The Experimental Research On The Repair Of Articular Cartilage Deffects By The Complex Of Autologous Chondrocyte-bone Matrix Gelatin

Objective To investigate the effect of transplantation of autologous "two-phase" bone matrix gelatin(BMG) engineered with autologous chondrocyte of the repair of rabbit articular cartilage defects . To compare the ability of the repairg articular cartilage defects with autologous and allogenic chondrocyte.Methods 1. The articular cartilage cells were digested from of rabbit' s knee joint and cultured in vitro. The digestion method with 0. 2%type IICollagenase would be used only.2. It was easy to get enough quantity of the chondrocytes throughtransferring in vitro, we observed its morphology and drew the growth curve.3. Identify chondrocyte through immunohistochemistry and Mallory Toluidine blue stainin.4. Getting out the part of the rabbit's ilium, according to the method of modified Urist, the "two-phase" BMG was acquired from degreasing, taking off the calcium and getting out the protein, and was cut as the certain size and shape as needed. It could be conserved after sterilizing within a low temperature.5. Inoculated autologous or allogenic chondrocyte into autologous BMG to set up a composite of the cell-carrier, observed it under the electric-microscope after culturing three days.6.Made the model of articular cartilage defects of rabbits, divided it into three groups: the experimental group was implanted the autologous chondrocyte composite, the experimental control group was implanted the allogenic chondrocyte composite, the blank group was implanted nothing, the specimen was observed roughly in the eighth week and the twelfth week respectively, HE staining and immunohistochemistry were applied, and then graded score with Wakitani histological grading system.Results 1.The method of digestion with 0.2% type II Collagenase and c ultured in vitro can acquire the large quantity of pure isolated a rticular cartilage cells.2. The chondrocyte were verified by the positive results of collagen II through immunohistochemistry, and the results of Mallory Toluidine blue stainin were blue.3. Observed the BMG and the composite of the cell-carrier by the electric-microscope, the surface of the BMG was rough and had many small openings, the size of the small openings was 100um-800um, the cells were observed growing in the BMG well, and the cells were also growing inside the wall of the small openings.4. Cartilage defects were repaired by hyaline-like cartilage in the experimental group and experimental control group on the eighth week and the twelfth week, the blank group were fiber repair at the same time. There was difference between the experimental group and the experimental control group on the eighth week and the twelfth week. But there was not an obvious statistical difference on with Wakitani histological grading system. They were better than the blank group.Conclusions Autologous chondrocytes and its composite with autologous "two-phase" BMG had the function to repair articular cartilage defects, the ability was superior to the allogenic chondrocytes and its composite with autologous "two-phase" BMG group,but they were better than that of nothing transplanted.