title: State of the art in non-animal approaches for skin sensitization testing: from individual test methods towards testing strategies.
abstract: The hazard assessment of skin sensitizers relies mainly on animal testing, but much progress is made in the development, validation and regulatory acceptance and implementation of non-animal predictive approaches. In this review, we provide an update on the available computational tools and animal-free test methods for the prediction of skin sensitization hazard. These individual test methods address mostly one mechanistic step of the process of skin sensitization induction. The adverse outcome pathway (AOP) for skin sensitization describes the key events (KEs) that lead to skin sensitization. In our review, we have clustered the available test methods according to the KE they inform: the molecular initiating event (MIE/KE1)-protein binding, KE2-keratinocyte activation, KE3-dendritic cell activation and KE4-T cell activation and proliferation. In recent years, most progress has been made in the development and validation of in vitro assays that address KE2 and KE3. No standardized in vitro assays for T cell activation are available; thus, KE4 cannot be measured in vitro. Three non-animal test methods, addressing either the MIE, KE2 or KE3, are accepted as OECD test guidelines, and this has accelerated the development of integrated or defined approaches for testing and assessment (e.g. testing strategies). The majority of these approaches are mechanism-based, since they combine results from multiple test methods and/or computational tools that address different KEs of the AOP to estimate skin sensitization potential and sometimes potency. Other approaches are based on statistical tools. Until now, eleven different testing strategies have been published, the majority using the same individual information sources. Our review shows that some of the defined approaches to testing and assessment are able to accurately predict skin sensitization hazard, sometimes even more accurate than the currently used animal test. A few defined approaches are developed to provide an estimate of the potency sub-category of a skin sensitizer as well, but these approaches need further independent evaluation with a new dataset of chemicals. To conclude, this update shows that the field of non-animal approaches for skin sensitization has evolved greatly in recent years and that it is possible to predict skin sensitization hazard without animal testing.
authors.affiliation: Department of Innovative Testing Strategies, Center for Health Protection, National Institute for Public Health and the Environment (RIVM), PO Box 1, 3720 BA, Bilthoven, The Netherlands. Janine.Ezendam@rivm.nl.
authors.forename: Janine
authors.identifier: http://orcid.org/0000-0002-8677-1986
authors.initials: J
authors.lastname: Ezendam
authors.affiliation: Department of Innovative Testing Strategies, Center for Health Protection, National Institute for Public Health and the Environment (RIVM), PO Box 1, 3720 BA, Bilthoven, The Netherlands.
authors.forename: Hedwig M
authors.initials: HM
authors.lastname: Braakhuis
authors.affiliation: Department of Innovative Testing Strategies, Center for Health Protection, National Institute for Public Health and the Environment (RIVM), PO Box 1, 3720 BA, Bilthoven, The Netherlands.
authors.forename: Rob J
authors.initials: RJ
authors.lastname: Vandebriel
mesh_terms: D000826:Animal Testing Alternatives / Q000639:trends; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D035843:Biomedical Research / Q000639:trends; D001711:Biotransformation; D002460:Cell Line; D002478:Cells, Cultured; D019295:Computational Biology; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D015507:Drugs, Investigational / Q000009:adverse effects* / Q000378:metabolism; D005103:Expert Systems; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation / Q000187:drug effects; D008954:Models, Biological; D012107:Research Design; D012867:Skin / Q000166:cytology / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D012869:Skin Absorption; D012882:Skin Tests / Q000592:standards / Q000639:trends; D015262:Xenobiotics / Q000378:metabolism / Q000633:toxicity*
chemical_list: D015415:Biomarkers; D015507:Drugs, Investigational; D015262:Xenobiotics
keywords: Adverse outcome pathway; Alternatives to animal testing; Hazard assessment; Skin sensitization; Testing strategies