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目的 :探索体外Chitosan对关节软骨细胞、滑膜细胞的转染。方法 :分离培养兔正常滑膜细胞、软骨细胞 ,使用荧光质粒 pEGFP -C3作为报道基因 ,制备Chitosan -DNA超微颗粒转染兔正常软骨细胞、滑膜细胞 ,荧光显微镜观察绿色荧光蛋白的表达。结果 :Chitosan能将DNA带入胞内 ,该DNA超微颗粒随时间的延长可缓慢进入胞内 ,其包被的DNA也可以被缓慢释放 ,且对软骨细胞的转染能力强。结论 :Chitosan在体外可介导关节软骨细胞、滑膜细胞基因转移 ,为今后关节疾病非病毒基因转移的研究提供实验依据。
Objective: To explore the transfection of Chitosan in vitro on articular chondrocytes and synovial cells. Methods: Rabbit normal synoviocytes and chondrocytes were isolated and cultured. Fluorescent plasmid pEGFP-C3 was used as a reporter gene to prepare Chitosan-DNA ultrafine particles for transfection of rabbit normal chondrocytes and synovial cells. The expression of green fluorescent protein (GFP) was observed under fluorescence microscope. Results: Chitosan can bring DNA into the cells. The DNA ultra-fine particles can slowly enter into the cells with time. The coated DNA can also be slowly released, and its ability to transfect chondrocytes is strong. CONCLUSION: Chitosan can mediate the gene transfer of articular chondrocytes and synovial cells in vitro and provide experimental evidence for the future study of non-viral gene transfer in joint diseases.