Effect of particle size and dispersion status on cytotoxicity and genotoxicity of zinc oxide in human bronchial epithelial cells

• Nanosized and fine ZnO were cytotoxic within a similar, narrow dose range.
• Only nanosized ZnO, without BSA, increased DNA damage after 3-h and 6-h treatment.
• Both forms of ZnO, regardless of BSA, induced micronuclei after a 48-h treatment.
• Narrow dose range of ZnO genotoxicity is challenging for test reproducibility.

Data available on the genotoxicity of zinc oxide (ZnO) nanoparticles (NPs) are controversial. Here, we examined the effects of particle size and dispersion status on the cytotoxicity and genotoxicity of nanosized and fine ZnO, in the presence and absence of bovine serum albumin (BSA; 0.06%) in human bronchial epithelial BEAS-2B cells. Dynamic light scattering analysis showed the most homogenous dispersions in water alone for nanosized ZnO and in water with BSA for fine ZnO. After a 48-h treatment, both types of ZnO were cytotoxic within a similar, narrow dose range (1.5–3.0 μg/cm2) and induced micronuclei at a near toxic dose range (1.25–1.75 μg/cm2), both with and without BSA. In the comet assay, nanosized ZnO (1.25–1.5 μg/cm2), in the absence of BSA, caused a statistically significant increase in DNA damage after 3-h and 6-h treatments, while fine ZnO did not. Our findings may be explained by better uptake or faster intracellular dissolution of nanosized ZnO without BSA during short treatments (3–6 h; the comet assay), with less differences between the two ZnO forms after longer treatments (>48 h; the in vitro micronucleus test). As ZnO is genotoxic within a narrow dose range partly overlapping with cytotoxic doses, small experimental differences e.g. in the dispersion of ZnO particles may have a substantial effect on the genotoxicity of the nominal doses added to the cell culture.