State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin 130022, PR China.
Biomaterials. 2010 Mar;31(7):1850-7. doi: 10.1016/j.biomaterials.2009.11.027. Epub 2009 Nov 28.
The development of transfection enhancement of liposomes with attributes of high stability and easy handling in gene therapy is challenging. In this study, we report didodecyldimethylammonium bromide (DDAB, a cationic lipid) coated gold nanoparticles (DDAB-AuNPs), which can enhance the transfection efficiency generated by two kinds of commercially available cationic liposomes: Lipotap and DOTAP. It showed that DDAB-AuNPs at the optimal concentrations could produce more than 2 times increase when measuring the number of cells expressed green fluorescent protein and 48-fold increase for luciferase levels after transfection, respectively. The electrophoretic mobility shift assay (EMSA) and confocal laser scanning microscopy (CLSM) experiments showed that more DNA molecules binding to the lipoplexes after adding DDAB-AuNPs. In addition, the flow cytometry (FCM) results indicated that DDAB-AuNPs increased cellular uptake efficiency of DNA molecules, which might account for the enhancement of transfection efficiency. It has also been found that the DDAB-AuNPs could decrease the cytotoxicity of liposomes to the cells.
在基因治疗中，开发具有高稳定性和易于处理特性的脂质体转染增强剂是一项具有挑战性的工作。在这项研究中，我们报告了二油酰基磷脂酰乙醇胺（DOPE，一种阳离子脂质）包被的金纳米粒子（DDAB-AuNPs），它可以增强两种市售阳离子脂质体：Lipotap 和 DOTAP 产生的转染效率。结果表明，在最佳浓度下，当测量表达绿色荧光蛋白的细胞数量时，DDAB-AuNPs 可使转染效率提高 2 倍以上，而当测量荧光素酶水平时，转染效率可提高 48 倍。电泳迁移率变动分析（EMSA）和共聚焦激光扫描显微镜（CLSM）实验表明，加入 DDAB-AuNPs 后，更多的 DNA 分子与脂质体结合。此外，流式细胞术（FCM）结果表明，DDAB-AuNPs 提高了 DNA 分子的细胞摄取效率，这可能是转染效率增强的原因。此外，还发现 DDAB-AuNPs 可以降低脂质体对细胞的细胞毒性。
