Research Of Repairing Rabbit Knee Joint Cartilage Defect By Heparinized Collagen/Chitosan Scaffolds Binding With TGF-β1 And Adipose-derived Stem Cells

【Objective】In this study we investgate wether the full-thickness defects of knee articular cartilage in Rabbit can be repaired by heparinized collagen/chitosan scaffold binding with TGF—β1 and adipose-derived stem cells in vivo.【Methods】1.Adipose-derived stem cells were isolated from rabbit's adipose tissue,purified and cultured.Then the biological features are studied in vitro.2. Heparinized collagen/chitosan scaffold were constructed.The scaffold`s degradation and the impaction to TGF—β1 binding were studied 3. Adipose-derived stem cells were seeded onto the heparinized collagen/chitosan scaffold binding with TGF—β1 and kept them growing together for 2 days.Then they were planted into the rabbit full-thickness defects of knee articular cartilage.The defects are divided into three groups: the experimental group used binding with TGF—β1 heparinized collagen/chitosan scaffold and adipose-derived stem cells, the scaffold group only use binding with TGF—β1 heparinized collagen/chitosan scaffold and the blank control group used nothing.At 12 weeks after operation,the animals were sacrificed and the newly formed tissues were observed macroscopically and microscopically,graded histologically and analyzed statistically.【Results】1.The proliferation and expanding of the ADSCs are active in vitro. The generation time is about 3-5 days. The number of the ADSCs were on increase during the culture. 2. The scaffold`s degradation had no significant differentiation between heparinized collagen/chitosan scaffold and collagen/chitosan scaffold ,but the TGF—β1 binding ability was highly improved by the heparinizion. 3.In experimental group the cartilage defects were repared smoothly by the newly formed tissues and histological analysis of the newly formed tissues demonstrated new cartilage construction. The result was better than that in scaffold group and in control group.【Conclusion】These findings suggest that.the TGF—β1 binding ability can be improved by the heparinizion of collagen/chitosan and the full-thickness defects of articular cartilage using heparinized collagen/chitosan scaffolds binding with TGF—β1 and ADSCs is proved to be feasible.·...