分枝PEI修饰的PLGA纳米粒转染DNA的研究  

Surface Modification of PLGA Nanoparticles with Branches Polyethyleneimine for DNA Transfection Research

作　　者：季明辉[1,2] 胡贵方[1] 顾大勇[2] 谢伟东[3] 谈书勤[1] 徐华[4] 欧青叶[4] 

机构地区：[1]南方医科大学公共卫生与热带医学学院,广东广州515000 [2]深圳出入境检验检疫局疾病预防控制研究室,广东深圳518045 [3]清华大学深圳研究生院,广东深圳518045 [4]南通大学公共卫生学院,江苏南通226019

出　　处：《现代生物医学进展》2012年第26期5009-5014,共6页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81072680;30972827);质检总局项目(2010IK212);深港创新圈计划(联合资助)项目(ZYB200907090128A);深圳市科技计划切块项目(HZ0907004)

摘　　要：目的:建立基于聚(乳酸-羟基乙酸)纳米粒(PLGA)载DNA的基因转染体系,比较用空白聚(乳酸-羟基乙酸)纳米粒(PLG-A-E)吸附质粒DNA和用分枝PEI修饰后的PLGA纳米粒(PLGA-BPEI)吸附质粒DNA优缺点。方法:用乳化蒸发法制备纳米粒,对纳米粒进行表征研究,包括包封率、Zeta电位、粒径大小、稳定性,用荧光显微镜观察它们对NIH3T3和HEK293细胞的转染效率,用MTT检测对它们细胞的毒性。结果:制备了两种基于PLGA的纳米粒,PLGA-E和PLGA-BPEI粒径大小为200-270nm,zeta电位为0-30mV,在血清和不同的pH值时两者均较稳定,转染效率PLGA-BPEI较PLGA-E高,且释放时间早,但前者较后者对细胞毒性大。结论:这两种基于PLGA纳米粒均能有效转染质粒DNA,它们存在不同的优缺点,应根据不同需要进行选择。Objective： To build up two PLGA nanoparticle （NP）-based gene delivery systems, namely plasmid DNA （pDNA） encapsulated PLGA NPs （PLGA-E） and surface adsorbed pDNA on PLGA-BPEI NPs （PLGA-BPEI）, with respect to the extent of internalization and intracellular release of pDNA. Methods： Several formulations have also been evaluated systematically for determination of the optimal transfection efficiency. The zeta-potential, particle size measurements and DNase I protection assay established the importance of the BPEI chain length in regulating the effective loading and condensation of pDNA with PLGA-BPEI NPs and pDNA protection ability of PLGA-BPEI NPs. The stability of these formulations was also investigated as a function of serum concentration. In vitro time-dependent gene transfection efficiencies were studied in presence as well as in absence of serum for NIH3T3 and HEK293 cells. The cell viability were also investigated using MTTassay. Results： We build up two PLGA nanoparticle （NP）-based gene delivery systems namely, plasmid DNA （pDNA） encapsulated PLGA NPs （PLGA-E） and surthce adsorbed pDNA on PLGA-BPEI NPs （PLGA-BPEI）, the size of PLGA-E and PLGA-BPEI are between 200 to 270 nm, zeta are from 0 to 30mV, In serum and different pH both are stable, the transfection efficiency of PLGA-BPEI are higher than PLGA-E and release early, but the former have more toxic to the cell. Conclusion： We have developed two kinds of PLGA-based gene delivery systems and investigated their transfection efficiencies. Therefore it can be assumed that this kind of study should provide a strong basis in choosing appropriate delivery system for specific gene expression.

关 键 词：聚(乳酸-羟基乙酸) 纳米粒 基因转染 

分 类 号：Q782[生物学—分子生物学] Q75