Quantification of Candida albicans, effect of efflux pump inhibitors, N-acetylcystein and light-active micelles and validation of an in vivo model of denture stomatitis

Abstract

The aims of this research are: 1- to compare molecular and non-molecular methods of quantification of Candida albicans; 2- to evaluate the effect of efflux pump inhibitors on the inactivation of fluconazole-resistant C. albicans; 3- to develop a light-triggered micelles with curcumin and to evaluate its effect on antimicrobial photodynamic therapy (aPDT); 4- to evaluate the antibiofilm effect of N-acetyl cysteine (NAC) against C. albicans; and 5- to validate an in vivo model of denture stomatitis and to evaluate the expression of epithelial cytokeratins. In the study 1, quantification methods of C. albicans suspension by colonies forming units, cell counts in the In Cell Analyzer and quantitative polymerase chain reaction will be compared. In the study 2, verapamil and curcumin will be used as efflux pump inhibitors in order to reverse the fluconazole resistance of C. albicans and to potentiate aPDT in vitro and in vivo. In the study 3, light-triggered micelles with curcumin will be synthesized, characterized and its photodynamic effect against C. albicans, methicillin-resistant Staphylococcus aureus and Pseudomonas aeruginosa will be evaluated. In the study 4, the antifungal and antibiofilm concentrations of NAC will be established, their synergism with antifungals, and their effect on the matrix components of the fungal biofilm will be evaluated. In the study 5, individual acrylic devices will be adapted on palatal mucosa of rats and contaminated with different species of Candida, and the expression of epithelial cytokeratins will be assessed. Data will be analyzed by concordance, correlation and regression (study 1) and by parametric and/or non-parametric analysis of variance (studies 2, 3, 4 and 5), with ±=0.05. (AU)