PEG化壳聚糖质粒纳米粒的制备及其转染的研究

目的:制备壳聚糖纳米粒并构建聚乙二醇(PEG)化壳聚糖质粒纳米粒,研究其对大鼠主动脉内皮细胞的转染能力及细胞毒性.方法:采用离子交联法制备壳聚糖纳米粒,应用喷金扫描电子显微镜检测壳聚糖纳米粒粒径的分布与形态;通过静电吸附作用连接上pGenesil-1质粒(报告基岗);对壳聚糖质粒纳米粒进行PEG化的修饰;应用PEG化壳聚糖质粒纳米粒转染大鼠主动脉内皮细胞;采用噻唑蓝(MTT)法测定壳聚糖纳米粒对细胞的毒性作用.结果:喷金扫描电镜检测显示壳聚糖纳米粒呈均匀分散的球形颗粒,平均直径为5 nm;PEG化壳聚糖质粒纳米粒能转染大鼠主动脉内皮细胞;MTT结果显示壳聚糖纳米粒对细胞无毒性作用;壳聚糖质粒纳米粒对内皮细胞的转染效率为26%,PEG修饰壳聚糖质粒纳米粒转染细胞,转染率为63.4%.结论:对壳聚糖质粒纳米粒进行化学修饰不仅能提高其转染效率,且对细胞无毒性作用.

Preparation and transfection of PEG chitosan nanoparticles as gene vector

Objective To prepare chitosan/plamid nanoparticle (CS-NP) and assess its cytotoxicity and transgenic efficacy to endothelial cells from rat aorta. Methods Ion exchange method was used to prepare CS-NP. Diametric distribution and form of CS-NP were tested by puff gold scanning electron microscope (SEM). Con-neetion of pGenesil-1 with CS-NP was accomplished by electrostatic adsorption and CS-NP was chemically modi-fied by PEG. The PEG CS-NP was then used to transfect the endothelial cell from rat aorta. The toxic effects to endothelial cell were detected by MTT method. Results The results of SEM showed that CS-NP were of spheri-cal shape and well-dispersed, with average diameter of 5 nm. MTT test showed that PEG CS-NP was nontoxic to the aortic vascular cells and mediated a significantly higher transgene expression(63.4%) than CS-NP(26%). Conclusion The PEG CS-NP not only can improve the transfection efficiency, but also have no toxic effects on aortic vascular cells.