Effect of protein corona magnetite nanoparticles derived from bread in vitro digestion on Caco-2 cells morphology and uptake
Effect of protein corona magnetite nanoparticles derived from bread in vitro digestion on Caco-2 cells morphology and uptake
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DOI：
10.1016/j.biocel.2015.10.019
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发表时间：
2016-06-01
影响因子：
4
通讯作者：
Bombelli, Francesca Baldelli
中科院分区：
文献类型：
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作者：
Di Silvio, Desire;Rigby, Neil;Bombelli, Francesca Baldelli
生物流体中的纳米颗粒（NPs）立即与蛋白质相互作用，形成赋予其生物身份的生物分子冠（PC）。虽然已经报道了几项关于人血浆中PC形成的研究，但口服给药NP的PC研究较少，主要是在食物基质存在下。事实上，食物基质在消化时会发生几种动态变化，这些变化肯定会影响NP上形成的PC。缺乏对这一主题的研究显然与难以从如此复杂的环境中分离出代表性的PC NP有关。在这项工作中，磁铁矿纳米颗粒被添加到体外模拟消化的同时，面包和PC纳米颗粒分离后，胃和十二指肠阶段的蔗糖梯度超离心（UC）。根据大小和蛋白质组成对PC NP进行表征。然后在无血清环境中对Caco-2单层进行易位研究，并通过共聚焦显微镜表征细胞形态。从胃和十二指肠相分离的PC NPs在大小、表面电荷和蛋白质冠组成上不同。NP细胞摄取增强的消化PC诱导细胞单层的形态学变化。总的来说，在这项工作中，我们能够在食物基质的存在下从消化液中分离出PC NP，并研究它们对Caco-2细胞的生物学反应。(C)2015爱思唯尔有限公司版权所有。
Nanoparticles (NPs) in biological fluids immediately interact with proteins forming a biomolecular corona (PC) that imparts their biological identity. While several studies on the formation of the PC in human plasma have been reported, the PC of orally administrated NPs has been less investigated, mostly in the presence of a food matrix. In fact, food matrixes when digested are subject of several dynamic changes that will certainly affect the PC formed on the NPs. The lack of studies on this topic is clearly related to the difficulty in isolating representative PC NPs from such a complex environment. In this work magnetite NPs were added to in vitro simulated digestion simultaneously with bread and PC NPs were isolated after gastric and duodenal phases by sucrose gradient ultracentrifugation (UC). The PC NPs were characterized in terms of size and protein composition. Translocation studies were then performed on Caco-2 mono layers in a serum free environment and cell morphology was characterized by confocal microscopy. PC NPs isolated from gastric and duodenal phases were different in size, surface charge and protein corona composition. NP cellular uptake was enhanced by the digestive PC inducing morphology changes in the cell monolayer. Overall, in this work we were able to isolate PC NPs from digested fluids in the presence of a food matrix and study their biological response on Caco-2 cells. (C) 2015 Elsevier Ltd. All rights reserved.