Shannahan Jonathan H, Fritz Kristofer S, Raghavendra Achyut J, Podila Ramakrishna, Persaud Indushekar, Brown Jared M
*Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, The University of Colorado Anschutz Medical Campus, Aurora, Colorado 80045;
Department of Physics and Astronomy, Clemson University, Clemson, South Carolina, 29634; Clemson Nanomaterials Center and COMSET, Clemson University, Anderson, South Carolina, 29625.
Toxicol Sci. 2016 Aug;152(2):406-16. doi: 10.1093/toxsci/kfw097. Epub 2016 Jun 2.
Nanoparticle (NP) association with macromolecules in a physiological environment forms a biocorona (BC), which alters NP distribution, activity, and toxicity. While BC formation is dependent on NP physicochemical properties, little information exists on the influence of the physiological environment. Obese individuals and those with cardiovascular disease exist with altered serum chemistry, which is expected to influence BC formation and NP toxicity. We hypothesize that a BC formed on NPs following incubation in hyperlipidemic serum will result in altered NP-BC protein content, cellular association, and toxicity compared to normal serum conditions. We utilized Fe3O4 NPs, which are being developed as MRI contrast and tumor targeting agents to test our hypothesis. We used rat aortic endothelial cells (RAECs) within a dynamic flow in vitro exposure system to more accurately depict the in vivo environment. A BC was formed on 20nm PVP-suspended Fe3O4 NPs following incubation in water, 10% normal or hyperlipidemic rat serum. Addition of BCs resulted in increased hydrodynamic size and decreased surface charge. More cholesterol associated with Fe3O4 NPs after incubation in hyperlipidemic as compared with normal serum. Using quantitative proteomics, we identified unique differences in BC protein components between the 2 serum types. Under flow conditions, formation of a BC from both serum types reduced RAECs association of Fe3O4 NPs. Addition of BCs was found to exacerbate RAECs inflammatory gene responses to Fe3O4 NPs (Fe3O4-hyperlipidemic > Fe3O4-normal > Fe3O4) including increased expression of IL-6, TNF-α, Cxcl-2, VCAM-1, and ICAM-1. Overall, these findings demonstrate that disease-induced variations in physiological environments have a significant impact NP-BC formation, cellular association, and cell response.
在生理环境中，纳米颗粒（NP）与大分子结合形成生物冠（BC），这会改变NP的分布、活性和毒性。虽然BC的形成取决于NP的物理化学性质，但关于生理环境影响的信息却很少。肥胖个体和患有心血管疾病的人血清化学成分发生改变，预计这会影响BC的形成和NP毒性。我们假设，与正常血清条件相比，在高脂血症血清中孵育后在NP上形成的BC将导致NP - BC蛋白质含量、细胞结合及毒性发生改变。我们利用正被开发用作磁共振成像造影剂和肿瘤靶向剂的Fe3O4纳米颗粒来验证我们的假设。我们在动态流动体外暴露系统中使用大鼠主动脉内皮细胞（RAEC），以更准确地描绘体内环境。在水中、10%正常或高脂血症大鼠血清中孵育后，20nm聚乙烯吡咯烷酮（PVP）悬浮的Fe3O4纳米颗粒上形成了BC。添加BC会导致流体动力学尺寸增加和表面电荷减少。与正常血清相比，在高脂血症血清中孵育后，更多胆固醇与Fe3O4纳米颗粒结合。使用定量蛋白质组学，我们确定了两种血清类型之间BC蛋白质成分的独特差异。在流动条件下，两种血清类型形成的BC均降低了RAEC对Fe3O4纳米颗粒的结合。发现添加BC会加剧RAEC对Fe3O4纳米颗粒的炎症基因反应（Fe3O4 - 高脂血症＞Fe3O4 - 正常＞Fe3O4），包括白细胞介素 - 6（IL - 6）、肿瘤坏死因子 - α（TNF - α）、趋化因子配体2（Cxcl - 2）、血管细胞黏附分子 - 1（VCAM - 1）和细胞间黏附分子 - 1（ICAM - 1）表达增加。总体而言，这些发现表明，疾病引起的生理环境变化对NP - BC的形成、细胞结合及细胞反应有重大影响。
