壳聚糖微球转染人IL-1Ra与TGF-β1基因治疗兔骨关节炎

Transinfection of rabbit knee osteoarthritis models via chitosan microsphere as gene carriers with recombined human IL-1Ra gene and TGF-β1 gene

目的探讨壳聚糖微球转染人IL-1Ra与TGF-β1基因治疗兔膝关节早期骨关节炎的方法与效果。方法制备分别包裹IL-1Ra质粒DNA和TGF-β1质粒DNA的壳聚糖微球缓释系统，分组向兔膝关节早期骨关节炎模型关节腔注射含IL—1Ra基因和（或）TGF-β1基因的壳聚糖微球、不含上述基因的壳聚糖溶液，定期处死，使用ELISA检测关节腔灌洗液的IL-1Ra、TGF-β1浓度，取关节标本Mankin评分、苏木精一伊红（HE）染色、番红O染色及免疫组化检测。结果经过60d观察，壳聚糖微球转染基因组的IL-1Ra与TGF-β1的基因可持续表达，双基因组的软骨标本从外观、染色观察，损伤程度轻于单基因组。双基因组的Mankin评分明显低于单基因组（P〈0．05），单基因组的Mankin评分明显低于空白组（P〈0．05）。结论关节腔注射壳聚糖微球转染IL-1Ra与TGF-β1基因能抑制软骨的退变和促进软骨的修复。

Objective To explore the method and effect of transinfection of rabbit early knee os- teoarthritis models via ehitosan microsphere with gene of reeombined human IL-1Ra gene and TGF-β1 gene. Methods Chitosan mierospheres with plasmids of IL-1Ra gene and TGF-β1 gene, and rabbit early knee osteoarthritis models were prepared. Rabbits in different groups had intra-artieular injections of chitosan microsphere containing IL-1Ra gene and / or TGF-β1 gene, and ehitosan solution as control group before being executed regularly and randomly. The joint specimens were evaluated by HE staining, lycopene red O staining and immunohistochemical analysis and Mankin＇s score. ELISA was used for detection of IL-1Ra and TGF-β1 concentration of articular cavity fluid in each group. Results The control group was consistent with the pathological changes of early OA. In co-transinfection group, judging from the appearance and staining of cartilage, the OA damage of the specimens was less serious than other groups＇. Its Mankin＇s score was significantly lower than single-gene transinfection group （P 〈 0.05）, and the latters Mankin＇s score were significantly lower than control group （P 〈 0.05）. Conclusion Intra-artieular injection of chitosan mierospheres containing both IL- IRa gene and TGF-β1 gene could inhibit the degeneration of cartilage and promote cartilage repair.