| 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.分枝PEI 修饰的PLGA 纳米粒转染DNA 的研究*[J].,2012,12(26):5009-5014 |
| 分枝PEI 修饰的PLGA 纳米粒转染DNA 的研究* |
| Surface Modification of PLGA Nanoparticles with BranchesPolyethyleneimine for DNA Transfection Research* |
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| DOI： |
| 中文关键词: 聚（乳酸-羟基乙酸） 纳米粒 基因转染 |
| 英文关键词: PLGA Nanoparticles Gene transfection |
| 基金项目: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） |
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| 中文摘要: |
| 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（PLGA-
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。 |
| 英文摘要: |
| 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 surface 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. |
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