Pharmaceutical Development of Green Innovations Group, Faculty of Pharmacy, Silpakorn University, Nakhon Pathom, 73000, Thailand.
AAPS PharmSciTech. 2012 Dec;13(4):1302-8. doi: 10.1208/s12249-012-9857-5. Epub 2012 Sep 25.
The aim of this study was to investigate the transfection efficiency of cationic liposomes formulated with phosphatidylcholine (PC) and novel synthesized diethanolamine-based cationic lipids at a molar ratio of 5:1 in comparison with Lipofectamine™ 2000. Factors affecting transfection efficiency and cell viability, including the chemical structure of the cationic lipids, such as different amine head group (diamine and polyamine; and non-spermine and spermine) and acyl chain lengths (C14, C16, and C18) and the weight ratio of liposomes to DNA were evaluated on a human cervical carcinoma cell line (HeLa cells) using the pDNA encoding green fluorescent protein (pEGFP-C2). Characterizations of these lipoplexes in terms of size and charge measurement and agarose gel electrophoresis were performed. The results from this study revealed that almost no transfection was observed in the liposome formulations composed of cationic lipids with a non-spermine head group. In addition, the transfection efficiency of these cationic liposomes was in the following order: spermine-C14 > spermine-C16 > spermine-C18. The highest transfection efficiency was observed in the formulation of spermine-C14 liposomes at a weight ratio of 25; furthermore, this formulation was safe for use in vitro. In conclusion, cationic liposomes containing spermine head groups demonstrated promising potential as gene carriers.
本研究旨在比较以 5:1 摩尔比与 Lipofectamine™ 2000 形成的阳离子脂质体与磷脂酰胆碱（PC）和新型合成的二乙醇胺基阳离子脂质体的转染效率。影响转染效率和细胞活力的因素，包括阳离子脂质体的化学结构，如不同的胺头基（二胺和多胺；非亚精胺和精胺）和酰基链长度（C14、C16 和 C18）以及脂质体与 DNA 的重量比，在人宫颈癌细胞系（HeLa 细胞）上使用编码绿色荧光蛋白的 pDNA（pEGFP-C2）进行评估。这些脂质体在大小和电荷测量以及琼脂糖凝胶电泳方面的特性进行了表征。研究结果表明，由非精胺头基组成的阳离子脂质体的脂质体制剂几乎没有转染。此外，这些阳离子脂质体的转染效率依次为：精胺-C14＞精胺-C16＞精胺-C18。在重量比为 25 的精胺-C14 脂质体配方中观察到最高的转染效率，此外，该配方在体外使用是安全的。总之，含有精胺头基的阳离子脂质体作为基因载体具有很大的潜力。
