title: In vitro and in vivo genotoxicity investigations of differently sized amorphous SiO2 nanomaterials.
abstract: In vitro and in vivo genotoxic effects of differently sized amorphous SiO2 nanomaterials were investigated. In the alkaline Comet assay (with V79 cells), non-cytotoxic concentrations of 300 and 100-300μg/mL 15nm-SiO2 and 55nm-SiO2, respectively, relevant (at least 2-fold relative to the negative control) DNA damage. In the Alkaline unwinding assay (with V79 cells), only 15nm-SiO2 significantly increased DNA strand breaks (and only at 100μg/mL), whereas neither nanomaterial (up to 300μg/mL) increased Fpg (Formamidopyrimidine DNA glycosylase)-sensitive sites reflecting oxidative DNA base modifications. In the Comet assay using rat precision-cut lung slices, 15nm-SiO2 and 55nm-SiO2 induced significant DNA damage at ≥100μg/mL. In the Alkaline unwinding assay (with A549 cells), 30nm-SiO2 and 55nm-SiO2 (with larger primary particle size (PPS)) induced significant increases in DNA strand breaks at ≥50μg/mL, whereas 9nm-SiO2 and 15nm-SiO2 (with smaller PPS) induced significant DNA damage at higher concentrations. These two amorphous SiO2 also increased Fpg-sensitive sites (significant at 100μg/mL). In vivo, within 3 days after single intratracheal instillation of 360μg, neither 15nm-SiO2 nor 55nm-SiO2 caused genotoxic effects in the rat lung or in the bone marrow. However, pulmonary inflammation was observed in both test groups with findings being more pronounced upon treatment with 15nm-SiO2 than with 55nm-SiO2. Taken together, the study shows that colloidal amorphous SiO2 with different particle sizes may induce genotoxic effects in lung cells in vitro at comparatively high concentrations. However, the same materials elicited no genotoxic effects in the rat lung even though pronounced pulmonary inflammation evolved. This may be explained by the fact that a considerably lower dose reached the target cells in vivo than in vitro. Additionally, the different time points of investigation may provide more time for DNA damage repair after instillation.
authors.affiliation: Karlsruhe Institute of Technology (KIT), Institute of Applied Biosciences, Dept. of Food Chemistry and Toxicology, Karlsruhe, Germany.
authors.forename: Elena
authors.initials: E
authors.lastname: Maser
authors.affiliation: Experimental Toxicology and Ecology, BASF SE, Ludwigshafen, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Schulz
authors.affiliation: Scientific Consultancy-Animal Welfare, Neubiberg, Germany.
authors.forename: Ursula G
authors.initials: UG
authors.lastname: Sauer
authors.affiliation: IBE R&D gGmbH, Institute for Lung Health, Münster, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Wiemann
authors.affiliation: Experimental Toxicology and Ecology, BASF SE, Ludwigshafen, Germany.
authors.forename: Lan
authors.initials: L
authors.lastname: Ma-Hock
authors.affiliation: Experimental Toxicology and Ecology, BASF SE, Ludwigshafen, Germany; Material Physics, BASF SE, Ludwigshafen, Germany.
authors.forename: Wendel
authors.initials: W
authors.lastname: Wohlleben
authors.affiliation: Karlsruhe Institute of Technology (KIT), Institute of Applied Biosciences, Dept. of Food Chemistry and Toxicology, Karlsruhe, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Hartwig
authors.affiliation: Experimental Toxicology and Ecology, BASF SE, Ludwigshafen, Germany. Electronic address: robert.landsiedel@basf.com.
authors.forename: Robert
authors.initials: R
authors.lastname: Landsiedel
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D020552:Comet Assay; D004249:DNA Damage / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D005260:Female; D006801:Humans; D008168:Lung / Q000166:cytology / Q000187:drug effects; D008297:Male; D015162:Micronucleus Tests; D049329:Nanostructures / Q000737:chemistry / Q000633:toxicity*; D018384:Oxidative Stress / Q000187:drug effects; D010316:Particle Size; D051381:Rats; D017208:Rats, Wistar; D012822:Silicon Dioxide / Q000633:toxicity*
chemical_list: D012822:Silicon Dioxide
keywords: In vitro Alkaline unwinding assay; In vitro alkaline Comet assay; In vivo Comet assay; In vivo micronucleus test; Intratracheal instillation; Synthetic amorphous silica nanoparticles