Validation of the 3D Skin Comet assay using full thickness skin models: Transferability and reproducibility
| Autor: | V. Blatz, Frank Henkler, Kerstin Reisinger, Stefan Pfuhler, Astrid A. Reus, Anja Fischer, Cyrille Krul, Sebastian Hoffmann, Thomas R. Downs, Manfred Liebsch, Markus Schulz, Joep Brinkmann, Andreas Luch, Ralph Pirow |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Phenion®FT In vitro genotoxicity Health Toxicology and Mutagenesis Biomedical Innovation Human skin Cosmetics Gene mutation 0302 clinical medicine Fallic acid propyl ester Medicine Skin Cytotoxicity test Priority journal Micronucleus Tests Validation study Life Triskelion BV Reproducibility 7 12 dimethylbenz[a]anthracene Cross-Linking Reagents 030220 oncology & carcinogenesis Micronucleus test RAPID - Risk Analysis for Products in Development TARA - Toxicology and Risk Assessment Cyclohexanone Phthalic acid bis(2 ethylhexyl) ester Healthy Living Human DNA damage Mitomycin Predictive value Computational biology Dermal exposure 03 medical and health sciences Aphidicolin In vivo Genetics Humans Genotoxicity assay Human tissue Biology Comet assay Mutagenicity Tests business.industry Reproducibility of Results Cadmium chloride 030104 developmental biology Ethylnitrosourea Cell isolation ELSS - Earth Life and Social Sciences business Controlled study Mutagens |
| Zdroj: | Mutation Research-Genetic Toxicology and Environmental Mutagenesis, 827, 27-41 |
| Popis: | Recently revised OECD Testing Guidelines highlight the importance of considering the first site-of-contact when investigating the genotoxic hazard. Thus far, only in vivo approaches are available to address the dermal route of exposure. The 3D Skin Comet and Reconstructed Skin Micronucleus (RSMN) assays intend to close this gap in the in vitro genotoxicity toolbox by investigating DNA damage after topical application. This represents the most relevant route of exposure for a variety of compounds found in household products, cosmetics, and industrial chemicals. The comet assay methodology is able to detect both chromosomal damage and DNA lesions that may give rise to gene mutations, thereby complementing the RSMN which detects only chromosomal damage. Here, the comet assay was adapted to two reconstructed full thickness human skin models: the EpiDerm™- and Phenion® Full-Thickness Skin Models. First, tissue-specific protocols for the isolation of single cells and the general comet assay were transferred to European and US-American laboratories. After establishment of the assay, the protocol was then further optimized with appropriate cytotoxicity measurements and the use of aphidicolin, a DNA repair inhibitor, to improve the assay's sensitivity. In the first phase of an ongoing validation study eight chemicals were tested in three laboratories each using the Phenion® Full-Thickness Skin Model, informing several validation modules. Ultimately, the 3D Skin Comet assay demonstrated a high predictive capacity and good intra- and inter-laboratory reproducibility with four laboratories reaching a 100% predictivity and the fifth yielding 70%. The data are intended to demonstrate the use of the 3D Skin Comet assay as a new in vitro tool for following up on positive findings from the standard in vitro genotoxicity test battery for dermally applied chemicals, ultimately helping to drive the regulatory acceptance of the assay. To expand the database, the validation will continue by testing an additional 22 chemicals. © 2018 The Authors Chemicals/CAS: 7,12 dimethylbenz[a]anthracene, 57-97-6; aphidicolin, 38966-21-1; cadmium chloride, 10108-64-2; cyclohexanone, 108-94-1; ethylnitrosourea, 759-73-9; eugenol, 97-53-0; gallic acid propyl ester, 121-79-9; mitomycin, 1404-00-8, 50-07-7, 74349-48-7; phthalic acid bis(2 ethylhexyl) ester, 117-81-7 |
| Databáze: | OpenAIRE |
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