Center of Drug Metabolism and Pharmacokinetics, College of Pharmacy, China Pharmaceutical University, Nanjing 210009, PR China.
Colloids Surf B Biointerfaces. 2013 Jan 1;101:6-13. doi: 10.1016/j.colsurfb.2012.05.032. Epub 2012 Jun 9.
The ability of cationic liposomes composed of DC-Chol and cholesterol to carry pDNA into 293 T cells was investigated. Lipoplexes formed between DC-Chol/cholesterol liposomes and pDNA (encoding green fluorescent protein, GFP) were analyzed in terms of morphology observation, turbidity determination, particle size and zeta potential measurement, differential scanning calorimetry (DSC), gel retardation assay, cytotoxicity analysis in 293 T cells and transfection efficiency. The results showed that liposome preparation at or above 66.7 mol% cholesterol in formulation exhibited a calorimetric transition caused by anhydrous cholesterol domain at about 41°C. In comparison with the control, DOPE-containing liposomes, DC-Chol/cholesterol carriers showed more stable particle size, lower turbidity, higher activity for transfecting cells in the presence of high concentration serum (50% FBS), primarily due to the neutral domain formation by increasing mole ratios of cholesterol in formulation, as well as relatively lower cytotoxicity. Based on the results, it is suggested that incorporating high contents of cholesterol might be a potentially applicable method for various kinds of cationic lipids to obtain the gene carriers with high capability for in vivo transfection.
研究了由 DC-Chol 和胆固醇组成的阳离子脂质体将 pDNA 携带进入 293T 细胞的能力。用 DC-Chol/胆固醇脂质体和 pDNA（编码绿色荧光蛋白，GFP）形成的脂质体复合物，从形态观察、浊度测定、粒径和 Zeta 电位测量、差示扫描量热法（DSC）、凝胶阻滞分析、293T 细胞的细胞毒性分析和转染效率等方面进行了分析。结果表明，制剂中胆固醇摩尔分数等于或高于 66.7%的脂质体制备物在约 41°C 处表现出由无水胆固醇域引起的量热转变。与对照物 DOPE 脂质体相比，DC-Chol/胆固醇载体表现出更稳定的粒径、更低的浊度、在高浓度血清（50% FBS）存在下更高的转染细胞活性，这主要归因于通过增加制剂中胆固醇的摩尔比形成中性域，以及相对较低的细胞毒性。基于这些结果，建议掺入高含量的胆固醇可能是一种适用于各种阳离子脂质体的潜在方法，以获得具有高体内转染能力的基因载体。
