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Chitosan/CaneCPI-5 hybrid scaffold to prevent dental erosion: a preliminary study

Grant number: 19/25151-8
Support type:Scholarships abroad - Research Internship - Scientific Initiation
Effective date (Start): March 01, 2020
Effective date (End): June 30, 2020
Field of knowledge:Health Sciences - Dentistry
Principal Investigator:Marília Afonso Rabelo Buzalaf
Grantee:Marina Honorato Cardoso
Supervisor abroad: Prof Dr Dorothea Bruggemann
Home Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil
Local de pesquisa : University of Bremen, Germany  
Associated to the scholarship:19/09774-5 - Identification of acid resistant proteins in the acquired enamel pellicle in vivo after exposure to citric acid and hydrochloric acid as a function of variation of formation time: proteomic study, BP.IC

Abstract

Erosive tooth wear can be defined as irreversible mineral loss of dental hard tissues without microorganism involvement caused by intrinsic and extrinsic acids interference and mechanical abrasive forces, potentially increasing the tooth sensitivity. This clinical condition is caused mainly by gastroesophageal reflux disease, bulimia or anorexia nervosa (intrinsic origin), although the diet (extrinsic origin) plays a significant role due to the increasing consumption of acidic beverages in western societies. The pH of acidic beverages is higher than that of gastric acids, therefore the latter promote an increased destruction of the tooth structure. In contrast, the erosive process is counteracted by the acquired enamel pellicle (AEP), a thin layer predominantly formed from salivary proteins and their products through adsorption onto the enamel surface, but also includes non-salivary proteins, carbohydrates and lipids. According to previous studies, specific AEP proteins that strongly bind to enamel hydroxyapatite, such as a novel sugarcane cystatin called CaneCPI-5, might control erosion modulating calcium and phosphate concentrations within the oral cavity and acting as a mechanical barrier to the acids. Along with that, a polysaccharide derived from chitin known as Chitosan has been found to also protect the dental enamel against dental erosion. The association between this biopolymer and sugarcane cystatin forming a protective coating has a synergic potential and would be of great interest if applied in dental products development to improve their efficiency against acid erosion. Therefore, the objective of this study is to prepare a chitosan scaffold to study the modification with sugarcane cystatin CaneCPI-5. First, we will study the interaction with planar chitosan scaffolds and will subsequently prepare microfibrous chitosan scaffolds by wet-spinning to study the interaction between the two biopolmyers with an increased surface area. The topography and scaffold structure will be evaluated using Scanning Electronic Microscopy (SEM). The interaction between CaneCPI-5 and chitosan will be studied by Fluorescence Microscopy using fluorescently labelled CaneCPI-5 and Förster Resonance Energy Transfer (FRET) technique. Also, CaneCPI-5 concentration supernatants will be measured with UV/Vis-spectroscopy to indicate how much protein was adsorbed on the underlying chitosan scaffold. The obtained data will be analyzed by statistic tests after proper evaluation (p<0.05).