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Evaluation of novel salivary peptides for treatment of oral candidiasis

Grant number: 18/13904-9
Support type:Scholarships abroad - Research Internship - Doctorate
Effective date (Start): August 01, 2018
Effective date (End): May 03, 2019
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Janaina Habib Jorge
Grantee:Cláudia Viviane Guimarães Pellissari
Supervisor abroad: Walter Luiz Siqueira
Home Institution: Faculdade de Odontologia (FOAr). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
Local de pesquisa : Western University , Canada  
Associated to the scholarship:15/06016-1 - Antifungal activity and cytotoxicity of biomaterials coated with chitosan nanoparticles containing salivary proteins molecules, Histatin 1, Histatin 5 or DR9RR14 encapsulated, in co-culture, BP.DR

Abstract

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.