An in vitro liver model - assessing oxidative stress and genotoxicity following exposure of hepatocytes to a panel of engineered nanomaterials

BackgroundFollowing exposure via inhalation, intratracheal instillation or ingestion some nanomaterials (NM) have been shown to translocate to the liver. Since oxidative stress has been implicated as a possible mechanism for NM toxicity this study aimed to investigate the effects of various materials (five titanium dioxide (TiO2), two zinc oxide (ZnO), two multi-walled carbon nanotubes (MWCNT) and one silver (Ag) NM) on oxidative responses of C3A cell line as a model for potential detrimental properties of nanomaterials on the liver.ResultsWe noted a dose dependant decrease in the cellular glutathione content following exposure of the C3A cells to Ag, the ZnO and the MWCNTs. Intracellular ROS levels were also measured and shown to increase significantly following exposure of the C3A to the low toxicity NMs (MWCNT and TiO2). The antioxidant Trolox in part prevented the detrimental effect of NMs on cell viability, and decreased the NM induced IL8 production after exposure to all but the Ag particulate. Following 4 hr exposure of the C3A cells to sub-lethal levels of the NMs, the largest amount of DNA damage was induced by two of the TiO2 samples (7 nm and the positively charged 10 nm particles).ConclusionsAll ten NMs exhibited effects on the hepatocyte cell line that were at least in part ROS/oxidative stress mediated. These effects included mild genotoxicity and IL8 production for all NM except the Ag possibly due to its highly cytotoxic nature.

[1]  Z. Kmieć,et al.  Introduction — Morphology of the Liver Lobule , 2001 .

[2]  F. Hong,et al.  Hepatocyte apoptosis and its molecular mechanisms in mice caused by titanium dioxide nanoparticles. , 2010, Journal of hazardous materials.

[3]  E. Friedberg,et al.  Out of the shadows and into the light: the emergence of DNA repair. , 1995, Trends in biochemical sciences.

[4]  P. Tchounwou,et al.  Biochemical and histopathological evaluation of functionalized single‐walled carbon nanotubes in Swiss–Webster mice , 2011, Journal of applied toxicology : JAT.

[5]  Hassan S. Bazzi,et al.  Differences in subcellular distribution and toxicity of green and red emitting CdTe quantum dots , 2005, Journal of Molecular Medicine.

[6]  T. Xia,et al.  Toxic Potential of Materials at the Nanolevel , 2006, Science.

[7]  T. Dalton,et al.  Determining glutathione and glutathione disulfide using the fluorescence probe o-phthalaldehyde. , 2000, Analytical biochemistry.

[8]  Nicklas Raun Jacobsen,et al.  Biodistribution of gold nanoparticles in mouse lung following intratracheal instillation , 2009, Chemistry Central journal.

[9]  Ming Jiang,et al.  Synergistic genotoxicity caused by low concentration of titanium dioxide nanoparticles and p,p′‐DDT in human hepatocytes , 2009, Environmental and molecular mutagenesis.

[10]  M. Carrière,et al.  In vitro investigation of TiO2, Al2O3, Au nanoparticles and mutli-walled carbon nanotubes cyto- and genotoxicity on lung, kidney cells and hepatocytes , 2007 .

[11]  Ritesh K Shukla,et al.  ROS-mediated genotoxicity induced by titanium dioxide nanoparticles in human epidermal cells. , 2011, Toxicology in vitro : an international journal published in association with BIBRA.

[12]  P. Kuo,et al.  Nitric oxide and redox regulation in the liver: Part I. General considerations and redox biology in hepatitis. , 2010, The Journal of surgical research.

[13]  H. Karlsson,et al.  DNA damage induced by micro- and nanoparticles--interaction with FPG influences the detection of DNA oxidation in the comet assay. , 2012, Mutagenesis.

[14]  Marianne Geiser,et al.  Deposition and biokinetics of inhaled nanoparticles , 2010, Particle and Fibre Toxicology.

[15]  K. Kang,et al.  Mechanism of hepatic ischemia/reperfusion injury and protection against reperfusion injury. , 2002, Transplantation proceedings.

[16]  Stig Johan Wiklund,et al.  Aspects of design and statistical analysis in the Comet assay. , 2003, Mutagenesis.

[17]  Manuela Semmler-Behnke,et al.  Biodistribution of PEG-modified gold nanoparticles following intratracheal instillation and intravenous injection. , 2010, Biomaterials.

[18]  K. Paknikar,et al.  Interactions of silver nanoparticles with primary mouse fibroblasts and liver cells. , 2009, Toxicology and applied pharmacology.

[19]  Z. Kmieć,et al.  Cooperation of Liver Cells in Health and Disease , 2001, Advances in Anatomy Embryology and Cell Biology.

[20]  Manuela Semmler-Behnke,et al.  Biodistribution of 1.4- and 18-nm gold particles in rats. , 2008, Small.

[21]  W. Kreyling Lung deposition and biokinetics of inhaled nanoparticles , 2013 .

[22]  P. Hoet,et al.  Nanoparticles – known and unknown health risks , 2004, Journal of nanobiotechnology.

[23]  N. Holbrook,et al.  Cellular response to oxidative stress: Signaling for suicide and survival * , 2002, Journal of cellular physiology.

[24]  J. Gearhart,et al.  In vitro toxicity of nanoparticles in BRL 3A rat liver cells. , 2005, Toxicology in vitro : an international journal published in association with BIBRA.

[25]  Günter Speit,et al.  Sensitivity of the FPG protein towards alkylation damage in the comet assay. , 2004, Toxicology letters.

[26]  M. Ahamed,et al.  Zinc oxide nanoparticles selectively induce apoptosis in human cancer cells through reactive oxygen species , 2012, International journal of nanomedicine.

[27]  F. Hong,et al.  P38-Nrf-2 Signaling Pathway of Oxidative Stress in Mice Caused by Nanoparticulate TiO2 , 2011, Biological Trace Element Research.

[28]  David M. Brown,et al.  Calcium and ROS-mediated activation of transcription factors and TNF-alpha cytokine gene expression in macrophages exposed to ultrafine particles. , 2003, American journal of physiology. Lung cellular and molecular physiology.

[29]  K. Landfester,et al.  The effect of carboxydextran-coated superparamagnetic iron oxide nanoparticles on c-Jun N-terminal kinase-mediated apoptosis in human macrophages. , 2010, Biomaterials.

[30]  Jin Won Hyun,et al.  Silver nanoparticles induce oxidative cell damage in human liver cells through inhibition of reduced glutathione and induction of mitochondria-involved apoptosis. , 2011, Toxicology letters.

[31]  R. Abraham Cell cycle checkpoint signaling through the ATM and ATR kinases. , 2001, Genes & development.

[32]  A. Seifalian,et al.  The contemporary role of antioxidant therapy in attenuating liver ischemia‐reperfusion injury: A review , 2005, Liver transplantation : official publication of the American Association for the Study of Liver Diseases and the International Liver Transplantation Society.

[33]  D. Weiss,et al.  Human health implications of nanomaterial exposure , 2008 .

[34]  David M. Brown,et al.  An investigation into the potential for different surface-coated quantum dots to cause oxidative stress and affect macrophage cell signalling in vitro , 2010, Nanotoxicology.

[35]  Vicki Stone,et al.  The effects of PM10 particles and oxidative stress on macrophages and lung epithelial cells: modulating effects of calcium-signaling antagonists. , 2007, American journal of physiology. Lung cellular and molecular physiology.