The Experimental Research On The Repair Of Articular Cartilage Deffects By The Complex Of Autologous Chondrocyte-small Intest Inal Submuco Sa

Objective To investigate the effect of transplantation of small intest inal submuco sa(SIS)engineered with autologous chondrocytes of the repair of rabbit articular cartilage defects.Methods 1.The articular cartilage cells were digested from rabbit' s knee joint and cultured in vitro.The digestion method with 0.2%typeⅡcollagenase would be used only.2.It was easy to get enough quantity of the chondrocytes by isolated with single typeⅡcollagenase digestion and cultured in high density in vitro,we observed its morphology and drew the growth curve.3.Identify chondrocyte through immunohistochemical stain of collagen typeⅡand Mallory,Toluidine blue stain.4.the pig SIS was acquired from degreasing,taking off the cells ande protein,and getting out the muscle and m。5.Inoculated autologous chondorcytes into SIS 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 only implanted the scalfold,the blank group was implanted nothing,the specimen was observed roughly in the eighth week and the twelfth week respectively,HE staining and immunohistochemical stain of collagen type Ⅱwere applied,and then graded score with Wakitani histological grading system.Results 1.(1)The chondrocytes were completely isolated from the soli d matrix.The number of the harvested chondrocytes from every gram of wet cartilage was 40.3×106 on average and their viability rate was 6.8%on average.(2)The chondrocytes were proliferated rapidly dur ing the three generations and had triangle or multip-angle shape or e lliptic shape at the growing confluence.(3)The positive result was o bserved with Mallory,staining with toluidine blue and immunohistoche mical stain of collagen typeⅡ.2.The chondrocyte were verified by the positive results of collagenⅡthrough immunohistochemical stain of collagen typeⅡ,and the results of Mallory,Toluidine blue stain were blue.3.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.there was an obvious statistical difference on with Wakitani histological grading system and They all were better than the blank group.Conclusions 1.Enough number of good chondrocytes can be harvested by single collagenase digestion and confluent in vitro.2.Autologous chondrocytes and its composite with SIS had the function to repair articular cartilage defects,the ability was superior to the scalfold group,and they were better than that of nothing transplanted group.