一种用于高效非病毒基因传递的新型聚合物-脂质杂化纳米颗粒。
A novel polymer-lipid hybrid nanoparticle for efficient nonviral gene delivery.
机构信息
Institutes of Biomedical Sciences of Fudan University, Shanghai 200032, China.
出版信息
Acta Pharmacol Sin. 2010 Apr;31(4):509-14. doi: 10.1038/aps.2010.15. Epub 2010 Mar 29.
AIM
To develop a novel non-viral vector with high transfection efficiency and low cytotoxicity.
METHODS
Poly (ethylene glycol)-distearoylphosphatidylethanolamine (PEG-DSPE) was incorporated into polymer-lipid hybrid nanoparticles (PLN) to construct a PEG-DSPE modified long circulating PLN (L-PLN). The L-PLN was prepared by the emulsifying-solvent evaporation method, L-PLN and L-PLN/DNA complexes were characterized. Both HEK293 and MDA-MB-231 cells transfected by L-PLN/DNA complexes were observed under a fluorescence microscope. The transfection efficiency of the complexes to HEK293 cells was further evaluated by flow cytometry.
RESULTS
The GFP fluorescence intensity in HEK293 cells transfected by the L-PLN/DNA complexes (N/P=10) was about 37.2%, which was higher than those transfected by PLN alone or commercial Lipofectamine 2000. The L-PLN exhibited minimal toxicity at a low N/P ratio compared with other vectors.
CONCLUSION
L-PLN as a novel gene delivery system, has higher transfection efficiency and acceptable cytotoxicity compared to the corresponding PLN, which is beneficial for the development of non-viral gene transfer vectors and may offer an alternative strategy for the future gene therapy.
目的
开发一种具有高效转染效率和低细胞毒性的新型非病毒载体。
方法
聚乙二醇-二硬脂酰磷脂酰乙醇胺（PEG-DSPE）被整合到聚合物脂质混合纳米粒（PLN）中，构建了一种聚乙二醇-二硬脂酰磷脂酰乙醇胺修饰的长循环 PLN（L-PLN）。L-PLN 通过乳化-溶剂蒸发法制备，对 L-PLN 和 L-PLN/DNA 复合物进行了表征。通过荧光显微镜观察 L-PLN/DNA 复合物转染的 HEK293 和 MDA-MB-231 细胞。通过流式细胞术进一步评估了复合物对 HEK293 细胞的转染效率。
结果
L-PLN/DNA 复合物（N/P=10）转染的 HEK293 细胞中的 GFP 荧光强度约为 37.2%，高于单独转染 PLN 或商业 Lipofectamine 2000 的细胞。与其他载体相比，L-PLN 在低 N/P 比时表现出最小的毒性。
结论
与相应的 PLN 相比，L-PLN 作为一种新型基因传递系统具有更高的转染效率和可接受的细胞毒性，有利于非病毒基因传递载体的开发，并可能为未来的基因治疗提供一种替代策略。
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