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3D IN VITRO MODEL OF ATOPIC DERMATITIS IN RECONSTRUCTED HUMAN EPIDERMIS AND EXPOSURE TO PARTICULATE MATTER FROM ENVIRONMENTAL POLLUTANTS

Grant number: 22/09148-0
Support Opportunities:Scholarships in Brazil - Post-Doctoral
Effective date (Start): December 01, 2022
Effective date (End): November 30, 2024
Field of knowledge:Health Sciences - Pharmacy - Toxicological Analysis
Principal Investigator:Silvya Stuchi Maria-Engler
Grantee:Andressa Cristina Antunes Santos
Host Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

The skin is the largest organ in the human body and its main function is to act as a barrier against external agents. Atopic dermatitis (AD) is one of the most prevalent inflammatory skin diseases and is considered an expensive and difficult-to-control disease. It is estimated that at least 230 million people worldwide are affected by this condition. AD is a chronic disease that involves genetic and environmental factors with damage to the epidermal barrier, changes in the immune system and skin inflammation. A factor still little explored about this disease is its interaction with environmental pollutants such as particulate matter (PM). Recent data from our group showed that MP exposure to healthy epidermis can alter the expression of different proteins involved in the AD process, suggesting the MP potential action in the development of the disease. In this project, we aim to investigate the molecular and morphological changes caused by skin MP exposure in AD modulation using an in vitro 3D model. Therefore, histological and immunohistochemical techniques will be used for FLG (filagrin), LOR (loricrin), IVL (involucrin), CAS2 (carbonic anhydrase II) and CLDN 1 (claudin 1) markers. The evaluation of the permeability of the epidermal barrier will be evaluated by the transepithelial electrical resistance. To understand the changes in modulation, the cytokines Interleukin 1± (IL)-1±, IL6, IL8 and metalloproteinases (MMP) 1, MMP2 and MMP9 will be evaluated in the culture medium by ELISA. We will also evaluate gene expression by RT-qPCR of genes LOR, FLG1 and FLG 2, while the proteins aquaporin 3 (AQP3), FLG, LOR, NOTCH1 and aryl-hydrocarbon receptor (AhR) will be analyzed by Western Blotting. Finally, DNA fragmentation will be performed by TUNEL using flow cytometry. The data from this study will provide relevant information for the understanding of PM exposure pathophysiological aspects in the skin in vitro for the onset or worsening of AD. We will also contribute to the development and validation of biomimetic skin models to be used in safety and efficacy toxicological studies of new formulations with a protective effect against environmental pollution, in addition to suggesting possible targets for the treatment or regression of AD.

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