Genotoxicity of short single-wall and multi-wall carbon nanotubes in human bronchial epithelial and mesothelial cells in vitro

Although some types of carbon nanotubes (CNTs) have been described to induce mesothelioma in rodents and genotoxic effects in various cell systems, there are few previous studies on the genotoxicity of CNTs in mesothelial cells. Here, we examined in vitro DNA damage induction by short multi-wall CNTs (MWCNTs; 10-30 nm x 1-2 mu m) and single-wall CNTs (SWCNTs; >50% SWCNTs, similar to 40% other CNTs; <2 nm x 1-5 mu m) in human mesothelial (MeT-5A) cells and bronchial epithelial (BEAS 2B) cells, using the single cell gel electrophoresis (comet) assay and the immunoslot blot assay for the detection of malondialdehyde (M(1)dG) DNA adducts. In BEAS 2B cells, we also studied the induction of micronuclei (MN) by the CNTs using the cytokinesis-block method. The cells were exposed to the CNTs (5-200 mu g/cm(2), corresponding to 19-760 mu g/ml) for 24 and 48 h in the comet assay and for 48 and 72 h in the MN and M(1)dG assays. Transmission electron microscopy (TEM) showed more MWCNT fibres and SWCNT clusters in BEAS 2B than MeT-5A cells, but no significant differences were seen in intracellular dose expressed as area of SWCNT clusters between TEM sections of the cell lines. In MeT-5A cells, both CNTs caused a dose-dependent induction of DNA damage (% DNA in comet tail) in the 48-h treatment and SWCNTs additionally in the 24-h treatment, with a statistically significant increase at 40 mu g/cm(2) of SWCNTs and (after 48 h) 80 mu g/cm(2) of both CNTs. SWCNTs also elevated the level of M(1)dG DNA adducts at 1, 5, 10 and 40 mu g/cm(2) after the 48-h treatment, but both CNTs decreased M(1)dG adduct level at several doses after the 72-h treatment. In BEAS 2B cells, SWCNTs induced a statistically significant increase in DNA damage at 80 and 120 mu g/cm(2) after the 24-h treatment and in M(1)dG adduct level at 5 mu g/cm(2) after 48 h and 10 and 40 mu g/cm(2) after 72 h; MWCNTs did not affect the level of DNA damage but produced a decrease in M(1)dG adducts in the 72-h treatment. The CNTs did not affect the level of MN. In conclusion, MWCNTs and SWCNTs induced DNA damage in MeT-5A cells but showed a lower (SWCNTs) or no (MWCNTs) effect in BEAS 2B cells, suggesting that MeT-5A cells were more sensitive to the DNA-damaging effect of CNTs than BEAS 2B cells, despite the fact that more CNT fibres or clusters were seen in BEAS 2B than MeT-5A cells. M(1)dG DNA adducts were induced by SWCNTs but decreased after a 3-day exposure to MWCNTs and (in MeT-5A cells) SWCNTs, indicating that CNTs may lead to alterations in oxidative effects within the cells. Neither of the CNTs was able to produce chromosomal damage (MN). (C) 2012 Elsevier Ireland Ltd. All rights reserved.