摘要
目的探讨壳聚糖微球介导人IL-1Ra与TGF-β1基因转染兔软骨细胞。方法制备壳聚糖微球介导IL-1Ra质粒与TGF-β1质粒转染系统，检测其载药、体外释药、降解，转染体外培养的兔膝软骨细胞，荧光显微镜、荧光定量PCR、MTF检测。结果壳聚糖-IL-IRa DNA和壳聚糖-TGF-β1 DNA微球平均径粒（2．8±0．2）μm和（2．6±0．1）μm，包封率（88．3±4．1）％和（87．2±2．6）％；缓释分3个阶段。荧光显微镜观察、荧光定量PCR证实软骨细胞转染基因得到30d表达。MTY提示转染促进软骨细胞增殖。结论壳聚糖微球介导IL-1Ra与TGF-β1基因转染软骨细胞可获得较长期目的基因表达，可促进软骨细胞增殖，为用于基因治疗软骨退变和促进软骨修复提供基础。
Objective To explore transinfection of rabbit chondrocytes via chitosan microsphere with human IL-1Ra and TGF-β1 gene. Methods Chitosan-DNA microspheres carrying plasmids with IL- 1Ra and TGF-β1 genes were prepared. The encapsulation efficiency,DNA-released kinetics and lysozyme degradation in vitro were performed. Articular rabbit chondrocytes were co-transinfected with the plasmids with IL-1Ra and TGF-β1 genes via chitosan-DNA microsphere,evaluated by fluorescence microscope, TaqMan real-time PCR assay and MTF test. Results The chitosan microspheres with IL-1Ra and TGF-β1 genes were（2.8 ± 0.2）μm and （2.6 ± 0.1）μm in diameters respectively. The encapsulation efficiency were （88.3± 4.1）% and（87.2 ±2.6）%. During the degradation, significant morphological changes were noticed. The plasmids could be released in a muhiphasic fashion. Enhanced green fluorescent protein and Real-Time PCR analysis showed that genes were expressed in chondrocytes, lasting near 30 days. MTY indicated that the cotransinfection promoted the chondrocytes＇ proliferation. Conclusion IL-1Ra and TGF-β1 genes cotransfected into chondroeytes via ehitosan-DNA microsphere could be expressed in a long term and eotransinfection promoted the chondroeytes＇ proliferation, which is the base of inhibiting the degeneration of cartilage and oromote cartilage repair.
出处
《中华显微外科杂志》
CSCD
北大核心
2009年第6期471-473,F0003,共4页
Chinese Journal of Microsurgery
基金
国家自然科学基金（30600632）
关键词
壳聚糖
微球
白介素受体拮抗剂
转化生长因子Β
基因转染
软骨
Chitosan
Microsphere
Interleukin-1 receptor antagonist
Transforming growth factor β1
Gene transinfection
Cartilage
