dc.contributor.author
Pfuhler, Stefan
dc.contributor.author
Pirow, Ralph
dc.contributor.author
Downs, Thomas R.
dc.contributor.author
Haase, Andrea
dc.contributor.author
Hewitt, Nicola
dc.contributor.author
Luch, Andreas
dc.contributor.author
Merkel, Marion
dc.contributor.author
Petrick, Claudia
dc.contributor.author
Said, André
dc.contributor.author
Schäfer-Korting, Monika
dc.date.accessioned
2021-11-29T09:03:08Z
dc.date.available
2021-11-29T09:03:08Z
dc.identifier.uri
https://refubium.fu-berlin.de/handle/fub188/32881
dc.identifier.uri
http://dx.doi.org/10.17169/refubium-32607
dc.description.abstract
As part of the safety assessment process, all industrial sectors employ genotoxicity test batteries, starting with well-established in vitro assays. However, these batteries have limited predictive capacity for the in vivo situation, which may result in unnecessary follow-up in vivo testing or the loss of promising substances where animal tests are prohibited or not desired. To address this, a project involving regulators, academia and industry was established to develop and validate in vitro human skin-based genotoxicity assays for topically exposed substances, such as cosmetics ingredients. Here, we describe the validation of the 3D reconstructed skin (RS) Comet assay. In this multicenter study, chemicals were applied topically three times to the skin over 48 h. Isolated keratinocytes and fibroblasts were transferred to slides before electrophoresis and the resulting comet formation was recorded as % tail DNA. Before decoding, results of the validation exercise for 32 substances were evaluated by an independent statistician. There was a high predictive capacity of this assay when compared to in vivo outcomes, with a sensitivity of 77 (80)%, a specificity of 88 (97)% and an overall accuracy of 83 (92)%. The numbers reflect the calls of the performing laboratories in the coded phase, whereas those in parenthesis reflect calls according to the agreed evaluation criteria. Intra- and inter-laboratory reproducibility was also very good, with a concordance of 93 and 88%, respectively. These results generated with the Phenion® Full-Thickness skin model demonstrate its suitability for this assay, with reproducibly low background DNA damage and sufficient metabolic capacity to activate pro-mutagens. The validation outcome supports the use of the RS Comet assay to follow up positive results from standard in vitro genotoxicity assays when the expected route of exposure is dermal. Based on the available data, the assay was accepted recently into the OECD test guideline development program.
en
dc.format.extent
18 Seiten
dc.rights.uri
https://creativecommons.org/licenses/by-nc/4.0/
dc.subject.ddc
500 Naturwissenschaften und Mathematik::570 Biowissenschaften; Biologie::570 Biowissenschaften; Biologie
dc.title
Validation of the 3D reconstructed human skin Comet assay, an animal-free alternative for following-up positive results from standard in vitro genotoxicity assays
dc.type
Wissenschaftlicher Artikel
dcterms.bibliographicCitation.doi
10.1093/mutage/geaa009
dcterms.bibliographicCitation.journaltitle
Mutagenesis
dcterms.bibliographicCitation.number
1
dcterms.bibliographicCitation.pagestart
19
dcterms.bibliographicCitation.pageend
35
dcterms.bibliographicCitation.volume
36
dcterms.bibliographicCitation.url
https://doi.org/10.1093/mutage/geaa009
refubium.affiliation
Biologie, Chemie, Pharmazie
refubium.affiliation.other
Institut für Pharmazie
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refubium.resourceType.isindependentpub
no
dcterms.accessRights.openaire
open access
dcterms.isPartOf.eissn
1464-3804
refubium.resourceType.provider
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