Industrial grade 2D molybdenum disulphide (MoS2): an in vitro exploration of the impact on cellular uptake, cytotoxicity, and inflammation
Industrial grade 2D molybdenum disulphide (MoS2): an in vitro exploration of the impact on cellular uptake, cytotoxicity, and inflammation
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DOI：
10.1088/2053-1583/aa673f
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发表时间：
2017-06-01
期刊：
影响因子：
5.5
通讯作者：
McIntyre, Jennifer
中科院分区：
文献类型：
--
作者：
Moore, Caroline;Movia, Dania;McIntyre, Jennifer
自 2004 年对石墨烯进行液相单层分离和表征以来，最近石墨烯研究的激增带来了进步，加速了二硫化钼 (MoS2) 等替代二维材料的探索，其独特的物理化学性质可用于从尖端电子设备到纳米医学的各种应用。然而，为了评估对人类健康和环境的任何潜在影响，了解 MoS2 在细胞和亚细胞水平上的生物相互作用至关重要。值得注意的是，评估大规模生产技术生产的材料（而不是研究级材料）的潜在影响非常重要。本研究的目的是探索已建立的细胞系中的细胞毒性、细胞摄取和炎症反应，这些细胞系模拟不同潜在暴露途径（吸入，A549；摄入，AGS；单核细胞，THP-1），与不同尺寸（50 nm、117 nm和177 nm）的MoS2薄片一起孵育后，液相剥离。使用高内涵筛选 (HCS) 和活/死测定法，确定 1 μg ml(-1)（针对三种不同大小的 MoS2）不会对任何细胞系产生毒性作用。共焦显微镜图像显示所有情况下细胞形态均正常。透射电子显微镜 (TEM) 证实所有细胞系均吸收了所有 MoS2 纳米材料，MoS2 最终位于单膜囊泡中。然而，在这种亚致死剂量下，观察到炎症反应，这至少部分与纳米材料悬浮液和表面活性剂样品中脂多糖内毒素的存在有关。因此，使用 10 重 ELISA 检测细胞对 MoS2 或内毒素污染的炎症反应，该 ELISA 说明了细胞因子的产生。使用野生型和内毒素低反应性骨髓来源的树突状细胞进行的实验证实，炎症反应是由内毒素污染、MoS2纳米材料本身和稳定表面活性剂共同作用引起的。
The recent surge in graphene research, since its liquid phase monolayer isolation and characterization in 2004, has led to advancements which are accelerating the exploration of alternative 2D materials such as molybdenum disulphide (MoS2), whose unique physico-chemical properties can be exploited in applications ranging from cutting edge electronic devices to nanomedicine. However, to assess any potential impact on human health and the environment, the need to understand the bio-interaction of MoS2 at a cellular and sub-cellular level is critical. Notably, it is important to assess such potential impacts of materials which are produced by large scale production techniques, rather than research grade materials.The aim of this study was to explore cytotoxicity, cellular uptake and inflammatory responses in established cell-lines that mimic different potential exposure routes (inhalation, A549; ingestion, AGS; monocyte, THP-1) following incubation with MoS2 flakes of varying sizes (50 nm, 117 nm and 177 nm), produced by liquid phase exfoliation. Using high content screening (HCS) and Live/Dead assays, it was established that 1 mu g ml(-1) (for the three different MoS2 sizes) did not induce toxic effects on any of the cell-lines. Confocal microscopy images revealed a normal cellular morphology in all cases. Transmission electron microscopy (TEM) confirmed the uptake of all MoS2 nanomaterials in all the cell-lines, the MoS2 ultimately locating in single membrane vesicles. At such sub-lethal doses, inflammatory responses are observed, however, associated, at least partially, with the presence of lipopolysaccharide endotoxin in nanomaterial suspensions and surfactant samples. Therefore, the inflammatory response of the cells to the MoS2 or endotoxin contamination was interrogated using a 10-plex ELISA which illustrates cytokine production. The experiments carried out using wild-type and endotoxin hyporesponsive bone marrow derived dendritic cells confirmed that the inflammatory responses result from a combination of endotoxin contamination, the MoS2 nanomaterials themselves, and the stabilizing surfactant.