Centre de Biophysique Moléculaire, CNRS UPR4301, Inserm and University of Orléans, F-45071 Orléans cedex 02, France.
Laboratoire de Physique des Solides, CNRS, Univ. Paris-Sud, Université Paris-Saclay, 91405 Orsay Cedex, France.
Int J Pharm. 2019 Mar 25;559:58-67. doi: 10.1016/j.ijpharm.2019.01.003. Epub 2019 Jan 14.
Linear Polyethylenimine (lPEI) is an efficient cationic polymer for transfecting cells, both in vitro and in vivo, but poses concerns regarding cytotoxicity. Histidinylated lPEI (His-lPEI) exhibits also high transfection efficiency but lower cytotoxicity than lPEI. For the first time, we tested polyfection efficiency of polyplexes comprising both lPEI and His-lPEI. A series of pDNA polyplexes was prepared with mixtures of lPEI and His-lPEI and the amount of each polymer within His-lPEI/lPEI polyplexes was determined by flow cytometry. We show that His-lPEI/lPEI polyplexes exhibit properties similar to lPEI polyplexes in terms of size, morphology, assembly with pDNA, and polyplex stability while His-lPEI/lPEI polyplexes exhibit properties similar to His-lPEI polyplexes in terms of buffering capacity. Compared to polyplexes consisting only of lPEI or His-lPEI, the transfection profile reveals that His-lPEI/lPEI polyplexes containing 30% to 57% lPEI strongly increase polyfection efficiency of NIH3T3 fibroblasts and murine, as well as human skeletal muscle cell lines without cytotoxicity. Importantly, improved transfection of human dystrophin deficient skeletal muscle cell lines was obtained. These results indicate that His-lPEI/lPEI polyplexes are an improved non-viral vector for efficient transfection of dystrophin deficient skeletal muscle cell lines that should be tested on animals.
线性聚乙烯亚胺（lPEI）是一种有效的阳离子聚合物，可用于体外和体内转染细胞，但存在细胞毒性问题。组氨酸化 lPEI（His-lPEI）也表现出高转染效率，但比 lPEI 的细胞毒性低。我们首次测试了由 lPEI 和 His-lPEI 组成的多聚体的多转染效率。通过流式细胞术确定了 His-lPEI/lPEI 多聚体中每种聚合物的含量，用 lPEI 和 His-lPEI 的混合物制备了一系列 pDNA 多聚体。我们表明，His-lPEI/lPEI 多聚体在大小、形态、与 pDNA 的组装以及多聚体稳定性方面与 lPEI 多聚体具有相似的性质，而在缓冲能力方面与 His-lPEI 多聚体具有相似的性质。与仅由 lPEI 或 His-lPEI 组成的多聚体相比，转染谱表明，含有 30%至 57%lPEI 的 His-lPEI/lPEI 多聚体强烈增加了 NIH3T3 成纤维细胞和鼠以及人骨骼肌细胞系的多转染效率，而没有细胞毒性。重要的是，获得了人肌营养不良缺陷骨骼肌细胞系的转染改善。这些结果表明，His-lPEI/lPEI 多聚体是一种改良的非病毒载体，可有效转染肌营养不良缺陷的骨骼肌细胞系，应在动物身上进行测试。
