Quantification and visualization of cellular uptake of TiO2 and Ag nanoparticles: comparison of different ICP-MS techniques.
Quantification and visualization of cellular uptake of TiO2 and Ag nanoparticles: comparison of different ICP-MS techniques.
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DOI：
10.1186/s12951-016-0203-z
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发表时间：
2016-06-22
影响因子：
10.2
通讯作者：
Haase A
中科院分区：
文献类型：
--
作者：
Hsiao IL;Bierkandt FS;Reichardt P;Luch A;Huang YJ;Jakubowski N;Tentschert J;Haase A
纳米颗粒（NP）的安全性评估需要适用于量化组织和细胞对NP摄取的技术。为此目的最常用的技术是基于电感耦合等离子体质谱法（ICP-MS）。在这里，我们应用并比较三种不同的ICP-MS方法，以研究细胞摄取的TiO 2（直径7或20 nm，分别）和Ag（直径50或75 nm，分别）纳米粒子到分化的小鼠神经母细胞瘤细胞（神经-2a细胞）。将细胞与不同量的NP孵育。此后，通过激光烧蚀ICP-MS（LA-ICP-MS）直接分析它们，或裂解它们，并通过ICP-MS和通过单颗粒ICP-MS（SP-ICP-MS）分析裂解物。所有技术都证实，当将数值转换为基于NP数的剂量度量时，较小的颗粒被吸收到更高的程度。与ICP-MS和LA-ICP-MS相反，该测量已经通过SP-ICP-MS直接提供。通过SP-ICP-MS对细胞裂解物中NP尺寸分布的分析表明细胞内NP聚集体的形成。LA-ICP-MS成像显示，一些75 nm的Ag纳米颗粒似乎被吸附到细胞膜上，并且没有渗透到细胞中，而大多数50 nm的Ag纳米颗粒被内化。LA-ICP-MS证实了NP摄取的高细胞间变异性。基于我们的数据，我们建议联合收割机不同的ICP-MS技术，以可靠地确定平均NP质量和数量浓度，NP大小和大小分布模式，以及细胞间的NP摄取和细胞内定位的变化。本文的在线版本（doi：10.1186/s12951-016-0203-z）包含补充材料，可供授权用户使用。
Safety assessment of nanoparticles (NPs) requires techniques that are suitable to quantify tissue and cellular uptake of NPs. The most commonly applied techniques for this purpose are based on inductively coupled plasma mass spectrometry (ICP-MS). Here we apply and compare three different ICP-MS methods to investigate the cellular uptake of TiO2 (diameter 7 or 20 nm, respectively) and Ag (diameter 50 or 75 nm, respectively) NPs into differentiated mouse neuroblastoma cells (Neuro-2a cells). Cells were incubated with different amounts of the NPs. Thereafter they were either directly analyzed by laser ablation ICP-MS (LA-ICP-MS) or were lysed and lysates were analyzed by ICP-MS and by single particle ICP-MS (SP-ICP-MS). All techniques confirmed that smaller particles were taken up to a higher extent when values were converted in an NP number-based dose metric. In contrast to ICP-MS and LA-ICP-MS, this measure is already directly provided through SP-ICP-MS. Analysis of NP size distribution in cell lysates by SP-ICP-MS indicates the formation of NP agglomerates inside cells. LA-ICP-MS imaging shows that some of the 75 nm Ag NPs seemed to be adsorbed onto the cell membranes and were not penetrating into the cells, while most of the 50 nm Ag NPs were internalized. LA-ICP-MS confirms high cell-to-cell variability for NP uptake. Based on our data we propose to combine different ICP-MS techniques in order to reliably determine the average NP mass and number concentrations, NP sizes and size distribution patterns as well as cell-to-cell variations in NP uptake and intracellular localization. The online version of this article (doi:10.1186/s12951-016-0203-z) contains supplementary material, which is available to authorized users.