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Biocompatibility of ±-Ag2WO4, ²-Ag2MoO4 and ±-AgVO3 microcrystals: evaluation of proinflammatory cytokines and matrix metalloproteinases

Grant number: 18/01677-8
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): May 01, 2018
Effective date (End): September 30, 2020
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Carlos Eduardo Vergani
Grantee:Bruna Natalia Alves da Silva Pimentel
Home Institution: Faculdade de Odontologia (FOAr). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
Associated research grant:13/07296-2 - CDMF - Center for the Development of Functional Materials, AP.CEPID

Abstract

Fungal infections are important causes of morbidity and mortality. Currently there are different therapies with antifungal agents used to control these infections; however, the microbial tolerance to these agents has increased. For this reason, our group has evaluated the antifungal activity of silver-containing microcrystals as well as their cytotoxicity on human cells (human gingival fibroblast - FGH - and spontaneously immortalized Normal Oral Keratinocytes - NOK-si). The results obtained to date showed that the microcrystals tested (silver tungstate alpha, silver beta molybdate and alpha silver vanadate) showed fungistatic and fungistatic activity against Candida albicans and, at the same time, these doses showed no cytotoxicity on human cells (FGH and NOK-si). However, there are important gaps in the biocompatibility of microcrystals that should be carefully investigated as the ability to induce inflammatory response and degradation of matrix metalloproteinases (MMPs). The understanding of such responses is fundamental to ensure a better understanding of the mechanisms of inflammatory response and tissue regeneration facing these new materials. Thus, this project aims to evaluate the expression of proinflammatory cytokines (IL-1², IL-6, IL-8 and TNF-±) and MMPs (-8 and -9) in human cell cultures and reconstituted oral mucosa tissue (ROMT) after exposure to inhibitory concentrations and microcrystalline fungicides (±-Ag2WO4, ²-Ag2MoO4 and ±-AgVO3). To evaluate cell viability, the Alamar Blue assays will be performed. qPCR assays (for the evaluation of the gene expression of cytokines and MMPs) will be performed; flow cytometry (for quantification of soluble cytokines); Western Blot (for quantification of soluble MMPs); and zymography (to evaluate the enzymatic activity of MMPs). The results obtained will be submitted to statistical analysis using GraphPad Prism 5 software. A significance level of 5% will be adopted. (AU)