Control Study of Normal and Osteoarthritis Chondrocytes from Human in Vitro

Objective: To study the methods for isolation,culture and identification of articular chondrocytes from normal human and osteoarthritis patients in vitro,compare the biological characteristics and evaluate the biological activity between normal and osteoarthritis chondrocytes.Methods: Cartilage was harvested under sterile conditions from human traumatic knee joints and osteoarthritis knee joints in Total Knee Arthroplasty.Human articular chondrocytes were isolated by using two-step enzymatic digestion,and the cells were cultured and subcultured respectively in vitro.The cells were compared by observing cell morphology under phase-contrast microscope,drawing growth curve,measuring cell proliferation,toluidine blue staining and type II collagen immunohistochemistry staining.Results: The morphology of osteoarthritis chondrocytes was similar to fibroblasts,and the growth rate was significantly more slowly than that of normal chondrocytes.Measured cell proliferation by MTT,the proliferation rate of the 2nd,4th and 6th osteoarthritis chondrocytes was slower than that of the same generation normal chondrocytes at the same time points(P 0.05).Toluidine blue staining and type II collagen immunohistochemistry staining showed that the staining of osteoarthritis chondrocytes were lighter than that of the normal chondrocytes,and were almost no coloration after several passages.Conclusion: Within 5 generations,the normal chondrocytes grew well with obviously biological characteristics and turned to dedifferentiation after 5 generations.The osteoarthritis chondrocytes with slowly cell proliferation and early degeneration of biological characteristics were in accordance with the performance of degeneration of chondrocytes,providing experimental foundation for study of the osteoarthritis in the cellular level.