Antifungal effect of herbal formulation for mouthwash with pomegranate in mucosas infections caused by Candida albicans and Candida glabrata

Abstract

The objective of this work is to evaluate the antifungal action of a mouthwash formulation based on pomegranate peel extract, sodium trimetaphosphate and fluorine for application in the prevention and/or treatment of oral candidiasis. The effectiveness of the formulations on the growth inhibition of Candida albicans SC5314 and Candida glabrata CBS138 will be determined by the minimum inhibitory concentration by the microdilution method. In addition to reference strains, oral clinical strains will be evaluated. The biofilms of the strains of C. albicans and C. glabrata will be quantified using the Cell Viability Reagent (PrestoBlueTM) and microscopic images will be obtained to assess the percentage of cells with filament. To determine the effect of formulations on the balance between Candida, beneficial microorganisms (Lactobacillus) and epithelial cells, a co-culture system will be used, where cell viability will be determined by plating and by the XTT method. In order to have a more dynamic view of the process, a method based on confocal fluorescence microscopy using cell viability markers (e.g. SYTO9) will also be used. The cellular response at the level of the transcriptome of Candida cells to the formulations will be evaluated during plantonic growth and in preformed biofilms, based on DNA microarrays. The appropriate statistical tests will be applied with a 5% significance level.