title: No cytotoxicity or genotoxicity of graphene and graphene oxide in murine lung epithelial FE1 cells in vitro.
abstract: Graphene and graphene oxide receive much attention these years, because they add attractive properties to a wide range of applications and products. Several studies have shown toxicological effects of other carbon-based nanomaterials such as carbon black nanoparticles and carbon nanotubes in vitro and in vivo. Here, we report in-depth physicochemical characterization of three commercial graphene materials, one graphene oxide (GO) and two reduced graphene oxides (rGO) and assess cytotoxicity and genotoxicity in the murine lung epithelial cell line FE1. The studied GO and rGO mainly consisted of 2-3 graphene layers with lateral sizes of 1-2 µm. GO had almost equimolar content of C, O, and H while the two rGO materials had lower contents of oxygen with C/O and C/H ratios of 8 and 12.8, respectively. All materials had low levels of endotoxin and low levels of inorganic impurities, which were mainly sulphur, manganese, and silicon. GO generated more ROS than the two rGO materials, but none of the graphene materials influenced cytotoxicity in terms of cell viability and cell proliferation after 24 hr. Furthermore, no genotoxicity was observed using the alkaline comet assay following 3 or 24 hr of exposure. We demonstrate that chemically pure, few-layered GO and rGO with comparable lateral size (> 1 µm) do not induce significant cytotoxicity or genotoxicity in FE1 cells at relatively high doses (5-200 µg/ml). Environ. Mol. Mutagen. 57:469-482, 2016. © 2016 The Authors. Environmental and Molecular Mutagenesis Published by Wiley Periodicals, Inc.
authors.affiliation: National Research Centre for the Working Environment, Lersø Parkallé 105, DK-2100, Copenhagen Ø, Denmark.|Department of Science and Environment, Roskilde University, DK-4000, Roskilde, Denmark.
authors.forename: Stefan
authors.initials: S
authors.lastname: Bengtson
authors.affiliation: National Research Centre for the Working Environment, Lersø Parkallé 105, DK-2100, Copenhagen Ø, Denmark.
authors.forename: Kirsten
authors.initials: K
authors.lastname: Kling
authors.affiliation: National Research Centre for the Working Environment, Lersø Parkallé 105, DK-2100, Copenhagen Ø, Denmark.
authors.forename: Anne Mette
authors.initials: AM
authors.lastname: Madsen
authors.affiliation: National Research Centre for the Working Environment, Lersø Parkallé 105, DK-2100, Copenhagen Ø, Denmark.
authors.forename: Asger W
authors.initials: AW
authors.lastname: Noergaard
authors.affiliation: National Research Centre for the Working Environment, Lersø Parkallé 105, DK-2100, Copenhagen Ø, Denmark.
authors.forename: Nicklas Raun
authors.initials: NR
authors.lastname: Jacobsen
authors.affiliation: National Research Centre for the Working Environment, Lersø Parkallé 105, DK-2100, Copenhagen Ø, Denmark.
authors.forename: Per Axel
authors.initials: PA
authors.lastname: Clausen
authors.affiliation: R&D Department, Graphenea S.A, Tolosa Hiribidea 76, E-20018 Donostia, San Sebastian, Spain.
authors.forename: Beatriz
authors.initials: B
authors.lastname: Alonso
authors.affiliation: R&D Department, Graphenea S.A, Tolosa Hiribidea 76, E-20018 Donostia, San Sebastian, Spain.
authors.forename: Amaia
authors.initials: A
authors.lastname: Pesquera
authors.affiliation: R&D Department, Graphenea S.A, Tolosa Hiribidea 76, E-20018 Donostia, San Sebastian, Spain.
authors.forename: Amaia
authors.initials: A
authors.lastname: Zurutuza
authors.affiliation: CEA GRENOBLE, University Grenoble Alpes, F-38000 Grenoble, France.|Nanomaterials Technologies Department, CEA/LITEN/DTNM, 17 avenue des martyrs 38054 Grenoble cedex.
authors.forename: Raphael
authors.initials: R
authors.lastname: Ramos
authors.affiliation: CEA GRENOBLE, University Grenoble Alpes, F-38000 Grenoble, France.|Nanoscience and Cryogeny Institute CEA/DRF/INAC/MEM, 17 avenue des martyrs 38054 Grenoble cedex.
authors.forename: Hanako
authors.initials: H
authors.lastname: Okuno
authors.affiliation: CEA GRENOBLE, University Grenoble Alpes, F-38000 Grenoble, France.|Nanomaterials Technologies Department, CEA/LITEN/DTNM, 17 avenue des martyrs 38054 Grenoble cedex.
authors.forename: Jean
authors.initials: J
authors.lastname: Dijon
authors.affiliation: National Research Centre for the Working Environment, Lersø Parkallé 105, DK-2100, Copenhagen Ø, Denmark.|Department of Public Health, University of Copenhagen, DK-1014, Copenhagen K, Denmark.
authors.forename: Håkan
authors.initials: H
authors.lastname: Wallin
authors.affiliation: National Research Centre for the Working Environment, Lersø Parkallé 105, DK-2100, Copenhagen Ø, Denmark.|Department of Micro- and Nanotechnology, Technical University of Denmark, DK-2800 Kgs, Lyngby, Denmark.
authors.forename: Ulla
authors.initials: U
authors.lastname: Vogel
mesh_terms: D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D020552:Comet Assay; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D008401:Gas Chromatography-Mass Spectrometry; D006108:Graphite / Q000737:chemistry / Q000633:toxicity*; D051379:Mice; D017348:Microscopy, Electron, Scanning Transmission; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles / Q000737:chemistry / Q000633:toxicity*; D010087:Oxides / Q000737:chemistry / Q000633:toxicity; D010316:Particle Size; D017382:Reactive Oxygen Species / Q000378:metabolism; D013499:Surface Properties
chemical_list: D010087:Oxides; D017382:Reactive Oxygen Species; D006108:Graphite
keywords: comet assay; electron microscopy; elemental analysis; reactive oxygen species