期刊
ACS NANO
卷 12, 期 2, 页码 1373-1389出版社
AMER CHEMICAL SOC
DOI: 10.1021/acsnano.7b07734
关键词
2D materials; graphene; confocal live-cell imaging; inflammation; lungs; nanotoxicology
类别
资金
- European Union's 7th RTD Framework Program: Graphene Flagship project [FP7-ICT-2013-FET-F-604391]
- European Union's Horizon research and innovation program [696656]
- EPSRC-North West Nanoscience Doctoral Training Centre (NOWNANO DTC) [EP/G03737X/1]
- Medical Research Council (MRC)
- University of Manchester Strategic Fund
- Wellcome Trust
- Engineering and Physical Sciences Research Council (EPSRC) Centre for Doctoral Training in the Science and Applications of Graphene and Related Nanomaterials (Graphene NOWNANO CDT) [EP/L01548X/1]
- Biotechnology and Biological Sciences Research Council (BBSRC)
- EPSRC [EP/K005014/1] Funding Source: UKRI
- Engineering and Physical Sciences Research Council [1495511, 1353926] Funding Source: researchfish
The interest in graphene and its translation into commercial products has been expanding at a high pace. Based on previously described pulmonary safety concerns for carbon nanomaterials, there is a great need to define parameters guiding interactions between graphene-based materials and the pulmonary system. The aim of the present study was to determine the importance of two critical parameters: lateral dimensions of the material and coating with proteins in relation to each other and their pulmonary impact. Endotoxin-free materials with distinct lateral dimensions, s-GO (50-200 nm) and l-GO (5-15 mu m), were produced and thoroughly characterized. Exploiting intrinsic fluorescence of graphene oxide (GO) and using confocal live-cell imaging, the behavior of the cells in response to the material was visualized in real time. Although BEAS-2B cells internalized GO efficiently, l-GO was linked to higher plasma membrane interactions correlated with elevated reactive oxygen species (ROS) levels, pro-inflammatory response, and greater cytotoxicity, in agreement with the oxidative stress paradigm. For both GO types, the presence of serum alleviated lipid peroxidation of plasma membrane and decreased intracellular ROS levels. However, protein coating was not enough to entirely mitigate toxicity and inflammatory response induced by l-GO. In vitro results were validated in vivo, as l-GO was more prone to induce pulmonary granulomatous response in mice compared to s-GO. In conclusion, the lateral dimension of GO played a more important role than serum protein coating in determining biological responses to the material. It was also demonstrated that time-lapse imaging of live cells interacting with label-free GO sheets can be used as a tool to assess GO-induced cytotoxicity.
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