Transitions in collagen types during matrix-induced cartilage, bone, and bone marrow formation (original) (raw)
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Bone, 1988
Collagen turnover during rat long bone development and growth was investigated using immunofluorescence methods with specific polyclonal antibodies against native (triple helix) and denatured (breakdown products) forms of type I and II collagen. Labeling of cryostat sections with anti-native and denatured collagen type II antibodies resulted in a positive staining throughout the cartilage matrix of fetal and adult long bones. Likewise, native and denatured type I collagen could be detected in mineralized and non-mineralized bone matrix. Moreover, labeling with anti-denatured type I antibody evoked a strong intracellular staining of osteoblasts, but not of osteocytes. Denatured type I was also Irralized intra-pericellularly in the small chondrocytes comprising the primitive cartilage cores and the epiphyses of older long bones. On the other hand, apart from its localization in the cartilage matrix, denatured type II collagen was found specifically within the chondrocytes. These observations indicate that a continuous turnover of the major collagen types takes place in fetal and adult rat long bone tissue. Degradation of collagen apparently occurs intra-and extracellularly, and is mainly independent of the presence and activity of osteoclasts. The presence of denatured type I collagen in cartilage suggests that chondrocytes synthesize small amounts of type I collagen, which is immediately degraded to a denatured form .
Type XXVII collagen at the transition of cartilage to bone during skeletogenesis
Bone, 2007
COL27A1 is a member of the collagen fibrillar gene family and is expressed in cartilaginous tissues including the anlage of endochondral bone. To begin to understand its role in skeletogenesis, the temporospatial distributions of its RNA message and protein product, type XXVII collagen, were determined in developing human skeletal tissues. Laser capture microdissection and quantitative reverse-transcription polymerase chain reaction demonstrated that gene expression occurred throughout the growth plate, and that it was higher in the resting and proliferative zones than in hypertrophic cartilage. Immunohistochemical analyses showed that type XXVII collagen was most evident in hypertrophic cartilage at the primary ossification center and at the growth plate, and that it accumulated in the pericellular matrix. Synthesis of type XXVII collagen overlapped partly with that of type X collagen, a marker of chondrocyte hypertrophy, preceded the transition of cartilage to bone, and was associated with cartilage calcification. Immunogold electron microscopy of extracted ECM components from mouse growth plate showed that type XXVII collagen was a component of long non-banded fibrous structures, filamentous networks, and thin banded fibrils. The timing and location of synthesis suggest that type XXVII collagen plays a role during the calcification of cartilage and the transition of cartilage to bone.
Annals of Anatomy - Anatomischer Anzeiger, 2000
Hyaline cartilage has only a limited capacity of regeneration, thus, lesions of articular cartilage can lead to early osteoarthrosis. Current concepts in conservative orthopedic therapy do not always lead to satisfying results. As one new attempt to facilitate cartilage repair, autologous transplantation of articular chondrocytes is investigated in different assays. This study was designed to create a resistible and stable cell-matrix-biocomposite with viable and biosynthetically active human chondrocytes, osteoblasts or fibroblasts. This biocomposite might serve as an implant to treat deep osteochondral defects in the knee. We collected cartilage, spongiosa and skin probes from healthy patients undergoing hip-surgery and enzymatically liberated the chondrocytes, seeded them into culture flasks and cultured them until confluent. The spongiosa and the skin samples were also placed in culture flasks and cells cultured until confluent. After 4-6 weeks, cells were trypsinized and grown on a type I/III collagen matrix (Chondrogide TM, Geistlich Biomaterials, Wolhusen, Switzerland) for 7 days in standard Petri dishes and in a special perfusion chamber culture system. As controls, cells were seeded onto plastic surfaces. Then scaffolds were fixed and embedded for light microscopy and electron microscopy by routine methods.
Altered cartilage phenotype expressed during intramembranous bone formation
Journal of Bone and Mineral Research, 2009
The sequential phenotypic expression occurring during intramembranous bone formation was investigated using the tooth extraction socket created in rat alveolar bone in vivo. The differential expression of bone extracellular matrix genes, such as collagen I and osteocalcin, was confirmed by RNA transfer blot analysis and in situ hybridization during the active healing period of the bony socket. To clarify the possible involvement of the chondrogenic phenotype during the process of intramembranous bone formation, the expression of cartilage collagen I1 and IX was further examined in this model. It was found that both a,(II) and al(IX) mRNAs were present, but the al(IX) mRNA was a transcript from the downstream start site/promoter, which is a different site in the al(IX) gene from that used in hyaline cartilage. In situ hybridization indicated that the a,(IX) message was expressed by cells associated with bone matrix in the early formation stage. This finding led to the investigation of type IX collagen expression by osteogenic cells isolated from newborn rat calvariae, in which only the truncated form of al(IX) mRNA was indicated by RNA transfer analysis. The expression of collagen I1 and a truncated form of collagen I X may represent an early phenotypic feature of osteoblast differentiation.
Effect of Collagen Type I or Type II on Chondrogenesis by Cultured Human Articular Chondrocytes
Tissue Engineering Part A, 2013
Introduction: Current cartilage repair procedures using autologous chondrocytes rely on a variety of carriers for implantation. Collagen types I and II are frequently used and valuable properties of both were shown earlier in vitro, although a preference for either was not demonstrated. Recently, however, fibrillar collagens were shown to promote cartilage degradation. The goal of this study was to evaluate the effects of collagen type I and type II coating on chondrogenic properties of in vitro cultured human chondrocytes, and to investigate if collagen-mediated cartilage degradation occurs. Methods: Human chondrocytes of eight healthy cartilage donors were isolated, expanded, and cultured on culture well inserts coated with either collagen type I, type II, or no coating (control). After 28 days of redifferentiation culture, safranin O and immunohistochemical staining for collagen types I, II, X, and Runx2/ Cbfa1 were performed and glycosaminoglycan (GAG) and DNA content and release were examined. Further, expression of collagen type I, type II, type X, MMP13, Runx2/Cbfa1, DDR2, a2 and b1 integrin were examined by reverse transcriptase-polymerase chain reaction. Results: The matrix, created by chondrocytes grown on collagen type I-and II-coated membranes, resembled cartilage more than when grown on noncoated membranes as reflected by histological scoring. Immunohistochemical staining did not differ between the conditions. GAG content as well as GAG/DNA were higher for collagen type II-coated cartilage constructs than control. GAG release was also higher on collagen type I-and IIcoated constructs. Expression of collagen type X was higher of chondrocytes grown on collagen type II compared to controls, but no collagen X protein could be demonstrated by immunohistochemistry. No effects of collagen coating on DDR2 nor MMP-13 gene expression were found. No differences were observed between collagen types I and II. Conclusion: Chondrocyte culture on collagen type I or II promotes more active matrix production and turnover. No significant differences between collagen types I and II were observed, nor were hypertrophic changes more evident in either condition. The use of collagen type I or II coating for in vitro models, thus, seems a sound basis for in vivo repair procedures.
Biomaterials, 2002
The limited intrinsic repair capacity of articular cartilage has stimulated continuing efforts to develop tissue engineered analogues. Matrices composed of type II collagen and chondroitin sulfate (CS), the major constituents of hyaline cartilage, may create an appropriate environment for the generation of cartilage-like tissue. In this study, we prepared, characterized, and evaluated type II collagen matrices with and without CS. Type II collagen matrices were prepared using purified, pepsin-treated, type II collagen. Techniques applied to prepare type I collagen matrices were found unsuitable for type II collagen. Crosslinking of collagen and covalent attachment of CS was performed using 1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide. Porous matrices were prepared by freezing and lyophilization, and their physico-chemical characteristics (degree of crosslinking, denaturing temperature, collagenase-resistance, amount of CS incorporated) established. Matrices were evaluated for their capacity to sustain chondrocyte proliferation and differentiation in vitro. After 7 d of culture, chondrocytes were mainly located at the periphery of the matrices. In contrast to type I collagen, type II collagen supported the distribution of cells throughout the matrix. After 14 d of culture, matrices were surfaced with a cartilagenous-like layer, and occasionally clusters of chondrocytes were present inside the matrix. Chondrocytes proliferated and differentiated as indicated by biochemical analyses, ultrastructural observations, and reverse transcriptase PCR for collagen types I, II and X. No major differences were observed with respect to the presence or absence of CS in the matrices. r