Chitosan-DNA介导关节基因转移的体外研究

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.

Studies of in vitro Gene Transfer into Articular Mediated by Chitosan-DNA Nanoparticles

Purpose To study the Chitosan mediated reporter gene transfection of chondrocytes and synoviocytes of joint in vitro. Methods Chondrocytes and synoviocytes were separated and cultured from lapines' knee joint, Chitosan-DNA nanoparticles were prepared using fluorescent plasmid pEGFP-C 3 as a reporter gene, transfected to chondrocytes and synoviocytes and observed the expression of enhanced green fluorescent protein (EGFP) under a fluorescence microscope. Results The Chitosan-DNA nanoparticle system effectively delivered genes to the cells and the DNA can be send with a time lag, the results of the luminescence assay to detect the luciferase activity in the transfected cells indicated relatively high transfection efficiencies with chondrocytes. Conclusion Chitosan can mediate articular chondrocytes and synoviocytes gene transfer, this would provide experimental evidence for investigating non-viral gene transfer of joint disease.