Acquired enamel pellicle and dental biofilm engineering with CaneStat, an innovative hybrid protein: a new strategy for the control of dental caries

• Grant number: 24/15179-0
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: December 01, 2024
• End date: November 30, 2027
• Field of knowledge: Health Sciences - Dentistry - Social and Preventive Dentistry
• Principal Investigator: Marília Afonso Rabelo Buzalaf
• Grantee: Thais Francini Garbieri
• Host Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil
• Associated research grant: 19/26070-1 - Modulation of acquired enamel pellicle to control dental mineral loss: unravelling mechanisms to make therapies possible, AP.TEM

Abstract

The engineering of the Acquired Enamel Film (AEP) and dental biofilm allows the development of strategies aimed at protective effects in the control of erosive tooth wear and dental caries. Considering that the formation of dental biofilm occurs through the binding of adhesins on the surface of bacteria to complementary receptors for AEP proteins, the modification in the protein composition of this integument, due to enrichment with acid-resistant proteins, will have an impact on initial microbial colonization of tooth enamel, altering its entire structure. The objective of this study will be to evaluate the impact of modifying the proteomic profile of AEP formed in vivo, after treatment with a new recombinant hybrid protein, called CaneStat, as well as on bacterial biofilm and tooth enamel in vitro, ultimately aiming to prevent dental caries. Specifically, we will evaluate, after treatment with different concentrations of CaneStat gel (0.01, 0.025 and 0.05 mg/mL), its potential to 1) alter the proteomic profile of AEP in vivo; 2) alter the profile and viability of the biofilm and bacterial activity in vitro; 3) interfere with the enamel demineralization caused by the microcosm biofilm, also in vitro; and 4) alter the bacteria colonizing the biofilm, assessed by Checkerboard DNA-DNA Hybridization. After prophylaxis, volunteers will apply a mouthwash containing CaneStat, or control and after the experimental times, the AEP will be collected for proteomic analysis. For another phase, blocks of bovine enamel (4 x 4 mm) will be made, which will be divided into experimental groups treated with different concentrations of CaneStat, positive and negative controls. The blocks will be incubated in a human saliva pool for biofilm formation. The microcosm biofilm will be established using McBain's artificial saliva combined with pool of human saliva and subsequently, the blocks will be treated according to the experimental groups, for 5 days, with the treatment and/or culture medium, administered once a day. CFU count, confocal microscopy and TMRD-3D analyzes will be performed. For the last step, the microcosm biofilm will be cultivated with previously established treatments and then Checkerboard Hybridization analysis will be performed. This approach allows for new protection strategies involving AEP engineering, reinforcing its protective role, in addition to influencing the initial colonizers of the biofilm.