Interaction of Statherin-Derived Peptide with the Surface of Hydroxyapatite: Perspectives Based on Molecular Dynamics Simulations.

INTRODUCTION: Statherin-derived peptide (StatpSpS) has shown promise against erosive tooth wear. To elucidate its interaction with the hydroxyapatite (HAP) surface, the mechanism related to adsorption of this peptide with HAP was investigated through nanosecond-long all-atom molecular dynamics simulations. METHODS: StatpSpS was positioned parallel to the HAP surface in 2 orientations: 1 - neutral and negative residues facing the surface and 2 - positive residues facing the surface. A system containing StatpSpS without HAP was also simulated as control. In the case of systems with HAP, both partially restrained surface and unrestrained surface were constructed. Structural analysis, interaction pattern, and binding-free energy were calculated. RESULTS: In the peptide system without the HAP, there were some conformational changes during the simulation. In the presence of the surface, only moderate changes were observed. Many residues exhibited short and stable distances to the surface, indicating strong interaction. Specially, the residues ASP1 and SER2 have an important role to anchor the peptide to the surface, with positively charged residues, mainly arginine, playing a major role in the further stabilization of the peptide in an extended conformation, with close contacts to the HAP surface. CONCLUSION: The interaction between StatpSpS and HAP is strong, and the negative charged residues are important to the anchoring of the peptide in the surface, but after the initial placement the peptide rearranges itself to maximize the interactions between positive charged residues.

Statherin-Derived Peptide Protects against Intrinsic Erosive Enamel Wear in situ.

INTRODUCTION: This in situ study investigated the protective effect of a solution containing statherin-derived peptide (StatpSpS) against enamel intrinsic erosion (ERO). METHODS: Fifteen volunteers wore appliances containing 2 bovine specimens. The samples were subjected to ERO with HCl, mimicking dental ERO by intrinsic acid. The volunteers participated in 3 phases (double-blind and crossover): (1) deionized water (negative control); (2) commercial solution containing SnCl2/NaF/AmF (800 ppm Sn+2, 500 ppm F-, pH 4.5) (positive control); (3) solution containing 1.88 × 10-5m StatpSpS. Four times a day, the volunteers administered one drop of the solutions (50 µL, 1 min) on each specimen. After the treatment, erosive challenges were performed extraorally with 0.01 m HCl (pH 2.0, 4 times/day, 1 min, 150 mL). Enamel wear was assessed by profilometry. Data were analyzed by one-way RM-ANOVA/Bonferroni's tests (p < 0.05). RESULTS: In terms of the treatments, both the commercial solution (SnCl2/NaF/AmF) and StatpSpS significantly reduced the wear when compared to the negative control (p < 0.01), without significant differences between them (p > 0.05). CONCLUSION: The solution containing StatpSpS demonstrated protection against enamel intrinsic erosive wear. This study marks a significant advancement in the prevention of intrinsic erosive wear, utilizing StatpSpS in acquired pellicle engineering procedures.

Endodontic treatment of odontogenic infection in a patient at risk of medication-related osteonecrosis of the jaw: a case report and review of treatment criteria.

Medication-related osteonecrosis of the jaw (MRONJ) is an uncommon and severe condition in patients who are receiving treatment with antiresorptive and/or antiangiogenic drugs, which are frequently used to treat or prevent cancer metastases. The treatment of MRONJ is challenging because the choice of a specific dental treatment depends on several factors, including the systemic condition of the patient, type and dosage of medications, and clinical and radiographic characteristics of the dental lesions. This case report describes the conservative endodontic treatment of an odontogenic infection in a patient at risk of developing MRONJ owing to bisphosphonate treatment. Endodontic retreatment was performed to control the odontogenic infection and avoid tooth extraction. Certain factors favor the use of a conservative approach, such as a localized and small infection, the absence of systemic factors (such as metabolic disorders or medications) that can impair bone healing, and good oral hygiene.

Rinsing with Statherin-Derived Peptide Alters the Proteome of the Acquired Enamel Pellicle.

Changes in the proteomic profile of the acquired enamel pellicle (AEP) formed for 3 min or 2 h after rinsing with a peptide containing the 15 N-terminal residues of statherin, with serines 2 and 3 phosphorylated (StatpSpS), were evaluated. Nine volunteers participated in 2 consecutive days. Each day, after professional tooth cleaning, they rinsed for 1 min with 10 mL of phosphate buffer containing 1.88 × 10-5 M StatpSpS or phosphate buffer only (control). The acquired pellicle formed on enamel after 3 min or 2 h was collected with electrode filter papers soaked in 3% citric acid. After protein extraction, samples were analyzed by quantitative shotgun label-free proteomics. In the 3-min AEP, 19 and 131 proteins were uniquely identified in the StatpSpS and control groups, respectively. Proteins typically found in the AEP were only found in the latter. Only 2 proteins (neutrophil defensins) were increased upon treatment with StatpSpS, while 65 proteins (among which are several typical AEP proteins) were decreased. In the 2-h AEP, 50 and 108 proteins were uniquely found in StatpSpS and control groups, respectively. Hemoglobin subunits and isoforms of keratin were only found in the StatpSpS group, while cystatin-C, cathepsin D, and cathepsin G, isoforms of heat shock 70 and protocadherin were exclusively found in the control group. In addition, 23 proteins were increased upon treatment with StatpSpS, among which are histatin-1, serum albumin, and isoforms of neutrophil defensin and keratin, while 77 were decreased, most of them were typical AEP proteins. In both evaluated periods, rinsing with StatpSpS profoundly changed the proteomic profile of the AEP, which might impact the protective role of this integument against carious or erosive demineralization. This study provides important insights on the dynamics of the protein composition of the AEP along time, after rinsing with a solution containing StatpSpS.

Toothpaste containing TiF4 and chitosan against erosive tooth wear in situ.

OBJECTIVE: This study compared the protective effect of an experimental TiF4/Chitosan toothpaste with a commercial toothpaste on the prevention of erosive tooth wear (ETW) in situ. METHODS: Fifteen subjects took part in this crossover and double-blind study, in which they wore a palatal appliance containing 4 bovine enamel and 4 dentin in 3 phases (5 days each). Half of the samples were subjected to erosive challenges (90 s in 0.1 % citric acid, pH 2.5, 4 times/day), and the other half to erosive plus abrasive challenges (15 s plus 45 s of contact, 2 times/day). The phases corresponded to the application of the different toothpastes: 1) TiF4 (1400 ppm F-) plus Chitosan, 2) Elmex®, Erosion Protection (1400 ppm F-, Chitosan), and 3) Placebo (negative control). Tooth wear was measured using contact profilometry (µm) and submitted to two-way RM ANOVA/Tukey test (p < 0.05). RESULTS: No significant differences were detected between the experimental and commercial toothpastes, regardless of the challenge on both tissues. Both significantly reduce ETW compared to negative control (p < 0.0006). Tooth wear was increased by brushing only on eroded enamel (p < 0.01), but not on dentin (p = 0.6085). TiF4/Chitosan [erosion 2.98 ± 1.12 µm vs. erosion and abrasion 3.12 ± 1.33 µm] and Elmex® toothpastes [erosion 2.35 ± 0.93 µm vs. erosion and abrasion 2.98 ± 1.0 µm] minimized the impact of brushing compared to placebo on enamel [erosion 4.62 ± 1.48 µm vs. erosion and abrasion 5.15 ± 1.50 µm]. CONCLUSIONS: TiF4 plus chitosan toothpastes showed to be effective in minimizing the ETW as the commercial toothpaste is in situ. CLINICAL RELEVANCE: The experimental toothpaste has similar effect against ETW compared to the commercial toothpaste. Considering the increased ETW prevalence worldwide, this result supports clinical trials and a possible application of this experimental anti-erosive toothpaste in the future.

Effect of a metal artifact reduction algorithm on dehiscence and fenestration detection around zirconia implants with cone beam computed tomography.

OBJECTIVE: To assess the efficacy of the metal artifact reduction algorithm (MARA) of the Cranex 3D cone beam computed tomography (CBCT) device in the detection of peri-implant dehiscence and fenestration around zirconia implants. STUDY DESIGN: In total, 60 implants were placed in bovine ribs. Dehiscence and fenestration defects were created around the implants, after which 60 CBCT images were obtained with and 60 without activation of MARA. Three radiologists examined the images for the presence of defects. The area under the curve (AUC) from receiver operating characteristic analysis, sensitivity, and specificity were calculated to assess the ability to discriminate the presence vs absence of bone defects. One-way analysis of variance was employed to analyze outcome measures. The significance level was established at 5% (α = 0.05). RESULTS: AUC values indicated excellent discrimination of dehiscence on images with MARA activation and an excellent to outstanding range of discrimination with MARA deactivation. For fenestration, MARA activation and deactivation both led to outstanding discrimination. Sensitivity and specificity values revealed that activation of MARA was helpful in distinguishing the presence vs. absence of dehiscence, while both MARA conditions were helpful for fenestration. However, there were no statistically significant differences between MARA activation and deactivation for any outcome measure (P >.05). CONCLUSION: CBCT is suitable for detecting peri-implant defects, but MARA application does not significantly affect peri-implant dehiscence and fenestration detection.

Proteomic analysis of stimulated saliva in gastroesophageal reflux disease patients with and without erosive tooth wear: Observational study.

OBJECTIVE: To evaluate the difference in the proteomic profile of stimulated saliva in patients with gastroesophageal reflux disease (GERD) with (GE) and without (GNE) erosive tooth wear (ETW), regarding both human and bacterial proteins. METHODS: Stimulated saliva (SS) was collected from 16 patients (8/group). Samples were centrifuged at 4.500 g for 15 min under refrigeration to remove all debris. The supernatant from each saliva sample was taken and frozen at -80 °C. After extracting the proteins, they were submitted to reverse phase liquid chromatography and mass spectrometry (nLC-ESI-MS/MS). Label-free proteomic quantification was performed using Protein Lynx Global Service (PLGS) software (p < 0.05) for human and bacterial proteins. RESULTS: In total, 67 human proteins were common for GNE and GE groups. GNE group presented, compared to GE group, increase in proteins that confer antimicrobial and acid resistant properties, such as cystatins, histatin and immunoglobulins. However, GNE group had a marked decrease in subunits of hemoglobin (α, ß and delta). Regarding bacterial proteins, for SS, 7 and 10 unique proteins were identified in the GE and GNE groups, respectively. They are related to protein synthesis and energy metabolism and interact with human proteins typically found in saliva and supramolecular complexes of the acquired pellicle. CONCLUSIONS: Our data indicate that the stimulation of the salivary flow increases acid resistant and antimicrobial proteins in saliva, which might protect against ETW. CLINICAL SIGNIFICANCE: This pioneer study showed important differences in the human and bacterial proteome of SS in patients with GERD with or without ETW.

Effect of sweetener containing Stevia on the development of dental caries in enamel and dentin under a microcosm biofilm model.

OBJECTIVE: This study compared the effect of commercial and pure sweetener containing stevia to that of aspartame, to sucrose and xylitol on the development of dental caries. METHODS: 228 bovine enamel and root dentin were exposed to microcosm biofilm model using human saliva. From the 2nd to the 5th day, the samples were exposed daily to McBain saliva supplemented with 0.2% of the respective sweeteners/sugar, under 5% CO2 and 37 °C. The lactic acid and the colony-forming units (CFU) were quantified. The demineralization was analyzed by TMR. The data were compared statistically (Kruskal-Wallis/ Dunn, p<0.05). RESULTS: Pure stevia, pure aspartame, xylitol and control were able to significantly reduce 92% of lactate production compared to sucrose. Stevia finn, aspartame finn and sucrose showed similar production of lactic acid (around 0.45±0.12 g/L and 0.67±0.18 g/L, for enamel and dentin, p<0.0001). With respect to total lactobacilli and S. mutans/S. sobrinus CFU, xylitol and control did not show growth on enamel, while CFU numbers were found in stevia finn, aspartame finn and sucrose groups for both tissues. Enamel and dentin demineralization was significantly reduced for xylitol, control, pure stevia and pure aspartame (85% and 83% reduction, respectively) compared to stevia finn, aspartame finn and sucrose, which in turn did not differ from each other (sucrose ΔZ: 2913.7 ± 646.7 vol%.µm for enamel and 3543.3 ± 432.5 vol%.µm for dentin). CONCLUSIONS: Commercial sweeteners containing stevia and aspartame proved to be as cariogenic as sucrose, which may be due to the other components, since the pure forms were not cariogenic. CLINICAL RELEVANCE: Our study showed that some commercial sweeteners (aspartame and stevia) are as cariogenic as sucrose, which may be due to the presence of lactose. The population should be advice about the presence of lactose in such brand names, to avoid their consume.

Statherin-derived peptide protects against intrinsic erosion.

OBJECTIVES: In the present study, we used an in vitro initial intrinsic erosion model to evaluate: (experiment 1) the influence of the degree of serine (Ser) phosphorylation of peptides containing the 15 N-terminal residues of statherin and (experiment 2) the effect of different concentrations of the peptide with the best performance in experiment 1 on initial enamel erosion. DESIGN: Bovine enamel specimens were divided into 6 groups (n = 15/group) for each experiment. In experiment 1, the peptides evaluated (at 1.88 × 10-5 M) were: not phosphorylated (StatSS), phosphorylated in Ser2 (StatpSS), phosphorylated in Ser3 (StatSpS) phosphorylated in Ser2 and Ser3 (StatpSpS). Phosphate buffer and human recombinant statherin were used as negative and positive controls, respectively. In experiment 2, StatpSpS was evaluated at different concentrations: 0.94, 1.88, 3.76 and 7.52 × 10-5 M. Phosphate buffer and 0.1 mg/mL CaneCPI-5 were employed as negative and positive controls, respectively. In each experiment, the specimens were incubated with the solutions for 2 h, then the AEP was allowed to form (under human pooled saliva) for 2 h. The specimens were then challenged with 0.01 M HCl for 10 s. Demineralization was evaluated by percentage of surface hardness change (%SHC). Data were analyzed by ANOVA and Tukey's test (p < 0.05). RESULTS: In experiment 1, only StatpSpS significantly reduced the % SHC in comparison with control. In experiment 2, 1.88 × 10-5 M StatpSpS significantly reduced the %SHC in comparison with control. CONCLUSIONS: This is the first study showing that statherin-derived peptide might protect against intrinsic erosion.