title: Clastogenicity, photo-clastogenicity or pseudo-photo-clastogenicity: Genotoxic effects of zinc oxide in the dark, in pre-irradiated or simultaneously irradiated Chinese hamster ovary cells.
abstract: Zinc oxide (ZnO), a widely used ingredient in dermatological preparations and sunscreens, is clastogenic in vitro, but not in vivo. Given that ZnO has an approximately four-fold greater clastogenic potency in the presence of UV light when compared with that in the dark, it has been suggested to be photo-clastogenic. In order to clarify whether this increased potency is a genuine photo-genotoxic effect, we investigated the clastogenicity of ZnO (mean particle size, 100 nm) in Chinese hamster ovary (CHO) cells in the dark (D), in pre-irradiated (PI, i.e. UV irradiation of cells followed by treatment with ZnO) and in simultaneously irradiated (SI, i.e. ZnO treatment concurrent with UV irradiation) CHO cells at UV doses of 350 and 700 mJ/cm(2). The cytotoxicity of ZnO to CHO cells under the different irradiation conditions was as follows: SI>PI>D. In the dark, ZnO produced a concentration-related increase in chromosome aberrations (CA). In PI or SI CHO cells, ZnO was clastogenic at significantly lower concentrations (approximately two- to four-fold) when compared with effective concentrations in the dark, indicating an increased susceptibility of CHO cells to ZnO-mediated clastogenic effects due to UV irradiation per se. The incidence of CA in SI or PI cells was generally higher than that in the dark. At similar ZnO concentrations, SI conditions generally produced higher CA incidence than PI conditions. However, when ZnO concentrations producing similar cytotoxicity were compared, CA incidences under PI or SI conditions were nearly identical. The modest increase in the clastogenic potency of ZnO following UV irradiation contrasts with the results observed with genuine photo-clastogenic agents, such as 8-MOP, which may produce an increase in clastogenic potency of >15,000-fold under SI conditions. Our results provide evidence that, under conditions of in vitro photo-clastogenicity tests, UV irradiation of the cellular test system per se may produce a slight increase in the genotoxic potency of compounds that are clastogenic in the dark. In conclusion, our data suggest that minor increases in clastogenic potency under conditions of photo-genotoxicity testing do not necessarily represent a photo-genotoxic effect, but may occur due to an increased sensitivity of the test system subsequent to UV irradiation.
authors.affiliation: L'Oréal Research and Development, Worldwide Safety Department, 92600 Asnières, France. edufour@rd.loreal.com
authors.forename: Eric K
authors.initials: EK
authors.lastname: Dufour
authors.forename: Tirukalikundram
authors.initials: T
authors.lastname: Kumaravel
authors.forename: Gerhard J
authors.initials: GJ
authors.lastname: Nohynek
authors.forename: David
authors.initials: D
authors.lastname: Kirkland
authors.forename: Hervé
authors.initials: H
authors.lastname: Toutain
mesh_terms: D000818:Animals; D016466:CHO Cells; D002869:Chromosome Aberrations / Q000139:chemically induced / Q000528:radiation effects; D006224:Cricetinae; D004305:Dose-Response Relationship, Drug; D004307:Dose-Response Relationship, Radiation; D066298:In Vitro Techniques; D008027:Light; D008730:Methoxsalen / Q000528:radiation effects / Q000633:toxicity; D016296:Mutagenesis / Q000528:radiation effects; D009152:Mutagenicity Tests / Q000379:methods*; D009153:Mutagens / Q000633:toxicity*; D049329:Nanostructures; D010316:Particle Size; D014466:Ultraviolet Rays; D015034:Zinc Oxide / Q000737:chemistry / Q000633:toxicity*
chemical_list: D009153:Mutagens; D015034:Zinc Oxide; D008730:Methoxsalen