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In vitro study of antimicrobial peptides production by human palatal epithelial cell exposed to Candida albicans

Grant number: 11/17481-6
Support Opportunities:Regular Research Grants
Duration: December 01, 2011 - November 30, 2013
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Vanessa Soares Lara
Grantee:Vanessa Soares Lara
Host Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil
Associated researchers: Ana Regina Casaroto

Abstract

The presence of fungus Candida albicans in the microbial biofilm adherent underlying maxillary prosthesis is related to an inflammatory reaction of the palatal mucosa, known as denture stomatitis. Defensins and cathelicidins are antimicrobial peptides expressed by mucosal epithelial cells, both with activity against C. albicans, acting from the direct inhibition of pathogen to the modulation of innate and adaptive immune responses. Knowledge of joint secretion of defensins and cathelicidins is important for the detailed understanding of the pathology of inflammatory diseases, such as denture stomatitis. In vitro studies with human palatal epithelial cells exposed to C. albicans, comparatively evaluating the peptides production, were not found. The purpose of this study is to evaluate in vitro production of antimicrobial peptides ²-defensin (hBD-2) and cathelicidin (LL-37) by human palatal epithelial cells after challenge with C. albicans, correlating with apoptosis events and cell invasion caused by the fungus. Epithelial cells obtained from palatal tissue will be challenged by direct and indirect contact with viable C. albicans at 0, 12, 24 and 48 h. The apoptotic cells will be determined by fluorescent nuclear staining with Hoechst 33258. The cell invasion by the fungus will be assessed qualitatively by colorimetric method with acridini orange. hBD-2 and LL-37 secretion and gene expression will be determined, before and after challenge, by ELISA and real time RT-PCR. The results will be shown as mean ± standard deviation followed by factorial ANOVA, Contrast test and/or Mann-Whitney test (p<0,05). (AU)

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Scientific publications
(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)
CASAROTO, ANA REGINA; DA SILVA, RAFAELA ALVES; SALMERON, SAMIRA; RUBO DE REZENDE, MARIA LUCIA; DIONISIO, THIAGO JOSE; DOS SANTOS, CARLOS FERREIRA; PINKE, KAREN HENRIETTE; GUARIZO KLINGBEIL, MARIA FATIMA; SALOMAO, PRISCILA ARANDA; RIBEIRO LOPES, MARCELO MILANDA; et al. Candida albicans-Cell Interactions Activate Innate Immune Defense in Human Palate Epithelial Primary Cells via Nitric Oxide (NO) and (3-Defensin 2 (hBD-2). CELLS, v. 8, n. 7, . (11/17481-6, 15/03965-2)

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