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改良的纳米沉淀法制备载 DNA 的 PLGA 纳米粒。

Modified nanoprecipitation method to fabricate DNA-loaded PLGA nanoparticles.

机构信息

Department of Pharmacy, Shandong Medical College, Ji'nan, Shandong Province, PR China.

出版信息

Drug Dev Ind Pharm. 2009 Nov;35(11):1375-83. doi: 10.3109/03639040902939221.

Abstract

OBJECTIVE

The objective of this study was to formulate DNA-loaded poly(d,l-lactide-co-glycotide) (PLGA) nanoparticles by a modified nanoprecipitation method.

METHODS

DNA-loaded PLGA nanoparticles were prepared by the modified nanoprecipitation method and the double emulsion/solvent evaporation method. The characterizations of DNA-loaded nanoparticles such as entrapment efficiency, morphology, particle size, zeta potential, structural integrity of the loaded DNA, and stability of the loaded DNA in PLGA nanoparticles against DNase I, in vitro release, cell viability and in vitro transfection capability were investigated.

RESULTS

The resulted PLGA nanoparticles by the modified nanoprecipitation method had uniform spherical shape, narrow size distribution with average particles size near 200 nm, negative zeta potential of -12.6 mV at pH 7.4, and a sustained-release property in vitro. Plasmid DNA could be efficiently encapsulated into PLGA nanoparticles (> 95%) without affecting its intact conformation using this modified nanoprecipitation method, which was superior to the double emulsion/solvent evaporation method. The PLGA nanoparticles were much safer to A549 cell compared to commercial Lipofectamine 2000 and could successfully transfer plasmid-enhanced green fluorescent protein into A549 cells.

CONCLUSION

In conclusion, the modified nanoprecipitation method could be applied as an efficient way to fabricate DNA-loaded PLGA nanoparticles instead of the conventional double emulsion/solvent evaporation method.

摘要

目的

本研究的目的是通过改良的沉淀法制备载 DNA 的聚(丙交酯-共-乙交酯)(PLGA)纳米粒。

方法

通过改良的沉淀法和双乳液/溶剂蒸发法制备载 DNA 的 PLGA 纳米粒。考察了载 DNA 的纳米粒的特性,如包封效率、形态、粒径、Zeta 电位、载 DNA 的结构完整性以及载 DNA 在 PLGA 纳米粒中对 DNase I 的稳定性、体外释放、细胞活力和体外转染能力。

结果

改良沉淀法制备的 PLGA 纳米粒呈均匀的球形,粒径分布较窄,平均粒径接近 200nm,在 pH7.4 时 Zeta 电位为-12.6mV,具有体外缓释性能。使用改良的沉淀法可以有效地将质粒 DNA 包封到 PLGA 纳米粒中(>95%),而不会影响其完整构象,优于双乳液/溶剂蒸发法。与商业的 Lipofectamine 2000 相比,PLGA 纳米粒对 A549 细胞的安全性更高,并且可以成功地将质粒增强型绿色荧光蛋白转染到 A549 细胞中。

结论

总之,改良的沉淀法可以作为制备载 DNA 的 PLGA 纳米粒的有效方法,替代传统的双乳液/溶剂蒸发法。

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