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Effect of different experimental toothpastes and oral mouthrinses containing natural extracts on the relative metabolic activity of microcosm biofilm and on enamel demineralization prevention in vitro

Grant number: 18/26506-1
Support type:Scholarships abroad - Research Internship - Doctorate
Effective date (Start): March 13, 2019
Effective date (End): March 12, 2020
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Ana Carolina Magalhães
Grantee:Aline Silva Braga
Supervisor abroad: Marcella Esteves Oliveira
Home Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil
Local de pesquisa : Uniklinik RWTH Aachen, Germany  
Associated to the scholarship:17/17249-2 - Evaluation of the effect of different experimental and commercial natural extracts in the form of toothpastes and oral mouthrinses on the viability and activity of microcosm biofilm and on enamel demineralization prevention in vitro, BP.DR

Abstract

This work aims to evaluate the effect of experimental toothpastes and mouthrinses containing different natural extracts on the relative metabolic activity (%) of total bacteria, Streptococcus mutans, Streptococcus sobrinus, Lactobacilli (genus), Bifidobacterium dentium, Scardovia wiggsiae and Fusobacteria (genus) using analyses of qPCR and the enamel demineralization by TMR. For the microcosm biofilm formation, saliva will be collected from 10 healthy individuals, who must have not brushed their teeth for the last 24 h and must not have drunk or eaten for the last 2 h. The saliva will be diluted (70% saliva and 30% glycerol), and aliquots of 2 ml will be stored at -80ºC. A hundred and seventeen samples of bovine enamel (4 mm x 4 mm) will be prepared and distributed in groups according to the type of toothpaste or mounthrinse applied: mouthrinses - 1) 2.5 mg/mL Vochysia tucanorum Mart.; 2) 1.25 mg/mL Myrcia bella Cambess.; 3) 20 mg/mL Matricaria chamomilla L.; 4) Malvatricin® Plus (Daudt, Rio de Janeiro, Brazil); 5) PerioGard® (Colgate, Palmolive, São Paulo, Brazil) (positive control); and 6) PBS (negative control) (n=9) or toothpastes - 1) 2.5 mg/mL Vochysia tucanorum Mart.; 2) 1.25 mg/mL Myrcia bella Cambess.; 3) 20 mg/mL Matricaria chamomilla L.; 4) Propolis & Myrrh® (Tom's Maine, Kennebunk, EUA); 5) Colgate Total 12 Clean Mint® (Colgate-Palmolive, São Paulo, Brasil) (positive control); 6) Placebo; and 7) PBS (6 and 7 negative controls) (n=9). The enamel samples will be exposed to human saliva mixed with McBain saliva (1:50) containing 0.2% sucrose (v=1.5 mL/sample) for the microcosm biofilm formation, using 24-wells plates for 5 days. The pH will be monitored using electrode daily. On the last day, the DNA and RNA will be isolated using the extraction kit and the relative bacterial activity will be measured for different microorganisms by using qPCR, at RWTH Aachen University, Germany. The enamel demineralization will be quantified by using TMR at University of São Paulo, Brazil. The data will be submitted to the statistical analysis (p<0.05).Keyword: biofilms; phytotherapy; toothpastes; mouthwashes; PCR.