title: A panel of in vitro tests to evaluate genotoxic and morphological neoplastic transformation potential on Balb/3T3 cells by pristine and remediated titania and zirconia nanoparticles.
abstract: The FP7 Sanowork project was aimed to minimise occupational hazard and exposure to engineered nanomaterials (ENM) through the surface modification in order to prevent possible health effects. In this frame, a number of nanoparticles (NP) have been selected, among which zirconium (ZrO2) and titanium (TiO2) dioxide. In this study, we tested ZrO2 NP and TiO2 NP either in their pristine (uncoated) form, or modified with citrate and/or silica on their surface. As benchmark material, Aeroxide® P25 was used. We assessed cytotoxicity, genotoxicity and induction of morphological neoplastic transformation of NP by using a panel of in vitro assays in an established mammalian cell line of murine origin (Balb/3T3). Cell viability was evaluated by means of colony-forming efficiency assay (CFE). Genotoxicity was investigated by cytokinesis-block micronucleus cytome assay (CBMN cyt) and comet assay, and by the use of the restriction enzymes EndoIII and Fpg, oxidatively damaged DNA was detected; finally, the morphological neoplastic transformation of NP was assayed in vitro by cell transformation assay (CTA). Our results show that the surface remediation has not been effective in modifying cyto- and genotoxic properties of the nanomaterials tested; indeed, in the case of remediation of zirconia and titania with citrate, there is a tendency to emphasise the toxic effects. The use of a panel of assays, such as those we have employed, allowing the evaluation of multiple endpoints, including cell transformation, seems particularly advisable especially in the case of long-term exposure effects in the same cell type.
authors.affiliation: Laboratory of Medical Genetics, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, Via Roma 55, Pisa 56126, Italy, Doctoral School in GeneticsOncology and Clinical MedicineDepartment of Medical BiotechnologiesUniversity of Siena, 53100 Siena, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Stoccoro
authors.affiliation: Laboratory of Medical Genetics, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, Via Roma 55, Pisa 56126, Italy.
authors.forename: Sebastiano
authors.initials: S
authors.lastname: Di Bucchianico
authors.affiliation: Laboratory of Medical Genetics, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, Via Roma 55, Pisa 56126, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Uboldi
authors.affiliation: Laboratory of Medical Genetics, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, Via Roma 55, Pisa 56126, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Coppedè
authors.affiliation: European Commission, Joint Research Centre (JRC), Institute for Health and Consumer Protection (IHCP), Nanobiosciences (NBS) Unit, via E. Fermi 2749, 21027 Ispra (VA), Italy.
authors.forename: Jessica
authors.initials: J
authors.lastname: Ponti
authors.affiliation: Department of Pathology, Ospedale di Circolo and Department of Human Morphology, University of Insubria,via Ottorino Rossi 9, 21100 Varese, VA, Italy and.
authors.forename: Claudia
authors.initials: C
authors.lastname: Placidi
authors.affiliation: Institute of Science and Technology for Ceramics (CNR-ISTEC), National Research Council of Italy, Via Granarolo 64, 48018 Faenza, RA, Italy.
authors.forename: Magda
authors.initials: M
authors.lastname: Blosi
authors.affiliation: Institute of Science and Technology for Ceramics (CNR-ISTEC), National Research Council of Italy, Via Granarolo 64, 48018 Faenza, RA, Italy.
authors.forename: Simona
authors.initials: S
authors.lastname: Ortelli
authors.affiliation: Institute of Science and Technology for Ceramics (CNR-ISTEC), National Research Council of Italy, Via Granarolo 64, 48018 Faenza, RA, Italy.
authors.forename: Anna Luisa
authors.initials: AL
authors.lastname: Costa
authors.affiliation: Laboratory of Medical Genetics, Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, Via Roma 55, Pisa 56126, Italy, lucia.migliore@med.unipi.it.
authors.forename: Lucia
authors.initials: L
authors.lastname: Migliore
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D002471:Cell Transformation, Neoplastic / Q000139:chemically induced*; D004247:DNA / Q000187:drug effects; D004249:DNA Damage; D053768:Metal Nanoparticles / Q000737:chemistry / Q000633:toxicity*; D051379:Mice; D009152:Mutagenicity Tests; D018384:Oxidative Stress; D014025:Titanium / Q000494:pharmacology / Q000633:toxicity*; D015040:Zirconium / Q000494:pharmacology / Q000633:toxicity*
chemical_list: C009495:titanium dioxide; D004247:DNA; D015040:Zirconium; D014025:Titanium; C028541:zirconium oxide