Evaluation of titanium dioxide nanocrystal-induced genotoxicity by the cytokinesis-block micronucleus assay and the Drosophila wing spot test

•Anatase and rutile TiO2 NCs induce mutagenic activity in Drosophila melanogaster.•Anatase and rutile TiO2 NCs are mutagenic only in the HB crosses of SMART assay.•There are negative correlation between size and mutagenicity of anatase TiO2 NCs.•Only the smallest size of anatase TiO2 NCs induces micronucleus formation.

Titanium dioxide nanocrystals (TiO2 NCs) crystalline structures include anatase, rutile and brookite. This study evaluated the genotoxic effects of 3.4 and 6.2 nm anatase TiO2 NCs and 78.0 nm predominantly rutile TiO2 NCs through an in vitro micronucleus (MN) assay using V79 cells and an in vivo somatic mutation and recombination test in Drosophila wings. The MN assay was performed with nontoxic concentrations of TiO2 NCs. Only anatase (3.4 nm) at the highest concentration (120 μM) induced genotoxicity in V79 cells. In the in vivo test, Drosophila melanogaster larvae obtained from standard (ST) or high bioactivation (HB) crosses were treated with TiO2 NCs. In the ST cross, no mutagenic effects were observed. However, in the HB cross, TiO2 NCs (3.4 nm) were mutagenic at 1.5625 and 3.125 mM, while 78.0 nm NCs increased mutant spots at all concentrations tested except 3.125 mM. Only the smallest anatase TiO2 NCs induced mutagenic effects in vitro and in vivo. For rutile TiO2 NCs, no clastogenic/aneugenic effects were observed in the MN assay. However, they were mutagenic in Drosophila. Therefore, both anatase and rutile TiO2 NCs induced mutagenicity. Further research is necessary to clarify the TiO2 NCs genotoxic/mutagenic action mechanisms.

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