Evaluation of the Effect of a New Recombinant Hybrid Protein (CANEStat) on the Proteomic Profile of the Acquired Enamel Pellicle and Dental Erosion

• Grant number: 24/09851-8
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: September 01, 2024
• End date: August 31, 2025
• Field of knowledge: Health Sciences - Dentistry - Social and Preventive Dentistry
• Principal Investigator: Marília Afonso Rabelo Buzalaf
• Grantee: Caio Augusto Imai
• 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 presence of the acquired enamel pellicle (AEP) is one of the protective factors against dental erosion. Among the proteins of the acquired pellicle with potential to bind to enamel and acid-resistant properties, cystatin and statherin have been specifically identified. The modification of the proteomic profile of the acquired pellicle through the incorporation of proteins may affect its ability to protect against dental erosion. Therefore, this modification seems promising for the control of dental erosion. The present study aims to evaluate the impact of modifying the AEP using a new recombinant hybrid protein, named CaneStat, on its proteomic profile, ultimately aiming at the prevention against dental erosion. A crossover and triple-blind study will be conducted, consisting of 4 parallel phases, corresponding to treatments with CaneStat or with deionized water (negative control). In each phase, after prophylaxis, volunteers will rinse their mouths for 1 minute. After rinsing, the AEP will be formed for 3 minutes or 2 hours. AEP collection will be done using an electrode filter paper, and samples will be stored at -80 ºC. After the formation of the AEP, an adhesive tape containing a hole to demarcate an area will be fixed on the buccal surface of the upper right central incisor. On the demarcated area, an erosive challenge will be performed with 10 µL of citric acid 1% pH 2.5 (to simulate extrinsic erosion) for 10 seconds, with the acid being placed in contact with the enamel surface in the marked area by a micropipette. After 10 seconds, the acids will be aspirated and collected for analysis of calcium released from the enamel using the arsenazo III method. Subsequently, quantitative label-free proteomic analysis will be performed (Xevo Q-TOF, Waters) on the collected AEP samples. The PLGS software will be used to detect changes in protein expression among the different groups.