Nancy University, Department of Pharmaceutical Technology and Biopharmacy, EA 3452, Nancy, France.
Int J Pharm. 2011 Jan 17;403(1-2):276-84. doi: 10.1016/j.ijpharm.2010.10.040. Epub 2010 Oct 29.
The main objective of this study was to prepare two types of nanoparticles with poly(d,l-lactide-co-glycolide) (PLGA) and polyethylenimine (PEI) polymers. Plasmid DNA (pDNA) was adsorbed either on PLGA/PEI nanoparticles, or as PEI/DNA complex onto the surface of PLGA nanoparticles. Both types of nanoparticles were prepared by the double emulsion method. The nanoparticles were characterized by their size, zeta potential and pDNA or PEI/DNA complex adsorption. The PEI/DNA complex adsorption was confirmed with ethidium bromide assay. pDNA adsorption onto PLGA/PEI nanoparticles (PLGA/PEI-DNA) was studied by electrophoresis on agarose gel. Cytotoxicity and transfection efficiency of both types of nanoparticle and PEI/DNA complexes formulations were studied in head and neck squamous carcinoma cell line (FaDu). To improve endosomal release, photochemical internalization (PCI) was used. The zeta potential increased when the PEI/DNA complex adsorbed onto PLGA nanoparticles (PLGA-PEI/DNA). Optimal pDNA adsorption efficiency was achieved for nitrogen/phosphorous ratio≥20/1. In vitro transfection and cells viability on FaDu cells with or without PCI were found to be variable depending on the type and concentration of nanoparticles. The results showed that transfection efficiency for PLGA/PEI-DNA or PLGA-PEI/DNA nanoparticles ranged between 2 and 80%, respectively. PCI was found to slightly improve the transfection efficiency for all formulations.
本研究的主要目的是制备两种类型的纳米粒子,分别使用聚(丙交酯-共-乙交酯)(PLGA)和聚乙烯亚胺(PEI)聚合物。将质粒 DNA(pDNA)吸附到 PLGA/PEI 纳米粒子上,或者将其作为 PEI/DNA 复合物吸附到 PLGA 纳米粒子的表面。这两种类型的纳米粒子均通过双重乳液法制备。通过测量粒径、zeta 电位以及 pDNA 或 PEI/DNA 复合物的吸附量对纳米粒子进行了表征。通过溴化乙锭实验证实了 PEI/DNA 复合物的吸附。通过琼脂糖凝胶电泳研究了 pDNA 在 PLGA/PEI 纳米粒子(PLGA/PEI-DNA)上的吸附。在头颈部鳞状细胞癌细胞系(FaDu)中研究了两种类型的纳米粒子和 PEI/DNA 复合物制剂的细胞毒性和转染效率。为了提高内涵体释放,使用了光化学内化(PCI)。当 PEI/DNA 复合物吸附到 PLGA 纳米粒子上时,zeta 电位增加(PLGA-PEI/DNA)。当氮/磷比≥20/1 时,可实现最佳的 pDNA 吸附效率。在有无 PCI 的情况下,FaDu 细胞中的体外转染和细胞活力因纳米粒子的类型和浓度而异。结果表明,PLGA/PEI-DNA 或 PLGA-PEI/DNA 纳米粒子的转染效率分别在 2%至 80%之间。发现 PCI 略微提高了所有制剂的转染效率。