Assessment of cultured fish hepatocytes for studying cellular uptake and (eco)toxicity of nanoparticles

Environmental context. The production and application of engineered nanoparticles is rapidly increasing, and development of suitable models for screening nanoparticles for possible toxic effects is essential to protect aquatic organisms and support the sustainable development of the nanotechnology industry. Here, the suitability of isolated rainbow trout hepatocytes was assessed for high through-put toxicity screening of nanoparticles and for studying uptake of nanoparticles into cells. Abstract. Relatively little is known regarding the fate and possible toxic effects of engineered nanoparticles (ENPs) in the aquatic environment. We assessed the suitability of isolated trout hepatocytes for high throughput toxicity screening of ENPs, exposing them to a variety of metal and metal oxide nanoparticles and their bulk counterparts. We found no effects of the ENPs on cell viability, or on lipid peroxidation, with the exception of exposure to ZnO nanoparticles, or on glutathione-S-transferase (GST) levels, for exposure concentrations up to 500 μg mL–1. All ENPs, however, were internalised in the cultured hepatocytes, as shown by coherent anti-Stokes Raman scattering (CARS) as an imaging technique. Our findings suggest that fish hepatocyte cultures are suitable for studies investigating the cellular uptake of ENPs, but they do not appear to be sensitive to ENP exposure and thus not a good in vitro model for nanoparticle toxicity screening.

[1]  H. Jeng,et al.  Toxicity of Metal Oxide Nanoparticles in Mammalian Cells , 2006, Journal of environmental science and health. Part A, Toxic/hazardous substances & environmental engineering.

[2]  Robert N Grass,et al.  Oxide nanoparticle uptake in human lung fibroblasts: effects of particle size, agglomeration, and diffusion at low concentrations. , 2005, Environmental science & technology.

[3]  J. Cravedi,et al.  Glutathione-S-transferase subunits pattern in rainbow trout isolated hepatocytes , 1998 .

[4]  J. Cravedi,et al.  Maintenance of cytochrome P450 content and phase I and phase II enzyme activities in trout hepatocytes cultured as spheroidal aggregates , 1996 .

[5]  Awadhesh N Jha,et al.  Hydroxyl radicals (*OH) are associated with titanium dioxide (TiO(2)) nanoparticle-induced cytotoxicity and oxidative DNA damage in fish cells. , 2008, Mutation research.

[6]  F. Kargın,et al.  Antioxidant responses and metal accumulation in tissues of Nile tilapia Oreochromis niloticus under Zn, Cd and Zn + Cd exposures , 2009, Journal of applied toxicology : JAT.

[7]  S. Raisuddin,et al.  Effects of exposure to multiple trace metals on biochemical, histological and ultrastructural features of gills of a freshwater fish, Channa punctata Bloch. , 2008, Chemico-biological interactions.

[8]  M. Remškar,et al.  Effects of ingested nano‐sized titanium dioxide on terrestrial isopods (Porcellio scaber) , 2008, Environmental toxicology and chemistry.

[9]  Zhi Pan,et al.  Adverse effects of titanium dioxide nanoparticles on human dermal fibroblasts and how to protect cells. , 2009, Small.

[10]  G. E. Gadd,et al.  Comparative toxicity of nanoparticulate ZnO, bulk ZnO, and ZnCl2 to a freshwater microalga (Pseudokirchneriella subcapitata): the importance of particle solubility. , 2007, Environmental science & technology.

[11]  M. Pérez-López,et al.  Glutathione S-transferase cytosolic isoforms as biomarkers of polychlorinated biphenyl (Arochlor-1254) experimental contamination in rainbow trout. , 2002, Toxicology letters.

[12]  S. Lockett,et al.  Coherent anti‐stokes Raman scattering microscopy: A biological review , 2006, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[13]  Prakash D Nallathamby,et al.  In vivo imaging of transport and biocompatibility of single silver nanoparticles in early development of zebrafish embryos. , 2007, ACS nano.

[14]  A. Meister.  Glutathione metabolism and its selective modification. , 1988, The Journal of biological chemistry.

[15]  Wolfgang Kreyling,et al.  Ultrafine Particles Cross Cellular Membranes by Nonphagocytic Mechanisms in Lungs and in Cultured Cells , 2005, Environmental health perspectives.

[16]  R. L. Jones,et al.  Unique cellular interaction of silver nanoparticles: size-dependent generation of reactive oxygen species. , 2008, The journal of physical chemistry. B.

[17]  P. Midgley,et al.  Visualizing the uptake of C60 to the cytoplasm and nucleus of human monocyte-derived macrophage cells using energy-filtered transmission electron microscopy and electron tomography. , 2007, Environmental science & technology.

[18]  F. Gagné,et al.  Cytotoxicity of aged cadmium-telluride quantum dots to rainbow trout hepatocytes , 2008 .

[19]  J. Girard,et al.  Toxic Effects of Wastewaters Collected at Upstream and Downstream Sites of a Purification Station in Cultures of Rainbow Trout Hepatocytes , 2001, Archives of environmental contamination and toxicology.

[20]  Claudio Leonzio,et al.  Heavy metals and glutathione metabolism in mussel tissues , 1999 .

[21]  J. Flaws,et al.  Methoxychlor and estradiol induce oxidative stress DNA damage in the mouse ovarian surface epithelium. , 2008, Toxicological sciences : an official journal of the Society of Toxicology.

[22]  N. Aykin-Burns,et al.  Toxic metals and oxidative stress part I: mechanisms involved in metal-induced oxidative damage. , 2001, Current topics in medicinal chemistry.

[23]  Awadhesh N. Jha,et al.  Genotoxic and cytotoxic potential of titanium dioxide (TiO2) nanoparticles on fish cells in vitro , 2008, Ecotoxicology.

[24]  R. Wallace,et al.  Relation of Oxygen and Temperature in the Preservation of Tissues by Refrigeration , 1949, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[25]  K. Jan,et al.  Ultrafine titanium dioxide particles in the absence of photoactivation can induce oxidative damage to human bronchial epithelial cells. , 2005, Toxicology.

[26]  D. Bagchi,et al.  Oxidative mechanisms in the toxicity of metal ions. , 1995, Free radical biology & medicine.

[27]  C. Tyler,et al.  Fish hepatocyte cultures as an alternative to in vivo tests for screening oestrogen receptor active chemicals , 2007 .

[28]  C. Vulpe,et al.  Toxicogenomics of water chemistry influence on chronic lead exposure to the fathead minnow (Pimephales promelas). , 2008, Aquatic toxicology.

[29]  Anita Jemec,et al.  In vivo screening to determine hazards of nanoparticles: nanosized TiO2. , 2009, Environmental pollution.

[30]  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 .

[31]  Ritesh K Shukla,et al.  DNA damaging potential of zinc oxide nanoparticles in human epidermal cells. , 2009, Toxicology letters.

[32]  E. Verwey,et al.  Theory of the stability of lyophobic colloids. , 1955, The Journal of physical and colloid chemistry.

[33]  J. Girard,et al.  Purification and partial characterization of seven glutathione S-transferase isoforms from the clam Ruditapes decussatus. , 2002, European journal of biochemistry.

[34]  J. Heyder,et al.  Oxidative stress and lipid mediators induced in alveolar macrophages by ultrafine particles. , 2005, Free radical biology & medicine.

[35]  C. Tyler,et al.  Evaluation of a carp primary hepatocyte culture system for screening chemicals for oestrogenic activity. , 2009, Aquatic toxicology.

[36]  Mengling Liu,et al.  Vaginal delivery and serum markers of ischemia/reperfusion injury , 2006, International journal of gynaecology and obstetrics: the official organ of the International Federation of Gynaecology and Obstetrics.

[37]  Tung-Sheng Shih,et al.  The apoptotic effect of nanosilver is mediated by a ROS- and JNK-dependent mechanism involving the mitochondrial pathway in NIH3T3 cells. , 2008, Toxicology letters.

[38]  B. Derjaguin,et al.  Theory of the stability of strongly charged lyophobic sols and of the adhesion of strongly charged particles in solutions of electrolytes , 1993 .

[39]  C. Wood,et al.  The physiology of waterborne silver toxicity in freshwater rainbow trout (Oncorhynchus mykiss) 1. The effects of ionic Ag , 1996 .

[40]  Chao Liu,et al.  Comparative study of cytotoxicity, oxidative stress and genotoxicity induced by four typical nanomaterials: the role of particle size, shape and composition , 2009, Journal of applied toxicology : JAT.

[41]  Borys Shuter,et al.  Solvent-free atom transfer radical polymerization for the preparation of poly(poly(ethyleneglycol) monomethacrylate)-grafted Fe3O4 nanoparticles: synthesis, characterization and cellular uptake. , 2007, Biomaterials.

[42]  Joel G Pounds,et al.  Particokinetics in vitro: dosimetry considerations for in vitro nanoparticle toxicity assessments. , 2007, Toxicological sciences : an official journal of the Society of Toxicology.

[43]  Jinhee Choi,et al.  Oxidative stress induced by cerium oxide nanoparticles in cultured BEAS-2B cells. , 2008, Toxicology.

[44]  J. Cravedi,et al.  Xenobiotic metabolizing enzyme activities in aggregate culture of rainbow trout hepatocytes , 1995 .

[45]  T. Braunbeck,et al.  Cytological and biochemical effects of a mixture of 20 pollutants on isolated rainbow trout (Oncorhynchus mykiss) hepatocytes. , 2002, Ecotoxicology and environmental safety.

[46]  Enrique Navarro,et al.  Toxicity of silver nanoparticles to Chlamydomonas reinhardtii. , 2008, Environmental science & technology.

[47]  V. Morsch,et al.  Exposure to sublethal concentrations of Zn(II) and Cu(II) changes biochemical parameters in Leporinus obtusidens. , 2007, Chemosphere.

[48]  K. Paknikar,et al.  Cellular responses induced by silver nanoparticles: In vitro studies. , 2008, Toxicology letters.

[49]  Alke Petri-Fink,et al.  Effect of cell media on polymer coated superparamagnetic iron oxide nanoparticles (SPIONs): colloidal stability, cytotoxicity, and cellular uptake studies. , 2008, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[50]  Vicki Stone,et al.  Efficacy of Simple Short-Term in Vitro Assays for Predicting the Potential of Metal Oxide Nanoparticles to Cause Pulmonary Inflammation , 2008, Environmental health perspectives.

[51]  J J Schlager,et al.  Expression of genes related to oxidative stress in the mouse brain after exposure to silver-25 nanoparticles. , 2009, Toxicology letters.

[52]  G. Atli,et al.  Response of catalase activity to Ag+, Cd2+, Cr6+, Cu2+ and Zn2+ in five tissues of freshwater fish Oreochromis niloticus. , 2006, Comparative biochemistry and physiology. Toxicology & pharmacology : CBP.

[53]  J. Lead,et al.  High doses of intravenously administered titanium dioxide nanoparticles accumulate in the kidneys of rainbow trout but with no observable impairment of renal function. , 2009, Toxicological sciences : an official journal of the Society of Toxicology.

[54]  Xuezhi Zhang,et al.  The impact of ZnO nanoparticle aggregates on the embryonic development of zebrafish (Danio rerio) , 2009, Nanotechnology.

[55]  C. Hogstrand,et al.  Influence of culture conditions on metal-induced responses in a cultured rainbow trout gill epithelium. , 2007, Environmental science & technology.

[56]  Jongheop Yi,et al.  Ecotoxicity of silver nanoparticles on the soil nematode Caenorhabditis elegans using functional ecotoxicogenomics. , 2009, Environmental science & technology.

[57]  Shuk Han Cheng,et al.  Acute and long-term effects after single loading of functionalized multi-walled carbon nanotubes into zebrafish (Danio rerio). , 2009, Toxicology and applied pharmacology.

[58]  Y. Korchev,et al.  Immortalization of human alveolar epithelial cells to investigate nanoparticle uptake. , 2008, American journal of respiratory cell and molecular biology.

[59]  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.

[60]  Julian Moger,et al.  Imaging metal oxide nanoparticles in biological structures with CARS microscopy. , 2008, Optics express.

[61]  J. Narbonne,et al.  Acute effects of cadmium on liver phase I and phase II enzymes and metallothionein accumulation on sea bream Sparus aurata , 2008, Fish Physiology and Biochemistry.

[62]  J. Frazier,et al.  Isolated fish hepatocytes — model systems for toxicology research☆ , 1990 .

[63]  Nathalie Tufenkji,et al.  Aggregation of titanium dioxide nanoparticles: role of a fulvic acid. , 2009, Environmental science & technology.

[64]  Jongheop Yi,et al.  Oxidative stress and apoptosis induced by titanium dioxide nanoparticles in cultured BEAS-2B cells. , 2008, Toxicology letters.

[65]  Min-Kyeong Yeo,et al.  Effects of Nanometer Sized Silver Materials on Biological Toxicity During Zebrafish Embryogenesis , 2008 .

[66]  Lawrence E Murr,et al.  Comparative in vitro cytotoxicity assessment of some manufacturednanoparticulate materials characterized by transmissionelectron microscopy , 2005 .

[67]  Richard D Handy,et al.  Manufactured nanoparticles: their uptake and effects on fish—a mechanistic analysis , 2008, Ecotoxicology.

[68]  J. West,et al.  Correlating nanoscale titania structure with toxicity: a cytotoxicity and inflammatory response study with human dermal fibroblasts and human lung epithelial cells. , 2006, Toxicological sciences : an official journal of the Society of Toxicology.

[69]  K. Je.  Establishment of fish hepatocyte cultures for use in in vitro carcinogenicity studies. , 1984 .

[70]  R. Aitken,et al.  Manufacture and use of nanomaterials: current status in the UK and global trends. , 2006, Occupational medicine.