本文選題：非病毒基因載體 + 多聚精氨酸修飾的殼聚糖�。� 參考：《廣東醫(yī)學(xué)》2017年S1期

【摘要】：目的構(gòu)建多聚精氨酸-殼聚糖/p BMP-2納米粒,探討包封多聚精氨酸-殼聚糖/p BMP-2納米粒的PELA微球的體外成骨誘導(dǎo)效能。方法構(gòu)建多聚精氨酸修飾的殼聚糖多聚物,并與BMP-2質(zhì)粒基因凝聚成納米粒子,檢測其表征。熒光顯微鏡及Western blot分析其體外細(xì)胞轉(zhuǎn)染效能。進(jìn)而,將納米粒子包入PELA微球,與MC3T3-E1細(xì)胞共培養(yǎng),檢測BMP-2蛋白表達(dá),茜素紅染色分析其成骨誘導(dǎo)分化效能。結(jié)果多聚精氨酸-殼聚糖/p BMP-2納米粒粒徑(81.39±22.96)nm,能有效介導(dǎo)BMP-2基因轉(zhuǎn)染MC3T3-E1細(xì)胞,轉(zhuǎn)染率高于裸質(zhì)粒組及殼聚糖/p BMP-2納米粒組(P0.05)。PELA微球呈球形,粒徑為(43.34±15.24)μm,包封率達(dá)(67.49±0.85)%,42 d內(nèi)累積釋藥(56.87±5.09)%,與MC3T3-E1共培養(yǎng),第7天BMP-2分泌量達(dá)最高(64.348±2.396)pg/m L,21 d后分泌量仍能達(dá)到(51.763±1.789)pg/m L。茜素紅染色鈣結(jié)節(jié)形成數(shù)量明顯高于對照組。結(jié)論多聚精氨酸-殼聚糖能有效凝聚BMP-2基因成納米級粒子,促進(jìn)細(xì)胞轉(zhuǎn)染,并可通過PELA微球的包封實現(xiàn)納米粒的長效緩釋,促進(jìn)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 鹵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 鹵15.24 渭 m, the entrapment efficiency was 67.49 鹵0.85 渭 m, and the cumulative release rate was 56.87 鹵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 鹵2.396)pg/m / L on the 7th day, and still reached 51.763 鹵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.
【作者單位】： 華中科技大學(xué)同濟(jì)醫(yī)學(xué)院附屬荊州醫(yī)院骨科;華中科技大學(xué)同濟(jì)醫(yī)學(xué)院附屬荊州醫(yī)院檢驗科;
【基金】：荊州市科技計劃項目(編號:2012048)
【分類號】：R318.08;R68

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