A new reconstructed human epidermis for in vitro skin irritation testing. (August 2017)

• Record Type: Journal Article
• Authors: Pedrosa, Tatiana do Nascimento; Catarino, Carolina Motter; Pennacchi, Paula Comune; Assis, Sílvia Romano de; Gimenes, Fabrícia; Consolaro, Márcia Edilaine Lopes; Barros, Silvia Berlanga de Moraes; Maria-Engler, Silvya Stuchi
• Is Part Of: Toxicology in vitro. Volume 42(2017)
• Page Start: 31
• Page End: 37
• Publication Date: 2017-08
• Subjects: ANOVA one-way analysis of variance -- CAS Chemical Abstracts Service -- DMEM Dulbecco's Modified Eagle Medium -- CK10 cytokeratin 10 -- CK14 cytokeratin 14 -- PBS phosphate-buffered saline -- EC50 effective concentrations at 50% viability -- EGF Epidermal growth factor, ET50, effective time at 50% viability -- ELISA Enzyme Linked Immuno Sorbent Assay -- EU European Union -- HBV human hepatitis B virus -- HCV human hepatitis C virus -- HIV-1 human immunodeficiency virus-1 -- HIV-2 human immunodeficiency virus-2 -- HPV human papillomavirus -- HSV-1 herpes simplex virus 1 -- HSV-2 herpes simplex virus 2 -- HTLV-1 T-cell lymphotropic virus 1 -- HTLV-2 T-cell lymphotropic virus 2 -- HU/USP University of São Paulo Hospital -- MTT 3-(4, 5-dimethylthiazo-2-yl)-2, 5-diphenyltetrazoliumbromide -- NaCl sodium chloride -- NC negative control -- NHEK Normal human epidermal keratinocytes -- OD optical density -- OECD Organization for Economic Co-operation and Development -- PC positive control -- PCR polymerase chain reaction -- PS performance standards -- RGM Rheinwald and Green modified culture medium -- RHE reconstructed human epidermis -- RT-PCR Reverse transcription polymerase chain reaction -- SD standard deviation -- SDS sodium dodecyl sulfate -- TG testing guideline -- TGF-α transforming growth factor alpha -- USP-RHE reconstructed human epidermis developed by University of São Paulo -- VRM validated reference method; Irritation assays -- RHE -- OECD -- New skin models; Toxicity testing -- In vitro -- Periodicals; Toxicology -- Periodicals; 615.9
• DOI: 10.1016/j.tiv.2017.03.010
• Languages: English
• ISSNs: 0887-2333

Abstract

Abstract: Different models of reconstructed human epidermis (RHE) are currently validated to assess skin irritation in vitro and ultimately to the animal replacement of the Draize test. The development of a new RHE model is a challenge for many laboratories, representing a potential gain of autonomy and improvement of technological knowledge. The Organization for Economic Co-operation and Development (OECD) encourages the development of new models and, for this purpose, offers a thorough guideline on quality control parameters (OECD TG 439 performance standards). This work aimed to develop an RHE model ( i.e. USP-RHE) for in vitro skin irritation assays, following the OECD TG 439. The developed model presents a well-differentiated epidermis similar to the Validated Reference Methods (VRM) and to native human epidermis. Quality parameters, i.e. optical density of negative control, tissue integrity and barrier function, were similar to VRM and in accordance with OECD TG 439. Moreover, the USP-RHE model was shown to have 85, 7% of specificity (6/7), 100% of sensitivity (6/6) and 92, 3% of accuracy (12/13) when compared to in vivo UN GHS classification. The within-laboratory reproducibility was 92.3% (12/13). Thus, we demonstrated that USP-RHE model attends to all OECD TG 439 performance standards and is ready to be used by private and public laboratories and companies for future validation studies. Highlights: A new RHE model, USP-RHE, is presented here as open source protocol. The quality control parameters of USP-RHE were similar to VRM. The model presented 85, 7% of specificity, 100% of sensitivity and 92, 3% of accuracy.