Evaluation of the antifungal and immunomodulatory effects of inorganic nanoparticles associated with natural polyphenols on Candida albicans and Oral Candidiasis

Abstract

The present work aims to evaluate the antifungal and immunomodulatory effects of gold nanoparticles and iron nanoparticles (Np) associated with natural polyphenols (curcumin, gallic acid or resveratrol) against Candida albicans, using in vitro and in vivo study models. For this, the project will be distributed in eight stages: 1) Evaluation of the antifungal effects of Np associated with polyphenols against planktonic cultures of C. albicans by the minimum inhibitory concentration, minimum fungicidal concentration, and time-kill curve assays; 2) Analysis of the toxicity of Np with polyphenols on macrophages and keratinocytes cell lines; 3) Understanding the effects of these nanoformulations against C. albicans virulence factors, including filamentation capacity, and biofilm formation; 4) Assessing the physiological, and molecular effects of Np against fungal cells by investigating ergosterol content, oxidative radical production, lipid peroxidation, ultrastructural changes under transmission electron microscopy, and gene expression; 5) Study of the influence of nanoformulations on macrophage-Candida interaction by phagocytosis, oxidative radical production, macrophage fungicidal activity and intracellular proliferation capacity of C. albicans assays; 6) In vivo assays of the nanoformulations in the Galleria mellonella experimental model, using toxicity, hemocyte count, survival curve and quantification of C. albicans assays; 7) Selection of the nanoformulation with greater antifungal efficacy, and incorporation into a hydrogel matrix, followed by physical characterization, and antifungal activity assays against planktonic cultures; 8) Evaluation of the effects of the nanoformulation in hydrogel in the treatment of oral candidiasis using a murine model of denture stomatitis by the C. albicans burden, histological analysis, clinical inflammation analysis and enzymatic activity of myeloperoxidase and N-acetylglucosaminidase. The results will be submitted to an exploratory analysis for the selection of the most adequate statistical test for each experiment of this study, considering a significance level of 5%. (AU)