Evaluation of fibrin-based dermal-epidermal organotypic cultures for in vitro skin corrosion and irritation testing of chemicals according to OECD TG 431 and 439. (October 2016)

• Record Type: Journal Article
• Authors: Morales, Mariana; Pérez, David; Correa, Luis; Restrepo, Luz
• Is Part Of: Toxicology in vitro. Volume 36(2016)
• Page Start: 89
• Page End: 96
• Publication Date: 2016-10
• Subjects: RhE reconstructed human epidermis -- ORG organotypic skin culture -- H&E hematoxylin and eosin -- MTT 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide -- BD CBA Becton Dickinson Cytometric Bead Array -- OD optical density -- RH relative humidity -- RT room temperature -- SD standard deviation -- OECD Organisation for Economic Co-operation and Development -- TG test guideline -- UN GHS United Nations Globally Harmonized System; Fibrin-based matrix -- Dermal-epidermal organotypic culture -- Corrosive -- Irritant -- OECD test guideline 431 and 439 -- United Nations Globally Harmonized System -- Dermis -- Epidermis; Toxicity testing -- In vitro -- Periodicals; Toxicology -- Periodicals; 615.9
• DOI: 10.1016/j.tiv.2016.07.010
• Languages: English
• ISSNs: 0887-2333

Abstract

Abstract: Reconstructed human epidermis (RhE) models have been used for in vitro testing of the potential harmful effects of exposure to chemical compounds on health. In the past, skin irritation and corrosion were evaluated in animal models; however, in recent years, due to the bioethics implications of the method and, to minimize the use of experimental animals, alternative procedures have been proposed. The Organisation for Economic Co-operation and Development (OECD) in its test guidelines (TG) 431 and 439 indicates the requirements for validating new methods for the evaluation of skin corrosion and irritation, respectively. Here, we present an in-house human dermal-epidermal model, useful for the performance of these tests. Using the methods described in this work, it was possible to obtain human fibrin-based dermal-epidermal organotypic skin cultures (ORGs) displaying similar histological characteristics to native skin and expressing specific differentiation epithelial proteins. The end points to classify a substance as irritant or corrosive were cell viability evaluated by MTT assay, and cytokine release measured by BD CBA for human inflammatory cytokines. According to the MTT test, the ORGs correctly classified irritating and corrosive substances. Moreover, the cytokine release assay was difficult to interpret in the context of testing chemical hazard classification. Further experiments are needed to validate this new model for the evaluation of surfactants because the fibrin matrix was affected in the presence of these substances. Highlights: A human fibrin based dermal-epidermal organotypic skin culture (ORGs) was developed. Human primary keratinocytes and fibroblasts from atypical body region were used. All ORGs were built with high concentration of calcium (1.16 mM) throughout culture. ORGs were useful to perform the corrosion and irritation tests under OECD Test Guides.