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Sci Rep. 2017 Jan 12;7:40572. doi: 10.1038/srep40572.

Differential cytotoxic effects of graphene and graphene oxide on skin keratinocytes.

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Department of Life Sciences, University of Trieste, 34127 Trieste, Italy.
Department of Chemical and Pharmaceutical Sciences, University of Trieste, 34127 Trieste, Italy.
Department of Organic Chemistry, Facultad de Ciencias y Tecnologías Químicas-IRICA, University of Castilla-La Mancha, 13071 Ciudad Real, Spain.
CIC BiomaGUNE, Parque Tecnológico de San Sebastián, Paseo Miramón, 182, 20009 San Sebastián (Guipúzcoa), Spain.
Basque Foundation for Science, Ikerbasque, Bilbao 48013, Spain.


Impressive properties make graphene-based materials (GBMs) promising tools for nanoelectronics and biomedicine. However, safety concerns need to be cleared before mass production of GBMs starts. As skin, together with lungs, displays the highest exposure to GBMs, it is of fundamental importance to understand what happens when GBMs get in contact with skin cells. The present study was carried out on HaCaT keratinocytes, an in vitro model of skin toxicity, on which the effects of four GBMs were evaluated: a few layer graphene, prepared by ball-milling treatment (FLG), and three samples of graphene oxide (GOs, a research-grade GO1, and two commercial GOs, GO2 and GO3). Even though no significant effects were observed after 24 h, after 72 h the less oxidized compound (FLG) was the less cytotoxic, inducing mitochondrial and plasma-membrane damages with EC50s of 62.8 μg/mL (WST-8 assay) and 45.5 μg/mL (propidium iodide uptake), respectively. By contrast, the largest and most oxidized compound, GO3, was the most cytotoxic, inducing mitochondrial and plasma-membrane damages with EC50s of 5.4 and 2.9 μg/mL, respectively. These results suggest that only high concentrations and long exposure times to FLG and GOs could impair mitochondrial activity associated with plasma membrane damage, suggesting low cytotoxic effects at the skin level.

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