title: Comprehensive In Vitro Toxicity Testing of a Panel of Representative Oxide Nanomaterials: First Steps towards an Intelligent Testing Strategy.
abstract: Nanomaterials (NMs) display many unique and useful physico-chemical properties. However, reliable approaches are needed for risk assessment of NMs. The present study was performed in the FP7-MARINA project, with the objective to identify and evaluate in vitro test methods for toxicity assessment in order to facilitate the development of an intelligent testing strategy (ITS). Six representative oxide NMs provided by the EC-JRC Nanomaterials Repository were tested in nine laboratories. The in vitro toxicity of NMs was evaluated in 12 cellular models representing 6 different target organs/systems (immune system, respiratory system, gastrointestinal system, reproductive organs, kidney and embryonic tissues). The toxicity assessment was conducted using 10 different assays for cytotoxicity, embryotoxicity, epithelial integrity, cytokine secretion and oxidative stress. Thorough physico-chemical characterization was performed for all tested NMs. Commercially relevant NMs with different physico-chemical properties were selected: two TiO2 NMs with different surface chemistry - hydrophilic (NM-103) and hydrophobic (NM-104), two forms of ZnO - uncoated (NM-110) and coated with triethoxycapryl silane (NM-111) and two SiO2 NMs produced by two different manufacturing techniques - precipitated (NM-200) and pyrogenic (NM-203). Cell specific toxicity effects of all NMs were observed; macrophages were the most sensitive cell type after short-term exposures (24-72h) (ZnO>SiO2>TiO2). Longer term exposure (7 to 21 days) significantly affected the cell barrier integrity in the presence of ZnO, but not TiO2 and SiO2, while the embryonic stem cell test (EST) classified the TiO2 NMs as potentially 'weak-embryotoxic' and ZnO and SiO2 NMs as 'non-embryotoxic'. A hazard ranking could be established for the representative NMs tested (ZnO NM-110 > ZnO NM-111 > SiO2 NM-203 > SiO2 NM-200 > TiO2 NM-104 > TiO2 NM-103). This ranking was different in the case of embryonic tissues, for which TiO2 displayed higher toxicity compared with ZnO and SiO2. Importantly, the in vitro methodology applied could identify cell- and NM-specific responses, with a low variability observed between different test assays. Overall, this testing approach, based on a battery of cellular systems and test assays, complemented by an exhaustive physico-chemical characterization of NMs, could be deployed for the development of an ITS suitable for risk assessment of NMs. This study also provides a rich source of data for modeling of NM effects.
authors.affiliation: Division of Molecular Toxicology, Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Lucian
authors.initials: L
authors.lastname: Farcal
authors.affiliation: Division of Molecular Toxicology, Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Fernando
authors.initials: F
authors.lastname: Torres Andón
authors.affiliation: Department of Clinical and Experimental Medicine, University of Parma, Parma, Italy.
authors.forename: Luisana
authors.initials: L
authors.lastname: Di Cristo
authors.affiliation: Department of Biomedical, Biotechnological and Translational Sciences, University of Parma, Parma, Italy.
authors.forename: Bianca Maria
authors.initials: BM
authors.lastname: Rotoli
authors.affiliation: Department of Biomedical, Biotechnological and Translational Sciences, University of Parma, Parma, Italy.
authors.forename: Ovidio
authors.initials: O
authors.lastname: Bussolati
authors.affiliation: Department of Clinical and Experimental Medicine, University of Parma, Parma, Italy.
authors.forename: Enrico
authors.initials: E
authors.lastname: Bergamaschi
authors.affiliation: Nanobiosciences Unit, Institute for Health and Consumer Protection, European Commission-Joint Research Centre, Ispra, Italy.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Mech
authors.affiliation: Nanobiosciences Unit, Institute for Health and Consumer Protection, European Commission-Joint Research Centre, Ispra, Italy; Technical University of Denmark, Kongens Lyngby, Denmark.
authors.forename: Nanna B
authors.initials: NB
authors.lastname: Hartmann
authors.affiliation: Nanobiosciences Unit, Institute for Health and Consumer Protection, European Commission-Joint Research Centre, Ispra, Italy.
authors.forename: Kirsten
authors.initials: K
authors.lastname: Rasmussen
authors.affiliation: Nanobiosciences Unit, Institute for Health and Consumer Protection, European Commission-Joint Research Centre, Ispra, Italy.
authors.forename: Juan
authors.initials: J
authors.lastname: Riego-Sintes
authors.affiliation: Nanobiosciences Unit, Institute for Health and Consumer Protection, European Commission-Joint Research Centre, Ispra, Italy.
authors.forename: Jessica
authors.initials: J
authors.lastname: Ponti
authors.affiliation: Nanobiosciences Unit, Institute for Health and Consumer Protection, European Commission-Joint Research Centre, Ispra, Italy.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Kinsner-Ovaskainen
authors.affiliation: Nanobiosciences Unit, Institute for Health and Consumer Protection, European Commission-Joint Research Centre, Ispra, Italy.
authors.forename: François
authors.initials: F
authors.lastname: Rossi
authors.affiliation: National Institute for Public Health and the Environment, Bilthoven, The Netherlands.
authors.forename: Agnes
authors.initials: A
authors.lastname: Oomen
authors.affiliation: National Institute for Public Health and the Environment, Bilthoven, The Netherlands.
authors.forename: Peter
authors.initials: P
authors.lastname: Bos
authors.affiliation: Chinese Academy of Sciences Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, National Center for Nanoscience & Technology of China, Beijing, P. R. China.
authors.forename: Rui
authors.initials: R
authors.lastname: Chen
authors.affiliation: Chinese Academy of Sciences Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, National Center for Nanoscience & Technology of China, Beijing, P. R. China.
authors.forename: Ru
authors.initials: R
authors.lastname: Bai
authors.affiliation: Chinese Academy of Sciences Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, National Center for Nanoscience & Technology of China, Beijing, P. R. China.
authors.forename: Chunying
authors.initials: C
authors.lastname: Chen
authors.affiliation: Centre for BioNano Interactions, School of Chemistry and Chemical Biology, University College Dublin, Belfield, Dublin, Ireland.
authors.forename: Louise
authors.initials: L
authors.lastname: Rocks
authors.affiliation: Centre for Nano Safety, Edinburgh Napier University, Edinburgh, United Kingdom.
authors.forename: Norma
authors.initials: N
authors.lastname: Fulton
authors.affiliation: Centre for Nano Safety, Edinburgh Napier University, Edinburgh, United Kingdom; Institute of Occupational Medicine, Edinburgh, United Kingdom.
authors.forename: Bryony
authors.initials: B
authors.lastname: Ross
authors.affiliation: Centre for Nano Safety, Edinburgh Napier University, Edinburgh, United Kingdom.
authors.forename: Gary
authors.initials: G
authors.lastname: Hutchison
authors.affiliation: Institute of Occupational Medicine, Edinburgh, United Kingdom.
authors.forename: Lang
authors.initials: L
authors.lastname: Tran
authors.affiliation: Biomedizinisches Technologiezentrum, Westfälische Wilhelms-Universität, Münster, Germany.
authors.forename: Sarah
authors.initials: S
authors.lastname: Mues
authors.affiliation: Biomedizinisches Technologiezentrum, Westfälische Wilhelms-Universität, Münster, Germany.
authors.forename: Rainer
authors.initials: R
authors.lastname: Ossig
authors.affiliation: Biomedizinisches Technologiezentrum, Westfälische Wilhelms-Universität, Münster, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Schnekenburger
authors.affiliation: Department of Biomedicine and Prevention, University of Rome Tor Vergata, Rome, Italy.
authors.forename: Luisa
authors.initials: L
authors.lastname: Campagnolo
authors.affiliation: Department of Biomedicine and Prevention, University of Rome Tor Vergata, Rome, Italy.
authors.forename: Lucia
authors.initials: L
authors.lastname: Vecchione
authors.affiliation: Department of Biomedicine and Prevention, University of Rome Tor Vergata, Rome, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Pietroiusti
authors.affiliation: Division of Molecular Toxicology, Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Bengt
authors.initials: B
authors.lastname: Fadeel
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D053595:Embryonic Stem Cells / Q000187:drug effects; D004847:Epithelial Cells / Q000187:drug effects; D006801:Humans; D066298:In Vitro Techniques; D020128:Inhibitory Concentration 50; D007985:Leydig Cells / Q000187:drug effects; D008264:Macrophages / Q000187:drug effects; D008297:Male; D051379:Mice; D049329:Nanostructures / Q000737:chemistry* / Q000633:toxicity*; D010087:Oxides / Q000737:chemistry* / Q000633:toxicity*; D012708:Sertoli Cells / Q000187:drug effects; D012822:Silicon Dioxide; D014025:Titanium; D018675:Toxicity Tests; D015034:Zinc Oxide
chemical_list: D010087:Oxides; C009495:titanium dioxide; D012822:Silicon Dioxide; D014025:Titanium; D015034:Zinc Oxide