In vitro formation of mineralized cartilagenous tissue by articular chondrocytes
Study of the deep articular cartilage and adjacent calcified cartilage has been limited by the lack of an in vitro culture system which mimics this region of the cartilage. In this paper we describe a method to generate mineralized cartilagenous tissue in culture using chondrocytes obtained from the deep zone of bovine articular cartilage. The cells were plated on Millipore CMR filters. The chondrocytes in culture accumulated extracellular matrix and formed cartilagenous tissue which calcified when β-glycerophosphate was added to the culture medium. The cartilagenous tissue generated in vitro contains both type II and type X collagens, large sulfated proteoglycans, and alkaline phosphatase activity. Ultrastructurally, matrix vesicles were seen in the extracellular matrix. Selected area electron diffraction confirmed that the calcification was composed of hydroxyapatite crystals. The chondrocytes, as characterized thus far, appear to maintain their phenotype under these culture conditions which suggests that these cultures could be used as a model to examine the metabolism of cells from the deep zone of cartilage and mineralization of cartilagenous tissue in culture.
Key wordschondrocytes calcification articular cartilage
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- 7.Gibson, G. J.; Lin, D. L. Type X collagen morphology in calf growth plate and articular cartilage. Trans. Orthop. Res. Soc. 20:28; 1995.Google Scholar
- 14.Lovell, T. P.; Eyre, D. R. Unique biochemical characteristics of the calcified zone of articular cartilage. Trans. Orthop. Res. Soc. 13:511; 1988.Google Scholar
- 17.Oegema, T. R., Jr.; Thompson, R. C., Jr. The zone of calcified cartilage. Its role in osteoarthritis. In: Kuettner, K., ed. Articular cartilage and osteoarthritis. New York: Raven Press; 1992:319–331.Google Scholar
- 18.Oegema, T. R., Jr.; Thompson, R. C., Jr. Cartilage-bone interface (Tidemark). In: Brandt, K., ed. Cartilage changes in osteoarthritis. Indiana School of Medicine publication. Basel: Ciba-Geigy; 1990:43–52.Google Scholar
- 21.Poole, R. A. Cartilage in health and disease. In: McCarty, D.; Koopman, W., ed. Arthritis and allied conditions. A textbook of rheumatology. 12th ed. Malvern: Lea and Febiger; 1992:279–333.Google Scholar
- 27.Thompson, R. C.; Oegema, T. R., Jr.; Lewis, J. L., et al. Osteoarthrotic changes after acute transarticular load. J. Bone Joint Surg. 73A:990–1001; 1991.Google Scholar
- 29.Walker, G.; Carpenter, R. J.; Oegema, T. R., Jr., et al. Evidence for activity in the tidemark in normal articular cartilage. Trans. Orthop. Res. Soc. 15:182; 1990.Google Scholar
- 31.Whyte, M. P. Alkaline phosphatase: physiological role explored in hypophosphatasia. In: Peck, W. A., ed. Bone and mineral research. 6th ed. Amsterdam: Elsevier Science Publishers; 1989:175–218.Google Scholar
- 33.Wuthier, R. E. Mechanism of matrix vesicle mediated mineralization of cartilage. ISI Atlas Sci. Biochem. 1:231–241; 1988.Google Scholar
- 35.Yoon, K.; Golub, E. E.; Rodan, G. A. Alkaline phosphatase cDNA transfected cells promote calcium and phosphate deposition. In: Glimcher, M. J.; Lian, J. B., ed. Proceedings of the Third International Conference on the Chemistry and Biology of Mineralized Tissues. New York: Gordon and Breach Science Publishers; 1989:643–652.Google Scholar