title: Aneuploidogenic effects and DNA oxidation induced in vitro by differently sized gold nanoparticles.
abstract: Gold nanoparticles (Au NPs) are used in many fields, including biomedical applications; however, no conclusive information on their potential cytotoxicity and genotoxicity mechanisms is available. For this reason, experiments in human primary lymphocytes and murine macrophages (Raw264.7) were performed exposing cells to spherical citrate-capped Au NPs with two different nominal diameters (5 nm and 15 nm). The proliferative activity, mitotic, apoptotic, and necrotic markers, as well as chromosomal damage were assessed by the cytokinesis-block micronucleus cytome assay. Fluorescence in situ hybridization with human and murine pancentromeric probes was applied to distinguish between clastogenic and aneuploidogenic effects. Our results indicate that 5 nm and 15 nm Au NPs are able to inhibit cell proliferation by apoptosis and to induce chromosomal damage, in particular chromosome mis-segregation. DNA strand breaks were detected by comet assay, and the modified protocol using endonuclease-III and formamidopyrimidine-DNA glycosylase restriction enzymes showed that pyrimidines and purines were oxidatively damaged by Au NPs. Moreover, we show a size-independent correlation between the cytotoxicity of Au NPs and their tested mass concentration or absolute number, and genotoxic effects which were more severe for Au NP 15 nm compared to Au NP 5 nm. Results indicate that apoptosis, aneuploidy, and DNA oxidation play a pivotal role in the cytotoxicity and genotoxicity exerted by Au NPs in our cell models.
authors.affiliation: Department of Translational Research and New Technologies in Medicine and Surgery, Medical Genetics Unit, University of Pisa, Pisa, Italy.
authors.forename: Sebastiano
authors.initials: S
authors.lastname: Di Bucchianico
authors.affiliation: Department of Translational Research and New Technologies in Medicine and Surgery, Medical Genetics Unit, University of Pisa, Pisa, Italy.
authors.forename: Maria Rita
authors.initials: MR
authors.lastname: Fabbrizi
authors.affiliation: Department of Translational Research and New Technologies in Medicine and Surgery, Medical Genetics Unit, University of Pisa, Pisa, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Cirillo
authors.affiliation: Department of Translational Research and New Technologies in Medicine and Surgery, Medical Genetics Unit, University of Pisa, Pisa, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Uboldi
authors.affiliation: European Commission-Joint Research Centre, Institute for Health and Consumer Protection, NanoBioSciences Unit, Ispra, Italy.
authors.forename: Douglas
authors.initials: D
authors.lastname: Gilliland
authors.affiliation: School of Geography, Earth and Environmental Sciences, University of Birmingham, Edgbaston, Birmingham, UK ; Earth Sciences, Natural History Museum, Cromwell Road, London, UK.
authors.forename: Eugenia
authors.initials: E
authors.lastname: Valsami-Jones
authors.affiliation: Department of Translational Research and New Technologies in Medicine and Surgery, Medical Genetics Unit, University of Pisa, Pisa, Italy.
authors.forename: Lucia
authors.initials: L
authors.lastname: Migliore
mesh_terms: D024422:Aneugens / Q000494:pharmacology*; D000782:Aneuploidy; D000818:Animals; D002869:Chromosome Aberrations / Q000139:chemically induced*; D004249:DNA Damage / Q000502:physiology*; D006046:Gold / Q000494:pharmacology*; D006801:Humans; D008264:Macrophages / Q000187:drug effects / Q000502:physiology*; D008422:Materials Testing; D053768:Metal Nanoparticles / Q000008:administration & dosage*; D051379:Mice; D010084:Oxidation-Reduction / Q000187:drug effects*; D010316:Particle Size
chemical_list: D024422:Aneugens; D006046:Gold
keywords: Au nanoparticles; aneuploidy; cytotoxicity; micronuclei; oxidative DNA damage; particle size