Acquired pellicle and biofilm engineering with CaneCPI-5: Insights from proteomic and microbiomics analysis.

OBJECTIVE: In this in vivo proof-of-concept study, acquired pellicle engineering was implemented to promote alterations in the protein composition of the acquired enamel pellicle (AEP) and the bacterial composition of the dental biofilm after treatment with Sugarcane cystatin (CaneCPI-5). DESIGN: After prophylaxis, 10 volunteers rinsed (10 mL, 1 min) with the following solutions: 1) deionized water (H2O- negative control or 2) 0.1 mg/mL CaneCPI-5. The AEP and biofilm were formed along 2 or 3 h, respectively. The AEP was collected with electrode filter papers soaked in 3 % citric acid. After protein extraction, samples were analyzed by quantitative shotgun label-free proteomics. The biofilm microbiome was collected with a dental curette. The DNA was extracted, amplified, and analyzed by 16S-rRNA Next Generation Sequencing (NGS). RESULTS: Treatment with CaneCPI-5 increased several proteins with antimicrobial, acid-resistance, affinity for hydroxyapatite, structural and calcium binding properties, such as Cysteine-rich-3 (6-fold-p = 0.03), Cystatin-B (5.5-fold-p < 0.01), Neutrophil-defensin 1 (4.7-fold-p < 0.01), Mucin (3.9-fold-p < 0.01), Immunoglobulin-heavy-constant (3.8-fold-p < 0.01) and Lactotransferrin (2.8-fold-p < 0.01). Microbiome revealed that several commensal bacteria had their abundance increased after rinsing with CaneCPI-5, such as Corynebacterium and Neisseria, while Streptococcus and Prevotella nigrescens were decreased. The results indicate the efficiency of CaneCPI-5 in promoting beneficial changes in the AEP and biofilm, making this phytocystatin a potential target for incorporation into dental products. CONCLUSION: Cane demonstrated the capability to alter the protein composition of the acquired enamel pellicle (AEP) and the initial colonizers of the biofilm, enhancing the presence of proteins and bacteria crucial for dental protection.

In vivo modification of the enamel pellicle and saliva resveratrol levels after use of resveratrol-containing orodispersible capsules.

OBJECTIVES: To evaluate in vivo 1) the bioavailability of trans-resveratrol when administered through sublingual capsules; 2) the effect of resveratrol on the protein composition of the acquired enamel pellicle (AEP). DESIGN: Ten volunteers received a sublingual capsule containing 50 mg of trans-resveratrol. Unstimulated saliva was then collected after 0, 30, 60, and 120 min and AEP was collected after 120 min following administration of the capsule. In the next week, the volunteers received a placebo sublingual capsule, and saliva and AEP were collected again. Saliva samples were analyzed for free trans-resveratrol using high-performance liquid chromatopgraphy (HPLC), and AEP samples were subjected to proteomic analysis (nLC-ESI-MS/MS). RESULTS: Trans-resveratrol was detected in saliva at all the time points evaluated, with the peak at 30 min. A total of 242 proteins were identified in both groups. Ninety-six proteins were increased and 23 proteins were decreased in the Resveratrol group. Among the up-regulated proteins, isoforms of cystatins, PRPs, Mucin-7, Histatin-1, Lactotrasnferrin and Lysozyme-C were increased and the isoforms of Protein S100, Neutrophil defensins, Albumin, PRPs, and, Statherin were decreased in Resveratrol group. CONCLUSION: The sublingual capsule is effective at increasing the bioavailability of trans-resveratrol in saliva. Several proteins involved in important processes to maintain systemic and oral health homeostasis were identified. These proteins differently expressed due to the presence of trans-resveratrol deserve attention for future studies, since they have important functions, mainly related to antimicrobial action.

Acquired enamel pellicle and biofilm engineering with a combination of acid-resistant proteins (CaneCPI-5, StN15, and Hemoglobin) for enhanced protection against dental caries - in vivo and in vitro investigations.

OBJECTIVE: This study was designed in two-legs. In the in vivo, we explored the potential of a rinse solution containing a combination (Comb) of 0.1 mg/mL CaneCPI-5 (sugarcane-derive cystatin), 1.88 × 10- 5M StN15 (statherin-derived peptide) and 1.0 mg/mL hemoglobin (Hb) to change the protein profile of the acquired enamel pellicle(AEP) and the microbiome of the enamel biofilm. The in vitro, was designed to reveal the effects of Comb on the viability and bacterial composition of the microcosm biofilm, as well as on enamel demineralization. MATERIALS AND METHODS: In vivo study, 10 participants rinsed (10mL,1 min) with either deionized water (H2O-control) or Comb. AEP and biofilm were collected after 2 and 3 h, respectively, after rinsing. AEP samples underwent proteomics analysis, while biofilm microbiome was assessed via 16 S-rRNA Next Generation Sequencing(NGS). In vitro study, a microcosm biofilm protocol was employed. Ninety-six enamel specimens were treated with: 1)Phosphate-Buffered Solution-PBS(negative-control), 2)0.12%Chlorhexidine, 3)500ppmNaF and 4)Comb. Resazurin, colony-forming-units(CFU) and Transversal Microradiography(TMR) were performed. RESULTS: The proteomic results revealed higher quantity of proteins in the Comb compared to control associated with immune system response and oral microbial adhesion. Microbiome showed a significant increase in bacteria linked to a healthy microbiota, in the Comb group. In the in vitro study, Comb group was only efficient in reducing mineral-loss and lesion-depth compared to the PBS. CONCLUSIONS: The AEP modification altered the subsequent layers, affecting the initial process of bacterial adhesion of pathogenic and commensal bacteria, as well as enamel demineralization. CLINICAL RELEVANCE: Comb group shows promise in shaping oral health by potentially introducing innovative approaches to prevent enamel demineralization and deter tooth decay.

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.

Increase in plasma resveratrol levels and in acid-resistant proteins in the acquired enamel pellicle after use of resveratrol-containing orodispersible tablets.

OBJECTIVE: This study evaluated the effect of administration of trans-resveratrol-containing orodispersible tablets on the protein composition of the AEP and on blood plasma trans-resveratrol concentrations. METHODS: Ten volunteers participated in two crossover double-blind phases. In each phase, after dental prophylaxis, they received a trans-resveratrol (15 mg) orodispersible tablet, or a placebo tablet (without actives). The AEP formed after 120 min was collected with electrode filter papers soaked in 3 % citric acid. Blood samples were collected 30, 45, 60 and 120 min after the use of the tablet. After protein extraction, AEP samples were analyzed by shotgun labelfree quantitative proteomics and plasma samples were analyzed by high-performance liquid chromatography (HPLC). RESULTS: Eight hundred and two proteins were identified in the AEP. Among them, 336 and 213 were unique to the trans-resveratrol and control groups, respectively, while 253 were common to both groups. Proteins with important functions in the AEP had increased expression in the trans-resveratroltreated group, such as neutrophil defensins, S100 protein isoforms, lysozyme C, cystatin-D, mucin-7, alphaamylase, albumin, haptoglobin and statherin. Trans-resveratrol was detected in the plasma at all the times evaluated, with the peak at 30 min. CONCLUSIONS: The administration of trans-resveratrol in sublingual orodispersible tablets was effective both to increase the bioavailability of the polyphenol and the expression of antibacterial and acid-resistant proteins in the AEP, which might benefit oral and general health.

Hemoglobin Protects Enamel against Intrinsic Enamel Erosive Demineralization.

INTRODUCTION: This study investigated the changes in the acquired enamel pellicle (AEP) proteome when this integument is formed in vivo after treatment with sugarcane-derived cystatin (CaneCPI-5), hemoglobin (HB), and a statherin-derived peptide (StN15), or their combination and then exposed to an intrinsic acid challenge. The effectiveness of these treatments in preventing intrinsic erosion was also evaluated. METHODS: Ten volunteers, after prophylaxis, in 5 crossover phases, rinsed with the following solutions (10 mL, 1 min): control (deionized water-H2O) - group 1, 0.1 mg/mL CaneCPI-5 - group 2, 1.0 mg/mL HB - group 3, 1.88 × 10-5M StN15 - group 4, or a blend of these - group 5. Following this, AEP formation occurred (2 h) and an enamel biopsy (10 µL, 0.01 m HCl, pH 2.0, 10 s) was conducted on one incisor. The biopsy acid was then analyzed for calcium (Arsenazo method). The vestibular surfaces of the other teeth were treated with the same acid. Acid-resistant proteins in the residual AEP were then collected and analyzed quantitatively via proteomics. RESULTS: Compared to control, treatment with the proteins/peptide, mixed or isolated, markedly enhanced acid-resistant proteins in the AEP. Notable increases occurred in pyruvate kinase PKM (11-fold, CaneCPI-5), immunoglobulins and submaxillary gland androgen-regulated protein 3B (4-fold, StN15), Hb, and lysozyme C (2-fold, StN15). Additionally, a range of proteins not commonly identified in the AEP but known to bind calcium or other proteins were identified in groups treated with the tested proteins/peptide either in isolation or as a mixture. The mean (SD, mM) calcium concentrations released from enamel were 3.67 ± 1.48a, 3.11 ± 0.72a, 1.94 ± 0.57b, 2.37 ± 0.90a, and 2.38 ± 0.45a for groups 1-5, respectively (RM-ANOVA/Tukey, p < 0.05). CONCLUSIONS: Our findings demonstrate that all treatments, whether using a combination of proteins/peptides or in isolation, enhanced acid-resistant proteins in the AEP. However, only HB showed effectiveness in protecting against intrinsic erosive demineralization. These results pave the way for innovative preventive methods against intrinsic erosion, using "acquired pellicle engineering" techniques.

Legislation in Hospital Dentistry: Gaps, Perspectives and Desires

Abstract Objective: To analyze the existing state legislation, including not only the ordinary laws, but also their infralegal regulation and the state legislation on Hospital Dentistry. Material and Methods: A survey was carried out in the databases of the Legislative Assemblies of the Brazilian States and the Federal District, as well as the Regional and Federal Councils of Dentistry in Brazil. Subsequently, a survey was carried out in the databases of the Ministry of Health, State Dental Councils and Federal Dental Council in Brazil. Results: Only 8 Brazilian states have legislation in force regarding hospital dentistry, which represents 29.63% of the federative units. Among the Brazilian regions, the Midwest presented the highest prevalence of the laws found (37.50%), followed by the North (25%) and the other regions with the same coverage (12.50%). Also, an orientation and an ordinance from the Ministry of Health, six resolutions from the Federal Council of Dentistry, and a technical note from the National Health Surveillance Agency were found. Conclusion: Several States do not have rules on the subject, making it imperative to create a federal rule that not only imposes the presence of the dentist, but also regulates the proportion of the team, workload, and availability.

Desensitizing Agent Previously Applied During In-Office Bleaching: A Double-Blind Randomized Clinical Trial

Abstract Objective: To compare the clinical effect of two desensitizing agents used before the application of a bleaching gel based on 35% hydrogen peroxide (HP). Material and Methods: 30 patients were selected, and two desensitizing agents with different mechanisms of action were applied: Fluorine Neutral 2% (FN), which acts by blocking dentinal canaliculi while Potassium Nitrate 5% with 2% Sodium Fluoride (PN/SF) that acts in nerve transmission and blockade. Desensitizers were used before the application of 35% HP. For whitening, three clinical sessions were performed, with an interval of seven days, with three applications of the bleaching gel for 15 minutes, totaling 45 minutes/session. Tooth sensitivity (TS) was assessed with the numerical analog scale, and a spectrophotometer was used to obtain the color variation (ΔE). ΔE were submitted to ANOVA and Tukey test (p<0.05), and TS data were submitted to a two-way ANOVA analysis. Results: For sensitivity experience, the Tukey test indicated differences between PN/SF and the placebo I, but there was no statistically significant difference between FN and the placebo II. The TS was lower when the desensitizing gel was used during the bleaching procedure compared to after treatment, regardless of the desensitizing agents. Conclusion: PN/SF before in-office tooth bleaching can reduce TS intensity, and the use of desensitizing gel before bleaching did not affect the bleaching efficacy.

A new role for resveratrol: Protection of enamel against erosion.

OBJECTIVE: The aim of this study was to determine the effect of different concentrations of resveratrol in protecting enamel against initial dental erosion in vitro. METHODS: Ninety bovine enamel samples (4 × 4 mm) were divided into six groups: Phosphate buffered saline (negative control; PBS), Commercial solution (Elmex Erosion Protection™; positive control) and resveratrol at 4 different concentrations (1, 10, 100 or 400 µg/mL). Initially, the samples were incubated in saliva for the formation of the acquired pellicle (250 µL, 1 h, 37 °C, 250 rpm). Afterward, the samples were incubated in the respective treatments (250 µL, 1 min, 37 °C, 250 rpm) and then reincubated in saliva (250 µL, 1 h, 37 °C, 250 rpm). Finally, the samples were subjected to an erosive challenge by incubating in 1 % citric acid (1 mL, pH 3.5, 1 min, 25 °C, 250 rpm). The percentage surface microhardness change (% SMC) was assessed using a microhardness tester. Data were analyzed by Kruskal-Wallis and Dunn's tests (p < 0.05). RESULTS: The treatments with Elmex™ and resveratrol (1, 10 and 100 µg/mL) significantly protected enamel compared to the negative control, without significant differences among them. However, the group treated with the highest resveratrol concentration (400 µg/mL) did not show a significant difference from the negative control. CONCLUSIONS: Resveratrol at concentrations ranging from 1 to 100 µg/ml was effective in preventing loss of enamel surface microhardness. CLINICAL SIGNIFICANCE: This result suggests a potential new direction for the development of dental products based on resveratrol for the prevention of dental erosion.

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.

Gels containing statherin-derived peptide protect against enamel and dentin erosive tooth wear in vitro.

The effect of gels containing a statherin-derived peptide (Stn) on the protection against enamel and dentin erosive tooth wear (ETW) in vitro was evaluated. Bovine enamel and dentin specimens were divided into 2 groups (n = 15 and 18/group for enamel and dentin, respectively) that were treated with Chitosan or Carboxymethylcellulose (CMC) gels containing Stn15pSpS at 1.88 × 10-5 M or 3.76 × 10-5 M. Chitosan or CMC gels without active ingredients served as negative controls, while chitosan gel containing 1.23% F (as NaF) and acidulated phosphate fluoride gel (1.23% F) served as positive controls. The gels were applied on the specimens for 4 min. Stimulated saliva was collected from 3 donors and used to form a 2-h acquired pellicle on the specimens. Then, the specimens were submitted to an erosive pH cycling protocol 4 times/day for 7 days (0.01 M HCl pH 2.0/45 s, artificial saliva/2 h, and artificial saliva overnight). The gels were applied again during pH cycling, 2 times/day for 4 min after the first and last erosive challenges. Enamel and dentin loss (µm) were assessed by contact profilometry. Scanning electron microscopy (SEM) was analyzed using a cold field emission. Data were analyzed by two-way ANOVA (for chitosan and CMC gels, separately) and Tukey's multiple comparison test. SEM images showed changes to enamel topography after application oft the gels containing Stn or F. Regarding CMC-based gels, for enamel, none of the treatments significantly reduced ETW in comparison with placebo; for dentin, however, gels containing Stn, regardless the concentration, significantly reduced the ETW. Moreover, Chitosan-based gels, regardless the Stn concentration, were able to protect enamel and dentin against ETW. Gels containing Stn might be a new approach to protect against ETW.

Acquired enamel pellicle protects gastroesophageal reflux disease patients against erosive tooth wear

Abstract The objective of this study was to compare the protein profile of the acquired enamel pellicle (AEP) formed in vivo in patients with or without gastroesophageal reflux disease (GERD), and with or without erosive tooth wear (ETW). Twenty-four volunteers were divided into 3 groups: 1) GERD and ETW; 2) GERD without ETW; and 3) control (without GERD). The AEP formed 120 min after prophylaxis was collected from the lingual/palatal surfaces. The samples were subjected to mass spectrometry (nLC-ESI-MS/MS) and label-free quantification by Protein Lynx Global Service software. A total of 213 proteins were identified, or 119, 92 and 106 from each group, respectively. Group 2 showed a high number of phosphorylated and calcium-binding proteins. Twenty-three proteins were found in all the groups, including 14-3-3 protein zeta/delta and 1-phosphatidylinositol. Several intracellular proteins that join saliva after the exfoliation of oral mucosa cells might have the potential to bind hydroxyapatite, or participate in forming supramolecular aggregates that bind to precursor proteins in the AEP. Proteins might play a central role in protecting the dental surface against acid dissolution.

Health changes during Covid-19: a nationwide study with dental students

ABSTRACT OBJECTIVE To assess the changes in stress levels, social behavior, dietary and parafunctional habits, oral hygiene, among other conditions perceived by dental students in Brazil during the Covid-19 pandemic and evaluated the correlations between stress level and other variables. METHODS An online questionnaire was developed and validated. Undergraduates enrolled in private and public dental schools were recruited by convenience sampling. Data were collected on the perceived changes regarding stress levels, financial and social characteristics, dietary habits, oral hygiene, health conditions, and parafunctional habits. Quantitative variables were expressed as absolute and relative frequencies. Wilcoxon test evaluated comparisons between perceived changes, and correlations between changes in stress levels and other variables were analyzed by Spearman correlation (α = 0.05). RESULTS A total of 638 dental students, mean age of 22.95 ± 4.10 years, participated in the study. During the pandemic, the reported stress levels increased while household income decreased (p < 0.05). Late dinners and mindless eating increased in frequency, whereas oral hygiene decreased (p < 0.05). Most of the health conditions and parafunctional habits assessed changed (p < 0.05). Perceived stress levels showed poor negative correlations with household income (rS = −0.14), poor positive correlations with the pressure to contribute financially in the household (rS = 0.19), and poor positive correlations with food choice frequency (rS = 0.15) (p < 0.05). CONCLUSIONS Dental students reported perceived changes in stress levels, dietary habits, oral hygiene, health conditions, parafunctional habits, and social behavior. Moreover, the results showed poor correlations, as students with higher stress levels tended to have the lowest household income, feel pressured to contribute financially in the household, and present a high meal intake frequency.

Surface roughness evaluation and whitening efficiency on tooth enamel after using whitening toothpaste: a randomized double-blinded study

The aim of this randomized double-blinded study was to evaluate the enamel surface roughness and color change after one month of whitening toothpaste use and the color stability obtained 1 month after its interruption. 30 volunteers were divided into 3 groups (n = 10) corresponding to the dentifrices: 1) Colgate Total 12 Clean Mint (TD) (Control), 2) Colgate Luminous White (LW) and 3) Sensodyne Whitening Extra Fresh (SB). The volunteers were impression with addition silicone to obtain an epoxy resin replica of the upper central incisor for the initial surface roughness evaluation using a profilometer and the initial color of the incisors and canines was evaluated with a spectrophotometer after one week of wash-out. After 1 month, the color of the central incisors and canines was measured again, and the volunteers were molded to obtain a second replica to the final roughness analysis. Data were submitted to ANOVA-one way (p≤0.05). The results showed that there was no statistical difference between the dentifrices for color difference and surface roughness for all the studied conditions. It was possible to conclude that the whitening dentifrices used in this study were not able to alter the initial color of the teeth and did not cause changes in the surface roughness of enamel.

Ellagic acid on the quality of the adhesive interface of class I composite resin restorations after aging

To evaluate the effect of ellagic acid on the inhibition of matrix metalloproteinase by analyzing the quality of the adhesive interface with bond strength measures in periods of 24 hours and six months of storage. Method: 40 healthy human third molars were prepared with class I cavities (5x4x3mm). The teeth were divided into four experimental groups: Group 1- without application of ellagic acid and storage time of 24 hours; Group 2- with ellagic acid/24 hours; G3- without ellagic acid/six months; Group 4- with ellagic acid/six months. Then, the cavities were restored with Single Bond Universal adhesive and Z350 composite resin, with and without the previous application of ellagic acid. Subsequently, hourglass-shaped specimens were obtained and subjected to the bond strength (BS) test (n = 10) in a universal testing machine. The bond test was performed after 24 hours and six months of storage. For the standard evaluation (n = 3) the samples were infiltrated with silver nitrate and placed in a developing solution for analysis in a Scanning Electron Microscope (SEM). The data obtained were analyzed with the Kruskal-Wallis non-parametric test, showing a statistically significant difference. Results: The highest bond strength values were found for the 24-hour groups followed by the groups with six months of storage. For nano-infiltration, groups G1 and G2 showed lower infiltration than groups G3 and G4. Conclusion: The previous application of ellagic acid did not affect the BS of the adhesive interface of the adhesive system analyzed, regardless of storage time.

Solutions and Gels Containing a Sugarcane-Derived Cystatin (CaneCPI-5) Reduce Enamel and Dentin Erosion in vitro.

The effect of solutions and gels containing a sugarcane-derived cystatin (CaneCPI-5) on the protection against enamel and dentin erosion in vitro was evaluated. Bovine enamel and dentin specimens were divided into 2 groups (n = 135 and 153/group for enamel and dentin, respectively) that were treated with solutions or chitosan gels containing 0.1 or 0.25 mg/mL CaneCPI-5. The positive controls for solutions and gels were Elmex Erosion Protection™ solution and NaF gel (12,300 ppm F), respectively. Deionized water and chitosan gel served as controls, respectively. The solutions were first applied on the specimens for 1 min and the gels for 4 min. Stimulated saliva was collected from 3 donors and used to form a 2-h acquired pellicle on the specimens. Then, the specimens were submitted to an erosive pH cycling protocol 4 times/day for 7 days (0.1% citric acid pH 2.5/90 s, artificial saliva/2 h, and artificial saliva overnight). The solutions and gels were applied again during pH cycling, 2 times/day for 1 min and 4 min, respectively, after the first and last erosive challenges. Enamel and dentin losses (µm) were assessed by contact profilometry. Data were analyzed by 2-way ANOVA and Tukey's test (p < 0.05). All the treatments significantly reduced enamel and dentin loss in comparison with controls. Both CaneCPI-5 concentrations had a similar protective effect against enamel erosion, but only the higher concentration was as effective against dentin erosion as the positive control. Regarding the vehicles, only the 0.1 mg/mL gel performed worse than the positive control for dentin. CaneCPI-5 reduced enamel and dentin erosion to a similar extent as the fluoride-containing vehicles. However, dentin requires higher CaneCPI-5 concentrations, in the case of gels. Solutions or gels containing CaneCPI-5 might be a new approach to protect against dental erosion.

A sugarcane cystatin (CaneCPI-5) alters microcosm biofilm formation and reduces dental caries.

The antimicrobial and anticaries effects of CaneCPI-5 were evaluated. Ninety bovine enamel samples were treated for 60 s with either phosphate-buffered-saline (PBS), 0.12% chlorhexidine (CHX), 0.05 mg ml-1 CaneCPI-5, 0.1 mg ml-1 CaneCPI-5 or 0.5 mg ml-1 CaneCPI-5. They were incubated with inoculum (human saliva + McBain's saliva) for the first 8 h. From then until the end of the experiment, the enamel was exposed to McBain saliva with sucrose and, once a day, for 5 days, they were treated with the solutions. At the end of the experimental period, resazurin and viable plate count assays were performed. Enamel demineralization was also measured. All concentrations of CaneCPI-5 and CHX significantly reduced the activity of biofilms compared with PBS. For viable plate counts, all treatments similarly reduced the lactobacilli and total streptococci; for the mutans streptococci, 0.05 mg ml-1 CaneCPI-5 performed better than CHX. All CaneCPI-5 concentrations significantly reduced the integrated mineral loss. This study represents the first step regarding the use of CaneCPI-5 within the concept of acquired enamel pellicle and biofilm engineering to prevent dental caries.

Minimally invasive aesthetic treatment of white spots by dental fluorosis in children: case report

The presence of white spots due to dental fluorosis can cause aesthetic discomfort, being one of the factors for a search for dental treatment. Objective: Report a clinical case in which the aesthetic complaint was solved through the association of the dental bleaching to the microabrasion technique. Case report: Male patient, 13 years, attended to the Integrated Clinic of the School of Dentistry of the Federal University of Rio de Janeiro, with fluorosis, mainly in the antero-superior teeth and aesthetic complaint. Initially, it was made an immediate bleaching with hydrogen peroxide at 35%, in 3 sessions of 15 minutes each, to soften the disparity of shades between tooth-stain. The result was insufficient and the dental microabrasion technique was adopted. A pumice paste was applied together with gel of phosphoric acid at 37% under relative isolation, in a total of 8 applications (1 minute each). At the end of each application, it was made an abundant water washing, microengine mounted fine-grained disc polishing and topical application of neutral NaF2 in gel (4 minutes each application) to eliminate possible postoperative sensitivity. Conclusion: The dental bleaching and the microabrasion technique promoted satisfactory immediate clinical results that increased self-esteem of the patient in a minimally invasive way to dental structure.

Introdução: A presença de manchas brancas por fluorose dentária pode causar desconforto estético, sendo um dos fatores para busca de tratamento odontológico. Objetivo: Relatar um caso clínico em que a queixa estética foi resolvida através da associação do clareamento dentário à técnica de microabrasão. Relato do caso: Paciente do sexo masculino, 13 anos, compareceu a Clínica Integrada da Faculdade de Odontologia da UFRJ, com fluorose, principalmente nos dentes ântero-superiores e queixa estética. Inicialmente, realizou-se clareamento imediato com peróxido de hidrogênio a 35%, em 3 sessões de 15 minutos, para suavizar a disparidade de tons entre mancha-dente. O resultado obtido foi insuficiente e a técnica de microabrasão dentária foi adotada. Uma pasta de pedra pomes foi aplicada juntamente com gel de ácido fosfórico a 37% sob isolamento relativo, perfazendo um total de 8 aplicações (1 minuto cada). Ao final de cada aplicação, foi realizada lavagem abundante com água, polimento com disco de granulação fina montado em micromotor e aplicação tópica de NaF2 neutro em gel (4 minutos cada aplicação) para eliminar possível sensibilidade pós-operatória. Conclusão: O clareamento dentário e a técnica de microabrasão promoveram resultados clínicos imediatos satisfatórios que elevaram a autoestima do paciente, de maneira minimamente invasiva para estrutura dentária.

Acquired pellicle protein-based engineering protects against erosive demineralization.

OBJECTIVES: To evaluate, in vivo: 1) proteomic alterations in the acquired enamel pellicle (AEP) after treatment with sugarcane-derived cystatin (CaneCPI-5), hemoglobin (HB), statherin-derived peptide (StN15) or their combination before the formation of the AEP and subsequent erosive challenge; 2) the protection of these treatments against erosive demnineralization. MATERIALS AND METHODS: In 5 crossover phases, after prophylaxis, 10 volunteers rinsed (10 mL, 1 min) with: deionized water-1, 0.1 mg/mL CaneCPI-5-2, 1.0 mg/mL HB-3, 1.88 × 10-5 M StN15-4 or their combination-5. AEP was formed (2 h) and enamel biopsy (10 µL, 1%citric acid, pH 2.5, 10 s) was performed on one incisor for calcium analysis. The same acid was applied on the vestibular surfaces of the remaining teeth. The acid-resistant proteins within the remaining AEP were collected. Samples were quantitatively analyzed by label-free proteomics. RESULTS: Treatment with the proteins/peptide, isolated or combined, increased several acid-resistant proteins in the AEP, compared with control. The highest increases were seen for PRPs (32-fold, StN15), profilin (15-fold, combination), alpha-amylase (9-fold; StN15), keratins (8-fold, CaneCPI-5 and HB), Histatin-1 (7-fold, StN15), immunoglobulins (6.5-fold, StN15), lactotransferrin (4-fold, CaneCPI-5), cystatins, lysozyme, protein S-100-A9 and actins (3.5-fold, StN15), serum albumin (3.5-fold, CaneCPI-5 and HB) and hemoglobin (3-fold, StN15). Annexin, calmodulin, keratin, tubulin and cystatins were identified exclusively upon treatment with the proteins/peptide, alone or combined. Groups 2, 3 and 4 had significantly lower Ca released from enamel compared to group 1 (Kruskal-Wallis/Dunn's, p < 0.05). CONCLUSIONS: Treatment with CaneCPI-5, HB or StN15 remarkably increases acid-resistant proteins in the AEP, protecting against erosion. CLINICAL SIGNIFICANCE: Our results show, for the first time, that treatment with proteins/peptide remarkably increases acid-resistant proteins in the AEP, protecting against erosive demineralization. These findings open an avenue for a new preventive approach for erosive demineralization, employing acquired pellicle engineering procedures that may in the future be incorporated into dental products.