Tissue Engineered Cartilage Culture In Rabbits' Knee Cavity By Homologous Decalcified Bone Matrix Combined Bone Marrow Mesenchymal Stem Cell: An Experimental Research

Objective To comparison the superiority of cultivating tissue engineered cartilage by homologous decalcified bone matrix combine bone-marrow mesenchymal stem cell in rabbits'knee cavity with culture in vitro.Method Archiogeneration BMSC were isolated and purified by adhering to the culture glassware wall from neonatal New Zealand white rabbits, the third generation BMSC induce to chondrocyte by transforming growth factor TGF-β1, insulin-like growth factor IGF-1 and VitC. 7 days later,the cells combined with homologous decalcified bone matrix and culture in vitro(Group A), culture in rabbits'left knee cavity (Group B) and culture homologous decalcified bone matrix in rabbits'right knee cavity (Group C). Every 4 weeks after cellular transplant, all rabbits of group B,C were sacrificed and the specimens were made in paraffin sections. All the specimens were carried out with H-E stain , toluidine blue stain and typeⅡcollagen immunohistochemical staining with chromogen diaminobenzidine(DAB). Immunohistochemical optical density(OD) value were calculated by morphology analysis software. Measurement data were demonstrated with mean±standard, satistical analysis was performed by t t test, One-way ANOVA using SPSS 15.0 software.Results After cultivating for 4 weeks, H-E stain group A shows that chondrocytes diffused distribution on scaffold's surface.Group B shows that a great quantity chondrocytes in the scaffold,has found cartilage lacuna,matrix anachromasis and isogenous group. At the 8th week,group B's specimens maturated chondrocytes and isogenous groups arranged as columnar, surrounded by a pond of cellular matrix infiltrate interior the scaffolds. The decalcified bone matrix were absorbed partly. At the 12th week,all the scaffolds were absorbed obviously. Chondrocytes stuff in Group A's scaffolds ,but the arrangement in a mess . Group B's chondrocytes arranged such as the stress orientation.Compared the typeⅡcollagen immunohistochemical OD value between group A and B, in all groups, the typeⅡcollagen immunohistochemical OD value were calculated as mean±standard at 4th week, 8th week, and 12th week respectively, group B had significantly higher than group A.The OD value were analyzed using SPSS 15.0 software, the result show group B is better than Group A, the difference is significant.Conclusion Homologous decalcified bone matrix conbine induced BMSC ,which culture in rabbits'knee cavity could yield better histological feature tissue engineered cartilage.