Development of an artificial skin platform for risk assessment

• Grant number: 11/07441-7
• Support Opportunities: Regular Research Grants
• Start date: December 01, 2011
• End date: November 30, 2014
• Field of knowledge: Health Sciences - Pharmacy - Toxicological Analysis
• Principal Investigator: Silvya Stuchi Maria-Engler
• Grantee: Silvya Stuchi Maria-Engler
• Host Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• City of the host institution: São Paulo
• Associated researchers: Silvia Berlanga de Moraes Barros

Abstract

This project aims to assess the toxicity of novel substances using an in vitro two-tiered testing strategy in a technology transfer manner. This two-tiered assay is able to detect sensitizers and rank sensitizer potency based on its irritant properties. Besides that, this testing strategy provides information regarding chemical toxicity. This two-tiered assay is based on the use of MUTZ-LC and epidermal equivalents (EE). The secretion of IL-8 by MUTZ-LC is utilized as a parameter to identify sensitizers from a testing panel. Complementary information regarding the identification of sensitizers may be acquired with the investigation of MUTZ-LC maturation markers, such as CCR7 and CXCR4. EE will be used to investigate the potency of chemicals. The potency of a chemical is direct related to its toxic effect on the viable layers of the epidermis, which can be determined by MTT assay. The chemical concentration required to reduce EE metabolic activity to 50% of the maximum value compared to vehicle exposed cultures (EC50) will be used to rank chemical potency. Complementary information regarding potency of chemicals can be obtained determining IL-1a secretion in EE supernatants after chemical exposure. At first this project aims to implement the model in our lab (MUTZ and EE). Once the model is established, our goal is to determine whether the model is functional. Eight known substances (sensitizers: nitrobenzylbromide, cinnemaldehyde, formaldehyde and resorcinol; non-sensitizers: DMSO, phenol, isopropanol and SDS) will be tested at this stage. Novel substances will be tested only when the model responds in the same way as described in the literature. Pothomorphe umbellata, a native Brazilian plant, is popularly known to be effective in the treatment of skin lesions. This benefit is attributed to 4-nerolidylcatechol (4-NC), a compound extracted from this plant. Since melanomas show prominent resistance to apoptosis and exhibit extreme chemoresistance to multiple forms of therapy, novel compounds addressing induction of cell death are worth investigating. Brohen et al., have studied the effect of 4-NC on different melanoma cell lines and have shown that 4-NC showed cytotoxic activity for all melanoma cell lines tested associated with its capacity to induce apoptosis and inhibits cell invasiveness (due to G1 cell cycle arrest and inhibition of MMP-2 activity in melanoma cell lines). Besides that, 4-NC has also antioxidant and photoprotective properties, therefore it is a promising candidate for use in cosmetic and pharmaceutical formulations. Although the 4-NC potential applicability in cosmetic and pharmaceutical formulation mentioned above, it is still unknown weather this compound has a sensitizer effect on the skin. In order to be used in the cosmetic and pharmaceutical industries, 4-NC has to be incorporated in a formulation, which can have different forms, such as serums, gels, creams, emulsions etc. With the aid of the in vitro model developed in this project, we aim to investigate the toxic effect of different formulations containing 0.1% 4-NC (gel, gel-emulsion, emulsion) and the extract of P. umbellata (gel, gel-emulsion, emulsion) and determine whether 4-NC is a potential sensitizer. In this way, we expect to elucidate which formulations containing 4-NC or extract of P. umbellata would have the mildest effect on the skin when topically applied. (AU)

Scientific publications

(4)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

ZANONI, THALITA B.; TIAGO, MANOELA; FAIAO-FLORES, FERNANDA; DE MORAES BARROS, SILVIA B.; BAST, AALT; HAGEMAN, GEJA; DE OLIVEIRA, DANIELLE PALMA; MARIA-ENGLER, SILVYA S.. Basic Red 51, a permitted semi-permanent hair dye, is cytotoxic to human skin cells: Studies in monolayer and 3D skin model using human keratinocytes (HaCaT). Toxicology Letters, v. 227, n. 2, p. 139-149, JUN 5 2014. (10/07116-6, 11/07441-7)

PENNACCHI, PAULA COMUNE; SOARES DE ALMEIDA, MAIRA ESTANISLAU; ALVES GOMES, OCTAVIO LUIS; FAIAO-FLORES, FERNANDA; DE ARAUJO CREPALDI, MARIA CLARA; DOS SANTOS, MARINILCE FAGUNDES; DE MORAES BARROS, SILVIA BERLANGA; MARIA-ENGLER, SILVYA STUCHI. Glycated Reconstructed Human Skin as a Platform to Study the Pathogenesis of Skin Aging. TISSUE ENGINEERING PART A, v. 21, n. 17-18, p. 2417-2425, SEP 1 2015. (11/22812-1, 11/14327-6, 11/07441-7)

CATARINO, CAROLINA MOTTER; PEDROSA, TATIANA DO NASCIMENTO; PENNACCHI, PAULA COMUNE; DE ASSIS, SILVIA ROMANO; GIMENES, FABRICIA; LOPES CONSOLARO, MARCIA EDILAINE; DE MORAES BARROS, SILVIA BERLANGA; MARIA-ENGLER, SILVYA STUCHI. Skin corrosion test: a comparison between reconstructed human epidermis and full thickness skin models. EUROPEAN JOURNAL OF PHARMACEUTICS AND BIOPHARMACEUTICS, v. 125, p. 51-57, APR 2018. (13/00735-0, 11/07441-7, 11/14327-6, 11/22812-1, 13/12682-9)

PEDROSA, TATIANA DO NASCIMENTO; CATARINO, CAROLINA MOTTER; PENNACCHI, PAULA COMUNE; DE ASSIS, SILVIA ROMANO; GIMENES, FABRICIA; LOPES CONSOLARO, MARCIA EDILAINE; DE MORAES BARROS, SILVIA BERLANGA; MARIA-ENGLER, SILVYA STUCHI. A new reconstructed human epidermis for in vitro skin irritation testing. TOXICOLOGY IN VITRO, v. 42, p. 31-37, AUG 2017. (11/14327-6, 11/22812-1, 11/07441-7, 13/12682-9, 14/24400-0, 13/00735-0)