PEG化壳聚糖质粒纳米粒的制备及其转染的研究  

Preparation and transfection of PEG chitosan nanoparticles as gene vector

作　　者：赵惠萍[1] 边云飞[2] 高奋[2] 何军华[2] 马瑜[1] 刘改珍[2] 肖传实[2] 

机构地区：[1]山西医科大学第二临床医学院,太原030001 [2]山西医科大学第二医院心内科,太原030001

出　　处：《国际生物医学工程杂志》2009年第3期146-148,I0001,共4页International Journal of Biomedical Engineering

基　　金：山西省回国留学人员科研资助项目（2007重点项目-5）

摘　　要：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（报告基因）；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5nm；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。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）. Connection of pGenesil-1 with CS-NP was accomplished by electrostatic adsorption and CS-NP was chemically modified by PEG. The PEG CS-NP was then used to transfeet 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.

关 键 词：壳聚糖 纳米粒 质粒 结合 转染 

分 类 号：TB38[一般工业技术—材料科学与工程] R[医药卫生]