In vitro culture and identification of IL-1beta induced degeneration of cartilage cells in New Zealand white rabbits knee joint / 中国中西医结合杂志

ABSTRACT

<p><b>OBJECTIVE</b>To explore and identify the method for IL-1beta induced New Zealand rabbit knee chondrocyte degeneration, thus providing experimental bases for Chinese medical research on osteoarthritis from in vitro cultured chondrocytes.</p><p><b>METHODS</b>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-1beta induced model group (group M). No intervention was given to those in group Z. 10% FBS culture media containing 10 ng/mL IL-1beta was added to group M. All cells were passaged to the 3rd generation. They were compared using morphological observation, toluidine blue staining, type II collagen immunohistochemical staining, and flow cytometry.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>It was obviously seen that chondrocytes in IL-1beta induced New Zealand rabbit knee chondrocyte model obviously degenerated, which could be used in related experimental researches on osteoarthritis.</p>