Preparation, characterization, cytotoxicity and transfection efficiency of poly(DL-lactide-co-glycolide) and poly(DL-lactic acid) cationic nanoparticles for controlled delivery of plasmid DNA
Preparation, characterization, cytotoxicity and transfection efficiency of poly(DL-lactide-co-glycolide) and poly(DL-lactic acid) cationic nanoparticles for controlled delivery of plasmid DNA
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DOI：
10.1016/j.ijpharm.2007.05.023
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发表时间：
2007-10-01
影响因子：
5.8
通讯作者：
Singh, Jagdish
中科院分区：
文献类型：
--
作者：
Basarkar, Ashwin;Devineni, Dilip;Singh, Jagdish
本研究的目的是研究配方参数（即聚合物分子量和均化速度）对阳离子纳米粒子的各种物理化学和生物特性的影响。采用不同分子量的聚(DL-丙交酯-共-乙交酯)(PLGA)和聚(DL-乳酸)(PLA)，通过双乳液溶剂蒸发以两种不同的均质速度制备阳离子纳米颗粒，并在尺寸、表面电荷、形态、负载效率、质粒释放、质粒完整性、细胞毒性和转染效率方面进行表征。阳离子表面活性剂十六烷基三甲基溴化铵（CTAB）用于在纳米颗粒表面提供正电荷。通过孵育将报告质粒gWIZ (TM) Beta-gal负载到纳米粒子的表面。使用较高的均质速度和较低分子量的聚合物导致平均粒径减小、zeta 电位增加、质粒装载效率增加以及突发释放减少。从扫描电子显微照片可以明显看出，纳米颗粒显示出良好的形态。 MTT法体外细胞毒性研究表明毒性较低。纳米颗粒释放的 pDNA 的结构完整性得以保持。与用高分子量聚合物制备的纳米颗粒相比，用低分子量 PLGA 和 PLA 制备的纳米颗粒转染人胚胎肾 (HEK293) 细胞，导致 β-半乳糖苷酶的表达增加。我们的结果表明，PLGA 和 PLA 阳离子纳米颗粒可用于实现 pDNA 的延长释放，并且质粒释放速率和转染效率取决于配方变量。 (C) 2007 Elsevier B.V. 保留所有权利。
The objective of this study was to investigate the effect of formulation parameters (i.e. polymer molecular weight and homogenization speed) on various physicochemical and biological properties of cationic nanoparticles. Cationic nanoparticles were prepared using different molecular weights Of poly(DL-lactide-co-glycolide) (PLGA) and poly(DL-lactic acid) (PLA) by double emulsion solvent evaporation at two different homogenization speeds, and were characterized in terms of size, surface charge, morphology, loading efficiency, plasmid release, plasmid integrity, cytotoxicity, and transfection efficiency. Cationic surfactant, cetyltrimethylammonium bromide (CTAB), was used to provide positive charge on the surface of nanoparticles. Reporter plasmid gWIZ (TM) Beta-gal was loaded on the surface of nanoparticles by incubation. Use of higher homogenization speed and lower molecular weight polymer led to a decrease in mean particle size, increase in zeta potential, increase in plasmid loading efficiency, and a decrease in burst release. The nanoparticles displayed good morphology as evident from scanning electron micrographs. In vitro cytotoxicity study by MTT assay showed a low toxicity. Structural integrity of the pDNA released from nanoparticles was maintained. Transfecting human embryonic kidney (HEK293) cells with nanoparticles prepared from low molecular weight PLGA and PLA resulted in an increased expression of beta-galactosidase as compared to those prepared from high molecular weight polymer. Our results demonstrate that the PLGA and PLA cationic nanoparticles can be used to achieve prolonged release of pDNA, and the plasmid release rate and transfection efficiency are dependent on the formulation variables. (C) 2007 Elsevier B.V. All rights reserved.