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High Molecular Weight Poly(ethylenimine)-Based Water-Soluble Lipopolymer for Transfection of Cancer Cells.
Macromolecular Bioscience ( IF 4.6 ) Pub Date : 2020-05-25 , DOI: 10.1002/mabi.202000040
Heba Alaa Hosiny Abd Elhameed 1 , Ditta Ungor 2 , Nóra Igaz 3 , Mohana Krishna Gopisetty 3 , Mónika Kiricsi 3 , Edit Csapó 2, 4 , Béla Gyurcsik 1
Affiliation  

Over the past decade, search for novel materials for nucleic acid delivery has prompted a special interest in polymeric nanoparticles (NPs). In this study, the biological applicability of a water‐soluble cationic lipopolymer (WSLP) obtained by the modification of high molecular weight branched poly(ethylenimine) (PEI) with cholesteryl chloroformate is characterized and assessed for better cellular membrane permeability. To test the delivery efficiency of the produced lipopolymer, plasmid DNA (pDNA) encoding the enhanced green fluorescent protein and WSLP are mixed at different charge ratios. WSLP and WSLP/pDNA complexes are characterized by dynamic and static light scattering, particle charge detection, scanning electron microscopy, and transmission electron microscopy. The pDNA loading of WSLP is also verified by agarose gel electrophoresis. Cytotoxicity of PEI, WSLP, and of WSLP/pDNA is evaluated on human A549 and HeLa cells. A remarkable dependence of the toxicity on the dose, cholesterylation, and charge ratio is detected. Transfection is monitored by flow cytometry and by fluorescence microscopy. Importantly, cholesterylation decreases the toxicity of the polymer, while promoting high transfection efficiency in both cell lines. This work indicates a possible optimization mode of the high molecular weight PEI‐based WSLP rendering it a promising candidate for gene delivery.

中文翻译:

高分子量基于聚乙烯亚胺的水溶性脂聚合物,用于癌细胞的转染。

在过去的十年中,寻找用于核酸递送的新型材料引起了对聚合物纳米颗粒(NP)的特别关注。在这项研究中,通过对高分子量支链聚(乙烯亚胺)(PEI)改性的氯甲酸酯氯甲酸酯改性而获得的水溶性阳离子脂质聚合物(WSLP)的生物学适用性进行了表征,并评估了其对细胞膜的通透性。为了测试产生的脂聚合物的递送效率,将编码增强的绿色荧光蛋白的质粒DNA(pDNA)和WSLP以不同的电荷比率混合。WSLP和WSLP / pDNA复合物的特征在于动态和静态光散射,粒子电荷检测,扫描电子显微镜和透射电子显微镜。WSLP的pDNA负载量也通过琼脂糖凝胶电泳验证。在人A549和HeLa细胞上评估PEI,WSLP和WSLP / pDNA的细胞毒性。检测到毒性对剂量,胆甾醇化和电荷比的显着依赖性。通过流式细胞仪和荧光显微镜监测转染。重要的是,胆固醇酰化降低了聚合物的毒性,同时在两种细胞系中都促进了高转染效率。这项工作表明了基于高分子量PEI的WSLP的可能的优化模式,使其成为有希望的基因传递候选者。胆甾醇化降低了聚合物的毒性,同时在两种细胞系中均提高了转染效率。这项工作表明了基于高分子量PEI的WSLP的一种可能的优化模式,使其成为基因传递的有希望的候选者。胆甾醇化降低了聚合物的毒性,同时在两种细胞系中均提高了转染效率。这项工作表明了基于高分子量PEI的WSLP的可能的优化模式,使其成为有希望的基因传递候选者。
更新日期:2020-05-25
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