Evaluation effect of DNAse-associated antimicrobial photodynamic therapy in the treatment of Oral Candidosis in mice infected with Candida albicans

Abstract

The aim of the study will be to evaluate the efficacy of antimicrobial photodynamic therapy (aPDT) associated with DNase (DNase I) in the treatment of induced Candidosis Oral in mice infected with fluconazole-susceptible and resistant Candida albicans strains. Female mice (Swiss) approximately 5 weeks old will be immunosuppressed and inoculated with C. albicans (107 CFU/mL). The animals will be treated for 5 consecutive days. The animals of the aPDT group will be treated using Photodithazine 200 mg/L (PDZ) associated with 50 J/cm2 LED (group P+L+); in P+L- group only PDZ will be used; in group P-L+, the animals will be treated only LED; in DNase group, the enzyme will be applied for 5 minutes. In addition, the combination of therapies (DNAse/P+L+) will be evaluated. One group will only be inoculated with C. albicans (group P-L-) and another will consist of healthy animals (CNI group). After the treatments, C. albicans will be recovered by sterile swabs. Then serial dilutions will be performed and plated on Sabouraud Dextrose Agar (SDA). After 48 hours of incubation at 37°C, the number of CFU/mL will be determined. The mice will be sacrificed 24 hours and 7 days after the treatments. In addition to microbiological evaluation, macroscopic and histological analysis of the lesions will be performed. In addition, host immune response analysis (IL-6, IL-10, MCP-1, IFN-³, TNF, IL-12p-70 protein) will be performed on flow cytometry and expression of ²-defensins (mBD1 and mBD2) will be performed by RT-qPCR. Laser confocal microscopy techniques will be used for 3D structure marking of biofilm matrix components. To complement these analyzes, the expression of extracellular matrix related genes (BGL2 and PHR1), oxidative stress (SOD1) and host substrate adhesion (EFG1 and HWP1) will be evaluated. The results will be analyzed by the most appropriate statistical method. (AU)