Department of Pharmaceutical Sciences, Beijing Institute of Radiation Medicine, Beijing 100850, China; Anhui Medical University, Hefei 230001, China.
Department of Pharmaceutical Sciences, Beijing Institute of Radiation Medicine, Beijing 100850, China; Anhui Medical University, Hefei 230001, China.
Colloids Surf B Biointerfaces. 2015 Feb 1;126:257-64. doi: 10.1016/j.colsurfb.2014.12.032. Epub 2014 Dec 31.
Nonviral gene vectors are a hot topic for gene delivery. High cost and low transfection efficiency hinder the application of them. The aim of this study was to find out the optimal gene vectors with lower cost and more effective gene delivery than commonly used gene vectors. A cheap cationic lipid, didodecyldimethylammonium bromide (DDAB) was the basic component and the other components included oleic acid (OA), cholesterol (Chol), cholesteryl succinyl poly(ethylene glycol) 1500 (CHS-PEG), poly(D,L-lactide-co-glycolide)-methoxy-poly(ethylene glycol) (PLGA-PEG). The combinations of DDAB/OA/Chol, DDAB/OA/CHS-PEG and DDAB/PLGA-PEG were adopted to prepare the nanoassemblies named CNA, CPNA and PPNA, respectively. The optimal component ratios were screened out according to their Langmuir monolayer behavior. The optimal preparation method of nanoassemblies involved firstly compressing DNA or siRNA with the cationic lipid (DDAB) and secondly being coated with the helper lipids (OA and CHS-PEG) or the helper polymer (PLGA-PEG). The complexes of genes and cationic lipids were encapsulated into the core of CPNA and PPNA. The optimal gene vectors (CPNA and PPNA) with small sizes, low negative surface charges and non-exposure of cationic lipids were achieved. They had the advantages of no cytotoxicity, high transfection efficiency and low cost. More importantly, CPNA and PPNA were not sensitive to serum and showed the similar or higher transfection efficiency of pDNA and siRNA compared to Lipofectamine 2000. CPNA could mainly enter cell plasma based on endocytosis. The rational design method is useful for the design and optimization of DDAB-based gene carriers and other cationic lipid-based carriers.
非病毒基因载体是基因递送的热门话题。高成本和低转染效率阻碍了它们的应用。本研究旨在寻找成本更低、转染效率更高的基因载体，优于常用的基因载体。廉价的阳离子脂质体二油酰基二甲基溴化铵（DDAB）是基本成分，其他成分包括油酸（OA）、胆固醇（Chol）、琥珀酰化聚乙二醇 1500 胆固醇（CHS-PEG）、聚（D，L-乳酸-共-乙醇酸）-甲氧基聚乙二醇（PLGA-PEG）。采用 DDAB/OA/Chol、DDAB/OA/CHS-PEG 和 DDAB/PLGA-PEG 组合分别制备纳米组装体，命名为 CNA、CPNA 和 PPNA。根据其 Langmuir 单层行为筛选出最佳的组分比。纳米组装体的最佳制备方法包括首先用阳离子脂质体（DDAB）压缩 DNA 或 siRNA，其次用辅助脂质（OA 和 CHS-PEG）或辅助聚合物（PLGA-PEG）包被。基因和阳离子脂质复合物被包裹在 CPNA 和 PPNA 的核心中。最佳基因载体（CPNA 和 PPNA）具有较小的尺寸、较低的负表面电荷和非暴露的阳离子脂质，具有无细胞毒性、高转染效率和低成本的优点。更重要的是，CPNA 和 PPNA 对血清不敏感，与 Lipofectamine 2000 相比，对 pDNA 和 siRNA 的转染效率相似或更高。CPNA 主要通过内吞作用进入细胞浆。这种合理的设计方法有助于基于 DDAB 的基因载体和其他阳离子脂质载体的设计和优化。
