Dual nanocarrier of miconazole and cetylpyridinium chloride: assembly, physicochemical characterization, antibiofilm effect, cytotoxicity, and efficacy in the treatment of candidiasis in rats

• Grant number: 24/05943-5
• Support Opportunities: Regular Research Grants
• Start date: February 01, 2025
• End date: January 31, 2027
• Field of knowledge: Health Sciences - Dentistry - Dental Clinics
• Principal Investigator: Douglas Roberto Monteiro
• Grantee: Douglas Roberto Monteiro
• Host Institution: Faculdade de Odontologia (FOA). Universidade Estadual Paulista (UNESP). Campus de Araçatuba. Araçatuba , SP, Brazil
• Associated researchers: Carlos José Leopoldo Constantino ; Sandra Helena Penha de Oliveira

Abstract

This study aims to prepare and characterize a new dual nanocarrier of miconazole (MCZ) and cetylpyridinium chloride (CPC) based on chitosan(CS)-coated iron oxide nanoparticles (IONPs), as well as to evaluate its effect on oral candidiasis microcosm biofilm, its cytotoxicity on a reconstituted oral mucosal epithelium, and its efficacy in the treatment of oral candidiasis induced in rats. The nanocarrier IONPs-CS-MCZ-CPC will be obtained by mixing a solution of MCZ and CPC with a known weight of the IONPs-CS system (700 ¿g/mL). The physicochemical characterization of the dual nanocarrier will be performed by transmission electron microscopy, thermogravimetric analysis, X-ray diffraction, Fourier-transform infrared spectroscopy, dynamic light scattering, zeta potential, and MCZ-CPC release analysis. A saliva pool from five donors with oral candidiasis will be used as inoculum for the formation of microcosm biofilms, which will be formed during 72 hours on acrylic resin specimens positioned in the Amsterdam Active Attachment model. Afterward, the biofilms will be treated for 24 hours with different concentrations of the dual nanocarrier. The antibiofilm effect will be analyzed by quantifying cultivable cells, total biomass, metabolic activity, and lactic acid production. The structure and viability of biofilms will be evaluated by scanning electron microscopy and confocal laser scanning microscopy. The cytotoxic effect of the dual nanocarrier will be tested on a model of reconstituted oral mucosal epithelium, which will be grown in a specific culture medium and exposed to different concentrations of the nanocarrier for 24 hours. Next, cytokines (IL-6, IL-1¿, and stem cell factor) will be measured by enzyme-linked immunosorbent assay (ELISA), while cell viability will be analyzed by MTT and neutral red. To evaluate in vivo efficacy, a model of oral candidiasis induction in rats will be used. The animals will be immunosuppressed and infected with Candida albicans on the tongue dorsum. Three days after candidiasis induction, the rats will be treated for 7 consecutive days with topical applications of the dual nanocarrier. The animals will then be killed immediately before the first treatment, 24 hours after the third day of treatment and 24 hours after the last treatment. Next, the quantification of Candida on the tongue will be performed, in addition to histological evaluations to verify epithelial alterations and inflammatory response of the conjunctive tissue. Appropriate controls will be included in all biofilm, cytotoxicity, and in vivo efficacy assays. The data will be submitted to normality and homoscedasticity tests and, then, adequate statistical analysis will be applied for each result, with a significance level of 5%. (AU)