[In vitro culture and identification of IL-1beta induced degeneration of cartilage cells in New Zealand white rabbits knee joint].

Objective To explore and identify the method for IL-1β induced New Zealand rabbit knee chondrocyte degeneration, thus providing experimental bases for Chinese medical research on osteoarthritis from in vitro cultured chondrocytes. Methods Under aseptic conditions, bilateral knee joint cartilage was collected from 4-week old New Zealand rabbits. Chondrocytes were separated by type Ⅱ collagenase digestion and mechanical blowing method. They were randomly divided into two groups when passaged to the 2nd generation, the normal control group(group Z) and the IL-1β induced model group(group M). No intervention was given to those in group Z. 10% FBS culture media containing 10 ng/mL IL-1β was added to group M. All cells were passaged to the 3rd generation. They were compared using morphological observation, toluidin blue staining, type Ⅱ collagen immunohistochemical staining, and flow cytometry. Results Under inverted microscope, the second and the 3rd generation chondrocytes' phenotype of group Z was stable with good proliferation. Most cells turned into fusiform and slabstone shaped. In group M, most cells turned into long spindle shape or irregular shape. Results of toluidine blue staining and immunohistochemistry showed that the positive expression of chondrocytes after staining in group Z was superior to that in group M. Results of flow cytometry showed that there was statistical difference in the apoptosis rate of the second generation chondrocytes between group M and group Z(P 0. 01). Conclusion It was obviously seen that chondrocytes in IL-1β induced New Zealand rabbit knee chondrocyte model obviously degenerated, which could be used in related experimental researches on osteoarthritis.