharide breakdown is too small to account for vitamin A-inhibited sulfate incorporation. The effect of vitamin A is not readily reversed and chondrocytes become fibroblastic after a 24 hour treatment with 3 μg/ml of vitamin A. Because the incorporation of glucose into polysaccharides is inhibited faster than glucosamine or sulfate, it is suggested that vitamin A acts by an unknown mechanism to inhibit a step early in the pathway for mucopolysaccharide biosynthesis.
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  • 8
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    Calcified tissue international 16 (1974), S. 183-192 
    ISSN: 1432-0827
    Keywords: Mucopolysaccharides ; Spondyloepiphyseal dysplasia ; Cartilage ; Growth defects
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Iliac crest cartilage biopsies from five children with spondyloepiphyseal dysplasia congenita contained increased amounts of chondroitin-6-sulfate and keratan sulfate. The increase was similar to that observed in the cartilage of patients with Morquio's disease. However, unlike patients with Morquio's disease, those with spondyloepiphyseal dysplasia congenita did not excrete an abnormal amount of keratan sulfate in urine. Spondyloepiphyseal dysplasia congenita can be considered a mucopolysaccharidosis without mucopolysacchariduria.
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  • 9
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    Calcified tissue international 26 (1978), S. 47-49 
    ISSN: 1432-0827
    Keywords: Matrix vesicles ; Calcospherites ; Mineralization ; Bone ; Dentin ; Cartilage ; SEM
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Mineral particle cluster, corresponding to calcospherites or matrix vesicle-initiated clusters in calcifying cartilage and dentin and to collagen bundle-related mineralization in lamellar bone, have been isolated from NaOCl solutions used to dissolve the organic matrix in preparation of anorganic mineralizing fronts for scanning electron microscopy.
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  • 10
    ISSN: 1432-0827
    Keywords: Vitamin D ; Vitamin D deficiency ; Bone ; Cartilage ; Bone development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The role of vitamin D in early skeletal development was studied by measuring serum calcium and phosphorus, osseous tissue quantity and mineralization, and endochondral bone elongation in rat fetuses and pups from vitamin D-replete and vitamin D-deficient mothers. At the 20th day of pregnancy there was a slight, yet significant, increase in the amount of osteoid on trabecular bone surfaces in fetuses from vitamin D-deficient mothers. The fetal bones otherwise appeared normal in spite of severe skeletal changes in the vitamin D-deficient mothers. After parturition, the importance of vitamin D in skeletal development becomes progressively more obvious. Serum calcium levels were slightly, yet significantly, lower in vitamin D-deficient than in vitamin D-replete pups and these levels continued to fall in the vitamin D-deficient pups through lactation and after weaning. At 3 days postpartum, there was a small, yet significant, increase in the amount of osteoid on bone surfaces of the vitamin D-deficient pups. The relative amounts of osteoid in the vitamin D-deficient pups continued to increase through lactation and after weaning when compared with vitamin D-replete pups. By the 14th day of lactation and at later periods, there were significant reductions in metaphyseal mineralized tissues in the vitamin D-deficient pups when compared with the vitamin D-replete pups. At weaning and after weaning, there were substantial increases in growth plate thickness and decreases in longitudinal bone growth in the vitamin D-deficient pups when compared with the vitamin D-replete pups. The results from this study indicate that vitamin D does not appear to play a major role in fetal skeletal development. However, after birth, vitamin D becomes progressively more important with age for normal bone development, mineralization, and endochondral growth.
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  • 11
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    Calcified tissue international 4 (1969), S. 185-187 
    ISSN: 1432-0827
    Keywords: Cartilage ; Trauma ; Osteoporosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé La masse du mineral osseux du bout distal femoral a été mesuré dans 54 cas de rupture du cartilage semilunair médial 0–5 années après l'intervention chirurgique. La masse du mineral osseux était diminué. La masse du mineral osseux n'avait aucune tendance de restauration pendant le temps d'observation.
    Abstract: Zusammenfassung In 54 Fällen von medialen Meniscusschäden wurde die Skeletmasse in dem distalen Femurende gemessen 0–5 Jahre nach der Fortnehmung des geschädigten Meniscus. Die Skeletmasse war signifikant vermindert. Im Laufe der Zeit zeigte sich keine Tendenz zur Erhöhung der Skeletmasse.
    Notes: Abstract In 54 cases of ruptured medial semilunar cartilages of the knee joint the bone mineral mass in the distal end of the femur was measured 0–5 years following removal of the injured cartilage. The bone mineral mass was significantly decreased. There was no tendency of restoration of the bone mass with time.
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  • 12
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    Calcified tissue international 4 (1969), S. 260-268 
    ISSN: 1432-0827
    Keywords: Cartilage ; Histochemistry ; Staining ; Protein ; Polysaccharide ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des coupes de cartilage épiphysaire frais de jeunes rats, effectuées à la main, sont colorées à pH=4,5 dans des solutions à 0,01% de divers colorants cationiques, appartenant aux groupes de la thiazine, oxazine, azine, triphénylméthane, acridine, et phthallocyanine. Les granules intracellulaires métachromatiques, mises en évidence antérieurement par le bleu de toluidine, sont également identifiées à l'aide de l'azur A, le bleu de méthylène et le bleu de crésyl. Les granules se colorent moins bien à la thionine, le rouge neutre, la safranine O, le bleu de toluylène et l'acridine orange. Dans les conditions utilisées, la matrice de la zone de réserve et la matrice de la zone hypertrophique inférieure (en voie de calcification) se colorent, alors que les matrices des zones prolifératives et hypertrophiques supérieures ne prennent pas les colorants. La gallocyanine, le violet cristal, la fuchsine basique, l'azocarmin B, le bleu de gallamine et la bleu alcian ne se colorent pas ou donnent des réactions colorées différentes de celles décrites ci-dessus. Il semble que le pK et le poids moléculaire des colorants jouent un rôle important, mais ils ne paraissent pas être les seuls facteurs intervenant dans la coloration des granules. Un changement, lié à la calcification, semble intervenir au niveau du matériel métachromatique (probablement des polysaccharides protéiques), aussi bien dans la matrice que les cellules cartilagineuses épiphysaires.
    Abstract: Zusammenfassung Handpräparierte Schnitte von frischem Epiphysenknorpel junger Ratten wurden bei einem pH von 4,5 in 0,01% igen Lösungen verschiedener kationischer Farbstoffe folgender Klassen gefärbt: Thiazin, Oxazin, Azin, Triphenylmethan, Acridin und Phthalocyanin. Die intracellulären β-und γ-metachromatischen Granula, erstmals mit Toluidinblau im frischen Gewebe nachgewiesen, konnten auch gut mit Azur A, Methylenblau und Brillantkresylblau dargestellt werden. Die Granula konnten ebenfalls, aber weniger gut, mit Thionin, Neutralrot, Safranin D, Toluylenblau und Acridinorange gefärbt werden. Unter diesen Färbungsbedingungen werden die inaktive Matrixzone und die untere hypertrophische (verkalkende) Matrixzone angefärbt, während die proliferative und die obere hypertrophische Matrixzone sich nicht färben. Gallocyanin, Kristallviolett, basisches Fuchsin, Azokarmin B, Gallaminblau und Alzianblau färbten entweder gar nicht, oder gaben ein anderes als das obenbeschriebene Färbemuster. Es wird vorgeschlagen, daß das pK und das Molekulargewicht der Farbstoffe wichtig aber nicht unbedingt die einzigen Faktoren sind, die die Färbung der Granula bestimmen. Die Resultate zeigen, daß eine Veränderung im metachromatischen Material (vermutlich Proteinpolysaccharide) vorliegt, und zwar sowohl in der Matrix als in den Zellen des Epiphysenknorpels; diese Veränderung scheint im Zusammenhang mit der Verkalkung zu stehen.
    Notes: Abstract Hand-cut sections of fresh epiphyseal cartilage from young rats were stained at pH 4.5 in 0.01% solutions of various cationic dyes of the thiazine, oxazine, azine, triphenylmethane, acridine, and phthallocyanin classes. The intracellular β-and γ-metachromatic granules, previously demonstrated in fresh tissues with toluidine blue, were also demonstrated well with azure A, methylene blue, and brilliant cresyl blue. The granules were also demonstrated, but not as well, by thionin, neutral red, safranin O, toluylene blue, and acridine orange. Under the conditions of staining, the reserve zone matrix and the lower hypertrophic (calcifying) zone matrix stained, whereas the proliferative and upper hypertrophic zone matrix did not stain. Gallocyanin, crystal violet, basic fuchsin, azocarmine B, gallamine blue, and alcian blue either did not stain, or gave a different pattern of staining from that described above. It is suggested that the pK and molecular weight of the dyes are important, but not necessarily the only factors in determining the staining of the granules. The results indicate that there is a change in the metachromatic material (presumably proteinpolysaccharide) in both the matrix and cells of epiphyseal cartilage, which appears to be related to calcification.
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  • 13
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    Calcified tissue international 5 (1970), S. 85-90 
    ISSN: 1432-0827
    Keywords: Cartilage ; Electron transport ; Cytochrome oxidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Lorsque des lapins augmentent pondéralement de 165 à 2,205 grammes, l'activité en cytochrome oxydase de cartilage épiphysaire homogénéisé n'est pas modifiée, en utilisant des tests enzymatiques manométriques et spectrophotométriques; alors que l'activité en oxydase du cartilage xyphoide du sternum et de l'oreille homogénéisé décroit. Des spectres d'absorption optique des tissus cartilagineux, réalisés pour la première fois à l'aide de fragments d'oreilles de lapin et de cartilage embryonnaire de poulets, montrent un maximum d'absorption caractéristique pour le système respiratoire final des mitochondries, y compris les composés b, c et oxydasiques ainsi que les flavoprotéines.
    Abstract: Zusammenfassung Während Kaninchen ihr Körpergewicht von 165 auf 2205 g erhöhen, erfolgt keine Veränderung in der Cytochrom-Oxydase-Aktivität der epiphysären Knorpel-Homogenate in manometrischen und spectrophotometrischen Enzymversuchen, jedoch vermindern sich die Oxydase-Aktivitäten von xiphisternalen und von Ohrknorpel-Homogenaten. Lichtabsorptionsspektren von Knorpelgewebe wurden zum ersten Mal gemacht, indem Streifen von Kaninchenohrstücken und Stücke von Kükenembryoknorpel verwendet wurden. Solche Spektren zeigen charakteristische Absorptionsmaxima für den terminalen Atmungskomplex der Mitochondrien, einschließlich b, c und Oxydasekomponenten sowie Flavoprotein.
    Notes: Abstract As rabbits increase in size from 165 grams to 2,205 grams body weight, cytochrome oxidase activity of epiphyseal cartilage homogenates does not change in manometric and spectrophotometric enzyme assays, whereas the oxidase activities of xiphisternal and ear cartilage homogenates decline. Light absorption spectra of cartilage tissues were made for the first time, using strips of rabbit ear and pieces of chick embryo cartilage. Such spectra reveal characteristic absorption maxima for the terminal respiratory complex of mitochondria, including b, c and oxidase components and flavoprotein.
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  • 14
    ISSN: 1432-0827
    Keywords: Growth hormone ; Resorption ; Bone ; Cartilage ; Tartrate-resistant acid phosphatase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Young male Sprague-Dawley rats (5–7 weeks old, 80–120 g) were hypophysectomized (HX) and maintained on thyroxin and dexamethasone replacement therapies. Ten days after surgery, some HX rats received a single injection of human growth hormone (hGH), and others five daily injections of hGH. Tartrate-resistant acid phosphatase (TRAP) histochemistry was employed in order to evaluate the number of cells of resorptive potential in the metaphyseal bone of the proximal tibiae of HX rats and was compared with normal rats and HX rats that further received hGH replacement therapy. In normal rats, two populations of TRAP-positive cells were identified: multinuclear cells, which showed histological characteristics of osteoclasts, and small mononuclear cells, the number of which was overwhelming when compared with the number of TRAP-positive multinuclear cells. Both populations were reduced in the HX rat, but more so the mononuclear cells, which were assumed to represent the precursor pool of mature osteoclasts and chondroclasts (P〈0.005). Five daily injections of hGH to HX rats brought about a significant increase in the number of TRAP-positive multinuclear cells, the number of nuclei of these cells, and the number of mononuclear TRAP-positive cells, throughout the metaphyseal bone (P〈0.05). A single injection of hGH increased only the number of TRAP-positive multinuclear cells in the trabecula/bone marrow interface (P〈0.05), indicating a very rapid fusion of precursor cells into mature osteoclasts in that particular location. It was concluded that GH depletion caused a major reduction in the number of cells presenting resorption capacity and that a short hGH replacement regimen resulted in a gradual restoration of these cells n the metaphyseal bone of the proximal tibia of the HX rat.
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  • 15
    ISSN: 1432-0827
    Keywords: Insulin ; Cartilage ; Growth ; Condyle ; Mandible ; Mouse ; In vitro
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Condylar cartilages were cultured in the form of organ cultures on top of collagen sponges in medium containing 2% fetal calf serum and were treated with 3.5–350 nM insulin for 6 days. Doses of 175 nM of insulin caused a marked increase (+96%) in DNA synthesis and in proteoglycan production (+74%), features that manifested themselves structurally by a 60% increase in overall size of the cultured explants. Using a tissue culture system comprised of cartilage progenitor cells, insulin was found to enhance the differentiation of the progenitor cells so that by 6 days in culture and appreciable nodule of differentiated chondrocytes developed. The latter was surrounded by perichondrial cells whereas the extracellular matrix within the newly formed, insulin-induced, nodule reacted positively for cartilagespecific antigens (type II collagen and bone sialoprotein). It is suggested that insulin induces a direct stimulatory effect on progenitor cell proliferation, cartilage differentiation, and extracellular matrix deposition.
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  • 16
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    Calcified tissue international 7 (1971), S. 290-298 
    ISSN: 1432-0827
    Keywords: Proteinpolysaccharides ; Cartilage ; Calcification ; Binding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les protéines-polysaccharides (PP-L) de zones «au repos» et «en voie de calcification» de cartilage d'omoplate et de cloison nasale de veaux sont extraits à l'aide d'une solution aqueuse à 0.15 M KCl et fractionnés en une série de produits: PP-L3, PP-L4, PP-L5, PP-L6. Une fraction supplémentaire protéino-polysaccharidique, PP-L2, est extraite à partir des restes cartilagineux à l'aide d'hydroxylamine. Des différences de composition chimique parmi les fractions protéino-polysaccharides, obtenues à partir de ces trois sources de cartilage, son généralement faibles. La propriété de liaison du calcium des échantillons de PP-L, PP-L2, PP-L3, déterminée par la mesure de dialyse d'équilibre, semble principalement en rapport avec les contenus en acide uronique et en sulfate échangeable. L'application d'une technique d'échange cationique indique que le PP-L de zones «au repos» et «en voie de calcification» de cartilage de l'omoplate a des affinités identiques pour des quantités de calcium radioactif. Cependant, l'affinité de PP-L des deux zones de cartilage de l'omoplate pour des quantités de calcium radioactif est plus élevé que celle de PP-L de la cloison nasale. Les résultats obtenus ne montrent pas de différences suffisantes dans la composition chimique et l'affinité pour le calcium des protéines-polysaccharides de cartilage «au repos» et «en voie de calcification» de l'omoplate, pour rendre compte de l'enrichissement en calcium de la zone «en voie de calcification».
    Abstract: Zusammenfassung Proteinpolysaccharide (PP-L) von „ruhenden” und „verknöchernden” Zonen von Kalbs-Scapula-Knorpel und von Kalbs-Nasenseptum-Knorpel wurden mit wäßrigem 0,15 M KCl extrahiert und zu einer Serie von Produkten fraktioniert: PP-L3, PP-L4, PP-L5, PP-L6. Eine zusätzliche Proteinpolysaccharidfraktion, PP-L2, wurde mit Hydroxylamin aus den Knorpelrückständen extrahiert. Die Unterschiede in der chemischen Zusammensetzung zwischen den entsprechenden Proteinpolysacchariden von den drei Knorpel-Quellen waren im allgemeinen klein. Die Calciumbindungsfähigkeit der PP-L, PP-L2 und PP-L3-Proben, gemessen durch Gleichgewichtsdialyse, schien hauptsächlich deren Gehalt an Uronsäure und austauschbarem Sulfat wiederzugeben. Die Anwendung einer Kationenaustausch-Technik deutete darauf hin, daß PP-L von „ruhenden” und „verknöchernden” Zonen von Scapulaknorpel eine ähnliche Affinität für Tracermengen von Calcium hat. Jedoch war die Affinität von PP-L für Tracermengen von Calcium aus beiden Zonen von Scapulaknorpel größer als diejenige von PP-L aus dem Nasenseptum. Die aus dieser Studie hervorgehenden Daten zeigen nicht genügend Unterschiede an zwischen der chemischen Zusammensetzung und der Calcium-affinität von Proteinpolysacchariden aus „ruhendem” und „verknöcherndem” Scapulaknorpel, um die Calciumaufnahme in der „verknöchernden” Zone zu erklären.
    Notes: Abstract Proteinpolysaccharide (PP-L) of “resting” and “ossifying” zones of calf scapular cartilage and of calf nasal septum cartilage was extracted with aqueous 0.15 M KCl and fractionated into a series of products: PP-L3, PP-L4, PP-L5, PP-L6. An additional proteinpolysaccharide fraction, PP-L2, was extracted from the cartilage residues with hydroxylamine. Differences in chemical composition among corresponding proteinpolysaccharides obtained from the three cartilage sources were, in general, small. The calcium binding capacity of the PP-L, PP-L2, and PP-L3 samples, as measured by equlibrium dialysis, appeared to reflect principally their uronic acid and exchangeable sulfate contents. Application of a cation exchange technique indicated that PP-L from “resting” and “ossifying” zones of scapular cartilage had similar affinities for tracer quantities of calcium. However, the affinity of PP-L from both zones of scapular cartilage for tracer quantities of calcium was greater than that of PP-L from nasal septum. The data obtained from this study do not indicate sufficient differences between the chemical composition and calcium affinity of proteinpolysaccharides from “resting” and “ossifying” scapular cartilage to account for the calcium uptake in the “ossifying” zone.
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  • 17
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    Calcified tissue international 9 (1972), S. 140-151 
    ISSN: 1432-0827
    Keywords: Cartilage ; Epiphysis ; Cell ; Differentiation ; Endoplasmic reticulum ; Mitochondria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Une étude ultrastructurale de chondrocytes hypertrophiques est rendue possible par suite d'une meilleure méthode de fixation. Contrairement à l'opinion qui veut que les cellules hypertrophiques meurent, nos résultats suggèrent nettement une activité cellulaire allant du début de la croissance de la cellule au stade où la lacune s'ouvre. La séquence des altérations cellulaires est décrite, en allant de l'augmentation de l'ergastoplasme à la présence de ribosomes libres et de granules denses aux électrons dans les mitochondries. Les diverses fonctions possibles de la cellule sont décrites. Il est possible que les chondrocytes hypertrophiques se transforment en cellules osseuses.
    Abstract: Zusammenfassung Eine verbesserte Fixationsmethode ermöglichte die Untersuchung der Feinstruktur des hypertrophischen Chondrocyten. Im Gegensatz zur allgemeinen Annahme, daß die hypertrophischen Zellen absterben, weisen unsere Resultate deutlich darauf hin, daß die Zellaktivität vom Moment der Zellvergrößerung bis zum Stadium, in welchem die Lacunen aufbrechen, vorhanden ist. Der Ablauf der Veränderungen innerhalb der Zelle wurde beschrieben, wobei die hervorstechendsten Charakteristika die Zunahme des endoplasmatischen Reticulums, das Auftreten von vielen freien Ribosomen und das Vorkommen von elektronenmikroskopisch dichten Granula in den Mitochondrien waren. Die möglichen Funktionen der Zelle wurde besprochen. Es ist äußerst interessant, daß dadurch die Hypothese, wonach die hypertrophischen Chondrocyten schließlich zu Knochenzellen werden, unterstützt wird.
    Notes: Abstract A study of the ultrastructure of the hypertrophic chondrocyte has been made possible by an improved method of fixation. In contrast to the general belief that the hypertrophic cells die, our results strongly suggest cell activity from the beginning of enlargement of the cell up to the stage where the lacuna breaks open. The sequence of alterations within the cell have been described, with the increase of endoplasmic reticulum contents, the appearance of many free ribosomes and the occurrence of electron-dense granules in the mitochondria as the most prominent characteristics. The possible functions of the cell have been discussed. Most interesting is the support for the hypothesis that hypertrophic chondrocytes may ultimately become bone cells.
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  • 18
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    Calcified tissue international 9 (1972), S. 131-139 
    ISSN: 1432-0827
    Keywords: Cartilage ; Epiphyses ; Endoplasmic reticulum ; Golgi apparatus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Une méthode de fixation des cellules cartilagineuses de la métaphyse est mise au point. Une meilleure préservation des chondrocytes applatis montre de petites vésicules qui semblent naître dans l'ergastoplasme et se trouvent en grande quantité au niveau de l'appareil de Golgi. Différents types de vacuoles sont visibles dans cet appareil, à savoir des ≪vacuoles intermédiaires≫ et de ≪larges vacuoles≫ contenant des inclusions. Il parait probable que les vésicules et vacuoles jouent un rôle spécifique dans le transport et la production de polysaccharides de protéines sulfatées.
    Abstract: Zusammenfassung Es wurde eine Methode zur Fixation der Knorpelzellen aus der Epiphysenplatte entwickelt. die besser erhaltenen abgeflachten Chondrocyten zeigen kleine Bläschen, welche aus dem groben endoplasmatischen Reticulum zu stammen scheinen und in großen Mengen im Golgi-Bereich vorliegen. Verschiedene Arten von Vacuolen können im Golgi-Apparat unterschieden werden, z.B. “intermediate vacuoles” und “large vacuoles”, wobei die letztgenannten gewöhnlich Einschlüsse enthalten. Es ist wahrscheinlich, daß die Bläschen sowie die Vacuolen eine spezifische Rolle beim Transport und der Bildung von sulfatierten Proteinpolysacchariden spielen.
    Notes: Abstract A method of fixation has been developed for the cartilage cells of the epiphyseal plate. Improved preservation of the flattened chondrocytes reveals small vesicles that seem to originate in the rough endoplasmic reticulum and are found in large amounts in the Golgi area. Different types of vacuoles can be distinguished in the Golgi complex, e.g. “intermediate vacuoles” and “large vacuoles”, the latter usually containing inclusions. It seems likely that the vesicles and vacuoles each play a specific role in the transport and production of sulfated protein polysaccharides.
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  • 19
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    Development genes and evolution 200 (1991), S. 162-171 
    ISSN: 1432-041X
    Keywords: Retinoid ; Determination ; Muscle ; Cartilage
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The time of determination of cartilage and skeletal muscle was studied by making chimeric grafts or explants of small tissue pieces from several stages of early chick or quail embryos. Chondrogenesis was assessed by histology or with antibodies directed against type II collagen or cartilage proteoglycan, while myogenesis was detected immunohistochemically with antibodies directed against 3 different muscle markers, including muscle myosin. Grafts from Hensen's node, primitive streak and segmental plate of donor embryos of Stage 3–5 (Hamburger and Hamilton) were transplanted under the ectoderm in the extraembryonic area of Stage 12 host embryos. In addition, explants and mesodermal cells were cultured on glass in DMEM+F12 medium supplemented with 10% FCS. The results showed that determined myogenic cells could first be detected in Hensen's node and the primitive streak at Stage 3+−4 and that they developed from mesodermal cells located between the epiblast and hypoblast. Myogenic cells also appeared in grafted and explanted segmental plate with or without notochord from Stage 5 embryos. On the other hand, cartilage cells only formed in grafted and explanted segmental plate that also contained notochord. RA (1 ng/ml) could induce the formation of cartilage cells in the explanted primitive streak without Hensen's node or notochord taken from Stage 3–5 embryos and could also promote the differentiation of myogenic cells in primitive streak from Stage 3 embryo. Thus RA can substitute for Hensen's node or the notochord in the induction of cartilage cells and has some stimulatory effects on the differentiation of myogenic cells. Additional evidence indicates that the hypoblast might play an “inductive” role in the formation of the notochord which may subsequently promote the differentiation of cartilage cells.
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  • 20
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    Development genes and evolution 200 (1991), S. 202-207 
    ISSN: 1432-041X
    Keywords: Extracellular matrix ; Morphogenesis ; Cartilage ; Femur rudiment ; Proteoglycan turnover
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Turnover of extracellular matrix (ECM) proteoglycans was studied in chick cartilaginous femur rudiments grown in organ culture. Femora from six-day-old embryos showed nearly normal growth rates during the first few days in culture. By labeling the rudiment with 35S-sulfate or 14C-glucosamine, it was demonstrated that the cartilaginous ECM undergoes rapid turnover. It was also found that the metabolic fate of the proteoglycans is to be released as macromolecules into the culture medium. When a rudiment was cut to obtain two epiphyses it was observed that each part grows and synthesizes proteoglycans at nearly normal rates, which indicates that the isolated epiphyses, like the whole rudiment, behave as autonomous systems. We suggest that the turnover of ECM components is part of the continuous remodelling process rudiments undergo during their growth and development. In order to study cell-ECM interaction in morphogenesis, we made an attempt to prepare an intact cell-free ECM. Epiphyses were heated at 45.2° C for 1 h. The treatment caused complete cessation of growth and biosynthesis. When the cut surface of a live epiphysis was brought into apposition to a heat-treated epiphysis and the attached pair placed in organ culture, it was found that the heat-treated epiphysis begins to grow and reaches almost the same size as its live counterpart. We discuss the possible advantage of this new experimental system for studies on the role of ECM in morphogenesis.
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  • 21
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    Calcified tissue international 14 (1974), S. 251-255 
    ISSN: 1432-0827
    Keywords: Cartilage ; Bone ; Vitamin A ; Phosphatases
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Au cours d'une étude du cartilage épiphysaire de poulets normaux et déficients en vitamine A, la composition organique et l'activité phosphatasique du cartilage au repos, du cartilage en voie d'ossification et de l'os nouvellement formé ont été mesurées. Le cartilage en voie d'ossification et l'os nouvellement formé ont un contenu plus élevé en matériel inorganique, phosphate et collagène que le cartilage au repos. Une déficience en vitamine A provoque une augmentation du contenu en phospholipide de tous les trois tissus. Le cartilage au repos, en cas de déficience en vitamine A, présente, après homogénéisation et centrifugation, un surnageant dont l'activité en phosphatase alcaline et en glycérophosphatase est plus élevée que celle des témoins. Il semble que les effets enzymatiques de l'avitaminose A soient liés à des altérations de la membrane lysosomiale avec décharge de phosphatases. La minéralisation normale semble aussi faire intervenir une activité phosphatasique.
    Abstract: Zusammenfassung Diese Arbeit beschreibt Untersuchungen des Epiphysenknorpels bei normalen Küken und solchen mit Vitamin-A-Mangel. Die organische Zusammensetzung und die Phosphatase-Aktivität in ruhendem Knorpel, ossifizierendem Knorpel und neugebildetem Knochen wurden gemessen. Der ossifizierende Knorpel und der neugebildete Knochen hatten einen höheren Gehalt an anorganischem Material, an Phosphat und Kollagen als der ruhende Knorpel. Vitamin-A-Mangel führte zu einem erhöhten Phospholipidgehalt in allen drei Geweben. Nach Homogenisierung und Zentrifugierung fand sich im Überstand des ruhenden Knorpels von Vitamin-A-Mangelgewebe eine höhere alkalische Phosphatase- und Glycero-phosphatase-Aktivität als bei den Kontrollen. Es wird angenommen, daß die Wirkung des Vitamin A-Mangels auf die Enzyme im Zusammenhang steht mit der Schädigung der lysosomalen Membran, wobei Phosphatasen freigesetzt werden, und daß die normale Mineralisierung die Phosphatasen-Aktivität ebenfalls anregt.
    Notes: Abstract This paper reports studies of the epiphyseal cartilage in normal and vitamin A deficient chicks. The organic composition and the phosphatase activity in the resting cartilage, ossifying cartilage and new bone were measured. The ossifying cartilage and new bone had a higher content of inorganic material, phosphate and collagen than the resting cartilage. Vitamin A deficiency caused increase in the phospholipid content of all three tissues. The resting cartilage from vitamin A deficient tissue had, after homogenisation and centrifugation, a supernatant with an activity of alkaline phosphatase and glycerophosphatase higher than that in control samples. It is considered that effects of vitamin A deficiency on enzymes are related to defects of the lysosomal membrane with release of phosphatases, and that normal mineralisation also involves phosphatases activity.
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  • 22
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    Calcified tissue international 14 (1974), S. 275-281 
    ISSN: 1432-0827
    Keywords: Calcification ; Cartilage ; Lipids ; Membranes ; Vesicles
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les vésicules matricielles sont des particules extracellulaires, entourées par une membrane, qui sont nombreuses dans la matrice du cartilage en voie de calcification et qui sont en rapport morphologique avec le dépôt minéral. Les vésicules matricielles et les cellules de l'épiphyse d'os longs d'embryons de veaux sont libérées par digestion grâce à la collagénase et séparées par centrifugation différentielle. Les vésicules ainsi obtenues sont riches en lipides et contiennent significativement plus de sphingomyéline et phosphatidyl sérine que les fractions cellulaires. Le rapport moles de cholestérol sur moles de phospholipides totaux est plus élevé dans les vésicules que dans les cellules; ce résultat est en accord avec l'origine des vésicules au niveau des membranes cellulaires des chondrocytes.
    Abstract: Zusammenfassung Matrixvesikel sind extrazelluläre, membrangebundene Teilchen, welche in der Matrix von verkalkendem Knorpel gehäuft vorkommen und welche in morphologischem Zusammenhang mit der Mineralablagerung in diesem Gewebe stehen. Matrixvesikel und Epiphysenzellen aus Röhrenknochen von Kalbsembryonen wurden mittels Collagenaseverdauung freigesetzt und durch Differential-Zentrifugierung getrennt. Die Vesikelpräparate waren reich an Lipidien und enthielten bedeutend mehr Sphingomyelin und Phosphatidylserin als die Zellfraktionen. Das molare Verhältnis Cholesterol/Gesamtlipide war in den Vesikeln höher als in den Zellen; dieser Befund stimmt mit der Tatsache überein, daß die Vesikel ihren Ursprung in den Plasmamembranen der Chondrozyten haben.
    Notes: Abstract Matrix vesicles are extracellular, membrane-bounded particles which are abundant in the matrix of calcifying cartilage and which are morphologically related to the deposition of mineral in this tissue. The matrix vesicles and cells of the epiphyses of the long bones of fetal calves were liberated by digestion with collagenase and separated by differential centrifugation. Vesicle preparations were found to be lipid-rich and to contain significantly more sphingomyelin and phosphatidyl serine than cellular fractions. The ratio, moles cholesterol to moles total phospholipid, was higher in vesicles than in cells, a finding which is consistent with vesicles having their origins in the plasma membranes of the chondrocytes.
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  • 23
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    Calcified tissue international 10 (1972), S. 223-237 
    ISSN: 1432-0827
    Keywords: Ageing ; Cartilage ; Glycosaminoglycans ; Osteoarthritis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les glycosaminoglycanes du cartilage articulaire des condyles fémoraux de 119 cas d'autopsies humaines ont été isolés et caractérisés par le chlorure de cetylpyridinium. En utilisant la même méthode de colonne, avec micro-échelle, les glycosaminoglycanes sont estimés quantitativement dans le cartilage articulaire normal, de la naissance à l'âge de 95 ans, ainsi que dans des cartilages ostéo-arthritiques. Dans le cartilage normal, le chondroitine sulfate se rencontre sous la forme de chondroitine-4-sulfate et chondroitine-6-sulfate, dont le rapport décroit avec l'âge. Le chondroitine sulfate a un rapport sulfate/acide hexuronique en dessous de l'unité pour les moins de vingt ans et supérieur à l'unité dans les groupes d'âges plus élevés. Deux types de kératane sulfate et d'acide hyaluronique sont isolés dans tous les groupes. Les glycosaminoglycanes de cartilage ostéoarthritique ne montrent pas de différence avec le cartilage normal pour la quantité ou le rapport des isomères. Dans le cartilage normal, on note une décroissance significative des hexosamines totales de la naissance à l'âge de 11–20 par suite de la réduction de chondroitine sulfate. Chez l'adulte, un rapport en voie d'augmentation du kératosulfate plus la fraction glycoprotéique du chondroitine sulfate est observé. Le cartilage ostéo-arthritique montre une réduction de toutes les fractions glycosaminoglycanes sans changement de distribution. Le cartilage normal d'articulations ostéo-arthritiques est inchangé. Des courbes de solubilité indiquent une décroissance en P.M. du chondroitine sulfate dans le cartilage normal de la naissance à 21–30 ans, et une tendance vers des P.M. moins élevés dans le cartilage ostéoarthritique.
    Abstract: Zusammenfassung Die Glycosaminoglycane im Gelenkknorpel der femoralen Condylen von 119 menschlichen Autopsien wurden isoliert. Nachträglich wurden sie mit Hilfe von Cetylpyridinchlorid charakterisiert. Unter Verwendung derselben Säulentechnik als Mikromethode wurden die Glycosaminoglycane quantitativ im Gelenkknorpel von Gesunden von der Geburt bis zum 95. Altersjahr sowie in osteoarthritischem Knorpel gemessen. Im normalen Knorpel trat Chondroitinsulfat als Chondroitin-4-Sulfat und Chondroitin-6-Sulfat auf und das Verhältnis dieser beiden Substanzen nahm mit zunehmendem Alter ab. Unter dem 20. Altersjahr hatte das Chondroitinsulfat ein Sulfat/Hexuronsäure-Verhältnis kleiner als 1, in höheren Altersgruppen größer als 1. Zwei Typen von Keratansulfat und Hyaluronsäure wurden in allen Altersgruppen isoliert. Die Glycosaminoglycane von osteoarthritischem Knorpel zeigten keine Unterschiede zu denjenigen des normalen Knorpels in bezug auf Menge oder Verhältnis der Isomere. Im normalen Knorpel zeigte sich eine signifikante Abnahme der totalen Hexosamine von der Geburt bis zum 11–20. Altersjahr, welche auf eine Verminderung von Chondroitinsulfat zurückzuführen ist. Bei Erwachsenen wurde ein zunehmendes Verhältnis der Keratansulfat + Glycoproteinfraktion von Chondroitinsulfat beobachtet. Osteoarthritischer Knorpel zeigte einen Rückgang aller Glycosaminoglycanfraktionen ohne Änderung der Verteilung. Normaler Knorpel aus osteoarthritischen Gelenken war unverändert. Löslichkeitsprofile deuteten auf vermindertes Molekulargewicht von Chondroitinsulfat in normalem Knorpel von der Geburt bis zum 21.–30. Altersjahr und auf eine Tendenz zu niedrigerem Molekulargewicht in osteoarthritischem Knorpel.
    Notes: Abstract The glycosaminoglycans in articular cartilage from the femoral condyles of 119 human autopsy cases have been isolated and subsequently characterized using cetylpyridinium chloride. Using the same column method in microscale the glycosaminoglycans were estimated quantitatively in normal articular cartilage from birth to 95 years of age and in osteoarthritic cartilage. In normal cartilage chondroitinsulphate occurred as chondroitin-4-sulphate and chondroitin-6-sulphate, decreasing in ratio with age. The chondroitinsulphate had a sulphate/hexuronic acid ratio below unity, below 20 years, above unity in the higher age groups. Two types of keratansulphate and hyaluronic acid were isolated. The glycosaminoglycans isolated from osteoarthritic cartilage showed no differences from normal cartilage as to the amount of their moities of chondroitin sulphate isomers. In normal cartilage there was a significant decrease of total hexosamines from birth to 11–20 years, due to a reduction of chondroitinsulphate. In adults, an increasing ratio of the keratansulphate plus glycoprotein fraction to chondroitinsulphate was observed. Osteoarthritic cartilage showed a significant reduction of all glycosaminoglycan fractions without change of distribution. Normal cartilage taken from osteoarthritic joints was unaltered. Solubility profiles indicated decreases in molecular weight of chondroitin sulphate in normal cartilage from birth to 21–30 years, and a trend towards lower molecular weight in osteoarthritic cartilage.
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    Calcified tissue international 12 (1973), S. 16-29 
    ISSN: 1432-0827
    Keywords: Bone ; Cartilage ; Culture ; Glycosaminoglycans ; Sephadex ; Collagen ; Electron Microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des cultures de monocouches cellulaires d'épiphyses fémorales distales d'embryons de veaux sont étudiées immédiatement après la première sub-culture, réalisée après culture primaire. L'examen au microscope optique classique montre des dépôts disséminés de matériel métachromatique: en microscopie électronique, de fines fibrilles, considérées comme des fibres de collagène, en voie de développement, ont été observées. La nature des sécrétions cellulaires a été étudiée à l'aide de précurseurs radioactifs, fractionnés sur Sephadex G200, ainsi qu'à l'aide de chromatographie par échange d'ions. La digestion enzymatique par hyaluronidase et chondroitinase AC et ABC, bactérienne et testiculaire, démontre que les cellules synthétisent 70% de glycosaminoglycanes sulfatés et 30% de glycosaminoglycanes non sulfatés. Parmi les glycosaminoglycanes, 70% sont constitués par du chondroitine-4-sulfate, 20% par du chondroitine-6-sulfate et le reste probablement par du keratane-sulfate. Les études avec des acides aminés marqués indiquent que les cellules synthétisent une protéine de poids moléculaire élevé, contenant de l'hydroxyleproline, ainsi qu'une protéine non-collagénique, mise en évidence par incorporation de tryptophane.
    Abstract: Zusammenfassung Einschichtige Zellkulturen von der distalen Femurepiphyse von Kalbsembryonen wurden nach der ersten Subkultur untersucht, welche nach der Verschmelzung der primären Kulturen angesetzt wurde. Die lichtmikroskopische Untersuchung zeigte verstreute Ablagerungen von metachromatisch gefärbtem Material; bei der Elektronenmikroskopie wurden feine Fibrillen festgestellt, welche als Collagenvorstufe angesehen wurden. Nach einer Kultur von mehreren Tagen zeigte sich eine lacunäre Anordnung der Zellen. Die Beschaffenheit der Zellsekretionen wurde mittels radioaktiv markierten Verbindungen untersucht, welche auf Sephadex G200 und durch Ionenaustausch-Chromatographie fraktioniert wurden. Die enzymatische Verdauung mit bakterieller und testikulärer Hyaluronidase und Chondroitinase-AC und-ABC zeigte, daß die Zellen 70% sulfatierte und 30% nichtsulfatierte Glycosaminoglycane synthetisierten. Die sulfatierten Glycosaminoglycane enthielten 70% Chondroitin-4-Sulfat, 20% Chondroitin-6-Sulfat, und der Rest war wahrscheinlich Keratansulfat. Untersuchungen mit markierten Aminosäuren-Vorläufern deuteten darauf hin, daß die Zellen ein Hydroxyprolinhaltiges, hochmolekuläres Protein synthetisierten und dazu etwas Eiweiß, bei dem es sich auf Grund des Tryptophaneinbaues nicht um Collagen handeln kann.
    Notes: Abstract Monolayer cell cultures from the distal femoral epiphyses of embryo calves were studied following the first subculture, which was carried out after confluence in primary culture. Light microscopic examination revealed scattered deposits of metachromatic-staining material; on electron microscopy fine fibrils considered to be developmental collagen were seen. After several days in culture lacuna-like patterns of cells were seen. The nature of the cell secretions were studied by radioactive precursors, which were fractionated on Sephadex G200 and by ion exchange chromatography. Enzyme digestion with bacterial and testicular hyaluronidase and chondroitinase-AC and-ABC revealed that the cells synthesized 70% sulphated, and 30% non-sulphated glycosaminoglycans. Of the sulphated glycosaminoglycans 70% was chondroitin-4-sulphate, 20% chondroitin-6-sulphate, and the remainder probably keratansulphate. Studies were labelled amino acid precursors suggested that the cells synthesized a high-molecular weight protein containing hydroxyproline, as well as some non-collagenous protein, shown by tryptophan incorporation
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    Calcified tissue international 13 (1973), S. 83-92 
    ISSN: 1432-0827
    Keywords: Proteoglycan ; Collagen ; Cartilage ; Electron Microscopy ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé L'élimination de protéoglycans solubles de coupes de cartilage costal de boeuf, par extraction dans une solution de 4M d'hydrochlorure de guanidinium, permet de mettre en évidence des quantités abondantes de collagène dispersé et désagrégé dans la matrice. Les protéoglycanes, résistants à l'extraction, sont visibles sous forme de granules concentrés dans les régions périlacunaires. Les granulations plus importants des protéoglycanes semblent venir du chondrocyte. Dans la matrice, éloignée des chondrocytes, ces granules deviennent plus étroites. Un composant non granulaire “amorphe” masque les fibres de collagène, de telle sorte qu'elles sont difficilement visibles dans le cartilage intact.
    Abstract: Zusammenfassung Die löslichen Proteoglycane wurden mittels Extraktion in 4 M Guanidinhydrochlorid aus Rippenknorpelschnitten des Rindes entfernt. Dies erlaubte die Sichtbarmachung von großen Mengen von verstreuten und auseinandergerissenen Collagen in der Matrix. Die Protoglycane, welche sich nicht extrahieren lassen, erscheinen als kleine, in den perilacunären Regionen konzentrierte Körnchen. Die großen Proteoglycan-Körner scheinen in den Chondrocyten zu entstehen. Sobald sie sich in die Matrix, außerhalb der Chondrocyten, verlagern, werden die Körner kleiner. Ein nicht-granulärer, „amorpher” Bestandteil verhüllt die Collagenfasern, so daß diese im intakten Knorpel nicht deutlich gesehen werden können.
    Notes: Abstract Removal of the soluble proteoglycans from slices of bovine costal cartilage by extraction in 4 M guanidinium hydrochloride permitted the visualization of abundant amounts of dispersed and disaggregated collagen in the matrix. Proteoglycans which are resistant to extraction are seen as small granules which are concentrated in the perilacunar regions. Large proteoglycan granules appear to originate in the chondrocyte. As they come to occupy positions in the matrix distant from the chondrocyte, the granules become smaller. A non-granular, “amorphous” component masks the collagen fibers so that they cannot be readily seen in the intact cartilage.
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    Calcified tissue international 14 (1974), S. 31-48 
    ISSN: 1432-0827
    Keywords: Bone ; Cartilage ; Calcification ; Collagen ; Phosphate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Une méthode pour l'étude de la croissance des os longs de foetus de rat, en culture d'organe, dans un milieu chimiquement défini, a été mise au point. Les extrémités cartilagineuses et les parties centrales de l'os sont analysées séparément pour leur croissance et minéralisation en étudiant leur contenu en collagène, calcium et phosphate, poids sec, et incorporation de proline marquée en hydroxyproline. La croissance et la minéralisation des parties centrales osseuses sont plus lentes dans un milieu chimiquement défini qu'in vivo. La croissance peut être accélérée en ajoutant au milieu des acides aminés non essentiels, de l'albumine ou du sérum. Les extrémités cartilagineuses présentent une augmentation plus importante en poids et contenu en collagène que les parties centrales et l'adjonction de diverses substances a moins d'effet sur la croissance. La croissance et la minéralisation des parties centrales sont augmentées en élevant la concentration du milieu en phosphate de 1.5 à 4.5 mM, avec ou sans adjonction de sérum ou d'albumine. A une concentration faible de calcium (0.5 mM), la croissance et la minéralisation des parties centrales sont arrêtées. A une concentration faible en magnésium (0.5 mM), la minéralisation est augmentée, mais la croissance est arrêtée.
    Abstract: Zusammenfassung Es wird eine Methode beschrieben, mit welcher das Wachstum der Röhrenknochen von Rattenembryos in einem chemisch bestimmten Medium in Organkultur untersucht werden kann. Die Knorpelenden und Knochenschäfte wurden gesondert auf Wachstum und Mineralisation geprüft, indem Collagen-, Calcium- und Phosphatgehalt, das Trockengewicht und der Einbau von markiertem Prolin in Hydroxyprolin gemessen wurden. Wachstum und Mineralisation des Knochenschaftes waren langsamer in einem chemisch bestimmten Medium als in vivo. Das Wachstum konnte beschleunigt werden, indem dem Medium nicht-essentielle Aminosäuren, Albumin oder Serum beigegeben wurden. Die Knorpelenden zeigten eine viel stärkere Zunahme an Gewicht und Collagengehalt als die Schäfte, und Anreicherung des Mediums hatte weniger Wirkung auf ihr Wachstum. Das Wachstum und die Mineralisation der Knochenschäfte nahmen zu, wenn die Phosphatkonzentration im Medium zwischen 1,5 und 4,5 mM erhöht wurde, und zwar unabhängig davon, ob dem Medium Serum oder Albumin beigegeben wurde oder nicht. Bei niederer Calciumkonzentration (0,5 mM) im Medium wurden Wachstum und Mineralisation der Knochenschäfte beeinträchtigt. Bei niedriger Magnesiumkonzentration (0,5 mM) wurden die Mineralisation erhöht, das Wachstum hingegen gehemmt.
    Notes: Abstract A method for studying the growth of fetal rat long bones in a chemically defined medium in organ culture is described. Cartilage ends and bone shafts were analyzed separately for growth and mineralization by measuring the collagen, calcium, and phosphate content, dry weight, and incorporation of labeled proline into hydroxyproline. Growth and mineralization of the bone shaft were slower in a chemically defined medium thanin vivo. Growth could be enhanced by supplementation of the medium with non-essential amino acids, albumin or serum. Cartilage ends showed a greater increase in weight and collagen content than the shafts, and medium supplements had less effect on their growth. Bone shaft growth and mineralization were enhanced by increasing medium phosphate concentration over a range of 1.5 to 4.5 mM whether or not the medium was supplemented with serum or albumin. At a low medium calcium concentration (0.5 mM) bone shaft growth and mineralization were impaired. At a low magnesium concentration (0.5 mM) mineralization was enhanced, but growth was impaired.
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    Calcified tissue international 14 (1974), S. 49-58 
    ISSN: 1432-0827
    Keywords: Glycosaminoglycans ; Intracellular ; Chondrocytes ; Cartilage ; Embryonal ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les glycosaminoglycanes acides de chondrocytes isolés, en suspension, sont étudiés. La matrice extracellulaire est isolée par traitement successif à la trypsine et à la collagénase d'os d'embryons de poulet, âges de 15 à 17 jours. Après digestion à la papaïne et élimination des acides nucléïques par traitement à la DNAse et à la RNAse, les glycosaminoglycanes sont précipités par le CPC et isolés sous la forme de leur sel sodique. Les analyses des propriétés de solubilité du CP glycosaminoglycane par le microfractionnement d'Antonopoulos et coll. (1964) révèlent la présence de glycosaminoglycanes intracellulaires de poids moléculaire et/ou de densité de charge nettement plus faibles que ceux de la matrice extracellulaire. Sur des électrophorèses de microzones, une petite partie des glycosaminoglycanes intracellulaires isolées présente une mobilité similaire à celle des solutions témoins de sulfate de chondroitine, alors que la portion la plus importante ne se déplace qu'à mi-distance de l'acide hyaluronique et montre une large trainée, indiquant une faible charge négative. Ce fait semble dû à une perte de l'influence de régulation des composants matriciels éliminées, sur la synthèse des glycosaminoglycanes et au rejet dans le milieu de glycosaminoglycanes initialement synthétisé. Pour la première fois, une faible quantité d'acide hyaluronique intracellulaire est mise en évidence par une mobilité électrophorétique typique.
    Abstract: Zusammenfassung Saure Glykosaminoglycane von isolierten Chondrocyten in Suspension wurden untersucht, und die extracelluläre Matrix von 15–17 Tage alten Knochen von Kükenembryos wurde durch die aneinanderfolgende Behandlung mit Trypsin und Collagenase erhalten. Nach der Papain-Verdauung und Beseitigung der Nucleinsäuren mittels DNS- und RNSase-Behandlung wurden die Glykosaminoglycane mit CPC gefällt und als Natriumsalz isoliert. Untersuchungen der Löslichkeit der CP-Glykosaminoglycane mittels des Mikrofraktionierungsverfahrens von Antonopouloset al. (1964) zeigten intracelluläre Glykosaminoglycane mit meist niedrigerem Molekulargewicht und/oder niedrigerer Ladungsdichte als diejenigen der extracellulären Matrix. In der Mikro-Zonen-Elektrophorese zeigte nur ein kleiner Teil der isolierten intracellulären Glykosaminoglycane eine Mobilität, die dem Standard des Chondroitinsulfats entsprach. Der größte Teil legte nur die halbe Distanz der Hyaluronsäure zurück und zeigte breites “Tailing”, was auf eine niedrigere negative Ladung hindeutet. Die Ursache dafür wurde interpretiert als Verlust eines regulierenden Einflusses der entfernten Matrixkomponenten auf die Glykosaminoglycan-Synthese und als Abgabe ursprünglich synthetisierter Glykosaminoglycane in das Medium. Es wurde erstmals, anhand der typischen elektrophoretischen Beweglichkeit, eine kleine Menge von intracellulärer Hyaluronsäure nachgewiesen.
    Notes: Abstract Acid glycosaminoglycans were studied from isolated chondrocytes in suspension and the extracellular matrix obtained by sequential trypsin and collagenase treatment of 15- to 17-day-old embryonic chick bone. After papain digestion and removal of the nucleic acids by treatment with DNAse and RNAse, the glycosaminoglycans were precipitated by CPC and isolated as their sodium salt. Analyses of the CP-glycosaminoglycan solubility properties with the microfractionation procedure of Antonopouloset al. (1964) showed intracellular glycosaminoglycans of predominantly lower molecular weight and/or charge density than those of extracellular matrix. On micro-zone electrophoresis, only a minor part of the isolated intracellular glycosaminoglycans showed a mobility similar to that of the chondroitin sulphate standards while the major part moved only half the distance of hyaluronic acid and showed broad tailing, indicating a low negative charge. This was considered to be due to loss of a regulatory influence of the removed matrix components upon glycosaminoglycan synthesis and to release of originally synthesized glycosaminoglycans into the medium. For the first time, a small amount of intracellular hyaluronic acid was shown by typical electrophoretic mobility.
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    Calcified tissue international 2 (1968), S. 157-164 
    ISSN: 1432-0827
    Keywords: Protein-polysaccharide Synthesis ; Epiphyseal Plate ; Cartilage ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Une méthode de séparation des zones cellulaires de prolifération, de maturation et d'hypertrophie de la métaphyse épiphysaire du jeune lapin est mise au point, de façon à pouvoir étudier le métabolisme protéino-polysaccharidiquein vitro sur une base cellulaire. En utilisant l'incorporation de35SO4 comme index de la synthèse protéino-polysaccharidique et la concentration en hexosamine comme index du contenu en protéine-polysaccharide par zone, il apparait que la concentration en sulfate varie inversement avec le contenu en matrice protéino-polysaccharidique: elle est la plus élevée dans la couche des cellules hypertrophiques où le contenu en matrice est le plus bas. La signification possible de ce rapport est envisagée.
    Abstract: Zusammenfassung Es wird eine Methode zur Trennung der proliferativen, reifenden und hypertrophen Zellzonen der Epiphysenwachstumsplatte beim entwöhnten Kaninchen besprochen, die es ermöglicht, den Protein-Polysaccharid-Metabolismusin vitro für jeden Zelltyp gesondert zu untersuchen. Als Index der Protein-Polysaccharid-Synthese wurde die zeitbedingte Aufnahme von35SO4-Ionen verfolgt, während als Index des Protein-Polysaccharidgehaltes der Zonen die Hexosaminkonzentration gemessen wurde. Bei diesem Vorgehen wurde gefunden, daß die Sulfataufnahme entgegengesetzt zum Protein-Polysaccharidgehalt der Matrix variiert, indem sie die höchsten Werte in der hypertrophen Zellschicht auftweis, wo der Matrixgehalt am kleinsten ist. Die Bedeutung dieses Verhältnisses wird besprochen.
    Notes: Abstract A method is presented for separating the proliferative, maturing and hypertrophic cell zones of the epiphyseal growth plate of the weanling rabbit, in such a manner that protein-polysaccharide metabolism could be studiedin vitro on a per-cell basis. Using the timed incorporation of35SO4 as an index of protein-polysaccharide synthesis, and hexosamine concentration as an index of protein-polysaccharide content per zone, it was found that sulfate uptake varied inversely with matrix protein-polysaccharide content; being highest in the hypertrophic cell layer where matrix content was least. The possible significance of this relationship is discussed.
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    Calcified tissue international 22 (1977), S. 67-75 
    ISSN: 1432-0827
    Keywords: Cartilage ; Mineralization ; Hydroxyapative ; Invertebrates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract In vitro mineralization of the gill cartilage ofLimulus (horse-shoe crab) has been reported previously (Eilberg et al. 1975). The present study demonstrates that cranial cartilage ofLoligo (squid) and odontophore cartilage ofBusycon (marine snail) also mineralize in vitro in hydroxypatite-metastable media, but not in carbonate-metastable media. In all three of these cartilages, light phase-contrast microscopy revealed that the mineral phase occurred in the form of spherical or ovoid granules ranging between 2 and 9 μm in diameter. During mineralization, the granules appeared successively in the perichondrium, in the matrix, and finally, within chondrocytes. Mineralization occurred more rapidly inBusycon odontophore cartilage which has a significantly higher content of phosphatidyl serine than inLimulus gill orLoligo head cartilages. In all tissues the mineralization process is related to temperature, taking place most actively at 37°C and only weakly at 50°C.
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    Calcified tissue international 24 (1977), S. 37-39 
    ISSN: 1432-0827
    Keywords: Cartilage ; Matrix vesicles ; Mitochondria ; Respiratory enzymes
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    Topics: Biology , Medicine , Physics
    Notes: Summary This study attempted to detect evidence of mitochondrial terminal respiratory components in matrix vesicles isolated from rachitic rat tibial epiphyseal plates. Biochemical assays for cytochrome c oxidase, NAD isocitrate dehydrogenase, NADP isocitrate dehydrogenase and succinate-cytochrome c reductase were negative. Polarimetric determinations revealed that the addition of succinate to matrix vesicles in suspension did not cause any increase in oxygen utilization. Spectrophotometric tracings of deoxycholate-solubilized matrix vesicles showed no characteristic absorption peaks or maxima belonging to any of the cytochrome complex components. Attempts to prepare pyridine hemochromes of cytochrome prosthetic groups from the matrix vesicles were also unsuccessful. The above results indicate that key components of mitochondrial respiratory systems are not detectable in rachitic matrix vesicles. The results are compatible with the interpretation that such vesicles are not derived from mitochondria.
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    Calcified tissue international 27 (1979), S. 269-273 
    ISSN: 1432-0827
    Keywords: Cartilage ; Keratan sulfate ; Heparan sulfate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Biosynthesis of chondroitinase-resistant glycosaminoglycans as minor components was studied in the cartilages of chick embryos and of newborn chickens and rats. Sternal and knee cartilages were labeled in vitro with35SO 4 2− , and then35S-labeled glycosaminoglycans were analyzed. In rats up to 2 weeks old, only one glycosaminoglycan could be detected as heparan sulfate. In the chick embryos and the newborn chickens, however, keratan sulfate as well as heparan sulfate could be detected. As chondroitinase-sensitive glycosaminoglycans, large amounts of both chondroitin 4- and 6-sulfates were synthesized in the chick cartilage, but the synthesis of chondroitin 6-sulfate could scarcely be seen in the rat cartilage. The results seem to indicate that the biosynthesis of keratan sulfate has some relation to that of chondroitin 6-sulfate.
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    Calcified tissue international 27 (1979), S. 275-279 
    ISSN: 1432-0827
    Keywords: Proline ; Cartilage ; Bone
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    Notes: Summary Proline biosynthetic and degradative enzymes are unevenly distributed in differentiated mammalian tissues. Activities of the synthetic enzymes are relatively high in collagenous tissues, whereas activities of the degradative enzymes are high in noncollagenous tissues. In order to further characterize tissue-specific proline biosynthesis and degradation, we have determined proline enzyme activities during cartilage and bone formation induced by demineralized bone matrix. We can thus follow temporal changes in enzyme activity in a single tissue as different cell types develop. Ornithine aminotransferase and pyrroline-5-carboxylate reductase have peaks of activity which correlate with maximal type II collagen synthesis by chondrocytes. Both enzymes also are active during bone formation. In contrast, proline oxidase and pyrroline-5-carboxylate dehydrogenase are present at low levels and do not change as new cell types appear. Arginase activity peaks during the first 3 days and then rapidly decreases by the time cartilage and bone formation begin. These observations further substantiate the importance of proline biosynthesis in collagenous tissues. The close correlation between ornithine aminotransferase activity and type II collagen synthesis suggests that the pathway from ornithine to proline may be especially important during formation of type II collagen.
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    Calcified tissue international 5 (1970), S. 56-63 
    ISSN: 1432-0827
    Keywords: Epiphysis ; Cartilage ; Morphology ; Fluorescein ; Anatomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Court term d'incubation des cultures d'un organe du cartilage scapulaire chez le jeune chien dans un milieu raffiné avec la lysozyme ou la protamine produit des histologiques changements spécifiques dans les proprietés de la coloration du l'explant qui n'est pas produit par l'albumine ou par la lysozyme inactivée par les rayons ultraviolets. Ces modifications sont différent avec chacune des protéines. L'effet de la lysozyme, est précisement distributé tandis que la protamine est diffusée. Si on use la flouresceine avec les protéins, les même changements histologiques se produisent dans le cartilage et la flourescence c'est repandue précisement au même endroit, où les propriétés colorant du tissue ont été changé. L'observation qui montre que la lysozyme dans des spécifiques points anatomiques suggère qu'elle réagit chimiquement avec les constituants spécifiques de la matrice cartilagineuse, probablement avec des macromolécules négativement chargées.
    Abstract: Zusammenfassung lysozym und Protamin angereicherte Gewebekulturen des wachsenden Hundeschulterblattes erzeugen nach kurzer Zeit spezifische Veränderungen in der histologischen Anfärbbarkeit des Explantats, die jedoch nicht durch Albumin oder durch Lysozym, das durch Ultraviolettbestrahlung inaktiviert wurde, induziert werden können. Die hervorgerufenen Veränderungen sind für beide Proteine verschieden. Der Lysozymeffekt ist anatomisch genau verteilt, während der Protamineffekt zerstreut erscheint. Fluoresceinkonjugate dieser Proteine erzeugen identische Veränderungen im Knorpel, wobei die Verteilung der Fluorescenz mit der der Veränderungen der histologischen Anfärbbarkeit übereinstimmt. Die Beobachtungen zeigen, daß durch die Zugabe von Lysozym zu Gewebekulturen dieses Protein an speziellen anatomischen Stellen angereichert wird. Es kann daraus geschlossen werden, daß Lysozym chemisch sich mit spezifischen Komponenten des Knorpels verbindet, wahrscheinlich den negativ geladenen Makromolekülen.
    Notes: Abstract Short-term incubation of organ cultures of puppy scapulae in a medium enriched with lysozyme or protamine induces specific alterations in the histologic staining properties of the explant not induced by albumin or by ultraviolet-inactivated lysozyme. The induced changes are different for each two proteins. The lysozyme effect is anatomically distributed precisely while that of protamine is diffuse. If fluorescein conjugates of the proteins are used, identical histologic changes are induced in the cartilage and the distribution of fluorescence is precisely that of the altered tinctorial properties of the tissue. The observation, which shows that lysozyme, when added to cartilage, accumulates in specific anatomic sites suggests that it chemically interacts with specific components of cartilage matrix, presumably with negatively charged macromolecules.
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  • 34
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    Calcified tissue international 5 (1970), S. 170-182 
    ISSN: 1432-0827
    Keywords: Cartilage ; Bone ; Gonadectomy ; Histochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des coupes de tibias de rats mâles et femelles, gonadectomisés et témoins, sont soumises à divers examens histologiques et histochimiques pour étudier les effets de la gonadectomie au niveau du tissu osseux. Le cartilage épiphysaire proximal des rats castrés et des rattes, ayant subi l'ablation des ovaires, est oblitéré par de l'os 5 mois après l'intervention. En outre, la diaphyse présente une ostéoporose. La matrice intercellulaire du cartilage épiphysaire des rats gonadectomisés est fortement PAS positive, contrairement à la réaction fortement alcian positive de la matrice des animaux témoins. Les travées diaphysaires des rats gonadectomisés se colorent en rouge avec le PAS et contiennent des régions étendues d'ilôts cartilagineux calcifiés. En outre, le cartilage épiphysaire et les travées osseuses de ces animaux se colorent faiblement à la ninhydrine-Schiff. Les réactions colorées suggèrent que la gonadectomie induit une altération dans le degré d'agrégation protéino-mucopolysaccharidique de la matrice organique; ce qui interfère probablement avec la minéralisation osseuse.
    Abstract: Zusammenfassung Tibiaschnitte von unbehandelten und kastrierten männlichen und weiblichen Ratten wurden mit verschiedenen histologischen und histochemischen Techniken untersucht, um die Knochenveränderungen im Anschluß an die Gonadektomie zu bestimmen. Es wurde beobachtet, daß sowohl beim kastrierten Männchen wie beim ovarektomierten Weibchen der proximale Epiphysenknorpel nach dem fünften postoperativen Monat durch Knochen „eingekapselt” ist. Dazu wurde auch eine Osteoporese des Knochenschaftes beobachtet. Der Epiphysenknorpel der kastrierten Ratten zeigte eine intensiv PAS-positive interzelluläre Matrix, im Gegensatz zu der alcianophil reagierenden Matrix von Kontrolltieren. Die Diaphysentrabekel von kastrierten Ratten färbten sich mit PAS rot und enthielten große Bezirke von mineralisierten Knorpelkernen. Dazu färbten sich der Epiphysenknorpel und die Trabekel dieser Tiere nur schwach mit der Ninhydrin-Schiff-Reaktion. Die Resultate der verschiedenen Färbungen zeigten, daß nach Gonadektomie eine Veränderung im Aggregatzustand der Protein-Mucopolysaccharidkomponente der organischen Matrix stattfindet, was zu einer Interferenz im Knochenmineralisationsprozeß führen könnte.
    Notes: Abstract Sections of the tibias from both intact and gonadectomized male and female rats were subjected to various histological and histochemical methods to determine the changes in the bone following gonadectomy. It was observed that both in castrated males and in ovariectomized females, the proximal epiphyseal cartilage became “sealed off” by bone after the fifth post-surgical month. In addition osteoporosis of the shaft of the bone was also observed. The epiphyseal cartilage of the gonadectomized rats exhibited intense PAS positive intercellular matrix in contrast to the alcianophilic ature of the matrix from control animals. The diaphyseal trabeculae from gonadectomized rats stained red with PAS and contained wide regions of calcified cartilage cores. In addition, the epiphyseal cartilage and bony trabeculae from these animals stained faintly in the ninhydrin-Schiff reaction. The staining reactions indicated that in gonadectomy there was an alteration in the state of aggregation of protein-mucopolysaccharide component of the organic matrix, a situation which would result in an interference in the mineralization of the bone.
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  • 35
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    Calcified tissue international 55 (1994), S. 100-102 
    ISSN: 1432-0827
    Keywords: Synovial fluid ; Glycoprotein ; Cartilage ; BCP crystals
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract As cartilage matrix glycoprotein (CMGP) is a prominent matrix constituent, we analyzed the relationship of levels in plasma (CMGPP) and synovial fluid (CMGPS) to each other, to clinical diagnosis, and to degree of radiographic cartilage degeneration. CMGP was measured in matched synovial fluid and plasma specimens from 67 patients with various forms of arthritis using an ELISA technique. CMGPS consistently exceeded CMGPP, CMGPP levels correlated significantly with CMGPS levels, and CMGP retention in joint fluid, as calculated by the ratio CMGPS: CMGPP, was significantly higher in patients whose synovial fluids contain basic calcium phosphate crystals. No correlation of CMGPP or CMGPS with diagnosis or degree of catilage degeneration was observed. CMGP measurements are not useful diagnostically in patients with chronic arthritis and do not predict degree of radiographic degeneration. The association of basic calcium phosphate crystals with intraarticular retention of CMGP warrants further study.
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  • 36
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    Calcified tissue international 6 (1970), S. 113-119 
    ISSN: 1432-0827
    Keywords: Lipid ; Bone ; Cartilage
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Le contenu et la composition des lipides du tissue épiphysaire ont été étudiés chez le poulet. Les valeurs des lipides totaux, des triglycérides et des phospholipides sont, respectivement dans l'os, de 7,50, 1,81 et 1,02 mg/g et, dans le cartilage, de 5,20, 1,46 et 0,53 mg/g. Les principaux acides gras trouvés dans l'os aussi bien que dans le cartilage sont les acides palmitique et oléique, parmi les phospholipides, et les acides palmitique, oléique et linoléique parmi les triglycérides. Quant aux substrats étudiés, l'ordre d'incorporation dans les lipides totaux de l'os et du cartilage a été: palmitate 〉 glucose 〉 acétate 〉 citrate. L'acétate était le principal précurseur dans la synthèse des acides gras, alors que seule une faible protion de glucose a été trouvée dans les acides gras. L'estérification semble donc être la voie prédominante de la synthèse des lipides dans l'os et le cartilage du poulet.
    Abstract: Zusammenfassung Es wurde eine Untersuchung über den Gehalt und die Zusammensetzung der Lipide im Epiphysengewebe von Hühnchen durchgeführt. Es wurden 7,50 mg Gesamtlipide, 1,81 mg Triglyceride und 1,02 mg Phospholipide per Gramm Knochen gefunden. Die entsprechenden Werte per Gramm Knorpel waren 5,20, 1,46 und 0,53 mg. Die hauptsächlichsten Fettsäuren in den Phospholipiden von Knochen- und Knorpelgewebe waren Palmitin- und Ölsäure, in den Triglyceriden Palmitin-, Öl- und Linolsäure. Die untersuchten Verbindungen wurden in nachstehender Reihenfolge in Knochen- und Knorpel-Lipide eingebaut: Palmitin 〉 Glucose 〉 Acetat 〉 Citrat. Acetat war die hauptsächliche Ausgangssubstanz für die Fettsäure-Synthese, während nur ein unbeträchtlicher Teil der Glucose in den Fettsäuren vorgefunden wurde. Veresterung ist anscheinend der vorherrschende Weg der Fettsynthese in Knorpel- und Knochengewebe von Hühnchen.
    Notes: Abstract The content and composition of lipids of chick epiphyseal tissue were investigated. The amounts of total lipids, triglycerides and phospholipids in bone were 7.50, 7.81 and 1.02 mg/g, respectively, and in cartilage, 5.20, 1.46 and 0.53 mg/g, respectively. The main fatty acids of both bone and cartilage were palmitic and oleic in phospholipids, and palmitic, oleic and linoleic in triglycerides. For the substrates studied, the order of incorporation into total lipids of both bone and cartilage was palmitate 〉 glucose 〉 acetate 〉 citrate. Acetate was the main precursor for iatty acid synthesis whereas only a minor portion of glucose was found in the fatty acids. Esterification appeared to be the predominant pathway of lipid synthesis in chick bone and cartilage.
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  • 37
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    Calcified tissue international 9 (1972), S. 194-199 
    ISSN: 1432-0827
    Keywords: Growth plate ; Glycosaminoglycans ; Chondroitin sulfate ; Keratan sulfate ; Cartilage
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les concentrations de chondroitine-4-sulfate (C-4-S), chondroitine-6-sulfate (C-6-S) et kératane sulfate (KS) sont déterminées au niveau du cartilage métaphysaire provenant de lapins de 2 groupes d'âge, et de 3 zones métaphysaires de lapins d'âge identique. Avec l'augmentation de l'âge, la concentration de C-4-S augmente, C-6-S décroit et KS reste constant. Il n'y a pas de variation lorsque trois métaphyses différentes sont étudiées. Le radiosulfate qui se fixe comme chondroitane sulfate est aussi déterminé: on note ainsi un rapport inverse de la concentration en fonction de l'augmentation de l'âge.
    Abstract: Zusammenfassung Im Knorpel aus Wachstumsplatten von Kaninchen zweier verschiedener Alter wurden das Chondroitin-4-Sulfat (C-4-S), das Chondroitin-6-Sulfat (C-6-S) und das Keratansulfat (KS) bestimmt; die selben Bestimmungen wurden auch in drei verschiedenen Zonen der Wachstumsplatte bei Kaninchen von gleichem Alter ausgeführt. Bei zunehmendem Alter stieg die C-4-S-Konzentration, die C-6-S-Konzentration nahm ab und die KS-Konzentration blieb konstant. Innerhalb der drei an verschiedenen Zonen der Wachstumsplatte vorgenommenen Bestimmungen fanden sich keine Unterschiede. Die Aufnahme von Sulfat (mit35S markiert) als Chondroitinsulfat wurde ebenfalls bestimmt; diese zeigte ein entgegengesetztes Verhältnis zwischen Konzentration und zunehmendem Alter.
    Notes: Abstract Chondroitin-4-sulfate (C-4-S), chondroitin-6-sulfate (C-6-S) and keratan sulfate (KS) concentrations were measured in growth plate cartilage derived from rabbits of two different ages, and from three growth plate zones from rabbits of the same age. With increasing age, the concentration of C-4-S increased, C-6-S decreased and KS remained constant. There was no variation when three different growth plate zones were sampled. Radiosulfate uptake into chondroitan sulfate was also measured, and this showed an inverse relationship to concentration with increasing age.
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  • 38
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    Calcified tissue international 7 (1971), S. 139-149 
    ISSN: 1432-0827
    Keywords: Bone ; Cartilage ; Estrogen ; Ultrastructure ; Growth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Dix rats Holtzman mâles et sevrés sont sacrifiés injection intrapéritonéale d'oestradiol (Progynon, Schering) aqueux, à des doses quotiediennes de 1 μ g. par g de poids. Des témoins, ayant reçu une dose équivalente de liquide de dilution, sont sacrifiés à des intervalles de 1 heure à 6 jours, identiques aux temps de sacrifice des animaux injectés. Les cartilages épiphysaires supérieurs des tibias tibias (ECP) étudiés en microscopie électronique, montrent, dès trois heures après l'ionjection, une augmentation nette de 'activié sécrétoire, caractérisée, au niveau de la zone de sécrétion matricielle, par l'abondance dans les citernes golgiennes d'un matériel piqueté, constitué par des complexes protéino-polysaccharidiques. La désintégration de la membrane limitante de vésicules golgiennes individuelles est plus avancée après vingt quatre heures: après trois jours de traitement, seules quelques vésicules restent intactes et des plages d'un matériel initialement intravacuolaire sont visibles dans le cytoplasme. De longs filaments, rappelant les précurseurs ou les fibrilles primaires du collagène, sont visibles dans cette sécrétion. Après six jours, de grandes plages de cettre subestance remplissent les cellules de la couche pré-hypertrophieque, avec déplacement de l'ergastoplasme en périphérie. Des vacuoles cytoplasques, contenant un matériel semblable à celui qu'on retrouve dans la lacune, et présentant des filament finement moniliformes et disposés en rayons le long de la membrane limitante, sont visibles. Ces observations suggèrent une accélération initiale de l'activité sécrétoire chondrocytaire, suivie par un retard de transfert. La rétention consécutive et la polymérisation intracellulaire de produits précollagéniques accélèrent l'hypertrophie et favorisent ainsi la dégénérescence précoce des chondrocytes. Ces altérations ultrastructurales paraissent être spécifiques aux oestrog`enes.
    Abstract: Zusammenfassung Zehn männliche Hotlzmann-Ratten, die im Entwöhnungsstadium waren, erhielten täglich wässerige Oestradioldosen (Progynon, Schering) von 1 μ/g Körpergewicht i.p. Dann wurden sie gleichzeitig mit Kontrolltieren, welche die gleiche Menge Verdünnungsmittel erhalten hatten, in Intervallen von 1 Std bis zu 6 Tagen getötet. Platten des oberen tibialen Epiphysenknorples (ECP), welche für die Elektronenmikroskopie präpariert wurden, zeigtem, daß schon 3 Std nach der Injektion ein bemerkenswerte Erhöhung der sekretorischen Tätigkeit entsteht. Dies wurde in der Zone der Matrixausscheidung sichtbar, wo sich in den Golgi-Zisternen eine Anhäufung von punktiertem, aus Proteinpolysaccharid-Komplexen bestehendem Material zeigte. Der Zerfall der Membran, welche die einzelnen Golgi-Bläschen umgibt, nahm nach 24 Std zu; nach 3 Tagen Behandlung blieben nur wenige Gefäße intakt, und Ansammlungen von ursprünglich intravacuolörem Material konnten im Grundplasma beobachtet werden. Lange Fasern, welche auf primäre oder Prae-Kollagefibrillen hindeuteten, konnten in diesem Sekret gesehen werden. Nach 6 Tagen wurden die Zellen in der prähypertrophen Zone mit dieser Substanz richtiggehend überschwemmt, und das rauhe endoplasmatische Reticulum wurde anschließend gegen die Zellperipherie verlagert. Die oft beobachteten cytoplasmatischen Vacuolen enthielten ein Material, das dem in den Lacunen vorkommenden ähnlich ist und zeigten auf der ungebrenden Membran feinperlige, radial angeordnete Fasern. Unsere Beobachtungen deuten auf eine anfängliche Beschleuning der chondrocytischen sekretorischen Tätigkeit, mit nachfolgender Transportverlangsamung, hin. Die dadurch entstehende Retention und intrazelluläre Polymerisation von präkollagenen Produkten beschleunigt die Hypertrophie und begünstigt dadurch die frühe Degeneration von Chondrocyten. Diese ultrastrukturellen Veränderungen scheinen oestrogen-spezifisch zu sein.
    Notes: Abstract Ten male weanling Holtzman rats, injected intraperitoneally with aqueous estradiol (Progynon, Schering), in daily doses of 1 μg. per g body weight, were sacrificed, simultaneously with controls receiving an equivalent amount of diluent, at intervals ranging from one hour to six days. Upper tibial epiphyseal cartilage plates (ECP), procesed for electron microscopy, revealed, as early as three hours after injection, appreciable enhancement of secretory activity, evidenced, in the zone of matrix secretion, by the abundance in Golgi cisternae of stippled material representing proteinpolysaccharide complexes. Disintegration of the lining membrane of individual Golgi vesicles was advanced after twenty-four hours; following three days of treatment, few vesicles remained intact, and pools of initially intravacuolar material were observable in the gound plasm. Long filaments, suggestive of primary or precursor collagen fibrils were apparent in this secretion. After six days, virtual lakes of this substance filled cells in the zone of prehypertophy, with consequent displacement of the rough endoplasmic reticulum against the cell periphery. Cytoplasmic vacuoles, containing mateerial similar to that found in the lacunar moat, and displaying finely beaded, radially arrayed filaments on the lining membrane were frequently encountered. Our observations suggest an initial acclleration of chondrocytic secretory activity, with subsequent retardation of transport. The resultant retention and intracellular polymerization of precollagenous products accelerates hypertrophy, thereby promoting early degeneration of chondrocytes. These ultrastructural alterations are apparently estrogen-specific.
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  • 39
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    Calcified tissue international 7 (1971), S. 220-231 
    ISSN: 1432-0827
    Keywords: Bicarbonate ; Alkalosis ; Calcification ; Cartilage ; Carbonic anhydrase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Le mécanisme de l'élévation du (HCO 3 − ) dans les liquides extracellulaires du cartilage (Cfl) a été étudié au niveau, de métaphyses tibiales de Rat. Au cours d'étudesin vitro, les courbes pH- $$P_{CO_2 } $$ dans une lymphe synthétique ne sont pas modifiées de façon nette par des protéines-polysaccharides ou par une protéine cationique. (HCO 3 − ) de Cfl, aspiré à partir de pièces métaphysaires, incubées isolément, décroit rapidement en fonction du temps. Les résultats de ces deux expériences semblent infirmer un rôle des sécrétions cartilagineuses comme cause de gradients sang— Cfl (HCO 3 − ) in vivo. L'acétazolamide, administré à des ratsin vivo, réduit le gradient sang —Cfl (HCO 3 − ) jusqu'à, un seuil non dosable. Cette action ne peut être attribuée à l'acidose généralisée, produite par l'acétazolamide, étant donné que les rats témoins, ayant une acidose généralisée similaire, provoquée par un traitement à NH4Cl, présentent un gradient sang —Cfl (HCO 3 − ) net. La répartition de l'activité en anhydrase carbonique dans les tissus épiphysaires et métaphysaires de rats identiques est déterminée par micro-analyse. L'activité enzymatique n'est pas détectée dans des échantillons cartilagineux, mais est retrouvée, de façon significative, dans les structures adjacentes. L'activité en anhydrase carbonique, mesurée dans les structures adjacentes, est considérée comme les lieux de sécrétion d'HCO 3 − . Le rôle éventuel des capillaires épiphysaires et métaphysaires et des cellules osseuses dans la sécrétion d'HCO 3 − , est envisagé.
    Abstract: Zusammenfassung Der mechanismus, durch welchen (HCO 3 − ) in extrazellulären Knorpelflüssigkeiten (fl) der Wachstumsplatten von Rattentibiae erhöht ist, wurde untersucht. Beiin vitro Versuchen mit einer synthetischen Lymphe waren die pH- $$P_{CO_2 } $$ Kurven weder durch Proteinpolysaccharide noch durch kationisches Protein nachweisbar verändert. In Cfl, welche aus isolierten Inkubaten von Wachstumsknorpel entnommen wurden, nahm (HCO 3 − ) in Funktion der Zeit ebenfalls rasch ab. Die Resultate beider Experimente sprechen dagegen, daß die Knorpelsekretein vivo als Ursache der Blut-Cfl (HCO 3 − )- Gradienten in Betracht kommen. Acetazolamid, das Ratten verabreicht wurde, erniedrigte den Blut-Cfl (HCO 3 − )-Gradienten auf ein nicht mehr nachweisbares Niveau. Dieser Effekt konnte nicht einer durch Acetazolamid hervorgerufenen generalisierten Acidose zugeschrieben, werden, da Kontrollratten mit einem ähnlichen Grad von generalisierter Acidose, welche von einer NH4Cl-Behandlung herrührte, einen ansehnlichen Blut-Cfl (HCO 3 − )-Gradienten aufrechterhielten. Die Verteilung der Kohlensäureanhydrase-Aktivität in epiphysären und metaphysären Geweben von gleichartigen Ratten wurde durch Mikroanalyse bestimmt. Eine enzymatische Aktivität konnte in den Knorpelproben nicht nachgewiesen werden, wurde jedoch in signifikanten Mengen in den angrenzenden Geweben gefunden. Es wurde die Hypothese aufgestellt, daß die Stellen, wo diese Kohlensäureanhydrase-Aktivität in angrenzenden Geweben gemessen wurde, den Sekretionsstellen von HCO 3 − entspricht. Die mögliche Beteiligung von epiphysären und metaphysären Capillargefäßen und von Knochenzellen an, der HCO 3 − -Sekretion wird diskutiert.
    Notes: Abstract The mechanism by which (HCO 3 − ) is elevated in extracellular cartilage fluids (Cfl) of rat tibial growth plates was investigated. Inin vitro studies, the pH- $$P_{CO_2 } $$ curves in a synthetic lymph were not detectably altered by proteinpolysaccharides or by a cationic protein. Also, (HCO 3 − ) in Cfl aspirated from isolated incubates of growth cartilage decreased rapidly as a function of time. Results of both of these experiments mitigated against a role for cartilage secretions as the cause of blood-Cfl (HCO 3 − ) gradientsin vivo. Acetazolamide administered to ratsin vivo reduced the blood-Cfl (HCO 3 − ) gradient to an undetectable level. This effect could not be attributed to systemic acidosis produced by acetazolamide since control rats with a similar degree of systemic acidosis resulting from NH4Cl treatment, maintained a substantial blood-Cfl (HCO 3 − ) gradient. The distribution of carbonic anhydrase activity in epiphyseal and metaphyseal tissues of similar rats was determined by microassay. Enzymatic activity was not detected in cartilage samples, but was found in significant amounts in adjacent structures. This carbonic anhydrase activity measured in adjacent structures was hypothesized to represent sites of HCO 3 − secretion. The possible involvement in HCO 3 − secretion of epiphyseal or metaphyseal capillaries and bone cells is discussed.
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  • 40
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    Calcified tissue international 8 (1971), S. 24-35 
    ISSN: 1432-0827
    Keywords: Bone ; Cartilage ; Epiphysis ; Calcium ; Phosphorus ; Vitamin D
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les électrolytes ont été analysés dans le sérum et dans les tissues en voie de minéralisation, à des stades variables de calcification, chez des poulets et des pores normaux et rachitiques. L'humidité, les cendres et les matières organiques et déssèchées ont été analysées. Il apparait que d'autres électrolytes sériques, en plus du Ca, sont modifiés au cours du rachitisme. Le Mg et le P inorganique sont modifiés de façon variable au cours de l'avitaminose D. Des changements plus importants en contenu minéral des tissus sont observés dans les stades précoces de calcification que ceux que l'on a pu constater dans le sérum à des stades similaires, ce qui suggère un effet direct de l'avitaminose sur le tissu en voie de calcification. Quant au contenu du sérum, le dépôt de Ca est plus élevé dans les tissus d'animaux rachitiques par rapport aux témoins. Ceci n'est pourtant pas le cas du Mg ou du P inorganique, indiquant une affinité préférentielle du Ca pour le tissue rachitique. L'analyse gravimétrique de matières organiques et déssèchées des zones épiphysaires montre que les quantités de cartilage hypertrophique et en voie de dévelopment augmentent dans le cas de rachitisme, alors que le cartilage calcifié diminue, confirmant les observations histologiques antérieures. De manière inattendue, la proportion de cartilage au repos augmente au cours du rachitisme.
    Abstract: Zusammenfassung Die Elektrolyte im Serum und in mineralisierenden Geweben wurden zu verschiedenen Zeitpunkten der Verkalkung bei normalen Hühnern und Schweinen und bei solchen mit Vitamin D-Mangel analysiert. Wassergehalt, Asche, trockene und organische Substanz wurden ebenfalls gemessen. Die Ergebnisse zeigten, daß außer Ca auch andere Elektrolyte bei Rachitis verändert wurden. Mg und anorganischer P wurden durch den Vitaminmangel bei den beiden Gattungen verschiedenartig beeinflußt. In den Geweben wurde in den frühen Phasen der Verkalkung eine größere Änderung des Mineral-Spiegels gefunden, als dies zu übereinstimmenden Zeiten im Serum der Fall war, was auf eine direkte Wirkung des Vitaminmangels auf das verkalkende Gewebe schließen läßt. Im Verhältnis zum Serumspiegel wurde Ca in den Geweben der rachitischen Tiere zu einem größeren Grade als normal abgeschieden. Dies war nicht der Fall bei Mg oder anorganischem P, was einen Vorzug für Ca in den rachitischen Geweben anzeigt. Gravimetrische Analysen von organischer und trockener Substanz in epiphysischen Schichten zeigten, daß die Menge des proliferierenden und hypertrophischen Knorpels zunahm und daß verkalkter Knorpel bei Rachitis abnahm, gemäß klassischen histologischen Beobachtungen. achtungen. Ganz unerwartet war bei Rachitis eine verhältnismäßige Zunahme des ruhenden Knorpels festzustellen.
    Notes: Abstract Electrolytes were analyzed in serum, and in mineralizing tissues at varying stages of calcification, in normal and vitamin D-deficient chickens and pigs. Moisture, ash, and organic matter were also measured. Results revealed that in addition to Ca, other serum electrolytes were altered in rickets. Mg and inorganic P were diversely affected by the vitamin deficiency. Larger changes in mineral level were seen in the tissues in the early stages of calcification than were seen at corresponding times in the serum, suggesting a direct effect of the vitamin deficiency on the calcifying tissue itself. Relative to the serum level, Ca was deposited in the tissues of rachitic animals to a greater extent than in normal animals. This was not true for Mg or inorganic P, indicating a preferential affinity for Ca in the rachitic tissue. Gravimetric analyses of organic and dry matter in epiphyseal zones revealed that the amounts of proliferating and hypertrophic cartilage were increased, and calcified cartilage decreased, in rickets, in accord with previous histological observations. Unexpectedly, the proportion of resting cartilage was also increased.
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  • 41
    ISSN: 1432-0827
    Keywords: Morphology ; Glycosaminoglycans ; Cartilage ; Chemistry ; Audioradiography ; Healing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Une perte de substance ostéo-cartilagineuse, de taille limitée et identique, est réalisée chez le lapin adulte et la cicatrisation est étudiée histologiquement et par autoradiographie après marquagein vitro au35S-sulfate. Une analyse microchimique est pratiquée pour le contenu et la composition en glycosaminoglycanes. 1. Entre la première semaine et la 4ème et 8ème semaine, un tissu conjonctif non-métachromatique se différencie en un cartilage métachromatique et la quantité de sulfate de chondroitine augmente de façon significative aux dépens des glycoprotéines. 2. Jusqu'à la 4ème semaine, la perte de substance est surtout comblée par de l'os néoformé: après cette période, la région est comblée au delà de la limite de la surface articulaire. 3. Le cartilage hyalin, ressemblant morphologiquement, autoradiographiquement et chimiquement au cartilage articulaire, en ce qui concerne la distribution en glycosaminoglycanes, constitute la surface articulaire de la perte de substance comblée dans un tiers des cas après 8 semaines. Le cartilage hyalin s'observe surtout dans les régions où de l'os néoformé a comblé la cavité médullaire. 4. Dans les deux tiers des cas, après 8 semaines, les surfaces articulaires des zones comblées comportent, non seulement du cartilage, mais aussi du tissu fibreux se formant essentiellement sur les parties latérales et dans les régions, où la cavité médullaire, fliant face, à la surface articulaire, n'a pas été comblée par du tissue osseux. La fraction glycoprotéique augmente par rapport à la fraction chondroitine sulfate. 5. Dans la majorité des cas, après 20 semaines, le cartilage néoformé subit des phénomènes dégénératifs, qui se traduisent par une diminution en chondroitine sulfate.
    Abstract: Zusammenfassung Bei ausgewachsenen Kaninchen wurde ein begrenzter, standardisierter, osteochondraler Defekt hervorgerufen, und das regenerierte Gewebe wurde histologisch und autoradiographisch durch Markierung in vitro mit35S-Sulfat und durch mikrochemische Bestimmung des Gehaltes und der Zusammensetzung der Glykosaminglykane untersucht. Die wichtigsten Befunde waren: 1. Zwischen 1 und 4–8 Wochen veränderte sich nichtmetachromatisches Bindegewebe zu metachromatisch gefärbtem Knorpel, und der Anteil an Chondroitin-Sulfat nahm auf Kosten der Glykoproteine signifikant zu. 2. Bis zu 4 Wochen war der Hauptteil des defekten Gebietes mit neugebildetem Knochen gefüllt; nach dieser Zeit lag dieser Bezirk oberhalb der Verknöcherungsgrenze in Richtung der Gelenkoberfläche. 3. Nach 8 Wochen bestand die Gelenkoberfläche des defekten Gebietes in einem Drittel der Fälle aus hyalinem Knorpel, der morphologisch, autoradiographisch und chemisch dem Gelenkknorpel in Bezug auf die Verteilung von Glykosaminoglykanen glich. Hyaliner Knorpel wurde hauptsächlich an Stellen beobachtet, wo neugebildeter Knochen die Markhöhle geschlossen hatte. 3. Nach 8 Wochen bestand die Gelenkoberfläche des defekten Gebietes in einem Drittel der Fälle aus hyalinem Knorpel, der morphologisch, autoradiographisch und chemisch dem Gelenkknorpel in Bezug auf die Verteilung von Glykosaminoglykanen glich. Hyaliner Knorpel wurde hauptsächlich an Stellen beobachtet, wo neugebildeter Knochen die Markhöhle geschlossen hatte. 4. Nach 8 Wochen bestanden Teile der Gelenkoberfläche des Defektes in zwei Dritteln der Fälle nicht nur aus Knorpel, sondern auch aus fibrösem Gewebe, welches vor allem in den seitlichen Teilen des Defektes und an Stellen vorlag, wo die Markhöhle gegenüber der Gelenkoberfläche nicht mit Knochengewebe verschlossen worden war. Die Glykoproteinfraktion nahm im Vergleich zur Chondroitin-Sulfatfraktion zu. 5. Nach 20 Wochen zeigten sich in den meisten Fällen bei neugebildetem Knorpel degenerative Veränderungen, welche durch eine gewisse Abnahme des Chondroitin-Sulfats wiedergegeben wurden.
    Notes: Abstract A limited, standardized osteochondral defect was created in adult rabbits and the regenerated tissue was examined histologically and autoradiographically after labellingin vitro with35S-sulphate, and microchemically for its content and composition of glycosaminoglycans. The principal findings were: 1. Between 1 week and 4 to 8 weeks, non-metachromatic connective tissue differentiated to metachromatically stained cartilage, and the proportion of the chondroitin sulphate increased significantly at the expense of the glycoproteins. 2. Up to 4 weeks, the major part of the defect area was filled with newly formed bone; after this time, the area lay above the level of the “tidemark”, towards the articular surface. 3. Hyaline cartilage with morphological, autoradiographic and chemical resemblance to the articular cartilage in terms of the distribution of glycosaminoglycans constituted the articular surface of the defect area in one-third of the cases at observation times after 8 weeks. Hyaline cartilage was observed mainly in areas where newly formed bone had closed the medullary cavity. 4. In two-thirds of the cases, after 8 weeks, parts of the articular surface of the defect consisted not only of cartilage but also of fibrous tissue, occurring mainly in the lateral parts of the defect and in areas where the medullary cavity facing the articular surface had not been sealed by bone tissue. The glycoprotein fraction increased relative to the chondroitin sulphate fraction. 5. In most cases after 20 weeks, newly-formed cartilage underwent degenerative changes, which were reflected in some reduction of the chondroitin sulphate.
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  • 42
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    Calcified tissue international 8 (1971), S. 106-113 
    ISSN: 1432-0827
    Keywords: Iliac crest ; Proteinpolysaccharide ; Cartilage ; Achondroplasia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les protéines-polysaccharides (PP) ont été extraits par des méthodes de rupture et de dissociation, à partir de cartilage de crête iliaque de neuf enfants achondroplasiques, dont 8 filles et 1 garçon, âgés de huit à treize ans. A la fois l'homogénéisation à grande vitesse du tissu dans l'eau et l'incubation à l'aide de chlorure de guanidinium ont libéré des quantités normales de PP, à poids moléculaire élevé, du cartilage de jeunes enfants. Des échanitllons de cartilage d'enfants plus âgés ont libéré peu ou pas de PP après extraction par l'eau. Les PP, insolubles dans l'eau, se solubilisent dans du chlorure de guanidinium 4 M, des solutions non tamponnées de MgCl2 et du tampon phosphate 0.067 M. Alors que l'extrait à la guanidine contient, après dilution à 0,4 M du chlorure de guanidinium, un constituant à poids moléculaire élevé, des extraits de MgCl2, tamponnés au phosphate, ne contiennent que des PP dégradés de poids moléculaire peu élevé. L'insolubilité de PP dans l'eau, de cartilage d'enfants, âgés de plus de dix ans, est interprètée comme une manifestation secondaire liée soit aux changements mucoprotéiques, se déroulant dans le cartilage à cet âge ou à d'autres facteurs. L'anomalie biochimique primitive pourrait se situer au niveau de la macromolécule de PP qui, à un certain stade de maturation du tissu, se traduit soit par une altération de l'état d'agrégation, soit par un rapport anormal avec le collagène.
    Abstract: Zusammenfassung Proteinpolysaccharide (PP) wurden mittels Homogenisierungs- und Extraktionsmethoden aus dem Beckenkamm-Knorpel von 9 achondroplastischen Kindern (8 Mädchen und 1 Knabe) im Alter von 8 bis 13 Jahren extrahiert. Sowohl eine hochtourige Homogenisierung des Gewebes in Wasser, als auch eine Inkubation mit Guanidinchlorid setzten normale PP-Mengen von hohem Molekulargewicht aus dem Knorpel von kleineren Kindern frei. Dagegen ergab die Wasserextraktion aus den Knorpelproben von größeren Kindern nur wenig oder kein PP-Material. Die wasserunlöslichen PP waren in 4 M Guanidinchlorid, ungepufferten MgCl2-Lösungen und 0,067 M Phosphatpuffer löslich. Während der Guanidinextrakt nach Verdünnung des Guanidinchlorids zu einer 0,4 M-Konzentration eine Komponente mit hohem Molekulargewicht enthielt, fand sich in MgCl2- und Phosphatpufferextrakten nur abgebautes PP-Material von niedrigem Molekulargewicht. Die Wasserunlöslichkeit von Knorpel-PP von über 10jährigen Kindern wird als sekundäre Erscheinung interpretiert, die entweder in Zusammenhang mit Mucopolysaccharid-Veränderungen steht, welche während dieses Alters im Knorpel auftreten, oder die auf andere unbekannte Faktoren zurückzuführen ist. Der primäre biochemische Defekt kann auf einer Abnormalität des PP-Makromoleküls beruhen, welche, bei einem gewissen Reifungsgrad des Gewebes, entweder in einem veränderten Aggregatzustand oder in einem abnormalen Verhältnis zur Kollagenmatrix zum Ausdruck kommt.
    Notes: Abstract Proteinpolysaccharides (PP) have been extracted by disruptive and dissociative methods from iliac crest cartilage of nine achondroplastic children, eight girls and one boy, from eight to thirteen years of age. Both high speed homogenization of the tissue in water or incubation with guanidinium chloride released from the cartilage of younger children normal amounts of PP of high molecular weight. Cartilage samples obtained from older children released instead little or no PP material by water extraction. The water-insoluble PP were found soluble in 4 M guanidinium chloride, unbuffered MgCl2 solution and 0.067 M phosphate buffer. While the guanidine extract contained, after dilution of guanidinium chloride to 0.4 M, a high molecular weight component, MgCl2 and phosphate buffer extracts contained only degraded PP material of low molecular weight. The insolubility in water of PP of cartilage of children over ten years of age is interpreted as a secondary manifestation related either to mucopolysaccharide changes occurring in the cartilage at that age, or to other unknown factors. The primary biochemical defect could be an abnormality of the PP macromolecule which, at a certain stage of maturation of the tissue, is reflected either in an altered state of aggregation or in an abnormal relationship with the collagen matrix.
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  • 43
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    Calcified tissue international 8 (1971), S. 276-286 
    ISSN: 1432-0827
    Keywords: Bone ; Cartilage ; Differentiation ; Collagen ; β-aminopropionitrile
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé L'injection d'embryons de poulet, incubés pendant neuf jours, avec du β-aminopropionitrile (BAPN), à des doses supérieures à 0,312 mg par embryon, empêche la formation de cartilage, normalement présent au niveau des os de membrane à partir du llème jour de l'inculation. L'hitsogenese de cette inhibition est décrite. BAPN augmente la propertion de collagène et diminue la synthèse des mucopolysaccharides acides, contenant de l'hesoxamine, au niveau des os de membrane. Le rapport entre les chainons tranversaux du collagène, sa solubilité, la synthèse des mucopolysaccharides acides et la stimulation mécanique (le facteur extrinsèque déclenchant la chondrogenèse dans le système) est discuté sous l'angle de la formation cartilagineuse sur les os de membrane, à partir de cellules souches.
    Abstract: Zusammenfassung Wird β-Aminopropionitrol (BAPN) in Dosen von über 0,312 mg/Embryo in 9 Tage inkubierte Hühnerembryonen injiziert, so wird die Bildung des normalerweise vom 11. Inkubationstag an auf dem Deckknochen vorliegenden Knorpels verhindert. Die Histogenese dieser Hemmung wird beschrieben. BAPN erhöhte den Anteil des löslichen Kollagens, ohne die Synthese des Totalkollagens zu verändern; es verminderte die Synthese der hexosaminhaltigen sauren Mucopolysaccharide innerhalb der Deckknochen. Die Diskussion betrifft die Beziehung zwischen der Quervernetzung des Kollagens, der Kollagenlöslichkeit, der Synthese von sauren Mucopolysacchariden und der mechanischen Stimulation (wobei der Extrinsicfaktor die Chondrogenese in diesem System auslöst) bei der Knorpelbildung aus den bipotentialen Keimzellen der Deckknochen.
    Notes: Abstract Injection into chick embryos incubated for nine days, of β-aminopropionitrile (BAPN) at doses above 0.312 mg/embryo prevented the formation of the cartilage normally present on the membrane bones from the eleventh day of incubation onwards. The histogenesis of this inhibition is described. BAPN increased the proportion of soluble collagen without altering total collagen synthesis and decreased the synthesis of hexosamine-containing acid mucopolysaccharides within the membrane bones. The relationship between cross-linking of collagen, collagen solubility, synthesis of acid mucopolysaccharides and mechanical stimulation (the extrinsic factor initiating chondrogenesis in this system) in the formation of cartilage from bipotential germinal cells on the membrane bones is discussed.
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  • 44
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    Calcified tissue international 33 (1981), S. 57-69 
    ISSN: 1432-0827
    Keywords: Thallium ; Chondrodystrophy ; Cartilage ; Bone ; Matrix
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Using the transmission electron microscope, we sought to describe the morphology of thallium sulfate-induced chondrodystrophy in chick embryos. There was cell death and degeneration in all zones of growth cartilage, but the cells and matrix of the hypertrophic zone were the most severely affected. Ultrastructural changes of the hypertrophic chondrocytes consisted of alteration of the cytoplasmic contents and of the intercellular matrix; the cell membrane was smooth and without cytoplasmic extensions. The cytoplasm was filled with dilated rough endoplasmic reticulum, vacuoles of varying sizes and contents, and lipidlike bodies with electron-dense granules; mineral crystals, collagen, and degenerating mitochondria were present. The matrix showed only spotty calcification and a reduced number of dense bodies, vesicles, and granules. The cells appeared to have failed to exteriorize cell products across the plasmalemma. Failure to exteriorize cell products and to form cytoplasmic processes reduced the number of potential nucleation sites for calcification. The ultrastructure of osteocytes was much less affected.
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  • 45
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    Calcified tissue international 33 (1981), S. 89-99 
    ISSN: 1432-0827
    Keywords: Soluble proteoglycans ; Resistant proteoglycans ; Collagen ; Bone ; Cartilage
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Soluble proteoglycans (SPG) were extracted from bovine (BCC) and human (HCC) costal cartilages by the dissociative method using 4 M guanidinium chloride (GuHCl). Proteoglycans which are resistant to extraction (RPG) were obtained following collagenase digestion or hydroxylamine treatment of the cartilage residues. Similarly, SPG were extracted from bovine metaphyseal and cortical bone using EDTA. The RPG were extracted from the bones using hydroxylamine. Density gradient fractionation under dissociative conditions of cartilage SPG and RPG followed by chromatography on Sepharose 2B revealed that A1D1 RPG are smaller than the SPG. SPG reacted with either collagenase or hydroxylamine are also smaller than the parent SPG. A1D1 fractions obtained from BCC-SPG and RPG or from mixtures of SPG and acid-soluble collagen are free of hydroxyproline. Hydroxyproline is not completely separated from HCC-RPG. Density gradient fractionation of bone proteoglycans and Sepharose chromatography of the A1 and A1D1 fractions showed that those obtained from metaphysis are larger than those from cortical bone. This was attributed to the presence of calcified cartilage in metaphyseal bone. The A1D1 fractions of the metaphyseal proteoglycans seemed to undergo self-association since this fraction is larger than the A1 fraction from which it is derived. Cortical bone proteoglycans do not behave similarly. Density gradient purification under dissociative conditions failed to separate hydroxyproline from the proteoglycans obtained from bone. It is hypothesized that in bone proteoglycans and collagen might be linked.
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  • 46
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    Calcified tissue international 22 (1977), S. 231-241 
    ISSN: 1432-0827
    Keywords: Alkaline phosphatase ; Calcification ; Cartilage ; Enamel organ ; Odontoblasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract The enzyme alkaline phosphatase (AP) (EC 3.1.3.1) in three different calcification areas was studied by means of a spectrophotometric micro method using p-nitrophenylphosphate as a substrate. Rat maxillary incisor odontoblasts and enamel organ from the zones of matrix formation and maturation and tissue from rabbit metatarsal cartilage were allowed to react with the substrate in glycine-NaOH buffer at room temperature. The reaction was found to be linear for a minimum of 20 min. The pH optima for AP from these tissues were in the pH range of 10.0–10.3. In order to compare AP from the four calcification areas different parameters were studied. Heating at 56°C or 60°C for varying times revealed that the enzymes were almost completely inactivated after 10 min. Mg2+ ions activated the enzymes by about 25% at concentrations of 2.5 mM (enamel organ 1.25 mM); while only higher concentrations of Mg2+ had an inactivating effect, Ca2+ and PO 4 3− ions were inactivating at varying concentrations. F− ions show no effect on AP activity at concentrations below 250 mM (enamel organ 125 mM) but caused inactivation of the enzymes at about 50% at 1 M. EDTA was found to be a very effective AP inactivator at concentrations above 0.06 mM, whereas urea did not noticeably affect the enzyme reactions at concentrations below 1 M. At higher concentrations, inactivation was observed. In order to determine AP localization in the epiphyseal plate successive 40-μm-thick, freeze-sectioned slices were analyzed. The activity was highest nearest the zone of cartilage calcification and decreased towards the reserve cell zone. It was concluded that the same AP isoenzyme is present in these quite different calcification loci.
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  • 47
    ISSN: 1432-0827
    Keywords: Electrolytes ; Phospholipids ; Cartilage ; Matrix Vesicles ; Calcium-phosphate ; Phospholipid complexes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Electrolytes and phospholipids of cartilage fractions were partitioned by extraction with organic and aqueous solvents into six solubility groups: Electrolytes I, II and III, and Lipids I, II and III. Of the total Ca, only 4% was water soluble (Electrolytes I); 4–12% was complexed with lipids (Electrolytes II); while the majority (84–92%) was insoluble (Electrolytes III). In contrast, nearly half of the Mg and Pi were water soluble. Of theneutral phospholipid, 95% was not complexed with mineral ions (Lipids I), but 30–45% of theacidic phospholipid was (Lipids II). Ca/Pi ratios were extremely low in the water-soluble phase, but were in the range of amorphous calcium phosphate (ACP) in the insoluble. Molar ratios of the lipid-mineral complex were: Ca∶Mg∶Pi:acidic phospholipid, 4∶3∶2∶2. Mg/Ca ratios in the soluble fraction were high (5.5–8.9), sufficient to stabilize ACP. Kinetic studies revealed rapid turnover of soluble Ca, insoluble turning over much more slowly. Labeling of lipid-complexed Ca was rapid in cells, but occurred later in matrix vesicles, suggesting transfer. While lipid-Ca-Pi complexes can nucleate apatite in vitro, those present in vivo inside matrix vesicles apparently do not because of the excess Mg. We conclude therefore, that in vesicle-mediated calcification, lysis of the membrane may be essential to allow release of internal Mg.
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  • 48
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    Calcified tissue international 28 (1979), S. 201-208 
    ISSN: 1432-0827
    Keywords: Cartilage ; Proteoglycans ; Ascorbic acid ; Alpha-tocopherol ; Arylsulfatase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Previous studies conducted in cell culture utilizing chondrocytes isolated from human articular cartilage showed that inclusion of ascorbic acid in the nutrient medium resulted in an increase in the net biosynthesis of sulfated proteoglycans concomitant with a reduction in the activities of arylsulfatases A and B. This suggested a possible connection between the stability of the sulfated macromolecules and the activity of these desulfating enzymes. To examine whether this relationship also was operative in a more native milieu, the effect of ascorbate on these parameters was measured in an organ culture system using slices of human articular cartilage where the structural extracellular framework surrounding the chondrocytes is comparable to that present in vivo. In addition, since numerous studies have shown that vitamin E inhibits lysosomal enzymes, the effect of alpha-tocopherol on these parameters was also investigated. Concurrent with an inhibition of arylsulfatase A and B activities, an increase in sulfated proteoglycan biosynthesis per unit of DNA as reflected in35S-sulfate uptake was present in all cartilage specimens examined 6 days after the introduction of vitamin C in the culture fluid. A larger fraction of the newly synthesized sulfated macromolecules was incorporated into the matrix of the tissue in samples maintained in the vitamin-supplemented cultures. Supplementation of the organ culture medium with vitamin E resulted in the inhibition of arylsulfatase A and no change in arylsulfatase B activities. In contrast to the action of vitamin C which stimulated acid phosphatase activity, a potent inhibition of this enzyme was effected by vitamin E. Sulfated proteoglycan content was greatly enhanced by alpha-tocopherol, although the vitamin did not alter the distribution of the newly synthesized molecules between the tissue and medium fractions. Inhibition of acid phosphatase and arylsulfatase A by alpha-tocopherol was evident in tissue lysates and partially purified enzyme preparations. This implies a direct interaction between the enzymes and vitamin E. The actions of alpha-tocopherol and ascorbate on cartilage enzymes and structural components were dissimilar. Both vitamins, however, appeared to enhance the stability of sulfated proteoglycans in the complex structure comprising articular cartilage.
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  • 49
    ISSN: 1432-0827
    Keywords: Diphosphonates ; Cartilage ; Collagen ; Erbium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The bone-seeking agent99mTc-labeled 1-hydroxyethylidene-1,1-diphosphonic acid (HEDP) unexpectedly binds to particles of human articular cartilage as well as cortical bone in vitro. Collagen also sequesters this compound, suggesting that collagen contributes to the uptake of99mTc-HEDP by cartilage and bone. Particles of the bone mineral calcium hydroxyapatite also bind99mTc-HEDP in vitro. Pretreatment of particles with Er3+ stimulates binding in each case. Lowering the pH of incubation to pH 2 has this effect for bone, cartilage, and collagen, but not for calcium hydroxyapatite. Mechanisms additional to the simple ionic attraction between the phosphonate groups of HEDP and metal cations such as Ca2+ are responsible for the uptake of99mTc-HEDP by body tissues.
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  • 50
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    Calcified tissue international 35 (1983), S. 526-532 
    ISSN: 1432-0827
    Keywords: Cartilage ; PTH ; in vitro ; Growth ; cAMP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Addition of N6-monobutyryl cyclic AMP (BtcAMP) or methylisobutylxanthine to embryonic chick pelvic cartilages incubated in a serum-free medium for 3 days stimulates the cartilages to grow predominately through increasing cellular hyperplasia [1]. The present study investigated whether parathyroid hormone (PTH) added to the serum-free organ culture system would mimic the growth-promoting effects of BtcAMP through increasing endogenous cartilage cyclic AMP content. Parathyroid hormone 10 nM stimulatesin vitro pelvic cartilage growth as compared with cartilage grown in medium alone, as estimated by increases in wet weight (64%), dry weight (91%), total soluble protein (19%), and deoxyribo-nucleic acid (DNA) content (51%). Parathyroid hormone (0.1–10nM) in a concentration-dependent manner produces increases in wet weight and radiolabeled precursor incorporation of [3H]thymidine and [14C]-L-leucine over cartilage incubated in medium alone. Both PTH-treated and BtcAMP-treated cartilage show histological changes of increased cellularity as well as raised DNA content and elevated DNA/protein ratios. PTH increases cartilage cyclic AMP content, with the maximum rise occurring after 30 min incubation, and a return to levels found in pelvic cartilages incubated in medium alone by 4 hours. Consequently, cartilages incubated in medium containing PTH for 4 hours and then incubated in medium alone for the remaining 3 days, grew as well as cartilages that had been incubated in medium containing PTH for the entire 72-hour period. Thus, PTH, by raising intracellular cyclic AMP, triggers cartilage growthin vitro.
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    Calcified tissue international 36 (1984), S. 576-579 
    ISSN: 1432-0827
    Keywords: Insulinlike growth factors ; Multiplication-stimulating activity ; Chondrocytes ; Cartilage
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Multiplication-stimulating activity (125I-MSA) has been shown to bind to isolated rabbit chondrocytes, the binding being dependent on time, temperature, and cell density. Nonspecific binding was approximately 15%. Unlabelled MSA at 100 ng/ml inhibited125I-MSA binding by 50%. Porcine insulin (0.5–10 µg/ml) did not compete with MSA but resulted in a 10–15% increase in125I-MSA binding. The data suggest that normal chondrocytes carry IGF2-type receptors as well as the IGF1 type previously described.
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  • 52
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    Calcified tissue international 5 (1970), S. 64-69 
    ISSN: 1432-0827
    Keywords: Epiphysis ; Cartilage ; Resorption ; Enzymes ; Implantation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des fragments d'omoplates, prélevés chez de jeunes rats, sont décalcifiés, puis soumis à la pronase, papaïne, hyaluronidase ou trypsine. Les cartilage épiphysaires d'omoplates témoins se colorent au bleu Alcian, mais ceux, traités à la pronase, papaïne et hyaluronidase, ne se colorent pas: la trypsine ne supprime pas la coloration. Lorsque les os sont réimplantés chez les animaux au niveau desquels ils ont été prélevés, le cartilage épiphysaire de ceux traités avec les 3 premiers enzymes est rapidement envahi par des capillaires et de petites cellules rondes. Les cartilages des témoins, l'os non traité et l'os traité avec de la trypsine ne sont pas envahis ainsi. Il semble que l'extraction des glycosaminoglycanes acides du cartilage puisse être un des facteurs controlant l'invasion capillaire au cours de l'ossification enchondrale.
    Abstract: Zusammenfassung Scapulastücke wurden von jungen Ratten entfernt, entkalkt und mit Pronase, Papain, Hyaluronidase oder Trypsin behandelt. Die epiphysären Knorpel von Kontroll-Scapula nahmen die Alcianblaufärbung an, die mit Pronase, Papain oder Hyaluronidase behandelten dagegen nicht; Trypsin hob die Färbungsreaktion nicht auf. Wenn die Knochen subcutan auf die Tiere implantiert wurden, von welchen sie entnommen worden waren, wurden die mit den ersten drei Enzymen vorbehandelten epiphysären Knorpel rasch von Capillaren und kleinen runden Zellen überflutet. Die Knorpel der unbehandelten Kontrollknochen und diejenigen von Trypsin-behandelten Knochen wurden nicht in dieser Weise überflutet. Es wird postuliert, daß die Entfernung von sauren Glucosaminoglycanen aus dem Knorpel einer der Faktoren sein kann, welche die Capillareninvasion auslösen, die in der endochondralen Calcifikation auftritt.
    Notes: Abstract Pieces of scapula were removed from young rats, decalcified, and treated with pronase, papain, hyaluronidase or trypsin. The epiphyseal cartilages of control scapulae took up the Alcian blue stain, but those treated with pronase, papain or hyaluronidase did not; trypsin did not abolish the staining reaction. When the bones were implanted subcutaneously into the animals from which they had been taken, the epiphyseal cartilages of those treated with the first three enzymes were rapidly invaded by capillaries and small round cells. The cartilages of the control, untreated bones and those from bones treated with trypsin were not invaded in this way. It is postulated that the removal of acid glycosaminoglycans from cartilage may be one of the factors that trigger the capillary invasion that occurs in endochondral calcification.
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  • 53
    ISSN: 1432-0827
    Keywords: Glycosaminoglycans ; Cartilage ; Transplant ; Microchemistry ; Defect
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Un transplant dans une perte de substance ostéochondrale, mesurant 3×4×3 mm, a été réalisé au niveau de la tête fémorale de lapins adultes. On a transplanté du cartilage costal autologue, qui a été primitivement implanté pendant 4–5 semaines dans le tissu musculaire, et du cartilage costal autologue frais. Après des intervales de temps, variant de 2 à 52 semaines, les animaux sont sacrifiés. Les régions transplantées et les tissus voisins sont étudiés, après microdissection de coupes obtenues après congélation-dessication, à l'aide d'une technique micro-chimique quantitative, permettant d'analyser leur contenu en glycosaminoglycanes et en calcium. Le contenu en hexosamine (en % de poids sec organique) est généralement plus élevé dans les transplants que dans le cartilage articulaire après 2 et 6 semaines: après cette période, ce contenu est identique. La distribution des différentes fractions de glycosaminoglycanes est différente dans les transplants et le cartilage articulaire. Ainsi la fraction de sulfate de glycoprotéine et/ou de kératane est plus faible au niveau du transplant que dans le cartilage articulaire: l'inverse est vrai pour le sulfate de chondroitine. Après des intervalles de temps plus élévés, ces fractions ont tendance à s'égaliser dans le transplant et le cartilage articulaire. Les courbes de solubilité du sulfate de chondroitine présentent une grande similitude. Le contenu en calcium est généralement plus élevé dans le transplant que dans le cartilage articulaire. A partir de 12 semaines, une décroissance nette est observée par rapport au tissu témoi n. Les résultats chimiques indiquent que le cartilage de la région transplanté ne subit pas d'altérations dégénératives nettes.
    Abstract: Zusammenfassung Am Hüftgelenkskopf von erwachsenen Kaninchen wurde ein 3×4×3 mm großer osteochondraler Defekt geschaffen. In diesen wurde autologer Rippenknorpel, welcher zuerst während 4–5 Wochen intramuskulär implantier war, und frischer autologer Rippenknorpel transplantiert. An 6 verschiedenen Beobachtungszeitpunkten zwischen 2 und 52 Wochen wurden die Tiere getötet. Das Ausgangsmaterial, das Gewebe an der Transplantationsstelle auf der Gelenkfläche und der Gelenkknorpel oberhalb der “tidemark” wurden von jedem Tier für sich, nach Mikrodissektion an gefriergetrockneten Gefrierschnitten untersucht. Der Gehalt und die Zusammensetzung der Glykosaminoglykane in diesen Materialien wurde mit einer quantitativen mikrochemischen Technik und der Kalkgehalt mittels einer Ionenaustauschmethode untersucht. Der Hexosamingehalt, ausgedrückt in Prozent des Trockengewichtes, war allgemein etwas höher in den Transplantaten als im Gelenkknorpel an den Beobachtungszeiten 2 und 6 Wochen, danach war dieser sehr gleich. Die gegenseitige Verteilung der verschiedenen Glykosaminoglykan-Fraktionen war unterschiedlich in den Transplantaten und im Gelenkknorpel. Die Glykoprotein- und/oder Keratansulfatfraktion war kleiner in den Transplantaten als im Gelenkknorpel, während das Gegenteil für das Chondroitinsulfat zutraf. Mit steigenden Beobachtungszeiten wurde eine größere Ähnlichkeit zwischen Transplantat und Gelenkknorpel mit Hinsicht auf die relativen Größen dieser Fraktionen offenbar. Die Löslichkeitsprofile des Chondroitinsulfates zeigten weitgehende Gleichheit. Der Kalkgehalt in den Transplantaten war allgemein höher in den Transplantaten als im Gelenkknorpel. Nach Versuchszeiten von 12 Wochen zeigten doch die Transplantate im Vergleich zu dem Ausgangsmaterial einen eindeutigen Abfall der Kalkwerte, wonach diese ziemlich konstant verblieben.
    Notes: Abstract In adult rabbits, a transplant was made to an osteochondral defect on the femoral head measuring 3×4×3 mm of autologous costal cartilage which had first been implanted intramuscularly for 4–5 weeks and fresh autologous costal cartilage. After 6 different observation times between 2 and 52 weeks the animals were killed. The original material, the tissue in the transplant area on the articular surface and the articular cartilage above the “tidemark” were analysed in each individual animal, after microdissection of freeze-dried sections of these structures, by a quantitative microchemical technique for their content and composition of glycosaminoglycans and for their calcium content. The hexosamine content (% per organic dry weight) was generally higher in the transplants than in the articular cartilage at observation times of 2 and 6 weeks, after which time it was similar. The distribution of the different glycosaminoglycan fractions was different in the transplants compared with the articular cartilage. Thus the glycoprotein and/or keratan sulphate fraction was smaller in the transplant than in the articular cartilage, while the opposite held for the chondroitin sulphate. With increasing observation times, some resemblance became evident between the transplant and articular cartilage with respect to the relative size of these fractions. The solubility profiles of the chondroitin sulphate were similar. The calcium content was generally higher in the transplant than in the articular cartilage, after 12 weeks it showed a pronounced decrease in comparison with the original material. The chemical findings indicate that the cartilage in the transplant area did not undergo any marked degenerative changes.
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  • 54
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    Calcified tissue international 5 (1970), S. 30-38 
    ISSN: 1432-0827
    Keywords: Phospholipids ; Bone ; Cartilage ; Mucopolysaccharides
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les tissus squelettiques de poissons téléostéens (Scomber scombrus, etGadus callaria) et d'élasmobranches (Raja batis etScyllium canicula) sont analysés pour leur contenu en lipides. Plus de phospholipides sont extraits des tissus osseux de téléostéens que des tissus cartilagineux d'élasmobranches. Outre la proportion lipidique plus élevée de l'os de téléostéens, des différences de composition lipidique ont été notées entre les deux tissus. Avant démineralisation, les extraits deGadus et Scomber sont surtout constitués de phospholipides neutres. Par contre, les phosphatides des tissus cartilagineux contiennent des quantités importantes de phospholipides acides. Après déminéralisation de l'os et du cartilage de poissions, les solvants pour lipides neutres éluent á la fois les phosphatides acides et neutres de ces tissus. Après extraction des lipides, le traitement des tissus déminéralisés par des solvant acides extrait à nouveau des lipides. Dans cette fraction finale, un grand nombre de phospholipides ont été retrouvés. A ce stade, d'autres phosphatides, que ceux mis en évidence chez les poissons précités, sont extraits à partir de l'os déminéralisé deScomber. Le principal phosphatide de la fraction finale est constitué par la cardiolipine dont des quantités plus importantes, que celles notées au niveau des autres tissus durs, sont retrouvées dans l'os deScomber.
    Abstract: Zusammenfassung Die Skeletgewebe der Knochenfische (Scomber scombrus undGadus callaria) und der Knorpelfische (Raja batis undScyllium canicula) wurden auf ihren Lipidgehalt untersucht. Es ergab sich, daß mehr Phospholipide aus dem Knochengewebe der Knochenfische extrahiert werden konnte, als dies aus dem Knorpelgewebe der Knorpelfische möglich war. Es wurde nicht nur eine größere Lipidmenge im Knochen der Knochenfische festgestellt, sondern auch noch Unterschiede in der Lipidzusammensetzung der beiden Gewebe beobachtet. Die vor der Demineralisation hergestellten Extrakte von Gadus und Scomber bestanden vorwiegend aus neutralen Phospholipiden. Dagegen enthielten die Phosphatide der Knorpelgewebe ansehnliche Mengen saurer Phospholipide. Nach der Demineralisation von Fischknochen und-knorpel wurden durch neutrale Lipidlösungsmittel sowohl saure als auch neutrale Phosphatide aus diesen Geweben eluiert. Wenn die demineralisierten Gewebe, denen bereits Lipide entzogen wurden, noch mit angesäuerten Lösungsmitteln behandelt wurden, konnten erneut Lipide aus Knochen und Knorpel extrahiert werden. Im letzten Extrakt konnten die verschiedensten Phospholipide nachgewiesen werden. In diesem Stadium konnten verhältnismäßig mehr Phosphatide aus dem demineralisierten Knochen von Scomber entzogen werden, als dies bei anderen Fischarten möglich war. Das wichtigste Phosphatid dieses letzten Extraktes war Cardiolipin. Tatsächlich war mehr Cardiolipin in den Knochen von Scomber vorhanden als in allen andern Hartgeweben.
    Notes: Abstract The skeletal tissues of teleost (Scomber scombrus andGadus callaria) and elasmobranch (Raja batis andScyllium canicula) fish were examined for the presence of lipids. It was found that more phospholipid was removed from the bony tissues of the teleosts than was extractable from the cartilaginous tissues of the elasmobranchs. Apart from there being a greater proportion of lipid associated with teleost bone, differences were also noted in the lipid composition of the two tissues. Before demineralization, inGadus andScomber, the extracts were predominantly of neutral phospholipids. In contrast, the phosphatides of the cartilaginous tissues contained appreciable quantities of acidic phospholipids. After demineralization of piscine bone and cartilage, neutral lipid solvents eluted both acidic and neutral phosphatides from these tissues. When the demineralized lipid extracted tissues were treated with acidified solvents, lipid was again removed from the bone and cartilage. In this final extract, a wide variety of phospholipids was detected. At this stage, more phosphatide was extracted from the demineralized bone ofScomber, compared with the other piscine species. The principal phosphatide of this final extract was cardiolipin. More cardiolipin was present in the bone ofScomber than in any other hard tissue.
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  • 55
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    Calcified tissue international 5 (1970), S. 270-276 
    ISSN: 1432-0827
    Keywords: Ultrastructure ; Cartilage ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Une solution de sulfate de chrome est utilisée à la fois comme fixateur, colorant et agent de déminéralisation pour l'étude ultrastructurale de cartilage, en voie de minéralisation. Cette technique permet de mettre en évidence un “fantôme cristallin” organique, en rapport avec chaque cristal. L'intérêt du sulfate de chrome comme agent de déminéralisation est souligné.
    Abstract: Zusammenfassung Bei Ultrastrukturuntersuchungen von mineralisierendem Knorpel wurde eine Chromsulfatlösung als Agens zur kombinierten Fixation, Färbung und Demineralisierung verwendet. Diese Technik zeigte das Vorhandensein eines organischen “Kristallschattens”, der jedem Kriställchen zugehört. Die Tauglichkeit von Chromsulfat als demineralisierendes Agens wird besprochen.
    Notes: Abstract A solution of chromium sulphate was used as a combined fixative, stain and demineralizing agent for the ultrastructural study of mineralizing cartilage. This technique revealed the presence of an organic ‘crystal ghost’ associated with each crystallite. The effectiveness of chromium sulphate as a demineralizing agent is discussed.
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  • 56
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    Calcified tissue international 54 (1994), S. 499-504 
    ISSN: 1432-0827
    Keywords: Mechanical factors ; Cartilage ; Growth and development ; pH ; Osmotic pressure ; Proteoglycan ; Mandibular condyle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Cartilage growth and remodeling are known to be influenced by the biochemical and mechanical environment of the tissue. Previous investigators have shown that chemical factors that are relevant to mechanical loading, such as osmotic pressure and pH, induce changes in cartilage metabolism in vitro. Using a neonatal rat mandibular condyle culture system, the objectives of the work reported here were to determine (1) how the growth is influenced by osmotically applied mechanical loads; and (2) whether changes in intratissue osmotic pressure or pH cause metabolic changes in the cartilage which are then reflected by altered growth behavior. High molecular weight (MW) uncharged macromolecules polyvinylpyrrolidone (PVP) and Ficoll (presumed unable to penetrate the tissue matrix) were used to examine the effect of osmotic loading on tissue growth; concentrations corresponding to osmotic pressures of up to 100 kPa resulted in a dose-dependent depression in growth and matrix accumulation. Raffinose (which can penetrate the matrix but not the cells) had no significant effect on growth for osmotic pressures of up to 87 kPa, suggesting that compression-induced changes in intratissue osmotic pressure are unlikely to provide a signal by which cells sense and respond to mechanical compression. By contrast, changes in medium pH resulted in dose-dependent changes in growth behavior. Specifically, slight alkalinity (acidity) greatly enhanced (diminished) growth and matrix accumulation; the sensitivity to pH suggests that intratissue pH could provide a mechanism for cells to sense local glycosaminoglycan concentration and mechanical compression. These data are also significant in that, while previous investigators have shown that osmotic forces and pH affect biosynthetic rates in short-term culture, the present studies indicate that resultant changes in biosynthesis lead to altered growth rates.
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  • 57
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    Calcified tissue international 7 (1971), S. 12-22 
    ISSN: 1432-0827
    Keywords: Bone ; Cartilage ; Testosterone ; Ultrastructure ; Growth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Bien que la clinique et l'expérimentation semblent démontrer que des doses élevées de testostérone provoquent un arrêt prématuré de la croissance, le mécanisme exact et le lieu précis de son action sur l'appareil de croissance des os longs restent indéterminés. Au cours de cette étude, des rats máles de 200 g sont injectés à l'aide de doses supra-physiologiques de testostérone pour observer les effects sub-microscopiques sur les diverses zones du cartilage épiphysaire. Au niveau de la zone de division cellulaire, on note une augmentation des cellules en division. Les cellules, en voie de maturation, présentent plus de produits de sécrétion, à un stade plus précoce de leur cycle d'évolution, et semblent subir une hypertrophie plus rapide. Dans la zone pré-hypertrophique, la matrice intercellulaire présente des foyers de calcification précoce, ainsi que des fibres collagènes plus longues et plus épaisses que chez les témoins. Il apparait que, chez l'animal entier, des doses même élevées de testostérone provoquent initialement une stimulation de la prolifération chondrocytaire, avant de favoriser les processus de maturation.
    Abstract: Zusammenfassung Obwohl experimentelle und klinische Erfahrung darauf hinweisen, daß hohe Dosen von Testosteron zu einem frühzeitigen Wachstumsabschluß führen, sind der genaue Mechanismus und der eigentliche Wirkungsort dieses Hormons im Wachstumsapparat der Röhrenknochen unbekannt geblieben. In diesem Experiment wurden 200 g schweren männlichen Ratten supraphysiologische Testosterondosen injiziert, um die submikroskopischen Auswirkungen auf die verschiedenen Zonen des Epiphysenknorpels zu beobachten. In der Zone der Zellmitosen fand sich eine erhöhte Anzahl von sich teilenden Zellen. Die reifenden Zellen häuften im Frühstadium ihres Lebenscyclus größere Mengen von Sekretionsprodukten an und schienen eine abruptere Hypertrophie durchzumachen. In der prähypertrophen Zone enthielt die interterritoriale Matrix Herde von früher und verfrühter Verkalkung, sowie dickere und längere Kollagenfasern als vergleichsweise in Kontrolltieren. Daraus wird geschlossen, daß bei unbehandelten Tieren sogar große Testosterondosen anfänglich eine Stimulation der Chondrocytenproliferation verursachen, bevor sie die Reifungsprozesse veranlassen.
    Notes: Abstract Although experimental and clinical experience indicates that large doses of testosterone lead to premature cessation of growth, the exact mechanism and precise site of action of this hormone on the growth apparatus of long bones remain unknown. In this study, plateaued male rats were injected with supraphysiologic doses of testosterone to observe the submicroscopic effects on the various zones of the epiphyseal cartilage. In the zone of cell division there were increased numbers of dividing cells. The maturing cells accumulated larger amounts of secretory products at earlier stages of their life cycle, and appeared to undergo a more abrupt hypertrophy. In the zone of prehypertrophy, the interterritorial matrix contained foci of early and premature calcification and thicker and longer collagen fibers than at comparable levels in controls. It is concluded that in intact animals, even large doses of testosterone initially cause a stimulation of chondrocyte proliferation, prior to promoting maturation processes.
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  • 58
    ISSN: 1432-0827
    Keywords: Cartilage ; Glycosaminoglycans ; Chromatography ; Ions ; Ageing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Articular cartilage from the lower femoral epiphysis of human autopsy cases was collected in ten age groups from birth to 95 years and in osteoarthrosis of two grades of severity. By chromatography on Ecteola cellulose columns, the different glycosaminoglycans were separated and keratan sulphate was fractionated. The results are consistent with earlier studies using CPC-cellulose column technique, i.e. a higher content of total hexosamine and chondroitin sulphate was found in early childhood. Furthermore, normal articular cartilage in adults showed a higher content and a higher degree of heterogeneity of keratan sulphate than in childhood. In osteoarthrosis, a decreased content of total hexosamine due to a decrease both of chondroitin sulphate and keratan sulphate was found.
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  • 59
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    Calcified tissue international 10 (1972), S. 49-57 
    ISSN: 1432-0827
    Keywords: Histochemistry ; Cartilage ; Enzymes ; Vitamin D ; Starvation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé 1. La distribution quantitative de phosphatase alcaline et de déshydrogénas eglucose-6-phosphate dans les zones hypertrophiques, en voie de prolifération du cartilage de rats rachitiques, pendant la cicatrisation induite par l'administration de vitamin D ou par jeun prolongé, est étudiée. 2. L'activité en phosphatase alcaline de la zone hypertrophique est environ triple de celle de la zone de prolifération du rat sevré normal et environ quadruple de celle des rats rachitiques. 3. Apres administration d'une seule dose de vitamine D (8000 unités), il se produit une augmentation significative de l'activité en phosphatase alcaline de la a zone en voie de prolifération. Une augmentation moins nette est observée dans la zone de prolifération de rats ayant jeuné 48 heures. 4. On observe une diminution significative de l'activité en phosphatase alcaline de la partie centrale de la zone hypertrophique de cartilage aprés traitement a la vitamine D. Une telle diminution n'a pas été observée chez le rat qui jeûne. 5. Chez le rat sevré normal, l'activité en déshydrogénase glucose-6-phosphate (G-6-PDH) est plus élevée dans la zone de prolifération que dans la zone hypertrophique. Chez le rat rachitique non traité, le contenu en enzyme est à peu pres identique dans les deux zones. 6. Apres traitement a la vitamine D, on note une décroissance nette de l'activité en G-6-PDH. Une telle décroissance n'est pas observée chez le rat qui jeûne.
    Abstract: Zusammenfassung 1. Diese Arbeit beschreibt die quantitative Verteilung der alkalischen Phosphatase und der Glucose-6-Phosphat-Dehydrogenase in proliferierenden und hypertrophischen Zonen von rachitischem Rattenknorpel während der durch Vitamin D-Gabe hervorgerufenen Heilung oder während der Mangeldiät. 2. Die Aktivität der alkalischen Phosphatase in der hypertrophischen Zone war etwa dreimal so groß wie in der proliferierende Zone der normalen entwöhnten Ratte und etwa viermal so groß bei den rachitischen Ratten. 3. Nach einer einmaligen Vitamin D-Gabe (8000 i.E) erfolgte eine signifikante Erhöhung der alkalischen Phosphatasen-Aktivität in der proliferierenden Zone. Eine weniger signifikante Erhöhung wurde in der proliferierenden Zone von Ratten festgestellt, welche 48 Std gefastet hatten. 4. Nach Vitamin D-Behandlung erfolgte eine signifikante Abnahme der alkalischen Phosphatasen-Aktivität in der mittleren Region der hypertrophischen Zone des Knorpels. Bei den fastenden Ratten wurde keine solche Abnahme beobachtet. 5. Bei der normalen entwöhnten Ratte war die Aktivität der Glucose-6-Phosphat-Dehydrogenase (G-6-PDH) größer in der proliferierenden als in der hypertrophischen Zone. Bei der unbehandelten rachitischen Ratte war der Enzymgehalt in beiden Zonen etwa gleich. 6. Nach Vitamin D-Behandlung erfolgte eine merkliche Abnahme der G-6-PDH-Aktivität. Bei der fastenden Ratte wurde keine solche Abnahme beobachtet.
    Notes: Abstract 1. The quantitative distribution of alkaline phosphatase and glucose-6-phosphate dehydrogenase in proliferating and hypertrophic zones of rachitic rat cartilage during healing induced by vitamin D administration or starvation is presented. 2. Alkaline phosphatase activity in the hypertrophic zone is about three times greater than that of the proliferating zone of the normal weanling rat and approximately four times greater in the rachitic rats. 3. Following a single dose of vitamin D (8000 I. U.) there was a significant elevation in alkaline phosphatase activity of the proliferating zone. A less significant increase was observed in the proliferating zone of rats which had breen fasted 48 hours. 4. There was a significant decrease in the alkaline phosphatase activity of the central region of hypertrophic zone of cartilage after vitamin D treatment. No such decrease was observed in the fasted rats. 5. In the normal weanling rat glucose-6-phosphate dehydrogenase (G-6-PDH) activity was greater in the proliferating zone as compared to the hypertrophic zone. In the untreated rachitic rat the enzyme content was about the same in the two zones. 6. Following vitamin D treatment there was an appreciable decrease in the G-6-PDH activity. No such decrease was observed in the fasted rat.
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  • 60
    ISSN: 1432-0827
    Keywords: Joint ; Arthritis ; Pyrophosphates ; Cartilage ; Calcinosis ; Glycosaminoglycans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les glycosaminoglycanes du cartilage articulaire de 4 patients, atteints de la maladie de dépot de cristaux CPPD, et de 13 sujets témoins sont fractionnés sur des colonnes de CPC cellulose et ECTEOLA-cellulose. Aucune différence dans la quantité totale d'hexosamines n'a été trouvée dans les deux groupes. Une augmentation de la concentration relative de sulfate de kératane et une décroissance du rapport chondroitine sulfate 4-sulfate et 6-sulfate sont observées chez les sujets atteints. Les courbes de solubilité du chondroitine sulfate sont en augmentation dans les subfractions de faible poids moléculaire et/ou contenant du sulfate dans la maladie de dépot des cristaux CPPD. Les courbes de solubilité du kératane sulfate indiquent un contenu de poids moléculaire élevé et/ou en sulfate dans le cartilage pathologique. Ces modifications cliniques peuvent être partiellement dues à un trouble métabolique des glycosaminoglycanes, contemporain ou précédant le dépot du cristal. Les altérations sont différentes de celles observées au cours de l'ostéoarthrite et dans le cartilage normal en voie de minéralisation.
    Abstract: Zusammenfassung Die Glycosaminoglycane aus Gelenkknorpel von 4 Patienten mit der CPPD (Calcium-Pyrophosphat-Dihydrat)-Kristallablagerunskrankheit und von 13 Kontrollen wurden in CPC-Cellulose- und ECTEOLA-Cellulose-Säulen fraktioniert. Die beiden Gruppen zeigten keinen Unterschied in der Gesamtmenge der Hexosamine. In der Patientengruppe stellte man eine Zunahme der relativen Konzentration von Keratansulfat und eine Abnahme des Chondroitin-4-Sulfat/Chondroitin-6-Sulfat-Verhältnisses fest. Bei der CPPD-Kristallablagerungskrankheit zeigten die Löslichkeitsprofile von Chondroitinsulfat eine Zunahme an Subfraktionen mit einem niederen Molekulargewicht und/oder einem niederen Sulfatgehalt. Andererseits deuteten die Löslichkeitsprofile von Keratansulfat auf hohes Molekulargewicht und/oder hohen Sulfatgehalt im kranken Knorpel. Diese chemischen Veränderungen könnten zum Teil das Ergebnis eines geschädigten Stoffwechsels der Glycosaminoglycane sein und könnten mit der Kristallablagerung einhergehen oder möglicherweise schon vor der Ablagerung eintreten. Die Veränderungen unterscheiden sich von denjenigen in der Osteoarthrose und von normal mineralisierendem Knorpel.
    Notes: Abstract The glycosaminoglycans of articular cartilage in 4 patients with CPPD crystal-deposition disease and 13 controls were fractionated on CPC-cellulose and ECTEOLA-cellulose columns. No difference in the total amount of hexosamines was found between the two groups. An increase in the relative concentration of keratan sulphate and a decrease of the ratio of 4 sulphated and 6-sulphated chondroitin sulphate were encountered in the patient group. Solubility profiles of chondroitin sulphate showed an increase in subfractions representing low molecular weight and/or sulphate content in CPPD crystal-deposition disease. On the other hand, the solubility profiles of keratan sulphate indicated high molecular weight and/or sulphate content in the diseased cartilage. These chemical changes might partly be a result of impaired metabolism of the glycosaminoglycans concomitant with, and possibly also preceding, crystal deposition. The changes are different from those found in osteoarthrosis and from normally mineralizing cartilage.
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  • 61
    ISSN: 1432-0827
    Keywords: Cartilage ; Mineralization ; Histochemistry ; Matrix vesicles
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les expériences portent sur la minéralisation de la plaque épiphysaire tibiale du rat de souche Long-Evans, étudiée après traitement à la cortisone, propylthiouracile ou après jeûn prolongé. Dans des conditions normales, le calcium et le phosphate augmentent au niveau de la matrice extracellulaire, alors que les mucopolysaccharides sulfonés diminuent. Par contre, les vésicules de la matrice au niveau desquels se forment les cristaux d'hydroxyapatite, augmentent. Dans les rats ayant subi un traitement à la propylthiouracile, les cristaux d'hydroxyapatite sont très apparents. Ceci est du à une augmentation du dépôt en calcium, et à une diminution des granules des mitochondries qui contiennent probablement du calcium et du phosphate. En outre, une augmentation du nombre des vésicules de la matrice est visible ainsi qu'une décroissance de la quantité des mucopolysaccharides sulfonés. Dans les rats traités à la cortisone, les cristaux d'hydroxyapatite sont présents, mais dans une quantité moindre que dans les rats ayant subi l'effet du propylthiouracile. Le dépôt en calcium est légèrement réduit; les granules des mitochondries sont plus nombreuses que dans les groupes précédents, le nombre des vesicules de la matrice est plus faible, et les mucopolysaccharides sulfonés sont plus apparents que dans les rats traités à la propylthiouracile. Dans les rats ayant subi l'effet du jeûn, les cristaux d'hydroxyapatite sont fortement réduits ou entièrement absents. Ceci est du à une réduction de dépôt du calcium, une augmentation du nombre des granules des mitochondries (ce qui semble indiquer que les phénomènes de transport vers la matrice extracellulaire sont ralentis), alors que les vésicules de la matrice sont présentes dans des quantités réduites. Les mucopolysaccharides sont plus apparents que dans les animaux traités à la cortisone ou à la propylthiouracile.
    Abstract: Zusammenfassung Die Untersuchung beruht auf einem Vergleich der Mineralisation in der hypertrophischen Zone in der Epiphysealplatte von Long Evans Ratten die mit Kortison, Propylthiourazil oder einfachem Fasten behandelt wurden. Unter Normalbedingungen lassen sich in der extrazellulären Matrix der Calcifikationszone die folgenden Veränderungen beobachten: der Gehalt an Calcium und Phosphat nimmt zu, derjenige an Mukopolysacchariden nimmt ab, während die Matrixvesiclen, in denen sich die Bildung des Hydroxylapatits vollzieht, zunehmen. In Ratten die mit Propylthiourazil behandelt wurden, treten die Hydroxylapatitskristalle besonders hervor. Dies hängt mit einer Zunahme der Calciumablagerung zusamen sowie einer Abnahme der Mitochondriengranulation (in denen vermutlich Calcium und Phosphat enthalten sind). Ferner hängt damit zusammen eine numerische Zunahme der Matrixvesiceln sowie ein starke Abnahme des Gehalts an sulfonierten Mucopolysacchariden. In den mit Kortison behandelten Ratten sind Hydroxylapatikristalle nachweisbar, wenn auch weniger zahlreich als in den mit Propylthiourazil behandelten. Dem entspricht auch eine leicht reduzierte Calciumablagerung sowie eine Mitochondrialgranulation die derjenigen der anderen Ratten überlegen ist; Matrixvesiceln sind weniger zahlreich und sulfonierte Mucopolysaccharide sind deutlicher nachweisbar als in den Tieren, die Propylthiourazil erhielten. Fasten führt zu einem auffallenden Verlust an Hydroxylapatitkristallen. Diese können sogar nicht mehr zu erkennen sein. Dies hängt mit verminderter Calciumablagerung zusammen sowie einer Zunahme der Mitochondrialgranulation. Dies ist vermutlich Ausdruck einer Transportverzögerung zur extrazellularen Matrix. Nach Fasten ist auch die Anzahl der Matrixvesiceln auffallend herabgesetzt, und der Gehalt an sulfonierten Mucopolysacchariden ist größer als in den mit Kortison bzw. Propylthiourazil behandelten Tieren.
    Notes: Abstract Comparison of mineralization in the hypertrophic zone of the tibial epiphyseal plate in immature rats was carried out after treatment with cortisone, propylthiouracil, or after fasting. Under normal conditions, in the extracellular matrix at the calcification front, calcium and phosphate increased, sulfated mucopolysaccharides decreased, and matrix vesicles, which serve as the locus for the formation of hydroxyapatite crystals, increased. In propylthiouracil-treated rats, hydroxyapatite crystals were prominent, related to an increase in calcium deposition, a decrease of mitochondrial granules (thought to contain calcium and phosphate), an increase in the number of matrix vesicles, and to a marked decrease in the amount of sulfated mucopolysaccharide. In cortisone-treated rats, hydroxyapatite crystals were present but they were not as numerous as in the propylthiouracil-treated rats. Correspondingly, calcium deposition was slightly reduced, mitochondrial granules were more numerous than in the previous groups of rats, matrix vesicles were less numerous, and sulfated mucopolysaccharide were more prominent than in the propylthiouracil-treated rats. In fasted rats, hydroxyapatite crystals were markedly reduced or absent, and related to a decrease in calcium deposition, an increase in the number of mitochondrial granules (suggesting a delay in transport to the extracellular matrix). Matrix vesicles were markedly reduced in number, and sulfated mucopolysaccharide much more prominent than in either the cortisone or the propylthiouracil-treated rats.
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  • 62
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    Calcified tissue international 15 (1974), S. 253-267 
    ISSN: 1432-0827
    Keywords: Ageing ; Cartilage ; 35S-incorpotation ; Glycosaminoglycans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé L'incorporationin vitro de35S-sulfate en fractions chondroitine sulfate, étudiée quantitativement et après séparation, a été déterminée dans diverses couches de cartilage bovin en fonction de l'âge. Après incubationin vitro dans une solution de Tyrode avec du35S-sulfate, les specimens sont lavés pendant 12 heures dans une solution saturée de sulfate de sodium. Aucune perte en chondroitine sulfate n'est observée dans le cartilage articulaire jeune et adolescent après incubation et lavage: les protéoglycanes semblent fermement liés. Au contraire, des pertes importantes de chondroitine sulfate sont observées dans le cartilage épiphysaire. Chez la vache, on ne constate pas de perte de condroitine sulfate dans les couches superficielles, mais des pertes augmentées de chondroitine et kératane sulfates sont notées dans les couches moyenne et profonde. Quel que soit l'âge, la couche de surface articulaire présente des «activités spécifiques» élevées dans les diverses fractions de chondroitine sulfate et une incorporation de35S-sulfate dans le chondroitine sulfate de poids moléculaire et/ou de densité de charge élevès, suggérant une activité métabolique augmentée. Dans la matrice de cette couche, le chondroitine sulfate est surtout constitué de fractions à poids moléculaire faible et peu chargées Dans les autres couches, l'incorporation du35S s'effectue en fonction de la distribution du chondroitine sulfate. Ce fait suggère que la sulfatation s'effectue à l'intérieur des cellules. Les résultats semblent indiquer un taux de synthèse identique pour différents types de chondroitine sulfate. L'incorporation augmentée, du35S-sulfate en chondroitine sulfate à des âges plus avancés indiquent une synthèse plus rapide. probablement secondaire à une dégradation augmentée.
    Abstract: Zusammenfassung Der in-vitro-Einbau von35S-Sulfat in Chondroitinsulfat-Fraktionen wurde in verschiedenen Schichten von bovinem Gelenkknorpel in Zusammenhang mit dem Alter quantitativ bestimmt. Nach in-vitro-Inkubation in Tyrodelösung mit35S-Sulfat wurden die Proben 12 Std lang in gesättigter Natriumsulfatlösung gewaschen. In jungem und jugendlichem Gelenkknorpel erfolgte nach der Inkubation und dem Waschen kein Verlust von Chondroitinsulfat. Die Proteoglycane scheinen fest gebunden zu sein. Hingegen wurde beim Epiphysenknorpel ein großer Verlust von Chondroitinsulfat festgestellt. Bei der Kuh verloren die Oberflächenschichten kein Chondroitinsulfat, aber bei den mittleren und tiefen Schichten erfolgte ein zunehmender Verlust von Chondroitin- und Keratansulfat. Bei allen Altersstufen zeigte die 40 μ dicke Oberflächenschicht des Gelenkknorpels hohe „spezifische Aktivitäten” in den verschiedenen Chondroitinsulfat-Fraktionen und Einbau von35S-Sulfat in Chondroitinsulfat von vorwiegend hohem Molekulargewicht und/oder hoher Ladungsdichte, was einen erhöhten Umsatz vermuten läßt. In der Matrix dieser Schicht bestand das Chondroitinsulfat vorwiegend aus Fraktionen mit niederem Molekulargewicht und niederer Ladung. In den anderen Schichten entsprach das Bild des35S-Einbaus weitgehend der Verteilung des Chondroitinsulfats. Dies deutet darauf hin, daß die Sulfatierung innerhalb der Zellen vollzogen wird. Weiter ließen die Befunde ähnliche Umsatzraten für die verschiedenen Chondroitinsulfate vermuten. Der erhöhte Einbau von35S-Sulfat ins Chondroitinsulfat bei höherem Alter deutet auf eine erhöhte Synthese, wahrscheinlich sekundär zum erhöhten Abbau.
    Notes: Abstract The incorporationin vitro of35S-sulphate into chondroitin sulphate fractions, quantitatively assessed and separated, was studied in different layers of bovine articular cartilage in relation to age. After incubationin vitro in Tyrode solution with35S-sulphate the specimens were washed for 12 hours in saturated sodium sulphate solution. No loss of chondroitin sulphate occurred in the young and adolescent articular cartilage after the incubation and washing procedures. Thus, the proteoglycans appear to be firmly bound within this cartilage In contrast, large losses of chondroitin sulphate were found from the epiphyseal cartilage. In the cow, no chondroitin sulphate was lost from the superficial layers but increasing losses of both chondroitinand keratansulphate occurred from the middle to the deep layers. In all ages, the articular surface layer 40 μ in thickness showed high “specific activities” in the different chondroitin sulphate fractions and incorporation of35S-sulphate into chondroitin sulphate of predominantly high molecular weight and/or charge density suggesting an increased turnover rate of these fractions. In the matrix of this layer, the chondroitin sulphate consisted of predominantly low molecular weight/low charged fractions. In the other layers, a pattern of35S incorporation was found largely reflecting the actual chondroitin sulphate distribution pattern. This suggests that sulphation is completeted intracellularly. Further, the data suggested similar turnover rates for the different chondroitin sulphates. Increased incorporation of35S-sulphate into chondroitin sulphate at higher ages indicated increased rate of synthesis, probably secondary to increased degradation.
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  • 63
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    Calcified tissue international 2 (1968), S. 197-213 
    ISSN: 1432-0827
    Keywords: Ossification ; Epiphysis ; Cartilage ; Mucopolysaccharides ; Growth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé La crête iliaque est une épiphyse de traction, où s'insèrent de larges faisceaux de collagène. Chez l'enfant, elle est constituée par une couche importante de cartilage hyalin surmontant une plaque de croissance. Des divisions de cellules chondroblastiques, au niveau du cartilage hyalin et de la plaque de croissance, sont plus nombreuses pendant les deux periodes de croissance rapide du tronc, à savoir l'enfance et l'adolescence. L'ossification enchondrale, identique à celle des os longs, s'observe au niveau de la plaque de croissance de la crête iliaque, entre les régions où se produisent la calcification et l'ossification; elle semble identique à celle qui se produit au niveau de l'insertion de certains ligaments. Un centre d'ossification secondaire apparait au niveau de la partie médiane du cartilage de la crête pendant l'adolescence. L'os se forme, souvent, directement au-dessus du cartilage comme dans le cas de la reconstitution d'une nécrose osseuse aseptique. La concentration en galactosamine du cartilage de la crête iliaque d'enfants normaux décroit brutalement au cours des deux premières années de la vie. Puis elle décroit plus lentement jusqu'à l'âge de neuf à douze ans, pour augmenter à nouveau jusqu'à seize ans. Cette augmentation coïncide avec le début de la croissance de l'adolescence. La concentration en glucosamine reste constante. L'hydroxylproline augmente nettement pendant l'enfance, puis reste inchangée.
    Abstract: Zusammenfassung Der Beckenkamm ist eine Zugepiphyse, an welcher große Bündel von kollagenen Fasern ansetzen. Beim Kind besteht sie aus einer Masse von hyalinem Knorpel, der die Wachstumsplatte überlagert. Die Knorpelzellteilung im hyalinen Knorpel und in der Wachstumsplatte ist während der beiden raschen Wachstumsperioden des Rumpfes in der Kindheit und in der Adoleszenz gesteigert. Die endochondrale Verknöcherung, wie sie in den Röhrenknochen zu sehen ist, nimmt in der Wachstumsplatte des Beckenkammes ihren Verlauf zwischen Zonen in welchen Calcification und Ossification stattfinden, ähnlich jener an der Ansatzstelle gewisser Ligamente. Ein sekundäres Ossifikationszentrum entwickelt sich in der Mitte des Kammknorpels während der Adoleszenz. In manchen Bezirken bildet sich der Knochen direkt über dem Knorpel, indem er diesen ersetzt, ein Vorgang, der auch bei der Abheilung aseptischer Knochennekrosen in Form des sogenannten “kriechenden Ersatzes” beobachtet werden kann. Die Galaktosamin-Konzentration im Knorpel des Beckenkammes normaler Kinder sinkt beträchtlich während der zwei ersten Lebensjahre. Später nimmt sie allmählich weiter ab bis zum Alter von 9–12 Jahren, um anschließend bis zum 16. Lebensjahr wieder anzusteigen. Diese Zunahme fällt mit der Zeit des Wachstumsstoßes in der Adoleszenz zusammen. Die Konzentration des Glucosamins innerhalb der untersuchten Altersgruppe bleibt praktisch konstant. Das Hydroxyprolin nimmt im Kleinkindesalter stark zu, bleibt jedoch später konstant.
    Notes: Abstract The iliac crest is a traction epiphysis where large bundles of collagen fibers insert. In the child it is formed by a large mass of hyaline cartilage overlying a growth plate. Cartilage cell divisions in the hyaline cartilage and in the growth plate are more numerous during both periods of rapid trunk growth, infancy and adolescence. Enchondral ossification as seen in the long bones occurs in the iliac crest growth plate in between areas where calcification and ossification takes place resembling that at the insertions of certain ligaments. A secondary center of ossification develops in the middle of the crest cartilage during adolescence. In many areas bone forms directly over the cartilage, replacing it, resembling the “creeping replacement” observed in healing of aseptic bone necrosis. The galactosamine concentration in the iliac crest cartilage of normal children decreases sharply during the first two years of life. Then, it declines gradually until nine to twelve years of age, and then, again, increases until sixteen years of age. This increase takes place at the time when the adolescent growth spurt occurs. Glucosamine concentration remains nearly constant throughout the age range studied. Hydroxyproline increases sharply during infancy and stays unchanged thereafter.
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  • 64
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    Calcified tissue international 2 (1968), S. 253-261 
    ISSN: 1432-0827
    Keywords: Calcium ; Ion ; Binding ; Chondroitin ; Cartilage
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé La liaison du calcium par le sulfate de chondroitin C a été étudiée en utilisant des méthodes biologiques et physicochimiques pour la détermination quantitative du calcium ionique. Les valeurs moyennes pour logK j, le logarithme de la constante de formation non corrigée d'un complexe calcium-sulfate de chondroitin C, sont 1,64 par la technique de cœur de grenouille, 1,55 par la méthode de murexide, 1,39 par l'ultrafiltration et 1,04 par l'électrode sélective du calcium. L'ordre de grandeur de ces valeurs suggèrent que le sulfate de chondroitin C a une capacité de liaison pour calcium relativement élevée, mais les valeurs calculées pour un paramètre d'échangeK p indiquent que le calcium a une affinité faible pour le polysaccharide.
    Abstract: Zusammenfassung Die Bindung von Calcium mit Chondroitinsulfat C wurde untersucht mittels biologischer und physikalisch-chemischer Methoden für die quantitative Bestimmung des ionischen Calciums. Im Durchschnitt ergab logK j, der Logarithmus der unkorrigierten Formationskonstante für einen Calciumchondroitinsulfatkomplex, 1,64 mit der Technik des Froschherzens, 1,55 mit der Murexidmethode, 1,39 mittels Ultrafiltration und 1,04 mit den Calciumselektiven Elektroden. Die Größe dieser Werte zeigt, daß Chondroitinsulfat eine relativ hohe Bindungskapazität für Calcium hat, die berechneten Werte für einen AustauschparameterK p jedoch, daß Calcium eine kleine Affinität für Polysaccharide hat.
    Notes: Abstract The binding of calcium by chondroitin sulfate C was studied by employing biological and physicochemical methods for the quantitative determination of ionic calcium. Mean values for logK j, the logarithm of the uncorrected formation constant for a calcium-chondroitin sulfate complex, were 1.64 with the frog heart technique, 1.55 with the murexide method, 1.39 with ultrafiltration and 1.04 with the calcium selective electrode. The magnitude of these values suggests that chondroitin sulfate has a relatively high binding capacity for calcium, but calculated values for an exchange parameterK p indicate that calcium has a low affinity for the polysaccharide.
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    Calcified tissue international 15 (1974), S. 237-251 
    ISSN: 1432-0827
    Keywords: Cartilage ; Glycosaminoglycans ; Concentration ; Distribution ; Age
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les glycosaminoglycanes de diverses couches de cartilage articulaire bovin de veaux, génisses et vaches ont été étudiés quantitativement et qualitativement en utilisant une technique de microfractionnement, suivie d'electrophorèse en microzone des diverses fractions. Le contenu en eau, calcium et hydroxyproline a également été déterminé. Le chondroitine-4-sulfate constitue le glycosaminoglycane principal dans toutes les couches chez le veau et la génisse, alors que le chondroitine-6-sulfate prédomine chez la vache. Des fractions correspondantes des microcolonnes présentent une mobilité électrophorétique variable, aussi bien en fonction de la profondeur à partir de la surface articulaire qu'avec l'âge. En général, une concentration augmentée de glycosaminoglycanes est observée à tous les âges en s'éloignant de la surface articulaire. Chez les veaux et les génisses, le chondroitine sulfate est responsable de cette augmentation, alors que chez les vaches, ce sont le kératane sulfate et, vraisemblablement, les glycoprotéines qui sont les constituants majeurs, responsables des concentrations plus élevées en hexosamine observées dans les couches les plus profondes. A tout âge, le chondroitine sulfate montre une augmentation relative des fractions à poids moléculaire/densité de charge augmentés en fonction de la plus grande profondeur. Chez les veaux et les génisses, les fractions isolées présentent une homogénéité electrophorétique, alors que chez les vaches un chondroitine sulfate supplémentaire à faible mobilité est mis en évidence. Ce chondroitine sulfate de la surface articulaire est fort hétérogène en fonction de l'âge. Le rapport des fractions combinées kératane sulfate et glycoprotéines par rapport au chondroitine sulfate augmente notablement avec l'âge.
    Abstract: Zusammenfassung Die Glycosaminoglycane von verschiedenen Schichten aus bovinem Gelenkknorpel von Kälbern, jungen und älteren Kühen wurden quantitativ und qualitativ untersucht, und zwar mittels CPC-Mikrofraktionierung und darauffolgender Mikrozonen-Elektrophorese bei verschiedenen Fraktionen; auch der Gehalt an Wasser, Calcium und Hydroxyprolin wurde bestimmt. Chondroitin-4-Sulfat kam in allen Schichten des Knorpels von Kälbern und jungen Kühen am häufigsten vor, während Chondroitin-6-Sulfat bei älteren Kühen vorherrschte. Entsprechende Fraktionen von den Mikrosäulen zeigten unterschiedliche elektrophoretische Eigenschaften, je nach der Schichttiefe von der Gelenksoberfläche und nach dem Alter. Im allgemeinen wurde bei zunehmendem Abstand von der Gelenksoberfläche eine zunehmende Konzentration an Glycosaminoglycanen festgestellt. Bei Kälbern und jungen Kühen wurde diese Zunahme durch Chondroitinsulfat verursacht, während bei älteren Kühen Keratansulfat und möglicherweise Glycoproteine die Hauptkomponenten der erhöhten Hexosamin-Konzentrationen waren, welche bei zunehmender Schichttiefe festgestellt wurden. In allen drei Gruppen zeigte Chondroitinsulfat bei zunehmender Schichttiefe eine relative Zunahme von Fraktionen mit höherem Molekulargewicht und höherer Ladungsdichte. Bei Kälbern und jungen Kühen zeigten die einzelnen Fraktionen elektrophoretische Homogenität, während bei älteren Kühen ein zusätzliches Chondroitinsulfat mit äußerst langsamer Mobilität gefunden wurde. Das Chondroitinsulfat der Gelenksoberfläche zeigte bei allen Altersgruppen eine beträchtliche Heterogenität. Das Verhältnis der kombinierten Keratansulfat- und Glycoproteinfraktionen zum Chondroitinsulfat nahm mit zunehmendem Alter beträchtlich zu.
    Notes: Abstract The glycosaminoglycans in different layers of bovine articular cartilage from calves, heifers and cows were studied quantitatively and qualitatively using CPC microfractionation procedure and by subsequent microzone electrophoresis on different fractions; water, calcium and hydroxyproline were also estimated. Chondroitin-4-sulphate was the main glycosaminoglycan in all layers in calves and heifers while chondroitin-6-sulphate predominated in cows. Corresponding fractions from the microcolumns showed different electrophoretic mobility varying with the depth from the articular surface and with age. In general, increased concentration of glycosaminoglycans was found in all ages with increasing distance from the articular surface. In calves and heifers, chondroitin sulphate was responsible for this increase while in cows, keratan sulphate, and possibly glycoproteins, were the major components of the increased hexosamine concentrations found with increasing depth. In all ages chondroitin sulphate showed a relative increase of fractions with higher molecular weights/charge density with increasing depth. In the calves and heifers, the individual fractions showed electrophoretical homogeneity while in cows an additional chondroitin sulphate showing an extremely slow mobility was found. The chondroitin sulphate in the articular surface layer showed considerable heterogeneity in all ages. The ratio of the combined keratan sulphate and glycoprotein fractions to chondroitin sulphate increased significantly with increasing age.
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    Calcified tissue international 3 (1969), S. 107-124 
    ISSN: 1432-0827
    Keywords: Aspidin ; Bone ; Cartilage ; Dentine ; Evolution ; Fossil
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
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    Calcified tissue international 2 (1968), S. 214-228 
    ISSN: 1432-0827
    Keywords: Calcinosis ; Calcification ; Cartilage ; Collagen ; Mineral metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les facteurs, influençant la vitesse et l'intensité du phénomène d'association des ions calcium et phosphates avec des fibres contenant du collagène, et préparés à partir du tendon de boeuf par deux méthodes d'extraction différentes, ont été étudiés. Les fibres, obtenues par ces deux méthodes, nécessitent spécifiquement du phosphate pour absorber du calcium et vice versa. L'absorption ionique des deux préparations est inhibée par du Mg++, du pyrophosphate et un peptide acidique, isolé du sérum humain. Alors que les fibres contenant du collagène, préparées selon les deux méthodes, présentent une absorption ionique à des vitesses sensiblement identiques, seule une des méthodes donne une matrice réagissant positivement à la technique de coloration au nitrate d'argent de von Kossa. Etant donné que les deux critères de calcification sont intéressés de façon identique par des conditions de réaction et par des inhibiteurs, il apparait que les deux facteurs sont des manifestations de différents stades de calcification et que des études d'absorption ionique fournissent une base quantitative d'appréciation de la calcification, pouvant être d'importance pour l'étude du mécanisme et de contrôle de la minéralisation tissulaire.
    Abstract: Zusammenfassung Überprüft wurden die Faktoren, welche Geschwindigkeit und Ausmaß der Erscheinung beeinflussen, wobei Calcium- und Phosphationen sich mit den kollagenhaltigen, durch zwei verschiedene Extraktionsmethoden aus Rindersehnen gewonnenen Fasern eng zusammenbinden. Die mit beiden Methoden zubereiteten Fasern benötigen spezifisch Phosphat für die Calciumaufnahme und Calcium für die Phosphataufnahme. Die Ionenaufnahme beider Arten wird durch Mg++, Pyrophosphat und saure, aus dem menschlichen Serum isolierte Peptide gehemmt. Während die nach beiden Methoden präparierten kollagenhaltigen Fasern eine Ionenaufnahme von beinahe gleicher Geschwindigkeit verursachen, ergibt nur eine dieser Methoden eine Matrix, die mit der Silbernitratfärbung nach vonKossa positiv reagiert. Da beide Calcifikationskriterien gleicherweise durch Reaktionsbedingungen und Inhibitoren beeinflußt werden, wird daraus geschlossen, daß beide Erscheinungen verschiedener Stadien des Gesamtcalcifikationsprozesses sind. Untersuchungen über die Ionenaufnahme ergeben eine quantitative Angabe der Verkalkung, welche für die Erforschung des Mechanismus und der Kontrolle der Mineralisation der Gewebe wichtig sein könnte.
    Notes: Abstract Factors that influence the rate and extent of the phenomenon in which calcium and phosphate ions become firmly associated with collagen-containing fibers prepared from beef tendon by two different extraction methods have been investigated. The fibers produced by both methods specifically require phosphate for calcium uptake and calcium is required for phosphate uptake. Ion uptake by both types is inhibited by Mg++, pyrophosphate, and an acidic peptide isolated from human serum. Whereas the collagen-containing fibers prepared by both methods induce ion uptake at nearly identical rates, only one of the methods produced a matrix that gives a positive response to the silver nitrate staining technique of von Kossa. Since both criteria of calcification are similarly influenced by reaction conditions and inhibitors, it is concluded that both are manifestations of different stages of the overall calcification process and that studies of ion uptake provide a quantitative assessment of calcification which could be of importance for investigating the mechanism and control of tissue mineralization.
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    Calcified tissue international 30 (1980), S. 27-34 
    ISSN: 1432-0827
    Keywords: Ultrastructure ; Calcium ; Cartilage ; Vesicles
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    Topics: Biology , Medicine , Physics
    Notes: Summary The potassium pyroantimonate technique was utilized for the selective subcellular localization of calcium in the mandibular condylar cartilage of 1-day-old rats. Electron dense calcium pyroantimonate precipitates were localized principally in mitochondria and at the cell membrane of the chondrocytes. In addition, small intracellular vesicles 0.1–0.2µm in diameter were observed in proximity to the cell membrane of chondrocytes of the mid-hypertrophic zone. The results suggest that these vesicles were being extruded from the cell into the extracellular matrix. Energy-dispersive analysis by X-rays confirmed that calcium is the principal cation of the electron-dense precipitates.
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  • 69
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    Keywords: Mandible ; Cartilage ; Osteoid ; Differentiation ; In Vitro
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    Notes: Summary To evaluate the requirement of vascular and/or neurotrophic-derived factors on determination and differentiation of chondrogenic and osteogenic phenotypes, early embryonic quail and mouse mandibular processes were cultured using a modified Trowell method in a serumless, chemically-defined medium for 10 days. Quail HH stage 22 and mouse Theiler stage 16 mandibular processes formed cartilage and produced osteoid under these experimental conditions. Chondrogenic and osteogenic phenotypes were expressed without serum or other exogenous growth-promoting influences.
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  • 70
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    Calcified tissue international 36 (1984), S. 332-337 
    ISSN: 1432-0827
    Keywords: Proteolipid ; Phospholipids ; Vitamin D ; Calcification ; Cartilage
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Previous studies have shown thatin vitro calcification of chick epiphyseal cartilage matrix vesicles is proteolipid-dependent. The purpose of this research is to examine the role of proteolipid in cartilage calcificationin vivo by comparing the proteolipid concentration of normal and vitamin D-deficient chick epiphyseal cartilage, the relationship of proteolipid to other tissue lipids, and its ability to supportin vitro apatite formation. Proteolipid was isolated from the upper growth centers (reserve cell zone, upper proliferative zone) and lower growth centers (lower proliferative, hypertrophic, and calcified cartilage zones) of long-bone epiphyses from 3-week-old normal and rachitic male white leghorn chicks by Sephadex LH-20 chromatography of the crude phospholipid component of the total lipid extract. In both normal and rachitic tissue the proteolipid/dry weight and proteolipid/total lipid ratios were greater in the lower growth center than in the upper zones. No statistically significant change in the proteolipid/total lipid ratio in rachitic tissues relative to comparable cell zones in normal cartilages was observed. However, there was an increase in the nonproteolipid phospholipid content of rachitic tissues, altering the relative proteolipid/phospholipid composition. Whereas proteolipids from normal tissue supportedin vitro calcification, proteolipids from rachitic tissues did not, indicating a direct effect of vitamin D on proteolipid structure. These data support the hypothesis that failure of rachitic cartilage to calcifyin vivo may be due in part to alterations in phospholipid and proteolipid metabolism.
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    Calcified tissue international 36 (1984), S. 702-710 
    ISSN: 1432-0827
    Keywords: Corticosteroids ; Cartilage ; Organ culture ; Calcium ; Matrix vesicles
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary The present study examined the effects of various corticosteroid and noncorticosteroid hormones upon the ultrastructure of chondroprogenitor cells and chondroblasts in an organ-culture system of late fetal condylar cartilage. Corticosteroids, (triamcinolone acetonide, dexamethasone, corticosterone) at concentrations of 10−6–10−8M stimulated markedly a precocious formation of matrix vesicles by chondroblasts. This stimulation was accompanied by a significant accretion of calcium complexes intra- and extracellularly in both the chondroprogenitor cell population and chondroblastsin vitro, as well as in the newly induced matrix vesicles. Nonglucocorticoid steroids (progesterone, estradiol, testosterone, cortexolone) did not evoke similar effects. Progesterone and testosterone, however, seemed to adversely affect the ultrastructure of the cartilage cells, whereas estradiol appeared to have a favorable effect on the morphology of cultured condylar cartilage.
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    Calcified tissue international 5 (1970), S. 91-99 
    ISSN: 1432-0827
    Keywords: Rickets ; Cartilage ; Vitamin D ; Calcium ; Phosphate ; Mitochondria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Abstract Des métaphyses de rats rachitiques, alimentés par paires avec des rats témoins, normaux et traités à la vitamine D2 et des phosphates, ont été étudiées au microscope électronique. La répartition des granules mitochondriales est modifiée dans le tissu rachitique: des granules ne sont visibles que dans quelques cellules voisines de la région en voie de calcification. Les rats témoins présentent une répartition, en gradient, à travers toute la métaphyse. L'adjonction à ce régime cariogène de phosphate ou vitamine D2 ou les deux à la fois, permet de rétablir la répartition normale des granules, visible chez les témoins. Il semble qu'une des modifications spécifiques, induites dans les mitochondries des chondrocytes, par un régime pauvre en phosphate et déficient en vitamine D2, est la diminution de la formation de granules mitochondriales, contenant des produits inorganiques.
    Abstract: Zusammenfassung Wachstumszonen von rachitischen Ratten werden unter dem Elektronenmikroskop verglichen mit denjenigen von 3 Kontrollgruppen (normale Diät rachitogene Diät mit Vitamin D2 und Phosphatzusätzen und eingeschränkte Menge normaler Diät). Die Verteilung der Granula in den Mitochondrien war im rachitischen Gewebe verändert; Granula wurden nur in einigen Zellen festgestellt, welche an die Zone der provisorischen Kalzifikation angrenzten. Kontrollratten zeigten einen Gradienten von Granula auf der ganzen Wachstumsplatte. Ergänzung der rachitogenen Diät durch Phosphat, Vitamin D2 oder beide bewirkte die Wiederherstellung der Granula-Dichte und-Verteilung, welche die Kontrolltiere aufwiesen. Es wird vorgeschlagen, daß die verminderte Fähigkeit, mineralhaltige Mitochondrien-Granula zu bilden, eine spezifische Veränderung ist, welche in den Mitochondrien von Chondrocyten durch eine phosphatarme, Vitamin D2-defizitäre Diät herbeigeführt wird.
    Notes: Abstract Growth plates of rachitic, pair-fed control, normal and Vitamin D2- and phosphate-treated rachitic rats were studied with the electron microscope. Mitochondrial granule distribution was modified in the rachitic tissue; granules were noted only in a few cells adjacent to the zone of provisional calcification. Control rats demonstrated a gradient of granules throughout the growth plate. Supplementation of the rachitogenic diet with either phosphate, Vitamin D2, or both was able to re-establish the granule density and distribution found in control animals. It is suggested that one specific modification induced in mitochondria of chondrocytes by a low phosphate, Vitamin D2-deficient diet is the reduced ability to form mineral-containing mitochondrial granules.
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  • 73
    ISSN: 1432-0827
    Keywords: Endochondral mineralization ; Cartilage ; Alkaline phosphatase ; In vitro ; Levamisole
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Isolated mesenchymal limb bud cells from day-12 mouse embryos grown at high density in organoid culture at the medium/air interphase differentiate into chondrocytes and form cartilage nodules. Upon addition of β-glycerophosphate (β-GP), cartilage undergoes endochondral mineralization. This β-GP-induced mineralization was investigated by measuring the calcium content in the cultures and the activity of alkaline phosphatase (AP) in the cell mass and the medium. Calcium incorporation depended on the amount of β-GP added. After continuous treatment, mineralization began on day 8 of the culture period and increased linearly until day 15. In long-term cultures, periodical treatment for 6 days caused an increase in mineralization the older the cultures were, but the slope of increase was proportionately less steep. Treatment at the latest period on days 19–24 resulted in a markedly reduced mineralization. After short-term treatment (48 hours), mineralization increased also the older the cultures were and proceeded during further cultivation in β-GP-free medium. This kinetic behavior indicates a dependency of mineralization on cartilage maturation in this in vitro system. AP activity increased enormously and nearly logarithmically in the cell mass in β-GP-free medium, whereas β-GP treatment inhibited this drastic increase. In the medium, considerable activities of AP were also measurable from day 10 onward. It increased in β-GP-free medium up to day 14, but was diminished after mineralization had been induced. Levamisole inhibited AP activity dose dependently when added directly to the enzyme-containing medium (100% inhibition at 10-3 M). Added to the cultures from day 7 to 14, it partially inhibited AP activity and mineralization at 5×10-5 M; mineralization was totally inhibited at 10-3 M, but AP activity was still present. This high concentration was cytotoxic, as revealed ultrastructurally and by GAG estimation. This in vitro system comprises cartilage development and maturation, β-GP-inducible endochondral mineralization, and final degenerative changes; it may be an appropriate model for investigations on endochondral mineralization.
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    Calcified tissue international 52 (1993), S. 465-469 
    ISSN: 1432-0827
    Keywords: Type II collagen ; Mandibular condyle ; Cartilage ; Differentiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary An experiment was designed to mimic orthopedic functional appliances in order to investigate the spatial and temporal characteristics of type II collagen secretion as a marker of cartilage maturation in the mandibular condyle of young rabbits. The position of the glenoid fossa in relation to the condyle was altered so that articulation now took place more posteriorly. Histological sections of the condyles of 15-, 20- and 30-day-old experimental and control animals were stained with toluidine blue and with an anti-type II collagen antibody. A widened progenitor cell layer was found posteriorly in the experimental condyles and a narrow layer was found anteriorly to the articulating region. The chondroblast layer was also widened posteriorly, whereas the hypertrophic cell zone was narrower; the opposite was seen anteriorly. The effect was marked in 15- and 20-day-old animals and weak in 30-day-old animals. Type II collagen stain and strong toluidine blue metachromasia were not observed in the progenitor cell zone until the chondroblasts had acquired a flattened, slightly hypertrophic morphology, which was found deeper in the experimental condyles than in the controls. This is interpreted as a slowing down of the differentiation of chondroblasts as a result of the force applied. The effect of masticatory function may also be explained in terms of delayed differentiation of chondroblasts and increased growth.
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    Calcified tissue international 54 (1994), S. 296-303 
    ISSN: 1432-0827
    Keywords: Cartilage ; Chondrocytes ; Glycosaminoglycans ; Growth plate ; Sulfation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract We have used antibodies to chondroitin 4- and 6-sulfate and keratan sulfate along with Alcian blue staining of sulfated proteoglycans to investigate changes in content and sulfation within the avian growth plate. In normal chicks, chondroitin 4- and 6-sulfate content were similar in the proliferating and transitional zones but in the hypertrophic zone, chondroitin 4- and 6-sulfate were slightly lower (13% and 18%, respectively) and keratan sulfate was markedly lower (58%). Compared with the proliferative zone, Alcian blue staining of sulfated glycosaminoglycans was markedly lower in both the transitional (46%) and hypertrophic (22%) zones. In tibial dyschondroplasia, where chondrocyte maturation is arrested at the transitional zone, there was no difference in the chondroitin 4- and 6-sulfate or keratan sulfate staining between the proliferative and transitional zones, which were similar to normal birds. With Alcian blue staining there was no difference in the intensity of the staining within the proliferating zone compared with normal birds but stainign in the transitional chondrocytes was markedly higher (39%). These results suggest that in the early steps of chondrocyte maturation there may be a decrease in the degree of glycosaminoglycan sulfation without any alteration in glycosaminoglycan concentration, and that further maturation may be accompanied by a change in the nature of the proteoglycans which may also affect the level of sulfation.
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    Calcified tissue international 56 (1995), S. 376-381 
    ISSN: 1432-0827
    Keywords: Chondrocyte differentiation ; Cartilage ; TGF-β ; IGF-I osteochondrosis ; Pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Osteochondrosis and dyschondroplasia are common multifocal disturbances of endochondral ossification in many species of domestic animals, and are characterized by the retention of avascular cartilage. These cartilage disorders are characterized by a failure of chondrocyte differentiation, matrix mineralisation and its replacement by bone. Rabbit polyclonal antibodies to transforming growth factor-beta (TGF-β) and to insulin-like growth factor-I (IGF-I) were used to detect the two growth factors in normal and osteo-chondrotic porcine epiphyses. In the normal pig epiphyses IGF-I and TGF-β were present in the chondrocytes of the epiphyseal hyaline cartilage and IGF-I was readily localised to the hypertrophic chondrocytes in the growth cartilage adjacent to the epiphyseal ossification centre. Both growth factors were found to be deficient in chondrocytes at sites of osteochondrosis. Both these growth factors are thought to be involved in the cascade of events associated with chondrocyte function during endochondral ossification. Deficiencies in TGF-β and IGF-I demonstrated in porcine osteochondrosis and previously shown in avian dyschondroplasia suggest further similarities in the pathogenesis of these conditions.
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    Calcified tissue international 6 (1970), S. 55-69 
    ISSN: 1432-0827
    Keywords: Glycosaminoglycans ; Cartilage ; Embryo ; Limb ; Chick
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des données microchimiques quantitatives ont pu être obtenues sur les glycosaminoglycanes du cartilage embryonnaire de poulet, en voie de développement. Le glycosaminoglycane principal est constitué par le sulfate de chondroitine. Cette substance apparait en même temps que la différenciation histologique des membres et augmente rapidement en même temps que le cartilage des membres s'accroit. Des quantités significatives d'autres glycosaminoglycanes n'ont pu être retrouvées dans les membres totaux, les rudiments osseux ou dans des échantillons cartilagineux. Les caractéristiques du sulfate de chondroitine, obtenu par fractionnent au chlorure de cetylpyridinium, sont les mêmes pour tous les échantillons, y compris pour le cartilage qui diffère nettement par sa morphologie et sa réponse fonctionnelle. Des augmentations significatives en fonction de l'âge sont observées pour la concentration tissulaire de sulfate de chondroitine des rudiments osseux jeunes et de deux des trois zones histologiques de cartilage tibial. Les concentrations plus élevées, observées dans la zone hypertrophique, au cours des divers stades du développement du tibia, et l'augmentation marquée de la concentration dans cette zone au moment de la formation osseuse accélérée semblent indiquer que cette augmentation est liée à l'ossification en milieu cartilagineux.
    Abstract: Zusammenfassung Diese Untersuchung hat quantitative mikrochemische Resultate über den Gehalt an Glucosaminoglucanen im Knorpel des sich entwickelnden Hühnerembryos ergeben. Das hauptsächlich vorkommende Glucosaminoglucan war Chondroitinsulfat. Diese Substanz erschien gleichzeitig mit dem histologischen Auftreten von Knorpel in den Extremitäten und erhöhte sich rasch bei Zunahme des Knorpels in diesen. Signifikante Mengen anderer Glucosaminoglucane konnten weder in ganzen Extremitäten, noch in Knochenrudimenten oder mikroskopisch präparierten Knorpelproben nachgewiesen werden. Die mit der Cetylpyridinchloridfraktionierung gefundenen Charakteristika des Chondroitinsulfates waren für alle diese Proben dieselben, sogar wenn die Knorpelproben ausgesprochene Abweichungen in bezug auf Morphologie und bekannte funktionelle Eigenschaften zeigten. Eine signifikante, altersbedingte Zunahme der Chondroitinsulfatkonzentration im Gewebe konnte in jungen Gesamtknochenrudimenten und in 2 der 3 histologiscchen Zonen des Tibiaknorpels nachgewiesen werden. Die höheren Konzentrationen, die in den hypertrophischen Zonen in allen Stadien der Tibiaentwicklung beobachtet werden konnten und die deutliche Konzentrationszunahme in diesem Gebiet während der beschleunigten Knochenbildung liessen vermuten, daß diese Zunahme mit der endochondralen Knochenbildung in Zusammenhang steht.
    Notes: Abstract This study provides quantitative microchemical data on the glycosaminoglycans of developing chick embryo cartilage. The major glycosaminoglycan was chondroitin sulfate, which appeared co-incidentally with cartilage in the limbs and rapidly increased as the amount of cartilage increased. Significant amounts of other glycosaminoglycans were not found in whole limbs, whole bone rudiments or microscopically-dissected cartilage samples. The characteristics of the chondroitin sulfate as revealed by cetylpyridinium chloride fractionation were the same for all of these samples, even cartilage samples differing markedly in morphology and known functional response. Significant age-related increases in tissue concentration of chondroitin sulfate were demonstrated in early whole bone rudiments, and in two of the three histologic zones of tibial cartilage. The higher concentrations observed in the hypertrophic zone at all stages of tibial development and the marked increase in the concentration in this region at a time of accelerated bone formation suggested that this increase is related to endochondral bone formation.
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    Calcified tissue international 6 (1970), S. 316-328 
    ISSN: 1432-0827
    Keywords: Epiphysis ; Cartilage ; Bone ; Parathyroidectomy ; Histochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des coupes de tibias d'animaux entiers et parathyroidectomisés sont étudiées histologiquement et histochimiquement pour préciser les effets de la parathyroidectomie sur l'os. Les épiphyses cartilagineuses proximales montrent uneaugmentation relative de matrice intercellulaire, une diminution en cellules, et une désorganisation de l'arrangement cellulaire. Le cartilage épiphysaire des rats parathyroidectomisés montre une matrice intercellulaire intensément PAS positive, contrairement aux affinités à l'alcian de la matrice des animaux témoins. Les travées diaphysaires des animaux parathyroidectomisés se colorent en rouge au PAS et contiennent de nombreux ilôts étendus de cartilage calcifié. En outre, le cartilage épiphysaire et les travées osseuses des animaux parathyroidectomisés se colorent moins intensément à la réaction de ninhydrine—Schiff que l'os des témoins. L'étude par les diverses colorations histochimiques, après parathyroidectomie, indique une altération des constituants collagène-protéine et mucopolysaccharide. Ces changements sont susceptibles de produire un trouble de la calcification de l'os.
    Abstract: Zusammenfassung Tibiaschnitte von Kontrollratten und von parathyreoidektomierten Ratten wurden histologisch und histochemisch untersucht, um die nach Parathyreoidektomie aufgetretenen Knochenveränderungen festzustellen. Der proximale Epiphysenknorpel von parathyreoidektomierten Ratten zeigte einerelative Zunahme der intracellulären Matrix, eine Abnahme des Zellgehaltes und eine Veränderung der zellulären Anordnung. Die Epiphysenknorpel der parathyeoidektomierten Ratten wiesen eine intensiv PAS-positive intracelluläre Matrix auf, im Gegensatz zur alcianophilen Reaktion der Matrix der Kontrolltiere. Die Diaphysen-Trabekel der parathyreoidektomierten Tiere ließen sich mit PAS rot färben und enthielten breite Zonen von verkalkten knorpeligen Kernen. Ferner ließen sich die Epiphysenknorpel und die Knochen-Trabekel von parathyreoidektomierten Tieren im Vergleich mit den Kontrolltieren weniger intensiv mit der Ninhydrin-Schiff-Reaktion färben. Die Resultate der verschiedenen histochemischen Reaktionen zeigten, daß die Parathyreoidektomie eine Veränderung der Protein-Mucopolysaccharide des Kollagens zur Folge hat. Diese neuen Bedingungen könnten eine Veränderung der Knochenverkalkung hervorrufen.
    Notes: Abstract Sections of the tibias from intact and from aparathyroid animals were examined histologically and histochemically to determine the changes in bone following parathyroidectomy. The proximal epiphyseal cartilages of parathyroidectomized rats showed arelative increase of intercellular matrix, decrease in cellularity, and disorganization of the cellular arrangement. The epiphyseal cartilages of the aparathyroid rats exhibited an intense PAS-positive intercellular matrix in contrast to the alcianophilic nature of the matrix from control animals. The diaphyseal trabeculae from aparathyroid animals stained red with PAS and contained wide regions of calcified cartilaginous cores. In addition, the epiphyseal cartilages and bone trabeculae from the parathyroidectomized animals stained less intensely to the ninhydrin-Schiff reaction than did the bone from the intact animal. The response to the various histochemical reactions indicated that following parathyroidectomy there was a change in the collagen-protein mucopolysaccharide components. This condition would produce an alteration in the calcification of bone.
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  • 79
    ISSN: 1432-0827
    Keywords: Morphology ; Glycosaminoglycan ; Defect ; Cartilage ; Transplant
    Source: Springer Online Journal Archives 1860-2000
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    Description / Table of Contents: Résumé Des greffes autologues de cartilage costal ont été transplantées au niveau d'une perte de substance ostéo-cartilagineuse, réalisée au niveau de la tête fémorale chez 20 lapins adultes. Des examens histologiques et autoradiographiques (35S-sulfate) de ces régions, du cartilage articulaire et de régions témoins, 1 an et 1 an et demi après, ont montré que la surface articulaire et la région transplantée sont formées par du cartilage, présentant différents stades de dégénérescence. Du tissue fibreux est trouvé au niveau de la surface articulaire transplantée. Uni étude microchimique quantitative des glycosaminoglycanes, de l'hydroxyle-proline et du calcium est réalisée sur du matériel micro-disséqué après congélation-dissication. Les hexosamines totales et le sulfate de kératane et/ou la fraction glycoprotéique sont moins concentrés dans le transplant qu'au niveau de la surface articulaire d'articulations opérées, alors que le contraire est observé pour le sulfate de chondroitine. Le cartilage articulaire du côté opéré montre une réduction des hexosamines totales et de toutes les fractions de glycosaminoglycanes par comparaison au côté non opéré. Ces changements sont particulièrement prononcés dans les échantillons présentant des signes de dégénérescence au niveau de coupes colorées à l'Azur A, après congélation dessication. Il semble donc qu'au cours de la dégénérescence précoce du cartilage articulaire, les glycoprotéines et/ou le sulfate de kératane sont diminuées alors que le contenu en sulfate de chondroitine parait relativement inchangé.
    Abstract: Zusammenfassung Bei 20 ausgewachsenen Kaninchen wurde autologer Rippenknorpel zur Deckung eines osteochondralen Defektes am Femurkopf transplantiert. Nach 1 oder 1 1/2 Jahren wurde das Gewebe aus dem transplantierten Bereich und der articuläre Knorpel von den operierten und nicht operierten Gelenken untersucht. Die histologische und autoradiographische (35S-Sulfat) Untersuchung zeigte, daß die Gelenkoberfläche und der größte Anteil des transplantierten Bereiches hauptsächlich aus lebensfähigem Knorpelgewebe bestand, welches Degenerationszeichen in verschiedenen Graden aufwies. An der Gelenkoberfläche auf der transplantierten Seite wurde ebenfalls fibröses Gewebe gefunden. Die quantitative mikrochemische Untersuchung von Glykosaminoglukanen, Hydroxyprolin und Calcium wurde an lyophilisierten Gewebeschnitten vorgenommen. Die Gesamt-Hexosamine, die Keratansulfat-und/oder Glucoproteinfraktion waren im Transplantat niedriger als im Knorpel von den operierten Gelenken; für das Chondroitinsulfat wurde das Gegenteil festgestellt. Der Knorpel von den operierten Gelenken zeigte eine Verminderung der Gesamt-Hexosamine und aller Glykosaminoglucanfraktionen im Vergleich mit den nicht-operierten Gelenken. Diese Änderungen waren besonders ausgeprägt in lyophilisierten Gewebeschnitten, welche bei der Azur-A-Färbung eine Degeneration zeigten. Es wird vermutet, daß bei einer frühauftretenden, leichten oberflächlichen Gelenkknorpeldegeneration die Glucoprotein-und/oder die Keratansulfatfraktion vermindert ist, während das Chondroitinsulfat relativ unverändert bleibt.
    Notes: Abstract In 20 adult rabbits autologous costal cartilage was transplanted to an osteochondral defect on the femoral head. After 1, and 11/2 years the tissue of the transplant area and the articular cartilage from the operated and non-operated joints were studied. The histological and autoradiographic (35S-sulphate) study showed that the articular surface and the bulk of the transplant area consisted mainly of viable cartilaginous tissue showing signs of degeneration to varying degree. Also, fibrous tissue was found on the articular surface of the transplant area. The quantitative microchemical study of glycosaminoglycans, hydroxyproline and calcium was made on microdissected freeze-dried material. The total hexosamines and the keratan-sulphate and/or glycoprotein fraction were lower in the transplant than in the articular cartilage from the operated joints while the contrary was true for chondroitin sulphate. The articular cartilage from the operated joints showed a reduction of total hexosamines and of all glycosaminoglycan fractions compared to the non-operated joints. These changes were especially pronounced in samples showing signs of degeneration in Azur-Astained, freeze-dried sections. It is suggested that in early and slight superficial articular cartilage degeneration the glycoprotein and/or keratan-sulphate fraction is reduced while chondroitin-sulphate remains relatively unaltered.
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    Calcified tissue international 7 (1971), S. 307-317 
    ISSN: 1432-0827
    Keywords: Ultrastructure ; Cartilage ; Calcification ; Inorganic ; Organic
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé La phase organique (ou fantôme des cristaux) associéc à chaque cristal, ainsi que la substance de base associée à chaque cristal, ainsi que la substance de base associée à chaque amas cristallin, sont mises en évidence au niveau du cartilage calcifié en utilisant le sulfate de chrome basique comme agent de fixation, de coloration et de déminéralisation. Le traitement ultérieur du tissu, à l'aide de papaïne ou d'hyaluronidase, indique que les fantômes cristallins constitutent un complexe protéino-polysaccharidique et que la substance de base est formée par une protéine associée à un polysaccharide acide. Les rapports entre phases inorganique et organique sont discutés.
    Abstract: Zusammenfassung Die organische Phase (oder Kristallit-Schatten), die zu jedem Kristallit gehört, sowie das Hintergrundmaterial, das zu jeder Kristallitgruppe gehört, wurden in calcifiziertem Knorpel sichtbar gemacht. Zu diesem Zweck wurde basisches Chromsulfat als ein kombiniertes Fixierungs-, Färbe- und Demineralisierungsmittel verwendet. Nachfolgende Behandlung des Gewebes mit Papain oder Hyaluronidase läßt vermuten, daß die Kristallitschatten einen Proteinpolysaccharidkomplex darstellen und daß das Hintergrundmaterial hauptsächlich aus Protein mit einigen sauren Polysacchariden besteht. Die Beziehung zwischen anorganischen und organischen Phasen wird diskutiert.
    Notes: Abstract The organic phase (or crystallite ghost) associated with each crystallite, together with the background material associated with each crystallite cluster, was demonstrated in calcified cartilage using basic chromium sulphate as a combined fixative, stain, and demineralizing agent. Subsequent treatment of the tissue with papain, or with hyaluronidase, suggests that the crystallite ghosts represented a protein-polysaccharide complex and that the background material was principally protein together with some acid polysaccharide. The relationship between inorganic and organic phases is discussed.
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    Calcified tissue international 8 (1971), S. 36-53 
    ISSN: 1432-0827
    Keywords: Phospholipids ; Vitamin D ; Deficiency ; Epiphysis ; Cartilage ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les effets de divers degrés de rachitisme non compliqué, provoqué par avitaminose D, sur les concentrations, la composition, et l'extractibilité de phospholipides de tissus moux et en voie de calcification ont été étudiés chez le poulet et le porc. Le contenu en phospholipides des tissus en voie de calcification est modifié au cours du rachitisme; cependant, on n'a constaté que peu de changements dans les tissus moux. Les effets quantitatifs sur la concentration en lipides des tissus rachitiques calcifiés varient de zone en zone et semblent indiquer desvariations dans d'autres constituants tissulaires. Cependant les différences qualitatives indiquent un effet direct sur des phospholipides spécifiques. Les phospholipides acides présentent les changements de pourcentage les plus élevés bien que de nombreuses modifications statisstiquement significatives dans les phospholipides neutres ont pu être observés. Les zones stisulaires présentant le plus grand changement de composition phospholipidique au cours du rachitisme, sont constituées par le cartilage hypertrophique et en voie de prolifération et par l'os spongieux, alors que le cartilage calcifié est le moins affecté. Le rachitisme diminue aussi l'extractibilité de certains phospholipides neutres et acides des zones de minéralisation précoce. Ces résultats semblent indiquer la présence, dans le tissu rachitique, de concentrations élevées de phospholipide complexé avec le phosphate de calcium, probablement causé par l'inhibition de la minéralisation normale.
    Abstract: Zusammenfassung Die Auswirkung von verschiedenen Graden von Rachitis (verursacht durch Vitamin D-Mangel) auf die Mengen, die Zusammensetzung und auf die Extrahierbarkeit der Phospholipide von verkalkenden und weichen Geweben wurde an Hühnern und Schweinen untersucht. Es wurden verschiedene Wirkungen von Rachitis auf die Phospholipid-Zusammensetzung der verkalkenden Gewebe gefunden, in den weichen Geweben jedoch nur wenige. Der Grad dieser Wirkung auf die Lipid-Konzentration der rachitisch verknöchernden Gewebe änderte sich von Schicht zu Schicht und schien die Änderungen in anderen Gewebebestandteilen wiederzugeben. Die qualitativen Unterschiede zeigten jedoch eine direkte Wirkung auf spezifische Phospholipide an. Saure Phospholipide zeigten den größten prozentualen Unterschied, obgleich zahlreiche statistisch signifikante Unterschiede auch in den neutralen Phospholipiden gefunden wurden. Die Gewebeschichten, welche bei Rachitis die größte Änderung in der Phospholipidzusammensetzung zeigten, waren proliferierender und hypertrophischer Knorpel und spongiöser Knochen, während verkalkter Knorpel die geringste Änderung zeigte. Rachitis verminderte auch die Extrahierbarkeit von gewissen sauren und neutralen Phospholipiden aus den Schichten der frühen Mineralisierung. Diese Ergebnisse weisen auf das Vorhandensein von erhöhten Phospholipidmengen im rachitischen Gewebe hin, welche Calciumphosphat binden; dies wird wahrscheinlich durch die Blockierung der normalen Mineralisierung verursacht.
    Notes: Abstract The effect of various degrees of simple, vitamin D-deficient rickets on the levels, composition and extractability of phospholipids from calcifying and soft tissues was studied in chickens and pigs. A variety of effects of rickets on the phospholipid composition of the calcifying tissues were detected; there was little change in the soft tissues. The quantitative effects on the lipid concentration of rachitic hard tissues were variable from zone to zone and appeared to reflect variations in other tissue constituents. However, the qualitative differences indicated a direct effect on specific phospholipids. Acidic phospholipids displayed the greatest percentage change, although numerous statistically-significant changes were also seen in the neutral phospholipids. Tissue zones displaying the greatest change in phospholipid composition in rickets were proliferating and hypertrophic cartilage, and cancellous bone, with calcified cartilage showing the least effect. Rickets also reduced the extractability of certain acidic and neutral phospholipids from zones of early mineralization. These findings are interpreted as indicating the presence of elevated levels of phospholipid complexed with calcium phosphate in rachitic tissue, presumably due to the blockage of normal mineralization.
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    Calcified tissue international 8 (1971), S. 96-105 
    ISSN: 1432-0827
    Keywords: Iliac ; Proteinpolysaccharides ; Cartilage ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les protéines-polysaccharides (PP) ont été extraits de la crête iliaque cartilagineuse d'enfants à squelettes normaux, âgés de quelques jours à 16 ans, à l'aide de technique par rupture et par dissociation. L'homogénéisation du cartilage dans l'eau ou l'agitation dans du chlorure de guanidinium permet de recueillir un matériel PP semblable à poids moléculaire élevé. Les extraits aqueux contiennent des quantités considérables de collagène qu'on ne trouve qu'à l'état de trace dans les extraits à la guanidine. Des produits dégradés, probablement liés à l'action d'enzymes protéolytiques, sont obtenus après extraction du tissu avec un tampon phosphate 0,067 M de pH 7,4 à 30°; 0,5 M ou 3,0 M MgCl2 à 10°.
    Abstract: Zusammenfassung Proteinpolysaccharide wurden aus dem Beckenkamm-Knorpel von Kindern im Alter von wenigen Tagen bis 16 Jahren mit normalem Skelett extrahiert, und zwar sowohl durch Homogenisierung als auch durch Extraktionsmethoden. Die Homogensierung des Knorpels in Wasser ergab Proteinpolysaccharide von hohem Molekulargewicht und gleichen Eigenschaften wie nach Rühren mit Guanidinchlorid. Die wässerigen Extrakte enthielten ansehnliche Kollagenmengen, während in den Guanidinextrakten nur Spuren vorlagen. Abbauprodukte, die wahrscheinlich durch Einwirkung von proteolytischen Enzymen entstanden, wurden durch Extraktion dieses Gewebes mit 0,067 M Phosphatpuffer von pH 7,4 bei 30°, ferner mit 0,5 oder 3,0 M MgCl2 bei 10° gewonnen.
    Notes: Abstract Proteinpolysaccharides (PP) have been extracted from the iliac crest cartilage of skeletally normal children varying in age between a few days and sixteen years, with both disruptive and dissociative methods. Homogenization of the cartilage in water or stirring with guanidinium chloride yielded high molecular weight PP material of similar properties. The water extracts contained considerable amounts of collagen which was present only in traces in the guanidine extracts. Degraded products, probably due to the action of proteolytic enzymes, were obtained by extraction of the tissue with 0.067 M phosphate buffer pH 7.4 at 30°; 0.5 M or 3.0 M MgCl2 at 10°.
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    Cell & tissue research 122 (1971), S. 254-272 
    ISSN: 1432-0878
    Keywords: Cartilage ; Guinea pig ; Epiphyseal plate ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The fine structure of the upper tibial epiphyseal plate of the normal guinea pig has been investigated. On morphological and functional grounds this cartilage can be divided into four zones. In all zones the chondrocytes are distributed in an electronlucent organic matrix composed of thin collagenous fibrils and small non-membrane-bounded granules representing proteoglycans. 1. The reserve cell zone consists of a primitive type of hyaline cartilage. The cells have a very high nucleo-cytoplasmic ratio. The endoplasmic reticulum is the most developed cellular organelle. 2. In the proliferative zone the flattened cells are arranged in distinct columns. The endoplasmic reticulum and the Golgi complex are prominent. Mitochondria and lysosomes are more numerous than in the reserve cells. Extracellularly, the collagen fibrils are brought together in longitudinal and transversal septa. 3. In the hypertrophying zone the cells are more rounded and the pericellular zones wider than in the preceding zones. The endoplasmic reticulum contains dilated cisternae. The Golgi complex includes many large vacuoles. Mitochondria and lysosomes are more abundant than previously. 4. There is a progressive mineralization of the longitudinal septa in the calcifying zone. Within the cells the amount of endoplasmic reticulum is decreased. Its cisternae mostly are short, rounded, and non-continuous. The Golgi apparatus is poorly developed, and the mitochondria few.
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  • 84
    ISSN: 1432-0878
    Keywords: Bone ; Ossification ; Cartilage ; Matrix ; Chondrocytes ; Complement ; Matrix metalloproteinase ; Immunocytochemistry ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The first component of complement $$C\bar 1s$$ has been shown to degrade type I and type II collagens (Yamaguchi et al. 1990), the latter of which is a major constituent of the cartilage matrix. In order to understand the physiological roles of $$C\bar 1s$$ in cartilage resorption, the expression of C1s was examined by immunohistochemistry in the primary ossification center where the matrix is removed and replaced by bone marrow. Hypertrophic chondrocytes, endothelium and hematogenous elements in the capillary buds were intensely stained by a monoclonal antibody against C1s. Matrix metalloproteinase 9 (MMP-9, 92kDa gelatinase/type IV collagenase) was also immunolocalized in hypertrophic chondrocytes, mesenchymal cells in the primitive bone marrow and the cartilage matrix adjacent to the marrow. In addition, $$C\bar 1s$$ was found to activate the zymogen of MMP-9. These observations suggest that $$C\bar 1s$$ and MMP-9 coordinately participate in matrix degradation in cartilage.
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    Cell & tissue research 147 (1974), S. 505-527 
    ISSN: 1432-0878
    Keywords: Cartilage ; Ageing ; Chondrocyte and matrix ; Light microscopy ; Histochemistry ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Histological, histochemical, and ultrastructural investigations have been carried out on ageing costal and tracheal cartilage of rats. The following age groups of animals have been studied: 1, 7, 14, 20, 30, 45, 75 days, 6 months, and 2 years. Ageing induces cellular changes which are represented by a reduction in the number of chondrocytes, a progressive increase in glycogen deposition, and processes of degeneration, the most frequent of which is the accumulation of lipidic material within large cytoplasmic vacuoles. Changes in the intercellular matrix become evident after 20 days in costal cartilage and after 30 days in tracheal cartilage. Chondroitin sulphate decreases while keratan sulphate, whose presence is limited to the territorial matrix, increases. Glycoproteins increase slightly in young animals and then remain constant; they decrease in the subperichondrial areas in old animals. Ultrastructurally, the matrix of cartilage of young animals contains thin collagen fibrils, most of which have no periodic banding. Roundish electron dense granules are associated with these fibrils. Irregular filaments associated with small electron-dense circular bodies are present around chondrocytes as well as within cytoplasmic vacuoles. With increasing age, and coincident with the reduction of chondroitin sulphate, the thickness of collagen fibrils increases, their period becomes evident, and the associated matrix granules decrease in number and size. Areas containing these fibrils undergo calcification, which frequently starts within roundish bodies of cellular origin. Collagen fibrils with a period of 640 Å but a highly variable thickness are often present in cartilage of adult and old rats. These fibrils seem to be due to an abnormal synthetic activity of chondrocytes.
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  • 86
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    Keywords: Glucocorticoids ; Cartilage ; Growth ; Histochemistry ; Ultrastructure ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effects of different doses of various steroids on growth, and on costal and epiphyseal chondrocytes, have been studied in prenatal, immature, and adult Long-Evans rats using histochemical techniques, and both light and electron microscopy. Both prenatal and postnatal treatments have been employed. The steroids used were cortisone (CA), betamethasome (BM), and, in the prenatal group only, dexamethasone (DM). Body weight is reduced in all treated rats (except the low dose of CA) by day 17 of gestation, with greater weight reductions occurring in rats receiving the higher dose level of each steroid. In rats treated prenatally or neonatally, and sacrificed postnatally on days 39–43 or days 116–127, body weights, and tibial and tail lengths, are less than in correspondingly aged controls, thus showing a persistence of the effects of treatment. Costal and epiphyseal cartilages in prenatal rats show cellular, synthetic, and ultrastructural alterations induced by treatment with glucocorticoids but the responses are not necessarily comparable. Except for the low dose of DM, the higher doses of each steroid are more effective in inhibiting, or altering, growth and cellular differentiation in the developing fetuses. Surprisingly, a low dose of DM has a more devastating effect on the cells and extracellular matrix of both costal and epiphyseal cartilage, than do higher dose-levels of the various steroids. Low doses of CA and BM are also effective in inhibiting or altering growth and cellular differentiation, but their effectiveness is largely limited to 17 days of gestation. The order of effect of the various doses of the different steroids on fetal cartilage, listed in decreasing order of severity, is as follows: 0.12 DM, 0.24 DM, 0.42 BM, 50 CA, with 25 CA and 0.18 BM being approximately equal and only slightly different from control cartilages. The effect of prenatal or neonatal glucocorticoid treatment on chondrocytes is minimal in the 30–43 day, or 116–127 day, postnatal groups. In immature and adult rats, cortisone affects the chondrocytes more deleteriously than does betamethasone, and a 5.0 mg dose of CA seems to affect chondrocytes, body weight, and tibial and tail lengths more than 0.2 or 7.5 mg doses.
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  • 87
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    Keywords: Glucocorticoids ; Cartilage ; Growth ; Cell culture ; Differentiation ; Mouse (ICR)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Measurements of 3H-thymidine incorporation, quantitative autoradiography and morphometry were used to evaluate cell behavior during the recovery of mandibular condylar cartilage cultures following short-term exposure to a corticosteroid hormone in vitro. Apical segments of mandibular condyles of newborn mice were initially incubated in the presence of the hormone triamcinolone acetonide (10-6 M) for 24 h and were thereafter cultured for additional 6 days in hormone-free medium. The present results indicated that the treatment led to a decrease in the rate of incorporation of 3H-thymidine, a feature that lasted for 48 h following the removal of the hormone. Quantitative 3H-thymidine autoradiography of explants that were labeled in the presence of the hormone further substantiated the initial suppressive effect of the hormone on cellular proliferation, a feature that was followed by a recovery. Differences were noted in the pattern of distribution of labeled cells: in control explants, labeled cells progressively moved from the chondroprogenitor compartment into the differentiated portion of the cartilage; in hormone-treated explants, 3H-thymidine labeled cells were confined to the progenitor layer up to 5 days after the treatment and only then appeared in the chondrocytic compartment. The hormone's adverse effect upon differentiation was manifested by both morphology, and by causing a significant increase in the size of the progenitor layer (up to 50.5% on 4th post-treatment day) along with a 70.5% reduction in the size of chondroblastic layer. We conclude that a short-term exposure to a glucocorticoid hormone in vitro interferes with proliferation of chondroprogenitor cells and their subsequent differentiative pathway. While the proliferative activity was restored within 48 h, the hormone's effect on differentiation lasted for a considerably longer period of time.
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    Cell & tissue research 273 (1993), S. 381-389 
    ISSN: 1432-0878
    Keywords: Vigilin ; Cartilage ; Bone ; Differentiation ; In-situ hybridization ; Chicken
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The expression of vigilin was followed during chick embryonal development by in situ hybridization. Vigilin mRNA is abundantly expressed in tissues of mesenchymal and ectomesenchymal origin. The mesenchymal primordial cells of cartilage and bone did not show any significant, expression of vigilin. As tissue differentiation proceeded, vigilin mRNA levels increased in hyaline cartilage and in both endochondral as well as intramembranous bone. The results suggest that the expression of vigilin mRNA in cartilage- and bone-forming cells chondrocytes and osteobalsts, is dependent on the stage of development and cellular differentiation, although not a unique process of bone formation. Most striking is the correlation of the maximum vigilin mRNA expression in osteoblasts and hypertrophic chondrocytes to periods when cell-specific genes were highly transcribed and substantially translated, e.g., synthesis of procollagen and formation of extracellular matrix in bone and cartilage.
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    Cell & tissue research 103 (1970), S. 192-217 
    ISSN: 1432-0878
    Keywords: Cartilage ; Calcification ; Calcifying globules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The cartilage matrix in which the early calcium salts are deposited has been studied in the tibial epiphyses and in the costo-chondral junctions of 30-day-old guinea pigs. The results may be summarized as follows: (1) Structures of globular shape (“globules”) are to be found throughout the entire epiphyseal plate. (2) They have a homogeneous matrix and are bounded by a membrane. (3) Early calcification occurs in globules. Calcification of collagen fibrils seems to occur later. (4) The earliest mineral deposited would seem to consist of tiny granules about 20 Å in diameter. Then apatite crystals are laid down, initially in small clusters and later filling the globules completely. (5) The globules are strongly osmiophilic. They seem to contain a fair amount of neutral polysaccharides, but no acid polysaccharides except a coating on their outer membrane. Hyaluronidase digestion does not affect globules. Papain digestion makes them more reactive to uranium and lead. (6) Globules are of cellular origin but they are almost certainly not pre-formed in the chondrocytes. Finally, the present paper advances the hypothesis that some globules derive from degenerating chondrocytes and others from the processes of normal chondrocytes.
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  • 90
    ISSN: 1432-0878
    Keywords: Cartilage ; Bone ; Organ culture ; Joints ; Man (Primates)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Mandibular condyles of human fetuses, 14–21 weeks in utero, were kept in an organ culture system for up to 60 days. After 6 days in culture, the cartilage of the mandibular condyle appeared to have maintained its inherent structural characteristics, including all its various layers: chondroprogenitor, chondroblastic, and hypertrophic. After 12 days in culture, no chondroblasts could be seen; instead, the entire cartilage was occupied by hypertrophic chondrocytes. At the same time, the mesenchymal cells in the vicinity of the chondroprogenitor zone differentiated into osteoblast-like cells that produced type I collagen. The progenitor cells were still actively incorporating 3H-thymidine. The newly formed osteoid-like tissue lacked both metachromatic reactivity and a response to antibodies against chondroitin sulfate. Instead, the tissue reacted positively for osteocalcin (bone gla-protein). The process of new bone formation further progressed and, by the 20th day in culture, the new bone reacted positively for type I collagen, osteonectin, and to a lesser extent for chondroitin sulfate. The osteoid also underwent mineralization as revealed by both the von Kossa stain and vital staining with tetracycline. The above feature appeared even more intense in 40-day-old cultures. After 60 days, the newly formed bone contained osteoblasts and osteocytes, whereas the extracellular matrix revealed a high degree of matrix polarization. The results of the present study recapitulate findings reported for organ cultures of mice mandibular condyles. However, the in vitro process of de novo bone formation in human specimens requires a 6-fold longer culture time than that needed for mice condyles.
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    Cell & tissue research 272 (1993), S. 155-160 
    ISSN: 1432-0878
    Keywords: Immunomorphology ; Collagen type VI ; Cartilage ; Mouse (NMRI)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Collagen type VI was demonstrated immunomorphologically in articular cartilage (distal femur) of young (2–8 weeks) and adult mice by fluorescence and electron microscopy (gold-labelled second antibody—sandwich method) using pre-and post-embedding techniques. This collagen type was mainly seen in the vicinity of chondrocytes, and in larger amounts in adult cartilage. Electron-microscopic inspection (pre-embedding technique) revealed labelling above plaques that were 40–160 nm in size, and from which up to 7 fine filaments (≤ 10 nm) per unit sectional plane radiated. Using the post-embedding technique, only labelled plaques could be demonstrated; fine filaments were not perceptible. This was partly a result of the low contrast. It is assumed that the globular ends of up to 20 of the fine type VI filaments are anchored in one plaque and that the antibodies bind to the non-collagenous globular domains. Filaments radiated from the plaques and formed a threedimensional network that stabilized the structures of the cartilaginous matrix. Antibodies against fibronectin also labelled similar plaques. The ends of the type VI filaments are possibly linked into the plaques by fibronectin.
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  • 92
    ISSN: 1432-0878
    Keywords: Key words: EGF ; Cartilage ; Growth plate ; Hypophysectomy ; Growth hormone ; Immunohistochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Epiphyseal growth plate cartilages from the proximal tibia of normal, hypophysectomized, and growth hormone (GH)-treated hypophysectomized rats were subjected to immunohistochemistry for detection of epidermal growth factor (EGF). In the normal growth plate, EGF was distributed mainly in the proliferative zone. Hypophysectomy resulted in considerable atrophy of the chondrocytes and the cartilage matrix (a decreased number of mature-type chondrocytes and a decreased ratio of proliferating to hypertrophic chondrocytes) and a significant diminution of EGF immunoreactivity. Treatment with GH reversed these effects of hypophysectomy, causing an increased thickness of the growth plate and EGF-reactive sites in all chondrocyte layers. The most intense immunostaining for EGF, however, was frequently seen in the nuclei of chondrocytes with flattened appearance. It appears that EGF could be incorporated or synthesized in chondrocytes having marked mitogenic activity. The present results, taken with previous data on EGF involvement in growth of cartilaginous tissue in vivo and in vitro, strongly suggest that EGF-immunoreactive chondrocytes are involved in cartilage proliferation and growth under the specific influence of GH.
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  • 93
    ISSN: 1432-0878
    Keywords: Key words: Bone ; Ossification ; Cartilage ; Chondrocytes ; Complement C1s ; Development ; Immunohistochemistry ; In situ hybridization ; Hamster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Expression of the first complement component (C1s) has been examined in chondrocytes of hamster epiphyseal cartilage during development and fracture healing. C1s is immunostained with anti-hamster C1s monoclonal antibody, PG11. The C1s staining increases in accordance with chondrocyte differentiation and reaches a maximal level in hypertrophic chondrocytes. This change is observed at both the tibia ossification center and at the callus in which the replacement of cartilage by bone marrow takes place. The concomitant increase of C1s and chondrocyte hypertrophy has been confirmed by RNA blot and by in situ hybridization. These results, in addition to previous findings on C1s collagenolytic and gelatinolytic activities, suggest C1s participation in cartilage remodeling.
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  • 94
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    Electronic Resource
    Springer
    Cell & tissue research 189 (1978), S. 67-89 
    ISSN: 1432-0878
    Keywords: Cartilage ; Cortisone ; Parabiosis ; Light and electron microscopy ; Histochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cortisone-treated Buffalo rats have been parabiosed with untreated controls of the same age. The optical and electron microscopy, including histochemistry, of costal cartilage of these rats has been compared with that in single cortisone treated rats, single controls, and control parabiosed with control rats, at 14 and 28 days after parabiosis. Single cortisone-treated rats, in comparison to controls, have shown the greatest alteration in cellular morphology and in the extracellular matrix both at 14 and at 28 days. Cortisone-treated parabiosed rats demonstrate a gradation of these alterations. Cellular alterations include enhancement of lipid and glycogen deposition concurrently with the presence of numerous large cytoplasmic vacuoles containing beaded irregularly-shaped filaments, banded or unbanded collagen-like fibrils, and/or electron dense lamellar bodies. In the extracellular matrix, matrix vesicles, amianthoid fibers, randomly oriented unbanded fibrillar materials, and filament-like materials are most prominent in the single cortisone-treated rats and they are progressively less prominent in the cortisone-treated parabiosed rats, and in the parabiosed and single controls. Calcification of the extracellular matrix follows a similar pattern and is observed initially in pericellular halos of the single cortisone and in cortisonetreated rats parabiosed with controls. Histochemical techniques have shown that chondroitin sulfate is less demonstrable in the single cortisone and in the cortisone-treated parabiosed rats than it is in the single or parabiosed controls at 14 days but, at 28 days, all untreated or treated rats, single or parabiosed are basically comparable. Glycoproteins are prominent in the single cortisone-treated rats both at 14 and at 28 days and, at these same times, they are progressively less prominent in the cortisone-treated parabiosed rats and in the single or parabiosed controls. Many of the cortisone induced alterations in costal cartilage are suggestive of enhancement of the aging process.
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  • 95
    ISSN: 1432-0878
    Keywords: Osteoblasts ; Osteoclasts ; Differentiation ; Bone ; Cartilage ; Chicken
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The resorbing region of uncalcified cartilage in the tibia of embryonic chick was studied using 3H-proline autoradiography, histochemistry, and horseradish-peroxidase tracers. At the cartilage-bone marrow interface, two kinds of cells (A and B) were identified. Type-A cells were elongated, contacted the matrix of the uncalcified cartilage directly, and possessed extensive rough endoplasmic reticulum, one or two juxtanuclear Golgi apparatus and cell membranes exhibiting prominent alkaline phosphatase activity. Type-B cells were round to oval, mononucleate (occasionally binucleate), and contained abundant mitochondria, vacuoles and vesicles, well-developed Golgi apparatus, and lysosomes. The lysosomes and the majority of vacuoles and Golgi lamellae of these cells showed prominent acid phosphatase activity. Type-B cells accumulated more horseradish-peroxidase reaction product in their vacuoles and vesicles than type-A cells. Thick, banded collagen fibrils were occasionally found in the matrix of the resorbing surface. 3H-proline autoradiography revealed small numbers of grains at the cartilage-bone marrow interface. These findings suggest that type-A cells have osteoblastic and type-B cells osteoclastic properties and are precursor cells of osteoblasts and osteoclasts, respectively. The appearance of a mineral phase in the resorbing cartilage is probably important for the differentiation of these cells.
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  • 96
    ISSN: 1432-0878
    Keywords: Chondrocytes ; Cartilage ; Dexamethasone ; Osteoblasts ; Glucocorticoids ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effects of dexamethasone on the ability of cells enzymatically isolated from 21-day fetal rat calvaria to produce cartilage in vitro has been investigated. Primary cultures of single-cell suspensions of rat calvaria were grown for up to 28 days in vitro in α-minimal essential medium containing 15% fetal bovine serum, 50 μ/ml ascorbic acid, 10 mM Na β-glycerophosphate and dexamethasone at concentrations of 1 μM to 1 nM. Two types of nodules were present in dexamethasone-containing cultures. One has been characterized previously as bone (Bellows et al. 1986). The second morphologically resembled hyaline cartilage, possessed a strong Alcian blue-positive matrix and contained type-II, but not type-I, collagen. Both bone and cartilaginous nodules were spatially distinct and developed in isolation from each other. Cartilaginous nodules were found in the highest number at a dexamethasone concentration of 100 nM. Time-course experiments revealed that while the number of bone nodules increased continuously at least to day 28, the number of cartilaginous nodules remained constant after cultures had reached confluency. When cells were isolated separately from frontal and parietal bones and suturai regions, the greatest number of cartilaginous nodules developed from parietal bones. Since 21-day fetal rat calvaria contains 2 distinct patches of cartilage at the periphery of the parietal bones, it seems likely that this cartilaginous tissue is the origin of the cartilage cells. The results demonstrate that cultures of rat calvaria cells contain chondrocytes and possibly chondroprogenitor cells that are distinct from osteoprogenitors. Results support previous data that 100 nM dexamethasone permits the expression of and maintains the phenotype of chondrocytes in serum-containing cultures in vitro.
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  • 97
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    Electronic Resource
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    Cell & tissue research 293 (1998), S. 357-364 
    ISSN: 1432-0878
    Keywords: Key words Thyroxine ; Cartilage ; Differentiation ; Type X collagen ; Organ culture ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Thyroid hormone plays a role in skeletal maturation. The present study was undertaken to investigate the effect of thyroid hormone on growth and maturation of the epiphyseal cartilage in the rat femur organ culture system. The femora increased in both weight and size over time in culture in the absence of serum. The growth rate was suppressed by thyroxine treatment. Thyroxine induced chondrocyte hypertrophy in the area adjacent to the articular surface in the femoral condyle. In this area, the expression of type X collagen, a marker of chondrocyte terminal differentiation, was detected after 21 days in culture by immunohistochemistry and reverse transcription/polymerase chain reaction. These data suggest that thyroxine suppresses cartilage growth by stimulating chondrocyte terminal differentiation.
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  • 98
    ISSN: 1432-0878
    Keywords: Key words: Complement Cls ; Neoantigen ; Cartilage ; Hypertrophic chondrocyte ; Immunohistochemistry ; Rheumatoid arthritis ; Decorin ; Syrian hamster ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The secondary ossification center of 14- to 16-day-old hamster tibiae was examined immunohistochemically with active and inactive Cls-specific antibodies, RK5 and RK4, respectively. At the ossification center, chondrocytes differentiate from proliferating and hypertrophic to degenerating stages, and their site is occupied by the bone marrow. Cls was strongly immunostained in hypertrophic chondrocytes. In order to discover whether Cls is activated at a particular site, the cartilage was immunostained with RK5 and RK4. RK5 mainly reacted with degrading matrix around invading vessels. In contrast, RK4 strongly stained hypertrophic chondrocytes. Immunoelectron microscopy revealed Cls on degrading fragments of chondrocytes and fibers of cartilage matrix. Decorin, one of the major matrix proteoglycans, was dose and time dependently degraded by Cls. Type II collagen and type I gelatin were also degraded. Articular cartilage from patients with rheumatoid arthritis was positively immunostained (11/12 cases) with an anti-Cls monoclonal antibody (mAb) PG11, whereas normal articular cartilage (5/5 cases) was negative, suggesting Cls participation in the etiology of rheumatoid arthritis.
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  • 99
    ISSN: 1432-0878
    Keywords: Parathyroid hormone ; Cartilage ; Chondrocytes ; Joints ; Organ culture ; Mouse (ICR)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary This investigation presents the structural changes in condylar cartilage incubated in the presence of human parathyroid hormone (1–34) in an organ culture system for 6 to 12 days. Control cultures maintained their cartilaginous characteristics whereas human parathyroid hormone (1–34)-treated cultures revealed the following modifications: (1) The chondroprogenitor cell zone at the apical region of the explant underwent a substantial enlargement. The cells changed from a mesenchyme-like morphology into polygonal, glycogen-rich cells that were tightly attached to each other by a fibrillar intercellular matrix, but even by 12 days the apical region was comprised of healthy cells. (2) The mineralizing zone in the hypertrophic cartilage revealed a change in its cellular population. Hypertrophic chondrocytes were replaced by cells with amoeboid extensions and large numbers of secretory granules or vesicles. Based upon the above findings it appears that the condroprogenitor cells that are initially stimulated to proliferate, are being suppressed from subsequent differentiation into chondroblasts; and that hypertrophic chondrocytes apparently undergo a dedifferentiation process followed by development into an as yet unknown cell population.
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  • 100
    ISSN: 1432-0878
    Keywords: EGF ; Cartilage ; Growth plate ; Hypophysectomy ; Growth hormone ; Immunohistochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Epiphyseal growth plate cartilages from the proximal tibia of normal, hypophysectomized, and growth hormone (GH)-treated hypophysectomized rats were subjected to immunohistochemistry for detection of epidermal growth factor (EGF). In the normal growth plate, EGF was distributed mainly in the proliferative zone. Hypophysectomy resulted in considerable atrophy of the chondrocytes and the cartilage matrix (a decreased number of mature-type chondrocytes and a decreased ratio of proliferating to hypertrophic chondrocytes) and a significant diminution of EGF immunoreactivity. Treatment with GH reversed these effects of hypophysectomy, causing an increased thickness of the growth plate and EGF-reactive sites in all chondrocyte layers. The most intense immunostaining for EGF, however, was frequently seen in the nuclei of chondrocytes with flattened appearance. It appears that EGF could be incorporated or synthesized in chondrocytes having marked mitogenic activity. The present results, taken with previous data on EGF involvement in growth of cartilaginous tissue in vivo and in vitro, strongly suggest that EGF-immunoreactive chondrocytes are involved in cartilage proliferation and growth under the specific influence of GH.
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