Title: Comet assay in reconstructed 3D human epidermal skin models – investigation of intra- and inter-laboratory reproducibility with coded chemicals
Authors: REUS AstridREISINGER KDOWNS ThomasCARR GregZELLER AndreasCORVI RaffaellaKRUL CyrillePFUHLER Stefan
Citation: MUTATION RESEARCH-GENETIC TOXICOLOGY AND ENVIRONMENTAL MUTAGENESIS vol. 28 no. 6 p. 709–720
Publisher: ELSEVIER SCIENCE BV
Publication Year: 2013
JRC N°: JRC81082
ISSN: 1383-5718
URI: http://mutage.oxfordjournals.org/content/28/6/709.abstract
http://publications.jrc.ec.europa.eu/repository/handle/JRC81082
DOI: 10.1093/mutage/get051
Type: Articles in periodicals and books
Abstract: Reconstructed 3D human epidermal skin models are increasingly being used for safety testing of chemicals. Based on EpiDerm™ tissues, an assay was developed in which the tissues were topically exposed to test chemicals for 3h followed by cell isolation and assessment of DNA damage using the comet assay. Inter-laboratory reproducibility of the 3D skin comet assay was initially demonstrated using two model genotoxic carcinogens, methyl methane sulfonate (MMS) and 4-nitroquinoline-n-oxide (4NQO), and the results showed good concordance among three different laboratories and with in vivo data. In Phase 2 of the project, intra- and inter-laboratory reproducibility was investigated with five coded compounds tested at three different laboratories. For MMS and N-ethyl-N-nitrosourea (ENU), all laboratories found a dose-related and statistically significant increase (p<0.05) in DNA damage in every experiment. For 2,4-diaminotoluene (2,4-DAT), the overall result from all laboratories showed a smaller, but significant genotoxic response (p<0.05). For cyclohexanone (CHN), an increase compared to the solvent control acetone was observed only in one laboratory. However, the response was not dose-related and CHN was judged negative overall, as was p-nitrophenol (p-NP), which was the only compound showing clear cytotoxic effects. For p-NP, significant DNA damage generally occurred only at doses that showed substantial cytotoxicity (>30% cell loss), and the overall response was comparable in all laboratories despite some differences in doses tested. The results of the collaborative study for the coded compounds were generally reproducible among the laboratories involved and intra-laboratory reproducibility was also good. These data indicate that the comet assay in EpiDerm™ skin models is a relevant model for the safety assessment of compounds with a dermal route of exposure.
JRC Directorate:Institute for Health and Consumer Protection Historical Collection

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