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Evaluation of the anti-inflammatory effect of nanoparticles of Punica granatum Linné on the human monocyte line THP-1

Grant number: 19/01380-8
Support type:Scholarships in Brazil - Post-Doctorate
Effective date (Start): April 01, 2019
Effective date (End): November 30, 2020
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Vanessa Soares Lara
Grantee:Priscila Maria Aranda Salomão
Home Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil
Associated research grant:15/03965-2 - Role of the renin-angiotensin system in different oral inflammatory models: an experimental interdisciplinary and clinical approach, AP.TEM

Abstract

This study will evaluate in vitro if the phytotherapeutic drug Punica granatum Linné, which affects the renin-angiotensin system (RAS), also has some influence on the immunopathological mechanisms triggered in candidoses, especially in the most frequent form of oral candidiasis, denture stomatitis. Specifically, this study aims to evaluate in vitro whether the phytotherapeutic mentioned, in the form of nanoparticles, interferes in the immunoinflammatory response of monocytes against infection with Candida albicans. Initially, the cytotoxicity of the human monocytes of the THP-1 ATCC® line challenged with different concentrations of phytotherapeutic nanoparticles will be evaluated by Alarmablue viability assay. The anti-inflammatory effect of the compound on the human monocytes, previously stimulated in vitro with C. albicans or LPS, will be evaluated by the production of nitric oxide (NO) and hydrogen peroxide (H2O2), using Griess's colorimetric method and by degradation of fluorescent substrates, respectively, as well as by quantification of cytokines and chemokines produced by cells by Milliplex® Multiplex assay. Furthermore, the determination of genes and proteins related to the inflammatory and anti-Candida responses will be evaluated by RT-qPCR and mass spectrometry techniques. The results obtained will be submitted to the analysis of normality by means of the Kolmogorov-Smirnov test and, later, will be evaluated using the T and Anova one-way tests when the data present a normal distribution, or through the Mann-Whitney tests and Kruskall-Wallis if the data were non-parametric (p <0.05).