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An improved protocol for ICP-MS-based assessment of the cellular uptake of metal-based nanoparticles

机译:基于ICP-MS的基于金属基纳米粒子蜂窝摄取的基于ICP-MS评估的改进方案

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摘要

Along with a growing interest in biomedical applications of metal-based nanoparticles, there is a compelling need in systematic information on their behavior in human body systems, preferably at the cellular level. However, in most of the in-vitro uptake experiments, the nanomaterial was applied in its native form that in reality can hardly reach the cell. In this work, we developed an improved procedure in which prior to addition to the cells the particles are converted into the protein conjugates by incubation in human serum. The procedure was tested for gold nanoparticles of different size, chosen as a representative nanomaterial on multifunctional medicinal use, and MCF-7 cell line. Using ICP-MS to measure intracellular metal concentration, it was shown that an original state has significant effect on particle internalization. The protein corona significantly inhibits the uptake amount by MCF-7 cells, with the greatest influence (a 15-fold decrease compared to uncoated particles) being exerted over the smallest, 5-nm particles (3 pg Au/cell). Conjugates of larger particles (20 and 50 nm) are taken up more effectively (45 and 34 pg Au/cell, respectively). The advanced protocol makes the uptake results more reliable and its implementation may accelerate the preclinical development of metal-based nanoparticles as a viable theranostic implement. (C) 2019 Elsevier B.V. All rights reserved.
机译:随着对金属基纳米粒子的生物医学应用的兴趣日益增长,在系统信息中有令人尖锐的需求,优选在人体系统中的行为,优选在细胞水平。然而,在大部分体外摄取实验中,纳米材料以其天然形式施加,即现实中可能几乎不能到达细胞。在这项工作中,我们开发了一种改进的程序,在此之前,通过在人血清中孵育,颗粒将颗粒转化为蛋白质缀合物。该方法对不同尺寸的金纳米颗粒测试,作为多功能药用的代表性纳米材料,以及MCF-7细胞系。使用ICP-MS测量细胞内金属浓度,显示出原始状态对颗粒内化具有显着影响。蛋白质电晕显着抑制MCF-7细胞的摄取量,最大的影响(与未涂覆的颗粒相比,15倍降低)施加在最小的5nm颗粒上(3 pg au / cell)。较大颗粒(20和50nm)的缀合物更有效地溶解(分别为45和34pg Au / cell)。先进的协议使得摄取结果更可靠,其实现可以加速金属基纳米粒子的临床前开发作为可行的治疗工具。 (c)2019 Elsevier B.v.保留所有权利。

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