Ortega M T, Riviere J E, Choi K, Monteiro-Riviere N A
Nanotechnology Innovation Center of Kansas State (NICKS), Kansas State University, Manhattan, KS, United States.
Nanotechnology Innovation Center of Kansas State (NICKS), Kansas State University, Manhattan, KS, United States.
Toxicol In Vitro. 2017 Aug;42:150-160. doi: 10.1016/j.tiv.2017.04.020. Epub 2017 Apr 19.
Gold nanoparticles (AuNP) adsorb macromolecules to form a protein corona (PC) after systemic delivery, to which the kidney as the primary excretory organ is constantly exposed. The role of the PC on AuNP cell uptake and toxicity was investigated in vitro in human proximal tubule cells (HPTC) using 40 and 80nm branched polyethylenimine (BPEI), lipoic acid (LA) and polyethylene glycol (PEG) coated AuNP with or without (bare) PCs composed of human plasma (HP) or human serum albumin (HSA) for 0.25 to 24h. Time-dependent intracellular uptake, assessed by ICP-MS showed PC modulated cell uptake and cytotoxicity; with bare 40nm BPEI-AuNP showing the greatest responses. All AuNP showed minimal to no cytokine release. At the nontoxic dose, 40nm bare BPEI-AuNP significantly modified gene expression related to immunotoxicity, steatosis, and mitochondrial metabolism; while at the high dose, pathways of DNA damage and repair, apoptosis, fatty acid metabolism and heat shock response were modulated. HP corona BPEI-AuNP response was comparable to control. These studies clearly showed reduced uptake and cytotoxicity, as well as differentiated gene expression of AuNP with PCs, questioning the utility of in vitro studies using bare NP to assess in vivo effects. Significantly, only cationic bare BPEI-AuNP had HPTC uptake or cytotoxicity suggesting the relative safety of PEG and LA-AuNP as nanomedicine constructs.
金纳米颗粒（AuNP）在全身给药后会吸附大分子形成蛋白质冠（PC），作为主要排泄器官的肾脏会持续暴露于该蛋白质冠。使用40和80纳米的支链聚乙烯亚胺（BPEI）、硫辛酸（LA）和聚乙二醇（PEG）包被的AuNP，在有无由人血浆（HP）或人血清白蛋白（HSA）组成的（裸）蛋白质冠的情况下，对人近端小管细胞（HPTC）进行体外研究，以探究蛋白质冠对AuNP细胞摄取和毒性的作用，作用时间为0.25至24小时。通过电感耦合等离子体质谱（ICP-MS）评估的时间依赖性细胞内摄取表明，蛋白质冠调节细胞摄取和细胞毒性；裸40纳米BPEI-AuNP的反应最为强烈。所有AuNP均显示出最小程度的细胞因子释放或无细胞因子释放。在无毒剂量下，40纳米裸BPEI-AuNP显著改变了与免疫毒性、脂肪变性和线粒体代谢相关的基因表达；而在高剂量下，DNA损伤与修复、细胞凋亡、脂肪酸代谢和热休克反应等途径受到调节。HP冠BPEI-AuNP的反应与对照组相当。这些研究清楚地表明，有蛋白质冠的AuNP的摄取和细胞毒性降低，以及基因表达存在差异，这对使用裸纳米颗粒进行体外研究以评估体内效应的实用性提出了质疑。重要的是，只有阳离子裸BPEI-AuNP具有HPTC摄取或细胞毒性，这表明PEG和LA-AuNP作为纳米药物构建体具有相对安全性。
