Characterization of genotoxic response to 15 multiwalled carbon nanotubes with variable physicochemical properties including surface functionalizations in the FE1‐Muta(TM) mouse lung epithelial cell line

Carbon nanotubes vary greatly in physicochemical properties. We compared cytotoxic and genotoxic response to 15 multiwalled carbon nanotubes (MWCNT) with varying physicochemical properties to identify drivers of toxic responses. The studied MWCNT included OECD Working Party on Manufactured Nanomaterials (WPMN) (NM‐401, NM‐402, and NM‐403), materials (NRCWE‐026 and MWCNT‐XNRI‐7), and three sets of surface‐modified MWCNT grouped by physical characteristics (thin, thick, and short I–III, respectively). Each Groups I–III included pristine, hydroxylated and carboxylated MWCNT. Group III also included an amino‐functionalized MWCNT. The level of surface functionalization of the MWCNT was low. The level and type of elemental impurities of the MWCNT varied by <2% of the weight, with exceptions. Based on dynamic light scattering data, the MWCNT were well‐dispersed in stock dispersion of nanopure water with 2% serum, but agglomerated and sedimented during exposure. FE1‐Muta(TM) Mouse lung epithelial cells were exposed for 24 hr. The levels of DNA strand breaks (SB) were evaluated using the comet assay, a screening assay suitable for genotoxicity testing of nanomaterials. Exposure to MWCNT (12.5–200 µg/ml) did not induce significant cytotoxicity (viability above 92%). Cell proliferation was reduced in highest doses of some MWCNT after 24 hr, and was associated with generation of reactive oxygen species and high surface area. Increased levels of DNA SB were only observed for Group II consisting of MWCNT with large diameters and high Fe2O3 and Ni content. Significantly, increased levels of SB were only observed at 200 µg/ml of MWCNT‐042. Overall, the MWCNT were not cytotoxic and weakly genotoxic after 24 hr exposure to doses up to 200 µg/ml. Environ. Mol. Mutagen. 56:183–203, 2015. © 2014 Wiley Periodicals, Inc.

[1]  Y. Liu,et al.  Understanding the toxicity of carbon nanotubes. , 2013, Accounts of chemical research.

[2]  Dongmei Wu,et al.  Transcriptomic Analysis Reveals Novel Mechanistic Insight into Murine Biological Responses to Multi-Walled Carbon Nanotubes in Lungs and Cultured Lung Epithelial Cells , 2013, PloS one.

[3]  Antonio Nunes,et al.  Length-dependent retention of carbon nanotubes in the pleural space of mice initiates sustained inflammation and progressive fibrosis on the parietal pleura. , 2011, The American journal of pathology.

[4]  Jacob S. Lamson,et al.  Particle-Induced Pulmonary Acute Phase Response Correlates with Neutrophil Influx Linking Inhaled Particles and Cardiovascular Risk , 2013, PloS one.

[5]  U. Vogel,et al.  Validation of freezing tissues and cells for analysis of DNA strand break levels by comet assay , 2013, Mutagenesis.

[6]  Nicklas Raun Jacobsen,et al.  Inflammatory and genotoxic effects of sanding dust generated from nanoparticle-containing paints and lacquers , 2012, Nanotoxicology.

[7]  Nicklas Raun Jacobsen,et al.  Lung inflammation and genotoxicity following pulmonary exposure to nanoparticles in ApoE-/- mice , 2009, Particle and Fibre Toxicology.

[8]  Stefano Bellucci,et al.  Multi‐walled carbon nanotubes induce cytotoxicity and genotoxicity in human lung epithelial cells , 2012, Journal of applied toxicology : JAT.

[9]  Craig A Poland,et al.  Length-dependent pleural inflammation and parietal pleural responses after deposition of carbon nanotubes in the pulmonary airspaces of mice , 2012, Nanotoxicology.

[10]  G. Giambastiani,et al.  Tunable Epoxidation of Single‐Walled Carbon Nanotubes by Isolated Methyl(trifluoromethyl)dioxirane , 2014 .

[11]  Antonio Marcomini,et al.  Genotoxicity, cytotoxicity, and reactive oxygen species induced by single‐walled carbon nanotubes and C60 fullerenes in the FE1‐Muta™Mouse lung epithelial cells , 2008, Environmental and molecular mutagenesis.

[12]  Craig A. Poland,et al.  Asbestos, carbon nanotubes and the pleural mesothelium: a review of the hypothesis regarding the role of long fibre retention in the parietal pleura, inflammation and mesothelioma , 2010, Particle and Fibre Toxicology.

[13]  S. Bellucci,et al.  Comparative cyto-genotoxicity assessment of functionalized and pristine multiwalled carbon nanotubes on human lung epithelial cells. , 2012, Toxicology in vitro : an international journal published in association with BIBRA.

[14]  Tatyana Chernova,et al.  Pulmonary toxicity of carbon nanotubes and asbestos - similarities and differences. , 2013, Advanced drug delivery reviews.

[15]  Julia Catalán,et al.  Genotoxicity of short single-wall and multi-wall carbon nanotubes in human bronchial epithelial and mesothelial cells in vitro. , 2013, Toxicology.

[16]  Jacob S. Lamson,et al.  Carbon black nanoparticle instillation induces sustained inflammation and genotoxicity in mouse lung and liver , 2012, Particle and Fibre Toxicology.

[17]  Jin Sik Kim,et al.  Aspect ratio has no effect on genotoxicity of multi-wall carbon nanotubes , 2011, Archives of Toxicology.

[18]  Nicklas Raun Jacobsen,et al.  Increased mutant frequency by carbon black, but not quartz, in the lacZ and cII transgenes of muta™mouse lung epithelial cells , 2007, Environmental and molecular mutagenesis.

[19]  H. Karlsson,et al.  Copper oxide nanoparticles are highly toxic: a comparison between metal oxide nanoparticles and carbon nanotubes. , 2008, Chemical research in toxicology.

[20]  Håkan Wallin,et al.  Particle-induced pulmonary acute phase response may be the causal link between particle inhalation and cardiovascular disease , 2014, Wiley interdisciplinary reviews. Nanomedicine and nanobiotechnology.

[21]  John Parthenios,et al.  Chemical oxidation of multiwalled carbon nanotubes , 2008 .

[22]  Maumita Bandyopadhyay,et al.  Multi-walled carbon nanotubes (MWCNT): induction of DNA damage in plant and mammalian cells. , 2011, Journal of hazardous materials.

[23]  U. Vogel,et al.  Diesel exhaust particles are mutagenic in FE1-MutaMouse lung epithelial cells. , 2008, Mutation research.

[24]  Craig A. Poland,et al.  Carbon nanotubes introduced into the abdominal cavity of mice show asbestos-like pathogenicity in a pilot study. , 2008, Nature nanotechnology.

[25]  A. T. Saber,et al.  Inflammatory and genotoxic effects of nanoparticles designed for inclusion in paints and lacquers , 2012, Nanotoxicology.

[26]  J. Kanno,et al.  Induction of mesothelioma in p53+/- mouse by intraperitoneal application of multi-wall carbon nanotube. , 2008, The Journal of toxicological sciences.

[27]  Chetti Prabhakar,et al.  A REVIEW ON CARBON NANOTUBES , 2011 .

[28]  G. Leitinger,et al.  Combination of small size and carboxyl functionalisation causes cytotoxicity of short carbon nanotubes , 2012, Nanotoxicology.

[29]  François Huaux,et al.  Absence of carcinogenic response to multiwall carbon nanotubes in a 2-year bioassay in the peritoneal cavity of the rat. , 2009, Toxicological sciences : an official journal of the Society of Toxicology.

[30]  The Nanotechnology Panel of the American Chemistry Council (ACC) is pleased to offer comments on the National Institute for Occupational Safety and Health’s (NIOSH) draft Current Intelligence Bulletin (CIB) Occupational Exposure to Carbon Nanotubes and Nanofibers , 2011 .

[31]  Nicklas Raun Jacobsen,et al.  Mutation spectrum in FE1‐MUTATMMouse lung epithelial cells exposed to nanoparticulate carbon black , 2011, Environmental and molecular mutagenesis.

[32]  A. Collins Investigating oxidative DNA damage and its repair using the comet assay. , 2009, Mutation research.

[33]  Nicklas Raun Jacobsen,et al.  Pulmonary exposure to carbon black by inhalation or instillation in pregnant mice: Effects on liver DNA strand breaks in dams and offspring , 2012, Nanotoxicology.

[34]  Maria Dusinska,et al.  Can Standard Genotoxicity Tests be Applied to Nanoparticles? , 2012, Journal of toxicology and environmental health. Part A.

[35]  Jürgen Pauluhn,et al.  Subchronic 13-week inhalation exposure of rats to multiwalled carbon nanotubes: toxic effects are determined by density of agglomerate structures, not fibrillar structures. , 2010, Toxicological sciences : an official journal of the Society of Toxicology.

[36]  C. Schulze Transport of metal oxide nanoparticles across the human air-blood barrier : interactions with physiologically relevant media and proteins , 2010 .

[37]  Håkan Wallin,et al.  DNA Damage Following Pulmonary Exposure by Instillation to Low Doses of Carbon Black (Printex 90) Nanoparticles in Mice , 2014, Environmental and molecular mutagenesis.

[38]  P. White,et al.  Development and characterization of a stable epithelial cell line from Muta™Mouse lung , 2003, Environmental and molecular mutagenesis.

[39]  Robert H. Hurt,et al.  Bioavailability of Nickel in Single‐Wall Carbon Nanotubes , 2007 .

[40]  B. van Ravenzwaay,et al.  Inhalation toxicity of multiwall carbon nanotubes in rats exposed for 3 months. , 2009, Toxicological sciences : an official journal of the Society of Toxicology.

[41]  D. Koziej,et al.  Impact of sonication pretreatment on carbon nanotubes: A transmission electron microscopy study , 2013 .

[42]  Andrew Williams,et al.  Maternal inhalation of surface-coated nanosized titanium dioxide (UV-Titan) in C57BL/6 mice: effects in prenatally exposed offspring on hepatic DNA damage and gene expression , 2013, Nanotoxicology.

[43]  Andrij Holian,et al.  Effect of MWCNT size, carboxylation, and purification on in vitro and in vivo toxicity, inflammation and lung pathology , 2013, Particle and Fibre Toxicology.

[44]  U. Vogel,et al.  Nanotitanium dioxide toxicity in mouse lung is reduced in sanding dust from paint , 2012, Particle and Fibre Toxicology.

[45]  J. Kanno,et al.  Dose-dependent mesothelioma induction by intraperitoneal administration of multi-wall carbon nanotubes in p53 heterozygous mice , 2012, Cancer science.

[46]  Helinor J Johnston,et al.  Review of carbon nanotubes toxicity and exposure—Appraisal of human health risk assessment based on open literature , 2010, Critical reviews in toxicology.