载BMP-2基因纳米粒缓释微球的制备及其成骨诱导效能的研究

发布时间：2018-04-28 05:56

本文选题：非病毒基因载体 + 多聚精氨酸修饰的壳聚糖　；参考：《广东医学》2017年S1期

【摘要】：目的构建多聚精氨酸-壳聚糖/p BMP-2纳米粒,探讨包封多聚精氨酸-壳聚糖/p BMP-2纳米粒的PELA微球的体外成骨诱导效能。方法构建多聚精氨酸修饰的壳聚糖多聚物,并与BMP-2质粒基因凝聚成纳米粒子,检测其表征。荧光显微镜及Western blot分析其体外细胞转染效能。进而,将纳米粒子包入PELA微球,与MC3T3-E1细胞共培养,检测BMP-2蛋白表达,茜素红染色分析其成骨诱导分化效能。结果多聚精氨酸-壳聚糖/p BMP-2纳米粒粒径(81.39±22.96)nm,能有效介导BMP-2基因转染MC3T3-E1细胞,转染率高于裸质粒组及壳聚糖/p BMP-2纳米粒组(P0.05)。PELA微球呈球形,粒径为(43.34±15.24)μm,包封率达(67.49±0.85)%,42 d内累积释药(56.87±5.09)%,与MC3T3-E1共培养,第7天BMP-2分泌量达最高(64.348±2.396)pg/m L,21 d后分泌量仍能达到(51.763±1.789)pg/m L。茜素红染色钙结节形成数量明显高于对照组。结论多聚精氨酸-壳聚糖能有效凝聚BMP-2基因成纳米级粒子,促进细胞转染,并可通过PELA微球的包封实现纳米粒的长效缓释,促进成骨分化。
[Abstract]:Objective to construct poly (arginine / chitosan / p BMP-2) nanoparticles and to investigate the osteogenic effect of PELA microspheres encapsulated with poly (arginine / chitosan / p BMP-2) nanoparticles in vitro. Methods Poly (arginine) modified chitosan polymer was constructed and condensed with BMP-2 plasmid gene to form nanoparticles. Fluorescence microscope and Western blot were used to analyze the transfection efficiency. Then, the nanoparticles were encapsulated into PELA microspheres and co-cultured with MC3T3-E1 cells. The expression of BMP-2 protein was detected, and the osteogenic differentiation efficiency was analyzed by alizarin red staining. Results Poly arginine-chitosan / p BMP-2 nanoparticles (81.39 卤22.96 nm) could effectively mediate the transfection of BMP-2 gene into MC3T3-E1 cells. The transfection rate was higher than that in bare plasmid group and chitosan / p BMP-2 group. The particle size was 43.34 卤15.24 渭 m, the entrapment efficiency was 67.49 卤0.85 渭 m, and the cumulative release rate was 56.87 卤5.09 渭 m within 42 days. The amount of BMP-2 secreted by co-cultured with MC3T3-E1 reached the highest level of 64.348 卤2.396)pg/m / L on the 7th day, and still reached 51.763 卤1.789)pg/m / L after 21 days. The number of calcium nodules in alizarin red staining was significantly higher than that in control group. Conclusion Poly (arginine-chitosan) can effectively agglomerate BMP-2 gene into nano-particles, promote cell transfection, and achieve long-term sustained release of PELA microspheres and promote osteogenic differentiation.
【作者单位】：华中科技大学同济医学院附属荆州医院骨科;华中科技大学同济医学院附属荆州医院检验科;
【基金】：荆州市科技计划项目(编号:2012048)
【分类号】：R318.08;R68

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