Acquired enamel film engineering for protection against tooth erosion: in vivo evaluation of the protective effect of sugarcane-derived cystatin (CaneCPI-5), hemoglobin and/or peptide derived from staterin

• Grant number: 19/16254-8
• Support Opportunities: Scholarships in Brazil - Doctorate
• Effective date (Start): November 01, 2019
• Effective date (End): May 31, 2023
• Field of knowledge: Health Sciences - Dentistry - Social and Preventive Dentistry
• Principal Investigator: Marília Afonso Rabelo Buzalaf
• Grantee: Thamyris de Souza Carvalho
• Host Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil
• Associated scholarship(s): 21/12130-2 - Design and characterization of new resins functionalized with methacrylate-DNA conjugates hybridized with sugarcane-derived cystatin (Cane CPI-5) and human hemoglobin (HB)-DNA, BE.EP.DR

Abstract

Dental erosion is characterized by an irreversible loss of hard dental tissue caused by acids of non-bacterial origin. The Presence of Acquired Enamel film (PAE) is one of the protective factors against tooth erosion. Because of this, recent studies have focused on enhancing PAE with proteins that have high affinity for hydroxyapatite and are resistant to acid removal in procedures that have been termed "acquired film engineering". Among the acquired film proteins with enamel-binding potential and acid-resistant properties, cystatin, hemoglobin and/or statin-derived peptide (StN15pS2pS3) have recently been identified by our group. Thus, the objectives of this research will be: 1) to evaluate, in vivo, the alterations in the protein composition of the PAE after treatment of the dental surface with cystatin derived from sugarcane (CaneCPI-5), hemoglobin and/or the peptide derived from the staterin (StN15pS2pS3) or the combination of the 3 proteins before the formation of PAE and subsequent intrinsic or extrinsic erosive challenge; 2) to evaluate the protective potential of these treatments against the intrinsic or extrinsic erosive demineralization of the enamel and 3) to discover the mechanisms by which the treatment of the surface of the enamel with the peptide derived from the staterin is able to protect against dental erosion by means of test using Quartz Crystal Microbalance (QCM-D), electrokinetic analysis and Transmission Electron Microscopy (TEM) - (BEPE). For this, 10 volunteers will participate in a cross-blind, triple-blind study, consisting of 10 phases. At each stage, after dental prophylaxis, with the aim of removing all ECP, volunteers will perform mouthwash (1 min; 10 mL) with deionized water, 0.1 mg/mL CaneCPI-5, 1 mg/mL hemoglobin, StN15pS2pS3 1.88 x 10 -5 M) or solution containing mixture of the above 3 proteins. After the mouthwash, the ECP will be formed for 2 h. After 2 h for formation of PAE, for each treatment, the teeth will be isolated with cotton rolls. An adhesive tape containing an orifice of 4.52 mm2 in diameter shall be attached to the upper right central incisor. On the vestibular surfaces of the other upper and lower teeth, two types of erosive challenges will be performed in two different phases: application of 0.01 M HCl pH 2 or application of 1% citric acid pH 2.5 for 10 sec. The application will be done with a pipette. After washing for acid removal, the PAE will be collected using a sequence of electrode filter papers, soaked in 3% citric acid. A pool will be done with the filter papers obtained from every 3-4 volunteers for each group. After extraction of the proteins, they will be subjected to reverse phase liquid chromatography interconnected to a mass spectrometer (nLC-ESI-MS/MS). Protein-free quantification of markers will be done using Protein Lynx Global Service (PLGS) software. The same acidic challenges will be made on the upper right central incisor surface, and after 10 s of application of the acids (5 ¼L), the drop will be collected for analysis of P. For the external stage, the experimental design and detailing methodological tests will be described later, at the request of the BEPE. The evaluated proteins may in the future be incorporated into dental products, such as dentifrices, mouthwashes or gels, aiming at the enrichment of PAE with them and, consequently, to increase their acid-resistant capacity. (AU)

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)

CARVALHO, THAMYRIS SOUZA; ARAUJO, TAMARA TEODORO; OLIVEIRA VENTURA, TALITA MENDES; DIONIZIO, ALINE; FRAZAO CAMARA, JOAO VICTOR; MORAES, SAMANTA MASCARENHAS; PELA, VINICIUS TAIOQUI; MARTINI, TATIANA; LEME, JULIA CHAPARRO; BOGAZ DERBOTOLLI, ANA LUIZA; et al. Acquired pellicle protein-based engineering protects against erosive demineralization. Journal of Dentistry, v. 102, NOV 2020. (18/12041-7, 19/16254-8)