Modulation of acquired pellicle and dental biofilm for control of dental mineral loss: use of an innovative gel with caneCPI-5 associated with vitamin e

• Grant number: 23/16774-7
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: October 01, 2024
• End date: September 30, 2027
• Field of knowledge: Health Sciences - Dentistry - Social and Preventive Dentistry
• Principal Investigator: Marília Afonso Rabelo Buzalaf
• Grantee: Tamara Teodoro Araujo
• 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 modification of the acquired enamel pellicle (AEP) through the incorporation of a sugar cane-derived cistatin has shown promising effects in controlling erosive tooth wear (ETW) and dental caries. However, its mechanism of action involves a reaction with hydroxyapatite, requiring professional prophylaxis for the initial application. Recently, the combination of vitamin E with CaneCPI-5 in separate applications has demonstrated a synergistic effect in preventing dental erosion regardless of the application sequence, whether before or after the formation of AEP. Vitamin E could facilitate the distribution of CaneCPI-5 to the basal layer of AEP. Nevertheless, performing two separate applications is not well-received by patients. Therefore, in this study, we propose the development of a nanostructured micellar gel containing CaneCPI-5 and vitamin E. This gel will be evaluated for its potential to: 1) alter the proteomic profile of the AEP after in vivo treatment (Subproject 1); 2) affect changes in the metabolites of the AEP after in vivo treatment (Subproject 1); 3) modify the microcosm enamel biofilm profile (colony-forming units - UFC - count) and bacterial activity (confocal laser microscopy) (Subproject 2a); 4) reduce enamel demineralization caused by the microcosm biofilm, as assessed by digital 3D transverse microradiography - 3D-DTMR (Subproject 2a); 5) alter the biofilm-colonizing bacteria, as assessed by DNA-DNA Checkerboard Hybridization (Subproject 2b). For the first stage, a triple-blind crossover study will be conducted, consisting of four parallel phases. After prophylaxis, volunteers will have the pellicle formed for 1 h, following which a gel containing CaneCPI-5 + vitamin E (together or separately) and/or placebo will be applied for 4 min and then gently removed. Subsequently, a 1-h wait will allow for the interaction of the gel's active principles with the AEP proteins. The pellicle will be collected and prepared for proteomic and metabolomic analyses. In the second phase, 4x4 mm bovine enamel blocks measuring will be prepared and divided into groups according to the treatment applied [gels containing: CaneCPI-5 + vitamin E (together or separately), 1.23% acidulated phosphate fluoride, Chlorhexidine, or placebo]. The blocks will be incubated in a pool of human saliva for 8 h to form the biofilm. The microcosm biofilm will be established using McBain artificial saliva combined with the human saliva pool and incubated for 16 h to reach the initial 24 h. Subsequently, the specimens will be treated according to the experimental group for 4 min, followed by rinsing with PBS. Fresh McBain + sucrose medium will then be added. The experiment will last for 5 days, with treatment and medium administered once daily. CFU counting, confocal microscopy, and 3D-DTMR analyses will be conducted for evaluation. For the final stage, the microcosm biofilm will be cultured with the previously established treatments, followed by Checkerboard Hybridization analysis. This novel approach could revolutionize our understanding and manipulation of AEP structure and properties, paving the way for new protective strategies involving pellicle engineering, enhancing its protective role, and influencing the initial biofilm colonizers.