聚氰基丙烯酸正丁酯纳米颗粒的表面修饰和转染

Surface modification and transfection of polybutyl cyanoacrylate nanoparticles

背景:聚氰基丙烯酸烷基酯在人体内极易生物降解,且对许多组织具有生物相容性,近年来一直受到国内外医药界的广泛关注。目的:比较不同表面修饰的聚氰基丙烯酸正丁酯纳米颗粒的物理化学性质,并考察它们体外基因转染活性。设计、时间及地点:对比观察实验,于2007-09/12在中南大学卫生部纳米生物技术重点实验室完成。材料:采用乳化聚合的方法制备表面分别修饰壳聚糖、葡聚糖和氨基化的聚乙二醇的聚氰基丙烯酸正丁酯纳米颗粒。方法:透射电镜观察粒径大小、纳米粒形态;Zetasizer Nanosystem电位分析仪测定Zeta电位;MTT法检测纳米粒的细胞毒性;用流式细胞仪考察纳米粒的体外转染效率。主要观察指标:纳米粒的表面电位、粒径、细胞毒性及其转染效率。结果:成功制备了粒径约200nm左右,表面电位分别为25.53,-17.42和0.2935mV的壳聚糖、葡聚糖和氨基化的聚乙二醇修饰的聚氰基丙烯酸正丁酯纳米颗粒,且0.1g/L的质量浓度对HepG2细胞无明显毒性,带正电的壳聚糖修饰的聚氰基丙烯酸正丁酯纳米颗粒的转染效率明显高于葡聚糖和氨基化的聚乙二醇修饰的纳米粒。结论:聚氰基丙烯酸正丁酯纳米颗粒经表面修饰不同物质后可以带上正电、负电及中性电荷,而壳聚糖修饰的纳米粒能有效地将基因递送到细胞内,并且报告基因能在细胞内表达。因此,可用做基因递送的载体。

BACKGROUND： During the last decade, poly （alkyl cyanoacryiates） nanoparticles have been widely studied as characterizing well biodegradation and biocompatibility. OBJECTIVE： Compared with chemical and physical characteristics of different kinds of polybutyl cyanoacrylate nanoparticles （PBCA-NPs）, to investigate the transfection efficiency of PBCA-NPs in vitro. DESIGN, TIME AND SETTING： A contrast study was performed at Key Laboratory of Nanobiotechnology, Ministry of Public Health, Central South University between September and December 2007. MATERIALS： PBCA-NPs were modified by chitosan, dextran and amino-methoxyl poly （ethylene glycol） （mPEG-NH2） using latex polymerization. METHODS： Transmission electron microscope was used to observe particle size and morphology, Zetasizer Nano system was used to measure Zeta potential, MTT assay was used to detect cytotoxicity, and flow cytometry was used to determine transfection efficiency in vitro. MAIN OUTCOME MEASURES： Surface potential, diameter, cytotoxicity, and transfection efficiency of PBCA-NPs. RESULTS： The particle diameter was about 200 nm. The surface potentials were 25.53, 17.42, 0.293 5 mV respectively modified by chitosan, dextran and amino-methoxyi poly （ethylene glycol） （mPEG-NH2）. NPs presented no toxicity to HepG2 cells at the concentration of 0.1 g/L. And the transfection efficiency of chitosan-coated PBCA-NPs was higher than that of dextran and mPEG-NH2-coated PBCA-NPs. CONCLUSION： We can prepare different kinds of PBCA nanoparticies with positive, negative and neutral zeta potential. The chitosan-coated PBCA-NPs can delivery and express the gene into cells. Therefore, it can be worth to study as gene medicine carriers.