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Protein corona on magnetite nanoparticles and internalization of nanoparticle-protein complexes into healthy and cancer cells

机译:磁铁矿纳米颗粒上的蛋白质电晕和纳米颗粒-蛋白质复合物内在化进入健康和癌细胞的过程

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Superparamagnetic magnetite nanoparticles (MNPs) of different surface properties are incubated in complicated living fluid, including fetal bovine serum solution, cell complete culture medium and cell culture system with/without serum, to investigate the alteration of protein corona and its impact on cell internalization. The MNPs prepared by co-precipitation method are functionalized with L-Lysine (Lys), Glucosamic acid (GA) to obtain amine, carboxyl and hydroxyl groups, separately. All the particles adsorb serum proteins to form MNPs-protein complexes with the surface charge changing into negative. 1D SDS/PAGE gel images analysis indicates that the composition and content of hard protein corona on the surface of NPs are related to their functional groups and agglomeration, and the total amount of protein in the medium. In cell culture system, particles not only adsorb serum proteins, but also associate with cytosolic proteins arising from HepG2 and L02 cells. GA modified MNPs (MNPs-GA) exhibit bovine serum albumin anti-adsorption capability because of the terminal hydroxyl and carboxyl groups. MNPs-GA also shows the highest cellular uptake and label efficiency compared with uncoated MNPs and Lys modified MNPs, due to larger aggregates formation and specific protein corona composition, rather than commonly approved electrostatic interaction between particles and cells. For the first time, our results provide visualized reports on previously neglected, but indispensable protein corona of the MNPs after interaction with both healthy and cancer cells, suggesting that cytosolic protein corona from cells and aggregation of particles are important factors needed to be account for on studying the nano-bio interface.
机译:将不同表面特性的超顺磁性磁铁矿纳米粒子(MNP)在复杂的活液中孵育,包括胎牛血清溶液,细胞完全培养基和有/无血清的细胞培养系统,以研究蛋白质电晕的变化及其对细胞内在化的影响。通过共沉淀法制备的MNP分别用L-赖氨酸(Lys),葡萄糖酸(GA)官能化,分别获得胺,羧基和羟基。所有颗粒均吸附血清蛋白以形成MNPs-蛋白复合物,表面电荷变为负电荷。一维SDS / PAGE凝胶图像分析表明,NPs表面硬蛋白电晕的组成和含量与它们的官能团和团聚以及介质中蛋白质的总量有关。在细胞培养系统中,颗粒不仅吸附血清蛋白,而且还与HepG2和L02细胞产生的胞质蛋白结合。由于末端羟基和羧基,GA修饰的MNP(MNPs-GA)具有牛血清白蛋白抗吸附能力。与未涂覆的MNP和Lys修饰的MNP相比,MNPs-GA还显示出最高的细胞摄取和标记效率,这是由于形成了较大的聚集体和特定的蛋白质电晕成分,而不是通常认可的颗粒与细胞之间的静电相互作用。我们的结果首次提供了可视化的报告,显示了与健康细胞和癌细胞相互作用后MNP先前被忽略但必不可少的蛋白电晕,这表明细胞中的胞质蛋白电晕和颗粒聚集是需要考虑的重要因素。研究纳米生物界面。

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