Guangdong Provincial Key Laboratory of Chinese Medicine Pharmaceutics, School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, PR China.
ACS Biomater Sci Eng. 2021 Nov 8;7(11):5048-5063. doi: 10.1021/acsbiomaterials.1c00919. Epub 2021 Oct 14.
Specific delivery of NCEH1 plasmid is a promising approach to boost the cholesterol removal from lipid-laden macrophages for antiatherosclerosis. Polyethylenimine (PEI) is one of the most efficient gene carriers among nonviral vectors. However, the high transfection activity of PEI is always accompanied by profound cytotoxicity. To tackle the paradox between transfection efficiency and safety, we constructed a novel ATP-responsive multifunctional supramolecular polymer by cross-linking functionalized low-molecular-weight PEI via a boronic ester bond for NCEH1 plasmid delivery. The supramolecular polymer could condense NCEH1 plasmids to form stable nanosized polyplexes when the w/w ratios of the polymer and gene were higher than 2. ATP-triggered degradation of the polymer and pDNA release were characterized by a series of studies, including H NMR, P NMR, XPS, agarose gel electrophoresis, and ethidium bromide exclusion tests. In addition, the changes in particle size and morphology were observed in the presence of ATP. Interestingly, the supramolecular polymer showed broad spectrum antioxidant activities by measuring the elimination rates of different reactive oxygen species. In addition, the supramolecular polymer displayed a high buffering capability and good cytocompatibility as demonstrated by the results of the buffering capacity, a hemolysis assay, and a cytotoxicity test. Importantly, it was revealed that the supramolecular polymer/NCEH1 plasmid polyplex formulated at a w/w ratio of 20 was most effective in enhancing cholesterol removal from lipid-laden macrophages and reducing the accumulation of lipid droplets as evidenced by transfection study, cholesterol efflux assay, and oil red O staining studies. Collectively, the ATP-responsive multifunctional supramolecular polymer holds great potential for safe and efficient gene delivery for antiatherosclerosis.
特定递送 NCEH1 质粒是一种很有前途的方法，可以促进富含脂质的巨噬细胞中胆固醇的清除，从而达到抗动脉粥样硬化的效果。聚乙烯亚胺（PEI）是所有非病毒载体中最有效的基因载体之一。然而，PEI 的高转染活性总是伴随着严重的细胞毒性。为了解决转染效率和安全性之间的矛盾，我们通过硼酸酯键交联功能化的低分子量 PEI 构建了一种新型的 ATP 响应多功能超分子聚合物，用于 NCEH1 质粒的递送。当聚合物和基因的 w/w 比高于 2 时，超分子聚合物可以将 NCEH1 质粒凝聚成稳定的纳米级聚阳离子。通过一系列研究，包括 H NMR、P NMR、XPS、琼脂糖凝胶电泳和溴化乙锭排除试验，对聚合物和 pDNA 的降解和释放进行了表征。此外，还观察到在存在 ATP 的情况下粒径和形态的变化。有趣的是，超分子聚合物通过测量不同活性氧物种的消除率表现出广谱抗氧化活性。此外，超分子聚合物还表现出高缓冲能力和良好的细胞相容性，这一点可以通过缓冲能力、溶血试验和细胞毒性试验的结果得到证明。重要的是，研究表明，当 w/w 比为 20 时，超分子聚合物/NCEH1 质粒的聚阳离子复合物在增强富含脂质的巨噬细胞中胆固醇的清除和减少脂质滴的积累方面最为有效，这一点可以通过转染研究、胆固醇外排测定和油红 O 染色研究得到证明。总的来说，ATP 响应多功能超分子聚合物具有很大的潜力，可用于安全有效地进行抗动脉粥样硬化的基因递送。
