Initial Study Of Using Autologous Bone Marrow Mesenchymal Stem Cells Combining Homogeneity Variant Decalcified Bone Matrix To Repair Cartilage Defects

Objective: To investigate autologous bone marrow mesenchymal stem cells (BMSCs) culture of rabbit in vitro and its transformation to chondrocyte phenotype in order to discuss the possibility of culturing cells with the homogeneity variant decalcified bone matrix (DBM) of rabbit. Be ready for using tissue-engineered cartilage to repair cartilage defects.Methods: BMSCs were harvested from the femur of rabbit, then isolated and purified in vitro. The third generation BMSCs were taken and divided into the test group and the control group. The test gronp was induced by LG-DMEM with 10% fetal calf serum,transforming growth factor-β₁ (TGF-β₁) and dexamethasone. The control group was induced by LG-DMEM with 10% fetal calf serum. Then toluidine blue dyeing was performed to test the secretion of proteoglycans, immunohistochemical assay was performed to test the secretion of collagen type II after four days,seven days and fourteen days and calculate masculine rate. Prepare the homogeneity variant DBM of rabbit. Test the interval porosity of the bracket material by liquid substitution method after decalcification of two days,three days and four days. Observe the ultramicrostructur of DBM through scanning electron microscope and the adhere of DBM and cells after cultivate three days.Results: BMSCs were harvested easily and grew stably in vitro. The BMSCs of the test group transform to chondrocyte phenotype. Cell toluidine blue dyeing was allotri-dyeing. The diference between the test group and control group of collagen type II immunohistochemical staining was significant statistically. The porous structure of DBM decalcificated for three days fitted the cell to adhere and generation. The DBM and cells adhered well in vitro .Conclusions: The rabbit autologous BMSCs were harvested easily and can be transformed to chondrocyte phenotype in vitro. The homogeneity variant DBM of rabbit was made convenient and had satisfactory characteristics. The combination of DBM and cells was good and provided a choice to repair cartilage defects.