In vitro formation and aggregation of heterotypic collagen I and III fibrils

doi:10.1016/0141-8130(93)90030-P

International Journal of Biological Macromolecules

Volume 15, Issue 5, October 1993, Pages 299-304

https://doi.org/10.1016/0141-8130(93)90030-P Get rights and content

Abstract

In vitro fibrillogenesis of solutions containing pepsin digested and acid soluble collagens I and III from human and bovine skin were investigated by turbidity-time measurements, dark-field and electron microscopy. The maximum turbidity of these solutions exhibited inversely proportional dependence on the collagen III content. Self-assembly was accelerated by collagen III. As a measure of mass per unit length, the maximum turbidity shows a mean decrease of 88% when comparing the absorbance at 313 nm for 0% and 50% collagen III in a composite solution of acid extracted collagen. In contrast to these findings, the diameter of fibrils from acid extracted fetal calf skin with 50% collagen III, determined from electron micrographs, was only 23% smaller than for pure collagen I. Correspondence with investigations oninvitro fibrillogenesis with dark-field microscopy and electron microscopy, this phenomenon apparently derives from the bundling of fibrils. This may be interpreted to mean that bundling of fibrils is already suppressed at low collagen III concentrations. A comparison of acid and pepsin extracted fetal calf skin yielded similar behaviour of collagen I and III mixtures, even though the pepsin extract displayed a turbidity reduction that was about 25% less than the acid extract. For pepsin digested collagen from human and bovine skin, differences were found for maximum turbidity and the ability to form bundles decreasing with the biological age of the donor.

References (31)

R.O. Hynes
Cell
(1992)
J.M. Seyer
Biochim. Biophys. Acta
(1980)
A. Albini et al.
J. Invest. Dermatol.
(1985)
D.E. Birk et al.
Arch. Biochem.
(1984)
A.M. Romanic et al.
J. Biol. Chem.
(1991)
E.J. Miller et al.
Methods Enzymol.
(1982)
B.R. Williams et al.
Biol. Chem.
(1978)
R.A. Gelman et al.
J. Biol. Chem.
(1979)
G.C. Na et al.
J. Biol. Chem.
(1986)
A. Torre-Blanco et al.
J. Biol. Chem.
(1992)

S. Amudeswari et al.

Collagen Rel. Res.

(1987)

K. Piez

J.E. Scott

Biochem. J.

(1988)

K.G. Vogel et al.

Biochem. J.

(1984)

R. Fleischmajer et al.

J. Histochem. Cytochem.

(1988)

Cited by (31)

Fibril matrices created with collagen from the marine fish barramundi for use in conventional three-dimensional cell culture
2022, International Journal of Biological Macromolecules
Citation Excerpt :
Therefore, it is necessary to confirm the absence of type III collagen in both BC and TC. SDS-PAGE bands of α(III) which appear between α(I) and β(I) [32] were not observed for both the collagens in Fig. 1, indicating that the faster fibril formation of BC was not attributed to the presence of type III collagen. The presence of collagen telopeptides is another important factor affecting fibril formation rate because telopeptides act as a nucleus of collagen fibril and accelerates formation of fibrils [33].
Collagen obtained from fish offal (skin, scales, and bones) is required from some religious and ethnic groups, thus indirectly increasing demands for fish collagen for biomedical applications. The limitation of fish collagen is its lower thermal stability compared to mammalian collagen. In this study, we focused on collagen extracted from scales of the marine fish barramundi (Lates calcarifer) and demonstrated the suitability for the collagen to be utilized in collagen fibril matrices (CFM). Collagen was extracted from the scales through pepsin-digestion and purified (designated as “BC”). The denaturation temperature (Td) for BC was determined to be 36.4 °C, one of the highest among fish collagens. BC formed CFM which was thermally stable at 37 °C, while Td was lower than 37 °C. This could be explained by the fast fibril formation, initiating at temperatures near 20 °C in a temperature-elevated process. As a result, the NIH3T3 cells were successfully encapsulated in the CFM of BC and cultured three-dimensionally for 7 d. The cells spread and exhibited well-developed pseudopodia in the CFM of BC as observed in the CFM of pig collagen matrices. This is the first report on fish CFM used for conventional 3-D cell culture.
Skeletal muscle
2022, Sturkie's Avian Physiology
Skeletal muscle is the largest tissue in the body, constituting about 40% of its mass. This chapter covers the development and growth of avian skeletal muscle, beginning with a discussion of its structural diversity in different bird species, muscle fiber types, and the mechanism of skeletal muscle contraction. An overview of embryonic and posthatch development is then provided, with a focus on satellite cells, the adult myoblast stem cells that are responsible for all posthatch muscle growth. This is followed by a description of the roles of myogenic transcriptional regulatory factors and growth factors in muscle myogenesis. New emerging areas in avian skeletal muscle biology are addressed, including satellite cell heterogeneity, myopathies in meat-type poultry, and the association of these diseases with extracellular matrix organization and regulation. New strategies to combat the myopathies are discussed.
The water soluble zinc based metal-organic frameworks (Zn-MOFs) as potential inhibitors for collagen fibrillogenesis
2021, International Journal of Biological Macromolecules
Citation Excerpt :
Fibrillogenesis involves a two-step process, i.e., (i) the aggregation of individual helical molecules into nuclei and (ii) the growth of nuclei into fibrils. The efficiency of crosslinking and fibrillation rate is generally monitored by a change in turbidity (due to growth of fibril formation) at 313 nm. [27,28] The effect of a small molecule on the fibril formation was determined by its final turbidity (Δh) and the time taken to reach half the value of its turbidity (t1/2).
Small molecules ranging from organic to inorganic systems have been reported as stabilizing agents for collagen. Various transition metal complexes have been utilized as tanning agent. However, as per the environmental norms issued by various regulatory agencies, the presence of certain metals such as Cr, Fe, Al, Zr and Ti in leather has been restricted to minimal amount (50 ppm), an unsurmountable task. To overcome the above issue and find an alternative tanning system, here in this study, we have reported the interaction of two water-soluble zinc-based metal-organic frameworks (MOFs), i.e., ZnPV (1) and ZnPA (2), with collagen using various spectroscopic techniques. Fibrillation kinetics studies showed that a significant delay in fibril formation with Zn-MOFs treated collagen was observed compared to the collagen untreated/ treated with individual ligands and metal salt. Circular dichroism studies show that at a low weight ratio (1:0.2 and 1:1::Collagen: MOF), no perturbation in the triple helical structure was observed, while at higher weight ratio (1:4), denaturation of collagen occurs. FT-IR studies showed that no perturbation was observed in the amide backbone in MOF-treated collagen. Differential scanning calorimetric data revealed that both Zn-MOFs increased the thermal denaturation temperature by 22 ± 2 °C compared to the collagen treated with individual entities. The viscosity of collagen rises with the increase in the concentration of Zn-MOFs. To the best of our knowledge, this is the first report on the use of the metal-organic framework as a stabilizing agent for collagen structure and might help in exploring the MOFs as potential tanning agents.
A comparative study of the properties and self-aggregation behavior of collagens from the scales and skin of grass carp (Ctenopharyngodon idella)
2018, International Journal of Biological Macromolecules
Citation Excerpt :
Type I collagen is the most abundant and most widely used collagen, and its triple-helical structure that consists of three polypeptide chains has a high tensile strength [14]. It has been reported that type I collagen molecules are capable of in vitro self-aggregation into fibrils in neutral solution at room temperature [15]. This self-aggregation improves the specific surface area (SSA), resistance to enzyme degradation, and thermal stability of collagen-based materials [16].
Collagens were extracted from the scales and skin of Ctenopharyngodon idella (C. idella) as raw materials using an acid-enzyme hybrid method. The structural properties of the extracted collagens were compared using ultraviolet-visible spectrophotometry, Fourier transform infrared spectroscopy, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and differential scanning calorimetry. Additionally, the in vitro self-aggregation behaviors of the two types of collagens (fish skin- and scale-derived collagens) were compared using turbidimetric assays, aggregation assays, and scanning electron microscopy (SEM). The results showed that both types of extracted collagen were typical type I collagen with two α chains and intact triple-helical structures. The denaturation temperatures of the collagens from fish scales and skin were 34.99 °C and 39.75 °C, respectively. Both types of collagens were capable of self-aggregation in neutral salt solution at 30 °C, with aggregation degrees of 28% and 27.33% for the scale and skin collagens, respectively. SEM analysis revealed that both types of collagens could self-aggregate into interwoven fibers, and the fish scale-derived collagen had a more pronounced reticular fiber structure with a striped periodic D-band pattern of collagen fibrils, whereas the collagen fibers from the self-aggregation of fish skin-derived collagen had a certain degree of disruption without any D-band pattern.
Characterization of Collagen Type i and II Blended Hydrogels for Articular Cartilage Tissue Engineering
2016, Biomacromolecules
Biomaterials that provide signals present in the native extracellular matrix have been proposed as scaffolds to support improved cartilage regeneration. This study harnesses the biological activity of collagen type II and the superior mechanical properties of collagen type I by characterizing gels made of collagen type I and II blends. The collagen blend hydrogels were able to incorporate both types of collagen and retained chondroitin sulfate and hyaluronic acid. Cryo-scanning electron microscopy images showed that the 3:1 ratio of collagen type I to type II gels had a lower void space percentage (36.4%) than the 1:1 gels (46.5%). The complex modulus was larger for the 3:1 gels (G* = 5.0 Pa) compared to the 1:1 gels (G* = 1.2 Pa). The 3:1 blend consistently formed gels with superior mechanical properties compared to the other blends and has the potential to be implemented as a scaffold for articular cartilage engineering.
Recombinant human type II collagens with low and high levels of hydroxylysine and its glycosylated forms show marked differences in fibrillogenesis in vitro
1999, Journal of Biological Chemistry
Type II collagen is the main structural component of hyaline cartilages where it forms networks of thin fibrils that differ in morphology from the much thicker fibrils of type I collagen. We studied here in vitro the formation of fibrils of pepsin-treated recombinant human type II collagen produced in insect cells. Two kinds of type II collagen preparation were used: low hydroxylysine collagen having 2.0 hydroxylysine residues/1,000 amino acids, including 1.3 glycosylated hydroxylysines; and high hydroxylysine collagen having 19 hydroxylysines/1,000 amino acids, including 8.9 glycosylated hydroxylysines. A marked difference in fibril formation was found between these two kinds of collagen preparation, in that the maximal turbidity of the former was reached within 5 min under the standard assay conditions, whereas the absorbance of the latter increased until about 600 min. The critical concentration with the latter was about 10-fold, and the absorbance/microgram collagen incorporated into the fibrils was about one-sixth. The morphology of the fibrils was also different, in that the high hydroxylysine collagen formed thin fibrils with essentially no interfibril interaction or aggregation, whereas the low hydroxylysine collagen formed thick fibrils on a background of thin ones. The data thus indicate that regulation of the extents of lysine hydroxylation and hydroxylysine glycosylation may play a major role in the regulation of collagen fibril formation and the morphology of the fibrils.

View all citing articles on Scopus

View full text

In vitro formation and aggregation of heterotypic collagen I and III fibrils

Abstract

Cell

Biochim. Biophys. Acta

J. Invest. Dermatol.

Arch. Biochem.

J. Biol. Chem.

Methods Enzymol.

Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Collagen Rel. Res.

Biochem. J.

Biochem. J.

J. Histochem. Cytochem.