Abstract
In the recent years, the demand for natural extracts that can be added to dental products has been increased for the purpose of controlling the most frequent oral problems. Accordingly, this work aims to evaluate the effect of experimental and commercial dentifrices and mouthrinses containing different natural extracts on the viability and activity of microcosm biofilm as well as in the prevention of enamel demineralization in vitro. Initially, the values of MIC and MBC will be determined for three extract from Vochysia tucanorum Mart. and Myrcia bella Cambess. leaves and Matricaria chamomilla L. flower in aqueous and hydroalcoholic solution (5% ethanol) on S. mutans (ATCC 25175) and total human salivary microorganisms. In the second step, these extracts will be tested in the forms of solution and dentifrice and compared with commercial products containing natural agents with respect to the antimicrobial and anticaries effect using a microcosm biofilm model. For this purpose, saliva pool from 10 health individuals (70% saliva and 30% glycerol) will be mixed with McBain artificial saliva (with 0.2% sucrose) for the biofilm formation. Bovine enamel samples (4 mm x 4 mm) will be submitted to microcosm biofilm formation for 5 days, which will be treated daily (1x60s / day) with solution or dentifrice. With respect to solutions, we will test three experimental and three commercial ones: 1. Vochysia tucanorum Mart., 2. Myrcia bella Cambess., 3. Matricaria chamomilla L, 4. Orgânico natural®-Contente/Uberlândia-Brazil, 5. Boni Natural Menta & Malaleuca®-Boni/São Bernardo do Campo-Brazil, 6. Malvatricin Plus®-Daudt/Rio de Janeiro-Brazil, 7. PerioGard®-Colgate-Palmolive/São Paulo-Brazil (positive control) and 8. PBS (negative control). With respect to dentifrices, we will test three experimental and three commercial ones: 1. Vochysia tucanorum Mart., 2. Myrcia bella Cambess., 3. Matricaria chamomilla L, 4. Orgânico natural®-Contente/Uberlândia-Brazil, 5. Boni Natural Menta & Malaleuca®-Boni/São Bernardo do Campo-Brazil, 6. Propolis & Myrrh®-Tom's Maine-Kennebunk-USA, 7. Colgate Total 12 Clean Mint®-Colgate-Palmolive/São Paulo-Brazil (positive control) and 8. placebo dentifrice (negative control). The response variables will be: 1) percentages of live and dead bacteria and the presence of extracellular polysaccharides by fluorescence using confocal microscopy; 2) acidogenicity of the biofilm by measuring the presence of lactic acid using enzymatic method; 3) colony forming units (CFUs) counting for total microorganisms, total streptococci, total lactobacilli, Streptococcus mutans and Streptococcus sobrinus or 4) detection and analysis of relative metabolic activity (%) of total bacteria, S. mutans, S. sobrinus, Lactobacillus (genus), Bifidobacterium dentium, Scardovia wiggsiae and Fusobacteria (genus) by qPCR; 5) quantification of enamel demineralization using transverse microradiography. The data will be submitted to the appropriate statistical analysis (parametric or non-parametric test), considering p <0.05.
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