Considering that host defense system and conventional antimicrobial treatments are not fully effective against pathogenic microorganisms, the use of engineered peptides is quite motivating. Thus, the porpoise of this study will be to assess the potential effect of naturally occurring peptides in saliva (RR14 and DR9) and one hybrid engineered peptide (DR9-RR14) in oral mucosa coating to protect epithelial cells against C. albicans colonization, in which we propose to analyze a innovative approach of treatment for oral candidiasis in human reconstructed tissue. A protein pellicle and further biofilm formation of 48h will be performed on reconstructed human oral epithelium (HOE), according with the experimental groups. Control groups will be incubated with PBS, and experimental groups with protein in a concentration of IC 50 according to killing assay for the pellicle. After the biofilm growth, the HOE tissues will be fixed immediately. Histological and morphological evaluations using hematoxylin and eosin (H&E) and Periodic Acid-Schiff (PAS) assays will be applied. To determine cells apoptosis, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) will be assayed fluorescently using in situ cell death detection kit (Roche, West Sussex, UK) and HOE sections will be imaged under the fluorescent microscope. Statistical tests will be defined based on variation factors of the experiments and on data distribution and homoscedasticity. The significance level will be set at 5%. Potential Outcome and Clinical Importance: To develop antimicrobial peptides with broad-spectrum characteristics is also especially attractive in that multiple pathogens could potentially be targeted with one treatment possessing antibacterial, antiviral, or antifungal activity. The multidrug target approach has been successful in improving the efficacy of and reducing the emergence of resistance to HIV therapies. Thus, in combination, peptides have the potential to ultimately reduce the rate of emergence of resistant microbes, since selective pressure deviates away from one specific molecular target.
News published in Agência FAPESP Newsletter about the scholarship: