Association of L-ascorbic acid and cold atmospheric pressure plasma in the treatment of oral candidiasis: in vitro and in vivo evaluation

• Grant number: 21/00046-7
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Effective date (Start): March 01, 2021
• Effective date (End): October 31, 2024
• Field of knowledge: Health Sciences - Dentistry - Dental Clinics
• Principal Investigator: Cristiane Yumi Koga Ito
• Grantee: Noala Vicensoto Moreira Milhan
• Host Institution: Instituto de Ciência e Tecnologia (ICT). Universidade Estadual Paulista (UNESP). Campus de São José dos Campos. São José dos Campos , SP, Brazil
• Associated research grant: 19/05856-7 - Use of low temperature atmospheric pressure plasma in dentistry: from laboratory bench to clinics, AP.TEM
• Associated scholarship(s): 22/04598-7 - The effect of synergistic treatment of vitamin C and low temperature plasma on candidiasis induced in reconstituted oral mucosa, BE.EP.PD

Abstract

The aim of this project is to evaluate the association between the ascorbic acid and cold atmospheric pressure plasma (CAP) for the treatment of oral candidiasis. The study begin with in vitro experiments to establish the ideal conditions of interaction. The effect of association of CAP and different concentrations of L-ascorbic acid will be evaluated on planktonic cells of reference strains and clinical isolates of Candida albicans. The initial test range will be 5 mM to 90 mM L-ascorbic acid. Considering the results for planktonic cells, the effect of the association on 24 and 48 h biofilms will be investigated. Besides, the immunomodulatory effect of the protocol will be evaluated in RAW 264.7 macrophages. After the effective protocol will be tested for cytotoxicity to fibroblasts and keratinocytes and genotoxicity. The effective and safe protocol will be validated in murine model of induced oral candidiasis. For the experiment, 72 male mice (Mus musculus Swiss) will be randomly divided into 3 groups: treated (n=24, treated with CAP+ascorbic acid), positive control (n=24, treated with nystatin solution 100.000 IU), and negative control (n=24, without treatment). Three cycles of treatment will be done and animals will be sacrificed after 24 and 48 h. Tongues will be analyzed microbiologically to detect the number of fungal cells by plating method. Histopathological analyses, invading hypha numbering will be also performed. Tissues were analyzed for TNF-alpha, IL (interleucina)- 1² and IL-10 gene expression. Immunohistochemistry analysis using Ki67 and VEGF will be used for study cell and vascular proliferation. (AU)