In vitro evaluation of the protective potential against initial dental erosion of the modification of the acquired enamel pellicle with solutions containing statherin or SERINE-PHOSPHORYLATED or not statherin-derived peptides
Recently, procedures involving "Acquired Pellicle Engineering" have been proposed as a new strategy for the control of dental erosion. Incorporation of actives into mouthwash solutions or gels of topical applications could favor the ability of acquired enamel pellicle (AEP) to protect against erosion. In preliminary experiments of our group, statherin was identified as an AEP protein resistant to HCl removal at both pH 1 and pH 2, even in short-term PAE (3 min). In addition, statherin is reduced by 35% in AEP of patients with dental erosion, which makes the protein a good candidate to be used in Acquired Pellicle Engineering procedures. The objective of this project is to evaluate the protective potential against initial tooth erosion in vitro of the modification of the AEP film with statherine or phosphorylated or not statherin-derived peptides. In this in vitro study, 105 blocks of bovine enamel (4 X 4 mm) will be made, which will be divided into 7 groups (n = 15 / group), a positive control group consisting of CaneCPI-5 at 0.1 mg / mL, a negative control group, consisting of phosphate buffer solution and 5 experimental groups, consisting of a solution containing: StN15 Peptide (DSSEEKFLRRIGRFG) 1.88 X 10-5 M; Peptide StN15 (DpSpSEEKFLRRIGRFG) 1.88 X 10-5 M; Peptide StN15 (DSpSEEKFLRRIGRFG) 1.88 X 10-5 M; Peptide StN15 (DpSSEEKFLRRIGRFG) 1.88 X 10-5 M; Human Recombinant Statherin 1.88 X 10-5 M. Treatment of the specimens with the solutions will be done for 2 h at 37C under agitation. Stimulated saliva will be collected from 3 volunteers for the formation of the AEP (for 2 h) on the specimens. Then the specimens will be incubated in 0.01 M hydrochloric acid solution (pH 2) for 10 s at 37°C under constant stirring. Each specimen will be treated once daily for 3 days. Surface microhardness (SHM) analyzes will be performed and changes in SMH (post-erosion SHM baseline) will be calculated daily. The data will be analyzed using GraphPad InStat and will be checked against normality (Kolmogorov-Smirnov test) and homogeneity (Bartlett's test), to select the appropriate statistical test. The level of significance will be 5%.
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