Effects of Surface Charge of Hyperbranched Polymers on Cytotoxicity, Dynamic Cellular Uptake and Localization, Hemotoxicity, and Pharmacokinetics in Mice
Effects of Surface Charge of Hyperbranched Polymers on Cytotoxicity, Dynamic Cellular Uptake and Localization, Hemotoxicity, and Pharmacokinetics in Mice
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DOI：
10.1021/acs.molpharmaceut.7b00611
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发表时间：
2017-12-01
影响因子：
4.9
通讯作者：
Thurecht, Kristofer J.
中科院分区：
文献类型：
--
作者：
Chen, Liyu;Simpson, Joshua D.;Thurecht, Kristofer J.
纳米聚合物材料越来越多地被研究作为药物产品，药物/基因递送载体或健康监测设备。表面电荷是调节纳米材料在体内行为的主要参数之一。在本文中，我们展示了如何控制化学合成允许操纵纳米颗粒表面电荷，这反过来又极大地影响了体内行为。以三种甲基丙烯酸酯/甲基丙烯酰胺单体为原料，通过可逆加成-断裂链转移（RAFT）聚合法合成了结构规整的超支化聚合物（HBP）。通过动态光散射（DLS）和透射电子显微镜（TEM）测定，每个HBP具有约5 nm的流体动力学直径。在聚合物纳米颗粒中掺入荧光部分可以确定电荷如何影响纳米材料的体内药代动力学行为以及对它们的生物反应。观察到表面电荷、细胞摄取和细胞毒性之间的直接相关性，其中阳离子HBPs表现出比它们的中性和阴离子对应物更高的细胞摄取和细胞毒性。不同电荷的HBP在巨噬细胞内的分布的评价表明，所有的HBP积累在细胞质中，但阳离子HBP也贩运，并积累在细胞核内。尽管阳离子HBP会引起轻微溶血，但这通常低于体内安全性的可接受水平。药代动力学行为分析显示，阳离子和阴离子HBPs的血液半衰期较短，分别为1.82 +/- 0.51和2.34 +/- 0.93 h，而中性HBPs为5.99 +/- 2.30 h。这是由于带正电荷的表面更容易被调理素蛋白覆盖，因此对吞噬细胞更明显。这得到了体外流式细胞术和定性活细胞成像研究的支持，这些研究表明，阳离子HBP往往比中性和阴离子颗粒更有效、更迅速地被巨噬细胞摄取。
Nanoscaled polymeric materials are increasingly being investigated as pharmaceutical products, drug/gene delivery vectors, or health-monitoring devices. Surface charge is one of the dominant parameters that regulates nanomaterial behavior in vivo. In this paper, we demonstrated how control over chemical synthesis allowed manipulation of nanoparticle surface charge, which in turn greatly influenced the in vivo behavior. Three methacrylate/methacrylamide-based monomers were used to synthesize well-defined hyperbranched polymers (HBP) by reversible addition-fragmentation chain transfer (RAFT) polymerization. Each HBP had a hydrodynamic diameter of approximately 5 nm as determined by dynamic light scattering (DLS) and transmission electron microscopy (TEM). Incorporation of a fluorescent moiety within the polymeric nanoparticles allowed determination of how charge affected the in vivo pharmacokinetic behavior of the nanomaterials and the biological response to them. A direct correlation between surface charge, cellular uptake, and cytotoxicity was observed, with cationic HBPs exhibiting higher cellular uptake and cytotoxicity than their neutral and anionic counterparts. Evaluation of the distribution of the differently charged HBPs within macrophages showed that all HBPs accumulated in the cytoplasm, but cationic HBPs also trafficked to, and accumulated within, the nucleus. Although cationic HBPs caused slight hemolysis, this was generally below accepted levels for in vivo safety. Analysis of pharmacokinetic behavior showed that cationic and anionic HBPs had short blood half-lives of 1.82 +/- 0.51 and 2.34 +/- 0.93 h respectively, compared with 5.99 +/- 2.30 h for neutral HBPs. This was attributed to the fact that positively charged surfaces are more readily covered with opsonin proteins and thus more visible to phagocytic cells. This was supported by in vitro flow cytometric and qualitative live cell imaging studies, which showed that cationic HBPs tended to be taken up by macrophages more effectively and rapidly than neutral and anionic particles.