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 statin

• Grant number: 18/24864-8
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: March 01, 2019
• End date: February 29, 2020
• Field of knowledge: Health Sciences - Dentistry - Social and Preventive Dentistry
• Principal Investigator: Marília Afonso Rabelo Buzalaf
• Grantee: Julia Chaparro Leme
• Host Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil

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 (StN15pSpS) have recently been identified by our group. Thus, the objectives of this research will be to evaluate, in vivo, the alterations in the protein composition of PAE after treatment of the dental surface with cystatin derived from sugarcane (CaneCPI-5), hemoglobin, StN15pSpS or the combination of the 3 proteins before of the formation of PAE and subsequent intrinsic or extrinsic erosive challenge, as well as to evaluate the protective potential of these treatments against the intrinsic or extrinsic erosive demineralization of the enamel. To do so, 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 ECPs, volunteers will have a mouthwash (1 min; 10 mL) with deionized water, 0.1 mg / mL CaneCPI-5, 1 mg / mL hemoglobin, StN15pSpS concentration to be defined) or solution containing mixture of the above 3 proteins. After the mouthwash, the PAE 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 two-electrode filter paper sequence, the first one being soaked with 4% sodium lauryl sulfate and the second soaked in 3% citric acid. A pool will be done with the filter papers obtained from every 3-4 volunteers for each group. The same acidic challenges will be done on the upper right central incisor surface, and after the 10 s of application of the acids (5 ¼L), the drop will be collected for analysis of P. The proteins evaluated 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 increasing their acid-resistant capacity.