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  • 1
    ISSN: 1432-0827
    Keywords: Osteogenesis ; Osteoinduction ; Bone ; Matrix ; Cell
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Bone matrix demineralized in 0.6 N HCl at 2° for 24 h and implanted in muscle in allogeneic rats possesses consistently reproducible bone morphogenetic activity. Experiments on implants of matrix, obtained from donors injected with3H-tyrosine or3H-tryptophan, or Na35SO4, suggest that bone morphogenetic property is a protein or apart of a protein that is (1) insoluble in buffer solutions, pH 3.6 and 5.0; (2) degraded in buffer solutions at pH 7.4 by an endogenous sulfhydryl-group neutral proteinase; (3) digested by trypsin at 15° within 8 h without solubilization of the helical regions, possibly even without degradation of the nonhelical ends of the bone collagen molecule, and without any loss of the periodic ultrastructure of the collagen fibrils; (4) degraded or removed by 0.1 N NaOH at 2° within 24 h without solubilization of collagen; (5) biologically active even after nitration of tyrosyl groups with tetranitromethane. The release of only one-third of the radioactivity with loss of nearly all yield of new bone by limited tryptic digestion of3H-borohydride-reduced matrix indicates that the bone morphogenetic response is the function of a non-collagenous component. Autoradiographs of implants of matrix with non-collagenous proteins labelled with3H-tryptophan,3H-tyrosine, or both3H-tyrosine and3H-phenyl-alanine demonstrate random dissemination of the radioactive constituents and no evidence of local transfer of labelled proteins or soluble protein derivatives. Hypothetically, the bone morphogenetic response is controlled by an insoluble acidic bone morphogenetic protein or polypeptide (BMP) and a soluble neutral proteinase (BMP-ase) resembling trypsin in activity except functionally more specific for BMP. Firmly bound but separable from bone collagen, BMP is one of many short-lived morphogenetic substances appearing and disappearing throughout embryonic development and persisting in postfetal life. Where the BMP receptor resides and how it activates cell mechanisms of differential repression and derepression of such genes as code for osteogenesis is unknown.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 7 (1971), S. 108-113 
    ISSN: 1432-0827
    Keywords: Bone-Induction ; Alkaline phosphatase ; Enzyme ; Matrix
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des implants de matrice osseuse, susceptible d'engendrer une néoformation osseuse, sont préparés en décalcifiant de l'os cortical dans 0,6 N HCl à 2° pendant 48 heures. Les propriétés d'induction ont pu être inactivées en décalcifiant dans 0,6 N HCl contenant 66,5% d'alcool ethylique. Après implantation musculaire, les implants actifs et inactifs sont envahis par des cellules mésenchymateuses Il n'existe pas de différence significative dans le contenu en DNA d'implants actifs et inactifs, cependant seuls les implants actifs induisent la synthèse de phosphatase alcaline par des cellules nouvellement formées. L'activité en phosphatase alcaline s'observe à 5 jours; elle atteint un pic à 20 jours, après implantation, et décroit à 15% de la valeur maximale au 40ème jour suivant l'implantation. Le dépôt de calcium dans les implants, contemporain de la formation d'os nouveau, débute environ 12 jours après l'implantation et diminue progressivement pendant les 40 jours de la période expérimentale. Ni recalcification de la matrice originale, ni néoformation d'os ne s'observent dans les implants inactivés par l'acide et l'alcool. L'induction de synthèse de phosphatase alcaline s'observe, dans la phase pré-osseuse de morphogène, 5 jours avant le dépôt de calcium et n'est pas directement en rapport avec le mécanisme de la calcification.
    Abstract: Zusammenfassung Zur Gewinnung von Knochenmatrix-Implantaten, welche neue Knochenbildung anregen können, wurde kortikaler Knochen während 48 Std bei 2° mit 0,6 N HCl entkalkt. Die Fähigkeit neuen Knochen zu bilden, konnte zerstört werden, wenn der zur Entkalkung verwendeten 0,6 N HCl 66,5% Äthylalkohol zugesetzt wurde. Im Muskel implantiert wurden sowohl die aktiven als auch die inaktiven Implantate von mesenschymalen Zellen überschwemmt. Es fand sich kein signifikanter Unterschied des DNS-Gehaltes zwischen aktiven und inaktiven Implantaten, jedoch wurde die Synthese von alkalischer Phosphatase durch eine neue Zellpopulation nur im aktiven Implantat hervorgerufen. Die Aktivität der alkalischen Phosphatase lag nach 5 Tagen vor, erreichte 20 Tage nach Implantation den Höhepunkt, um nach dem 40. Tag nach Implantation auf 15% des Maximalwertes abzusinken. Die Calciumablagerung in den Implantaten, welche mit der Differenzierung von neuem Knochen zusammenfällt, begann ungefähr 12 Tage nach der Implantation und nahm während der 40tägigen Experimentierphase stetig zu. Im mit Säure-Alkohol inaktivierten Implantat fand weder eine Mineralisation der alten Matrix, noch eine Differenzierung von Knochen statt. Die Synthese von alkalischer Phosphatase wird im Verlaufe der Morphogenesis in der Phase vor der Knochenbildung angeregt, d. h. 5 Tage vor dem Auftreten von Calciumablagerungen. Sie steht nicht in direktem Zusammenhang mit dem Mineralisationsmechanismus.
    Notes: Abstract Implants of bone matrix, capable of inducing new bone formation, were prepared by decalcifying cortical bone in 0.6 N HCl at 2°C for 48 hours. The inductive property could be inactivated by decalcification in 0.6N HCl containing 66.5% ethanol. When implanted into muscle, both the active and inactive implants were invaded by mesenchymal cells. There was no significant difference in the DNA content of active and inactive implants, but only the active implants induced the synthesis of alkaline phosphatase by a new cell population. Alkaline phosphatase activity was present at 5 days, reached a peak at 20 days after implantation, then declined to 15% of the maximum value by the 40th day after implantation. The deposition of calcium in the implants, coinciding with the differentiation of new bone, began about 12 days after implantation, and gradually increased throughout the 40-day experimental period. Neither recalcification of old matrix nor differentiation of bone occurred in the acid-alcohol-inactivated implants. Induction of alkaline phosphatase synthesis occurred in the pre-osseous phase of morphogenesis 5 days before the appearance of calcium deposits and was not directly correlated with the mechanism of calcification.
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