DNA Damage Following Pulmonary Exposure by Instillation to Low Doses of Carbon Black (Printex 90) Nanoparticles in Mice

We previously observed genotoxic effects of carbon black nanoparticles at low doses relative to the Danish Occupational Exposure Limit (3.5 mg/m3). Furthermore, DNA damage occurred in broncho‐alveolar lavage (BAL) cells in the absence of inflammation, indicating that inflammation is not required for the genotoxic effects of carbon black. In this study, we investigated inflammatory and acute phase response in addition to genotoxic effects occurring following exposure to nanoparticulate carbon black (NPCB) at even lower doses. C57BL/6JBomTac mice were examined 1, 3, and 28 days after a single instillation of 0.67, 2, 6, and 162 µg Printex 90 NPCB and vehicle. Cellular composition and protein concentration was evaluated in BAL fluid as markers of inflammatory response and cell damage. DNA strand breaks in BAL cells, lung, and liver tissue were assessed using the alkaline comet assay. The pulmonary acute phase response was analyzed by Saa3 mRNA real‐time quantitative PCR. Instillation of the low doses of NPCB induced a slight neutrophil influx one day after exposure. Pulmonary exposure to small doses of NPCB caused an increase in DNA strand breaks in BAL cells and lung tissue measured using the comet assay. We interpret the increased DNA strand breaks occurring following these low exposure doses of NPCB as DNA damage caused by primary genotoxicity in the absence of substantial inflammation, cell damage, and acute phase response. Environ. Mol. Mutagen. 56:41–49, 2015. © 2014 The Authors. Environmental and Molecular Mutagenesis published by Wiley Periodicals, Inc. on behalf of Environmental Mutagen Society

[1]  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.

[2]  Steffen Loft,et al.  Inflammatory and genotoxic effects of diesel particles in vitro and in vivo. , 2004, Mutation research.

[3]  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.

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

[5]  Amaya Azqueta,et al.  The essential comet assay: a comprehensive guide to measuring DNA damage and repair , 2013, Archives of Toxicology.

[6]  J. Nagy,et al.  Discovery of a potent nanoparticle P‐selectin antagonist with anti‐inflammatory effects in allergic airway disease , 2003, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[7]  J. Heyder,et al.  Pulmonary and systemic effects of short-term inhalation exposure to ultrafine carbon black particles. , 2004, Toxicology and applied pharmacology.

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

[9]  Håkan Wallin,et al.  In utero exposure to nanosized carbon black (Printex90) does not induce tandem repeat mutations in female murine germ cells. , 2013, Reproductive toxicology.

[10]  Robert Gelein,et al.  Equivalent titanium dioxide nanoparticle deposition by intratracheal instillation and whole body inhalation: the effect of dose rate on acute respiratory tract inflammation , 2014, Particle and Fibre Toxicology.

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

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

[13]  Kurt Straif,et al.  Carcinogenicity of carbon black, titanium dioxide, and talc. , 2006, The Lancet Oncology.

[14]  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.

[15]  Marianne van der Hagen,et al.  The Nordic Expert Group for Criteria Documentation of Health Risks from Chemicals , 2006 .

[16]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[17]  L. Gallo Cardiovascular Disease , 1995, GWUMC Department of Biochemistry Annual Spring Symposia.

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

[19]  U. Vogel,et al.  Lack of acute phase response in the livers of mice exposed to diesel exhaust particles or carbon black by inhalation , 2009, Particle and Fibre Toxicology.

[20]  U. Vogel,et al.  Modest effect on plaque progression and vasodilatory function in atherosclerosis-prone mice exposed to nanosized TiO2 , 2011, Particle and Fibre Toxicology.

[21]  A. Zuckerman,et al.  IARC Monographs on the Evaluation of Carcinogenic Risks to Humans , 1995, IARC monographs on the evaluation of carcinogenic risks to humans.

[22]  G. Taubes Does Inflammation Cut to the Heart of the Matter? , 2002, Science.

[23]  P. Ridker,et al.  C-reactive protein and other markers of inflammation in the prediction of cardiovascular disease in women. , 2000, The New England journal of medicine.

[24]  Steffen Loft,et al.  Cytokine expression in mice exposed to diesel exhaust particles by inhalation. Role of tumor necrosis factor , 2006, Particle and Fibre Toxicology.

[25]  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.

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

[27]  U. Vogel,et al.  An experimental protocol for maternal pulmonary exposure in developmental toxicology. , 2011, Basic & clinical pharmacology & toxicology.

[28]  Takehiko Nohmi,et al.  Genotoxicity of nano/microparticles in in vitro micronuclei, in vivo comet and mutation assay systems , 2009, Particle and Fibre Toxicology.

[29]  Dongmei Wu,et al.  Exposure of pregnant mice to carbon black by intratracheal instillation: toxicogenomic effects in dams and offspring. , 2012, Mutation research.

[30]  Håkan Wallin,et al.  Prenatal exposure to carbon black (printex 90): effects on sexual development and neurofunction. , 2011, Basic & clinical pharmacology & toxicology.

[31]  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.

[32]  J. Gallacher,et al.  C-reactive protein, fibrinogen, and cardiovascular disease prediction. , 2012, The New England journal of medicine.

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

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

[35]  U. Vogel,et al.  Tumor necrosis factor is not required for particle-induced genotoxicity and pulmonary inflammation , 2005, Archives of Toxicology.