Intentional formation of a protein corona on nanoparticles: Serum concentration affects protein corona mass, surface charge, and nanoparticle-cell interaction
Intentional formation of a protein corona on nanoparticles: Serum concentration affects protein corona mass, surface charge, and nanoparticle-cell interaction
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DOI：
10.1016/j.biocel.2015.11.005
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发表时间：
2016-06-01
影响因子：
4
通讯作者：
Dutz, Silvio
中科院分区：
文献类型：
--
作者：
Graefe, Christine;Weidner, Andreas;Dutz, Silvio
纳米粒子与生物系统接触后立即形成的蛋白质冠对纳米粒子的生物命运起着至关重要的作用。在这里提出的研究中，我们描述了一种策略来控制结合在颗粒表面的冠蛋白的量，以及这种蛋白质冠对颗粒-细胞相互作用的影响。为了形成电晕，将聚乙烯亚胺（PEI）涂层的磁性纳米颗粒（MNP）在由胎牛血清（FCS）和细胞培养基组成的培养基中孵育。为了调节与颗粒结合的蛋白质的量，根据FCS含量改变孵育培养基的组成。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE），估计蛋白质冠质量，并确定参与蛋白质的大小分布。此外，测量了孵育颗粒的ζ电位。人血脑屏障代表细胞系HBMEC用于体外孵育实验。为了研究FCS依赖性蛋白质冠形成对MNP和细胞相互作用的后果，使用流式细胞术和激光扫描显微镜。Zeta电位以及SDS-PAGE清楚地揭示了随着孵育培养基中FCS量的增加，MNP上冠蛋白的量增加。对于与较低FCS浓度孵育的MNP，尤其是分子量在30 kDa和100 kDa之间的中等大小的蛋白质，可以在蛋白质冠内发现，而对于在较高FCS浓度内孵育的MNP，30 kDa和更少的冠蛋白的分数增加。蛋白质冠的存在减少了PEI包被的MNP与HBMEC细胞在30分钟孵育内的相互作用。(C)2015爱思唯尔有限公司版权所有。
The protein corona, which immediately is formed after contact of nanoparticles and biological systems, plays a crucial role for the biological fate of nanoparticles. In the here presented study we describe a strategy to control the amount of corona proteins which bind on particle surface and the impact of such a protein corona on particle-cell interactions. For corona formation, polyethyleneimine (PEI) coated magnetic nanoparticles (MNP) were incubated in a medium consisting of fetal calf serum (FCS) and cell culture medium. To modulate the amount of proteins bind to particles, the composition of the incubation medium was varied with regard to the FCS content. The protein corona mass was estimated and the size distribution of the participating proteins was determined by means of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Additionally, the zeta potential of incubated particles was measured. Human blood-brain barrier-representing cell line HBMEC was used for in vitro incubation experiments. To investigate the consequences of the FCS dependent protein corona formation on the interaction of MNP and cells flow cytometry and laser scanning microscopy were used. Zeta potential as well as SDS-PAGE clearly reveal an increase in the amount of corona proteins on MNP with increasing amount of FCS in incubation medium. For MNP incubated with lower FCS concentrations especially medium-sized proteins of molecular weights between 30 kDa and 100 kDa could be found within the protein corona, whereas for MNP incubated within higher FCS concentrations the fraction of corona proteins of 30 kDa and less increased. The presence of the protein corona reduces the interaction of PEI-coated MNP with HBMEC cells within a 30 min-incubation. (C) 2015 Elsevier Ltd. All rights reserved.