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Acquired pellicle engineering for protection against caries and tooth erosion: effect of cystatin, statherin and hemoglobin on the proteomic profile of the acquired pellicle, bacterial biofilm and dental enamel

Grant number: 19/08032-5
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): July 01, 2019
Effective date (End): June 30, 2023
Field of knowledge:Health Sciences - Dentistry - Social and Preventive Dentistry
Principal researcher:Marília Afonso Rabelo Buzalaf
Grantee:Tamara Teodoro Araujo
Home Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil

Abstract

The presence of the Acquired Enamel Pellicle (AEP) is one of the protective factors against dental erosion and dental caries. Among the AEP proteins with enamel binding potential and acid-resistant properties, cystatin, hemoglobin (Hb) and statherin have recently been identified by our group. Modification of the protein profile of the AEP by the incorporation of some proteins can interfere in the initial bacterial colonization of the biofilm, changing its structure. Therefore, the modification of the AEP can be promising not only for the control of dental erosion, but also for the control of dental caries, which has been little studied. Thus, the objectives of this project will be: 1) to evaluate changes in the proteomic profile of the AEP after in vivo treatment with CaneCPI-5, statherin-derived peptide (Stn15pSpS) and hemoglobin, alone or in combination (subproject 1); 2) to evaluate changes in the enamel microcosm biofilm profile by counting Colony Forming Units (CFU) after treatment with different concentrations of CaneCPI-5, Stn15pSpS and hemoglobin, isolated and in combination (subproject 2); 3) to compare the percentage of live and dead bacteria present in the biofilm formed by the treatments described above, by fluorescence using confocal microscope (subproject 2); 4) to compare the demineralization of the enamel caused by the microcosm biofilm submitted to the treatments described above, by transverse microradiography (subproject 2); 5) to evaluate the changes in the initial colonizing bacteria of the microcosm biofilm after treatment of the enamel with CaneCPI-5, Stn15pSpS and hemoglobin, isolated or in combination, by microbiomic analysis (subproject 3). For the first step, a triple blind study will be conducted, consisting of 5 parallel phases in which the volunteers will make the mouthwash with a solution containing each of the proteins/peptide or the combination. The AEP will be collected and prepared for proteomic analysis. For the second stage of the study, blocks of bovine enamel will be made, which will be divided into groups according with the treatment (CaneCPI-5, Stn15pSpS, hemoglobin, chlorhexidine or PBS). The specimens will then be incubated in a pool of human saliva and McBain artificial saliva for microcosm biofilm formation. UFC, confocal microscopy and transverse microradiography analysis will be performed. For the last step, the biofilm will be cultured with the previously established treatments and then the microbiomic analysis will be performed. The evaluated proteins may in the future be incorporated into dental products, such as dentifrices, mouthwashes or gels, aiming at the enrichment of the PAE with them thus increasing their acid-resistant capacity. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
ARAUJO, TAMARA TEODORO; CAMILOTI, GABRIEL DOMINGUES; VALLE, ALINE DIONIZIO; SILVA, NATARA DIAS GOMES; SOUZA, BEATRIZ MARTINES; CARVALHO, THAMYRIS DE SOUZA; CAMARA, JOAO VICTOR FRAZAO; SHIBAO, PRISCILA YUMI TANAKA; HENRIQUE-SILVA, FLAVIO; CRUVINEL, THIAGO; MAGALHAES, ANA CAROLINA; BUZALAF, MARILIA AFONSO RABELO. A sugarcane cystatin (CaneCPI-5) alters microcosm biofilm formation and reduces dental caries. BIOFOULING, v. 37, n. 1 JAN 2021. Web of Science Citations: 0.
BUZALAF, MARILIA AFONSO RABELO; ORTIZ, ADRIANA DE CASSIA; CARVALHO, THAMYRIS SOUZA; FIDELES, SIMONE ORTIZ MOURA; ARAUJO, TAMARA TEODORO; MORAES, SAMANTA MASCARENHAS; BUZALAF, NATHALIA RABELO; REIS, FERNANDA NAVAS. Saliva as a diagnostic tool for dental caries, periodontal disease and cancer: is there a need for more biomarkers?. EXPERT REVIEW OF MOLECULAR DIAGNOSTICS, v. 20, n. 5 MAR 2020. Web of Science Citations: 0.

Please report errors in scientific publications list by writing to: cdi@fapesp.br.