Development of probiotic formulations for bucal candidiasis prevention: in vitro and in vivo study

• Grant number: 16/25544-1
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: November 01, 2017
• End date: December 31, 2019
• Field of knowledge: Health Sciences - Dentistry
• Principal Investigator: Juliana Campos Junqueira
• Grantee: Felipe de Camargo Ribeiro
• Host Institution: Instituto de Ciência e Tecnologia (ICT). Universidade Estadual Paulista (UNESP). Campus de São José dos Campos. São José dos Campos , SP, Brazil

Abstract

Candidiasis represents the main fungal infection of the oral cavity and increased resistance to antifungal by Candida spp. Has led to the search for alternative antimicrobial agents, such as probiotics. In a previous study of our research group, it was verified that among several strains of Lactobacillus isolated from the oral cavity, the Lactobacillus paracasei strain 28.4 showed the greatest capacity to inhibit C. albicans and to reduce the development of candidiasis. Thus, the objective of this project will be to develop probiotic formulations with the L. paracasei strain 28.4 for local application in the oral cavity and to test its potential as an agent for the prevention and control of candidiasis. For the formulations preparation, L. paracasei 28.4 will be incorporated in different concentrations of hydrogels using gellan gum. Thereafter, the formulations will be evaluated for the number of L. paracasei viable cells and the different storage forms, including lyophilization and storage temperatures. In addition, the ability of probiotic formulations to inhibit the in vitro growth of C. albicans will be assessed by agar diffusion, biofilm formation and filamentation assays. The agar diffusion test will be performed in culture medium yeast extract, peptone, dextrose (YPD). The biofilms will be formed in acrylic resin bodies and evaluated by means of viable cell counting, total biomass analysis and scanning electron microscopy. The filamentation of C. albicans will be induced in 24-well plates and quantified under an optical microscope. Next, the formulations will be tested in the oral cavity of healthy mice, aiming to evaluate the delivery system and oral colonization by L. paracasei 28.4, using UFC / mL recovery techniques and real time PCR quantification. The formulation that shows the best results in the in vitro studies and in the delivery system in the oral cavity of healthy mice will be tested in mice with experimental oral candidiasis induced by immunosuppression. The effects of the probiotic formulation on oral candidosis will be evaluated by counting viable cells of Candida spp. and Lactobacillus spp., quantification of L. paracasei 28.4 and C. albicans by real time PCR, determination of macroscopic lesions on the dorsum of the tongue and analysis of histological lesions. In addition, a monitoring study will be carried out on the development of oral candidiasis in real time, using the innovative bioluminescence imaging method in IVIS Lumina. The data obtained will be submitted to an exploratory analysis to select the most suitable statistical test for each experiment. In all tests will be used the program GraphPad Prism, considering a level of significance of 5%.

Scientific publications

(5)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

RIBEIRO, F. C.; ROSSONI, R. D.; DE BARROS, P. P.; SANTOS, J. D.; FUGISAKI, L. R. O.; LEAO, M. P. V.; JUNQUEIRA, J. C.. Action mechanisms of probiotics on Candida spp. and candidiasis prevention: an update. Journal of Applied Microbiology, v. 129, n. 2, NOV 2019. (17/02652-6, 17/19219-3, 16/25544-1)

DE ALVARENGA, JANAINA ARAUJO; DE BARROS, PATRICIA PIMENTEL; DE CAMARGO RIBEIRO, FELIPE; ROSSONI, RODNEI DENNIS; GARCIA, MAIRA TERRA; DOS SANTOS VELLOSO, MARISOL; SHUKLA, SHASHANK; FUCHS, BETH BURGWYN; SHUKLA, ANITA; MYLONAKIS, ELEFTHERIOS; et al. Probiotic Effects ofLactobacillus paracasei28.4 to InhibitStreptococcus mutansin a Gellan-Based Formulation. PROBIOTICS AND ANTIMICROBIAL PROTEINS, v. 13, n. 2, p. 506-517, APR 2021. (16/25544-1, 17/15529-8)

RIBEIRO, FELIPE DE CAMARGO; JUNQUEIRA, JULIANA CAMPOS; DOS SANTOS, JESSICA DIANE; DE BARROS, PATRICIA PIMENTEL; ROSSONI, RODNEI DENNIS; SHUKLA, SHASHANK; FUCHS, BETH BURGWYN; SHUKLA, ANITA; MYLONAKIS, ELEFTHERIOS. Development of Probiotic Formulations for Oral Candidiasis Prevention: Gellan Gum as a Carrier To Deliver Lactobacillus paracasei 28.4. Antimicrobial Agents and Chemotherapy, v. 64, n. 6, JUN 2020. (16/25544-1)

ROSSONI, RODNEI DENNIS; RIBEIRO, FELIPE DE CAMARGO; SANTANA DOS SANTOS, HANNA FLAVIA; DOS SANTOS, JESSICA DIANE; OLIVEIRA, NICASSIA DE SOUSA; DOS SANTOS DUTRA, MARIGNES THEOTONIO; BIAZZI DE LAPENA, SIMONE APARECIDA; JUNQUEIRA, JULIANA CAMPOS. Galleria mellonella as an experimental model to study human oral pathogens. ARCHIVES OF ORAL BIOLOGY, v. 101, p. 13-22, MAY 2019. (17/19219-3, 16/25544-1)

PEREIRA, THAIS CRISTINE; DE BARROS, PATRICIA PIMENTEL; DE OLIVEIRA FUGISAKI, LUCIANA RUANO; ROSSONI, RODNEI DENNIS; RIBEIRO, FELIPE DE CAMARGO; DE MENEZES, RAQUEL TELES; JUNQUEIRA, JULIANA CAMPOS; SCORZONI, LILIANA. Recent Advances in the Use of Galleria mellonella Model to Study Immune Responses against Human Pathogens. JOURNAL OF FUNGI, v. 4, n. 4, DEC 2018. (17/19219-3, 16/25544-1, 17/02652-6)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

RIBEIRO, Felipe de Camargo. Desenvolvimento de formulações probióticas para prevenção da candidose bucal: estudo in vitro e in vivo. 2019. Doctoral Thesis - Universidade Estadual Paulista (Unesp). Instituto de Ciência e Tecnologia. São José dos Campos São José dos Campos.