| Grant number: | 15/09770-9 |
| Support Opportunities: | Regular Research Grants |
| Start date: | November 01, 2015 |
| End date: | October 31, 2017 |
| Field of knowledge: | Health Sciences - Dentistry |
| Principal Investigator: | Juliana Campos Junqueira |
| Grantee: | Juliana Campos Junqueira |
| Host Institution: | Instituto de Ciência e Tecnologia (ICT). Universidade Estadual Paulista (UNESP). Campus de São José dos Campos. São José dos Campos , SP, Brazil |
| City of the host institution: | São José dos Campos |
Abstract
The objective of this study will be isolate and identify strains of Lactobacillus spp. from the oral cavity of individuals caries free and to evaluate their probiotic action in Candida albicans and Streptococcus mutans by in vitro and in vivo studies. Saliva samples will be collected and identified by the API50CHL system and confirmed by Polymerase Chain Reaction (PCR). Thirty strains identified as Lactobacillus will be submitted to the tests for the study of inhibitory action on C. albicans and S. mutans. For the C. albicans study, it will be evaluated the in vitro effects of Lactobacillus strains on biofilm formation and on the ability of C. albicans filamentation. Next, ten strains that having better inhibitory action on C. albicans will be selected for the in vivo study using host models of G. mellonella and C. elegans. In the G. mellonella model, the experimental infection will be analyzed by survival curves, quantification of fungal load in the hemolymph, and hemocyte density quantification. For the C. elegans model, the infection will be evaluated by survival curves, count of CFU/mL in the intestinal tract of nematodes and study of C. albicans filamentation. In addition, it will be performed the quantification of gene expression of EFG1, ALS1 and HWP1 in heterotypic and monotypic biofilms of C. albicans by testing Real Time PCR (qPCR). In relation to S. mutans study, the thirty Lactobacillus strains will be submitted to tests of antibacterial activity against S. mutans in planktonic growth. Hereafter, 10 strains that exhibit higher antibacterial activity will be selected for the study of antimicrobial effects on biofilms of S. mutans formed on hydroxyapatite discs. For in vivo study, one Lactobacillus strain with higher inhibitory effect on biofilm of S. mutans will be selected to assess its effects on the development of experimental caries in rats. After infection by S. mutans and Lactobacillus strain in the oral cavity of the rats, sampling of biofilm will be collected to count CFU/mL and analysis of the maxilla and mandible in stereomicroscope for quantification of occlusal caries and smooth surface will be performed. The results will be submitted to an exploratory analysis for selecting the most appropriate statistical test for each experiment of this study. (AU)
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