Effect of acyl chain length on transfection efficiency and toxicity of polyethylenimine
Effect of acyl chain length on transfection efficiency and toxicity of polyethylenimine
复制标题
DOI：
10.1016/j.ijpharm.2009.05.052
复制
发表时间：
2009-08-13
影响因子：
5.8
通讯作者：
Williams, Adrian C.
中科院分区：
文献类型：
--
作者：
Aravindan, Latha;Bicknell, Katrina A.;Williams, Adrian C.
聚乙烯亚胺（PEI）是一种高效的非病毒基因载体，因为它具有高缓冲能力和DNA浓缩能力。在我们的研究中，使用乙酸酐或丙酸酐酰化聚合物主链上的氨基，以改变质子化行为和聚合物的亲水/疏水平衡。酰化伯胺的浓度用三硝基苯磺酸测定法测定。结果表明，与PEI相比，我们的改性聚合物在溶液中具有较低的缓冲能力。将聚合物与编码增强型绿色荧光蛋白的质粒以三种不同的比例（1：1、1：2和1：10 w/w DNA与聚合物）复合以形成聚合复合物，并在HEK 293细胞中评价它们的毒性和转染效率。酰化减少了聚合物上伯胺的数量和表面电荷，改善了血液相容性并降低了细胞毒性。氨基浓度的降低有助于优化DNA压缩并促进细胞中的多聚复合物解离，与母体聚合物相比，这增加了改性聚合物的转染效率。相对于PEI，缓冲能力大于50%且小于80%的聚合物显示出比PEI更高的转染效率。丙酸酐修饰的聚合物与DNA具有适当的相互作用，其提供DNA压缩和聚合复合物解离。这些系统更好地与细胞膜相互作用，因为它们的亲脂性略高，并形成比乙酸酐改性聚合物的复合物细胞毒性小的复合物。在测试的载体中，1：2 w/w DNA：聚合物组合物中的1：0.3mol/mol PEI：丙酸酐提供了具有改善的转染效率和降低的细胞毒性的最佳转染系统。(C)2009 Elsevier B. V.保留所有权利。
Polyethylenimine (PEI) is an efficient nonviral gene delivery vector because of its high buffering capacity and DNA condensation ability. In our study, the amino groups on the polymeric backbone were acylated using acetic or propionic anhydride to alter the protonation behaviour and the hydrophilic/hydrophobic balance of the polymer. The concentration of acylated primary amines was determined using trinitrobenzene sulphonic acid assay. Results showed that our modified polymers had lower buffering capacities in solutions compared to PEI. The polymers were complexed with plasmid encoding enhanced green fluorescent protein at three different ratios (1: 1, 1:2 and 1: 10 w/w DNA to polymer) to form polyplexes and their toxicities and transfection efficiencies were evaluated in HEK 293 cells. Acylation reduced the number of primary amines on the polymer and the surface charge, improving haemocompatibility and reducing cytotoxicity. The reduction in the concentration of amino groups helped to optimise DNA compaction and facilitated polyplex dissociation in the cell, which increased transfection efficiency of the modified polymers compared to the parent polymer. Polymers with buffering capacities greater than 50% and less than 80% relative to PEI, showed higher transfection efficiencies than PEI. The propionic anhydride modified polymers had appropriate interactions with DNA which provided both DNA compaction and polyplex dissociation. These systems interacted better with the cell membrane because of their slightly higher lipophilicity and formed polyplexes which were less cytotoxic than polyplexes of acetic anhydride modified polymers. Among the vectors tested, 1:0.3 mol/mol PEI: propionic anhydride in a 1:2 w/w DNA:polymer composition provided the best transfection system with improved transfection efficiency and reduced cytotoxicity. (C) 2009 Elsevier B.V. All rights reserved.