Abstract Objectives This study aimed to assess the effect of proanthocyanidin, palm oil and vitamin E against erosive and erosive+abrasive challenges in vitro after enamel pellicle formation in situ. Methodology Bovine enamel blocks (n=84) were obtained and divided into the following treatment groups: negative control (NC) - deionized water; positive control (PC) - SnCl2/NaF/AmF-containing solution; palm oil (PO); 2% proanthocyanidin (P2); vitamin E (VitE); 2% proanthocyanidin+palm oil (P2PO); and 2% proanthocyanidin+vitamin E (P2VitE). For 5 days, one half of the sample from each group was subjected to erosion and the other half was subjected to erosion+abrasion. The acquired enamel pellicle (AEP) was pre-formed in situ for 30 minutes. The specimens were then treated in vitro with solutions (500 µl, 30s for each group). Subsequently, the blocks were left in the oral cavity for another hour to obtain the modified AEP. The blocks were immersed in 0.5% citric acid (pH=2.5) for 90s, 4×/day. AEP formation and treatment were carried out before the first and third erosive challenges, and after these challenges, abrasive cycles (15s) were performed on half of the samples. Enamel wear was quantified by profilometry and data were analyzed by two-way ANOVA and Tukey’s test (p<0.05). Results All groups showed higher wear when exposed to erosion+abrasion than when exposed to erosion alone (p=0.0001). PO, P2VitE, P2, and P2PO showed enamel wear similar to the PC group, but only PC, PO and P2VitE differed from the NC group. The other groups behaved similarly to NC. Conclusion It was concluded that the combination of proanthocyanidin and vitamin E was effective in reducing wear in the face of in vitro erosive and erosive+abrasive challenges. erosiveabrasive n=84 n84 n 84 (n=84 (NC water (PC SnCl2/NaF/AmFcontaining SnCl2NaFAmFcontaining SnClNaFAmFcontaining SnCl2/NaF/AmF containing SnCl2 NaF AmF SnCl solution (PO) 2 P2 P (P2) VitE (VitE) proanthocyanidinpalm PPO (P2PO) proanthocyanidinvitamin P2VitE. PVitE . (P2VitE) days erosionabrasion abrasion (AEP preformed pre formed 3 minutes 500 (50 µl s group) Subsequently 05 0 0.5 pH=2.5 pH25 pH (pH=2.5 90s 4day day 4 4×/day 15s (15s samples twoway two way Tukeys Tukey p<0.05. p005 p p<0.05 (p<0.05) p=0.0001. p00001 p=0.0001 0001 (p=0.0001) n=8 n8 8 (n=8 AmFcontaining NaFAmFcontaining SnCl2NaFAmF SnClNaFAmF (PO (P2 (VitE (P2PO (P2VitE 50 (5 0. pH=2. pH2 (pH=2. p00 p<0.0 (p<0.05 p0000 p=0.000 000 (p=0.0001 n= (n= NaFAmF (P ( pH=2 (pH=2 p0 p<0. (p<0.0 p000 p=0.00 00 (p=0.000 (n pH= (pH= p<0 (p<0. p=0.0 (p=0.00 (pH p< (p<0 p=0. (p=0.0 (p< p=0 (p=0. (p p= (p=0 (p=

ABSTRACT Objective: To evaluate the clinical and radiographic response of pulp-dentin complex after selective caries removal with or without pulp lining in primary teeth. Material and Methods: Twenty-four primary molars with deep occlusal caries lesions and without pulpal alterations were selected from children, both genders, aged between 5 and 9 years old. After selective caries removal, the teeth were divided into three groups: without cavity liner (Group I), calcium hydroxide cement - CH (Group II), and Mineral trioxide aggregate - MTA (Group III). The final restoration was performed with resin-modified glass ionomer cement. Clinical and radiographic assessments were conducted at 6-month follow-up. The Kappa test determined intraexaminer reliability. Fisher's exact test evaluated intergroup comparisons (p<0.05). Results: All teeth showed clinical and radiographic success at the 6-month follow-up without statistically significant differences (p>0.05). Conclusion: Selective caries removal without cavity lining was acceptable for deep caries lesions in primary teeth. Objective pulpdentin dentin Methods Twentyfour Twenty four children genders old groups Group I, I , I) II, II II) III. III . III) resinmodified resin modified 6month month 6 followup. followup follow up. up reliability Fishers Fisher s p<0.05. p005 p p<0.05 0 05 (p<0.05) Results p>0.05. p>0.05 (p>0.05) Conclusion p00 p<0.0 (p<0.05 p>0.0 (p>0.05 p0 p<0. (p<0.0 p>0. (p>0.0 p<0 (p<0. p>0 (p>0. p< (p<0 p> (p>0 (p< (p> (p

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. (AEP GERD, , (GERD) ETW. ETW . (ETW) Twentyfour Twenty four groups 1 GERD. GERD) 12 lingualpalatal lingual palatal surfaces nLCESIMS/MS nLCESIMSMS nLC ESI MS/MS MS (nLC-ESI-MS/MS labelfree label free software 21 identified 119 9 10 group respectively calciumbinding calcium binding Twentythree three 1433 14 14-3- zetadelta zeta delta 1phosphatidylinositol. 1phosphatidylinositol phosphatidylinositol phosphatidylinositol. 1-phosphatidylinositol hydroxyapatite dissolution (GERD (ETW nLCESIMS MSMS 11 143 14-3 14-

Abstract Objectives To analyze the effect of 5 toothpastes containing different percentages of S-PRG fillers compared to NaF toothpaste and NaF varnish on the dentin hydraulic conductance (Lp). Methodology Dentin disks (1.0±0.2 mm thickness) were cut from third molars, and their Lp values were evaluated using Flodec. The specimens were allocated into 7 groups (n=8). The minimum (smear layer) and the maximum (after acid etching) Lp values were recorded. Lp was also assessed after treatment with either a 0wt.%, 1wt.%, 5wt.%, 20wt.%, or 30wt.% S-PRG toothpaste, a NaF toothpaste, or a NaF varnish. Toothpastes were applied by brushing for 15 s, allowing it to settle for 1 min, and rinsing with deionized water. The NaF varnish was applied for 4 min and was removed with a probe. Specimens were exposed to citric acid (6%, pH 2.1, 1 min) and their final Lp was recorded. The pH of all products was recorded (n=3) and specimens from each group were analyzed by Laser Scanning Confocal Microscopy (LSCM). Data were subjected to 2-way repeated measures ANOVA and post-hoc Bonferroni (a=0.05). Results The highest Lp reduction was noticed for the 5wt.% S-PRG toothpaste, NaF toothpaste, and NaF varnish. However, the toothpastes containing 5wt.%, 20wt.%, and 30wt.% of S-PRG were similar to all toothpastes but differed from the NaF varnish. After erosion, all groups retrieved their maximum Lp values, except for the NaF varnish. The LSCM evidenced deposits on the surface of specimens treated with 5%, 20%, and 30% S-PRG-based toothpastes and NaF toothpaste. Even more deposits were observed for the NaF varnish. After the erosive challenge, the deposits were diminished in all groups. Conclusion Toothpastes containing 5wt.%, 20wt.%, and 30wt.% of S-PRG fillers behaved similarly to a conventional NaF toothpaste, even after an erosive challenge. The NaF varnish promoted better reduction of the Lp, but its effect was also diminished after erosion.

Abstract: This randomized three-armed controlled clinical trial compared the effect of titanium tetrafluoride (TiF4) and sodium fluoride (NaF) varnishes on caries control in smooth surfaces of permanent dentition and children's acceptability. Sixty children (6-8 y/o) were randomly divided into TiF4 (2.45% F–), NaF (2.26% F–) or placebo (control) groups. Varnishes were applied on permanent teeth once a week for the first 4 weeks and after the 6th and 12th months of the study. The variables were as follows: International Caries Detection and Assessment System (ICDAS) scores, quantitative fluorescence changes, visual plaque index (VPI) and degree of acceptability. Two-way RM-ANOVA, ANOVA/Tukey and χ2 tests were performed (p < 0.05). No differences were found between the treatments with respect to ICDAS scores (p = 0.32). Only TiF4 reduced the mean fluorescence loss significantly at 18 months compared to the baseline (p = 0.003). TiF4 showed a lower percentage of new caries lesions by tooth surface than the placebo, while NaF did not induce such a change (p < 0.014). Regardless of the treatment, more than 95% of the participants reported being satisfied. For all groups, the VPI decreased significantly at 3 months compared to the baseline value (p < 0.001), with no differences between the treatments (p = 0.17). TiF4 had a similar ability to control caries lesions as NaF; however, only TiF4 differed from the placebo (p = 0.004). The acceptability of TiF4 varnish was similar to that of NaF varnish.

Abstract Non-inferiority randomized clinical trials are indicated when it is intended to prove that an experimental group is not inferior to a control group by more than a margin of non-inferiority. However, this type of study differs from traditional randomized clinical trials (superiority studies) because they have particularities that impact on the formulation of hypothesis to be tested, experimental design (non-inferiority margin determination, adapted sample size calculation, sensitivity of the study and data final analysis) and also on the presentation of data when writing the manuscript. Therefore, this article aims to present and discuss the particularities of non-inferiority clinical studies, since these requirements are fundamental to guarantee the validity of the conclusions of this type of study.

Abstract The acquired enamel pellicle (AEP) is an organic film, bacteria-free, formed in vivo as a result of the selective adsorption of salivary proteins and glycoproteins to the solid surfaces exposed to the oral environment. Objective: This study aimed to compare the proteomic profile of AEP formed in situ on human and bovine enamel using a new intraoral device (Bauru in situ pellicle model - BISPM). Material and Methods: One hundred and eight samples of human and bovine enamel were prepared (4×4 mm). Nine subjects with good oral conditions wore a removable jaw appliance (BISPM) with 6 slabs of each substrate randomly allocated. The AEP was formed during the morning, for 120 minutes, and collected with an electrode filter paper soaked in 3% citric acid. This procedure was conducted in triplicate and the pellicle collected was processed for analysis by LC-ESI-MS/MS. The obtained mass spectrometry MS/MS spectra were searched against human protein database (SWISS-PROT). Results: The use of BISPM allowed the collection of enough proteins amount for proper analysis. A total of 51 proteins were found in the AEP collected from the substrates. Among them, 15 were common to both groups, 14 were exclusive of the bovine enamel, and 22 were exclusive of the human enamel. Proteins typically found in the AEP were identified, such as Histatin-1, Ig alpha-1, Ig alpha 2, Lysozyme C, Statherin and Submaxillary gland androgen-regulated protein 3B. Proteins not previously described in the AEP, such as metabolism, cell signaling, cell adhesion, cell division, transport, protein synthesis and degradation were also identified. Conclusion: These results demonstrate that the proteins typically found in the AEP appeared in both groups, regardless the substrate. The BISPM revealed to be a good device to be used in studies involving proteomic analysis of the AEP.

Abstract The standardization of in situ protocols for dental erosion is important to enable comparison between studies. Objective: Thus, the objectives of this study were to evaluate the influence of the location of in situ intraoral appliance (mandibular X palatal) on the extent of enamel loss induced by erosive challenges and to evaluate the comfort of the appliances. Material and Methods: One hundred and sixty bovine enamel blocks were selected according to their initial surface hardness and randomly divided into two groups: GI - palatal appliance and GII - mandibular appliance. Twenty volunteers wore simultaneously one palatal appliance (containing 4 enamel blocks) and two mandibular appliances (each one containing 2 enamel blocks). Four times per day during 5 days, the volunteers immersed their appliances in 0.01 M hydrochloric acid for 2 minutes, washed and reinserted them into the oral cavity for 2 hours until the next erosive challenge. After the end of the in situ phase, the volunteers answered a questionnaire regarding the comfort of the appliances. The loss of tissue in the enamel blocks was determined profilometrically. Data were statistically analyzed by paired t-test, Chi-square and Fisher's Exact Test (p<0.05). Results: The enamel blocks allocated in palatal appliances (GI) presented significantly higher erosive wear when compared to the blocks fixed in mandibular appliances (GII). The volunteers reported more comfort when using the palatal appliance. Conclusions: Therefore, the palatal appliance is more comfortable and resulted in higher enamel loss compared to the mandibular one.

Abstract Alternatives for the treatment of caries disease, such as minimally invasive approaches, have been developed in recent years. Objective: To carry out clinical and radiographic evaluations of three cavity liners after selective caries removal. Methodology: Thirty-six primary molars with deep occlusal caries lesions without pulp involvement (from children of both genders, aged between 5 and 8 years) were randomly divided into the following groups: calcium hydroxide cement (CHC) group; mineral trioxide aggregate (MTA) group and Portland cement with added zirconium oxide (PCZ) group. The following-up period was 6- and 12-month. The clinical and radiographic success rates were evaluated through chi-square test. The radiographic measurements were compared by ANOVA followed by Tukey's test (p<0.05). Results: Thirty-six patients were included, but thirty-four returned for 12-month follow-up. The overall success rate of the therapy for the three groups was 94.11% and no statistically significant differences occurred in the comparison among groups (p>0.05). Nineteen radiographs were selected to measure the dentin barrier thickness. The intragroup comparison presented a statistically significant increase of the dentin barrier for all groups, at 12-month follow-up. However, the MTA group showed increase of the dentin barrier, over time, 6- to 12-month follow-up. The intergroup comparison revealed no statistically significant differences (p>0.05). Conclusion: The clinical and radiographic data showed that all cavity liners provided effective treatment of primary teeth after selective caries removal.

Abstract Objective: To evaluate in situ the effect of toothpastes containing casein phosphopeptide - amorphous calcium phosphate (CPP-ACP) and casein phosphopeptide-amorphous calcium phosphate associated to fluoride (CPP-ACPF) on initial erosion prevention. Material and Methods: Bovine enamel blocks (n = 192) were randomly assigned into 4 phases according to the baseline surface hardness: GI: CPP-ACP Paste (MI Paste™), GII: CPP-ACPF Paste (MI Paste Plus™), GIII: Fluoridated paste and GIV: Placebo Paste. In each of the 4 crossover phases, twelve volunteers wore intraoral palatal appliances containing 4 enamel blocks for 2 hours, then the tested treatments were applied intraorally (3 min) and the appliance was maintained in the mouth for another 3 hours. After, the appliances were removed and immersed in hydrochloric acid (0.01 M, pH 2.3) for 30 seconds to promote erosive demineralization. The final surface hardness was evaluated and percentage of surface hardness loss was calculated. The data were analyzed by ANOVA and Tukey’s test (α = 5%). Results: The application of CPP-ACP paste, independent of fluoride content, resulted in significant lower enamel hardness loss (GI: 9.26% ±3.48 and GII: 9.14% ±1.73) compared to NaF (GIII: 15.5% ± 3.94) and placebo (GIV: 16.7% ± 4.07) pastes, which did not show difference between them. Conclusion: The CPP-ACP pastes were able to reduce initial erosive demineralization in relation to fluoride and placebo pastes. Nevertheless the formulation of CPP-ACP with fluoride did not provide an additional benefit.

Resumen El objetivo de la investigación fue explorar los diálogos y saberes sobre sexualidad expresados por padres o madres de adolescentes. La metodología fue cualitativa con diseño narrativo, y los grupos focales sirvieron como técnica de recolección. Los resultados evidencian que los padres o madres aprenden sobre sexualidad desde sus propias experiencias, por lo cual este aspecto está presente en la comunicación con sus hijos. Respecto a los temas, siguen prevaleciendo discursos preventivos en torno al embarazo, excluyendo las infecciones de transmisión sexual (ITS), pornografía y diversidades sexuales. Se concluye que en los diálogos sobre sexualidad en estos participantes coexisten múltiples discursos, y se requieren estrategias de intervención orientadas a promover espacios formativos y habilidades para la comunicación referente a la sexualidad.

Abstract The study aimed to explore the dialog and knowledge of sexuality expressed by fathers or mothers of adolescents. A qualitative methodology with narrative design was used, with focus groups used for data collection. The results demonstrate that the parents or mothers learn about sexuality from their own experiences, so this aspect is present in communication with their children. The most discussed topic was pregnancy, excluding sexually transmitted infections (sti), pornography and sexual diversity. The study concludes that dialogs with these participants about sexuality contain multiples discourses; intervention strategies oriented to promoting training spaces and communication skills regarding sexuality are required.

Resumo O objetivo desta pesquisa foi explorar os diálogos e saberes sobre sexualidade expressos por pais ou mães de adolescentes. A metodologia foi qualitativa com desenho narrativo, e os grupos focais serviram como técnica de coleta. Os resultados demonstram que os pais ou mães aprendem sobre sexualidade a partir de suas próprias experiências, razão pela qual esse aspecto está presente na comunicação com seus filhos. A respeito dos temas, continuam prevalecendo discursos preventivos sobre a gravidez e excluem-se os referentes a infecções sexualmente transmissíveis (1st), pornografia e diversidade sexual. Conclui-se que, nos diálogos sobre sexualidade nesses participantes, coexistem múltiplos discursos e requerem-se estratégias de intervenção orientadas a promover espaços de formação e habilidades para a comunicação referente à sexualidade.

Abstract Oral health problems can influence people's Quality of Life (QoL) because of pain, discomfort, limitations, and other esthetics problems, affecting their social life, feeding, daily activities, and the individual's well-being. Objective: To compare oral health-related quality of life (OHRQoL) of children with and without oral clefts and their families. Materials and Methods: 121 children aged from 2 to 6 years, from both sexes, enrolled in the treatment routine of the Pediatric Dentistry Clinics of a Dental School and a Hospital for Cleft Treatment were divided into two groups: Group 1 - children with cleft lip and palate; Group 2 - children without cleft lip and palate. The OHRQoL was assessed using the validated Portuguese version of the Early Childhood Oral Health Impact Scale (B-ECOHIS). The questionnaire was answered individually, only once, at a private place. Mann-Whitney U test was used to verify differences between groups. Spearman's Rho test was used to associate sex and age with quality of life. The level of significance was set at 5% (p<0.05). Results: According to the parents’ perception on the OHRQoL of children with and without cleft lip and palate, oral health of children with oral clefts (Group 1) had a statistically significant impact on OHRQoL. The correlation of sex with impact on OHRQoL did not show statistically significant differences. On the other hand, the higher the age the higher the impact on QoL. Conclusions: The group comparison revealed that the cleft lip and palate negatively impacted on OHRQoL of 2 to 6-year-old children and their parents.

Abstract Objective The prevalence of dental erosion has been recently increasing, requiring new preventive and therapeutic approaches. Vegetable oils have been studied in preventive dentistry because they come from a natural, edible, low-cost, and worldwide accessible source. This study aimed to evaluate the protective effect of different vegetable oils, applied in two concentrations, on initial enamel erosion. Material and Methods Initially, the acquired pellicle was formed in situ for 2 hours. Subsequently, the enamel blocks were treated in vitro according to the study group (n=12/per group): GP5 and GP100 – 5% and pure palm oil, respectively; GC5 and GC100 – 5% and pure coconut oil; GSa5 and GSa100 – 5% and pure safflower oil; GSu5 and GSu100 – 5% and pure sunflower oil; GO5 and GO100 – 5% and pure olive oil; CON− – Deionized Water (negative control) and CON+ – Commercial Mouthwash (Elmex® Erosion Protection Dental Rinse, GABA/positive control). Then, the enamel blocks were immersed in artificial saliva for 2 minutes and subjected to short-term acid exposure in 0.5% citric acid, pH 2.4, for 30 seconds, to promote enamel surface softening. The response variable was the percentage of surface hardness loss [((SHi - SHf) / SHf )×100]. Data were analyzed by one-way ANOVA and Tukey’s test (p<0.05). Results Enamel blocks of GP100 presented similar hardness loss to GSu100 (p>0.05) and less than the other groups (p<0.05). There was no difference between GP5, GC5, GC100, GSa5, GSu100, GSa100, GSu5, GO5, GO100, CON− and CON+. Conclusion Palm oil seems to be a promising alternative for preventing enamel erosion. However, further studies are necessary to evaluate a long-term erosive cycling.

Abstract Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) is able to increase salivary calcium and phosphate levels at an acidic pH. Previous studies demonstrated that a CPP-ACP chewing gum was able to enhance the re-hardening of erosion lesions, but could not diminish enamel hardness loss. Therefore, there is no consensus regarding the effectiveness of CPP-ACP on dental erosion. Objective This in situ study investigated the ability of a CPP-ACP chewing gum in preventing erosive enamel loss. Material and Methods: During three experimental crossover phases (one phase per group) of seven days each, eight volunteers wore palatal devices with human enamel blocks. The groups were: GI – Sugar free chewing gum with CPP-ACP; GII – Conventional sugar free chewing gum; and GIII – No chewing gum (control). Erosive challenge was extraorally performed by immersion of the enamel blocks in cola drink (5 min, 4x/day). After each challenge, in groups CPP and No CPP, volunteers chewed one unit of the corresponding chewing gum for 30 minutes. Quantitative analysis of enamel loss was performed by profilometry (µm). Data were analyzed by Repeated-Measures ANOVA and Tukey’s test (p<0.05). Results The use of chewing gum (CPP and No CPP) resulted in lower erosive enamel loss compared with the control group (p<0.05). CPP-ACP chewing gum (CPP) did not improve the protection against erosive enamel loss compared with conventional chewing gum (No CPP) (p>0.05). Conclusion The CPP-ACP chewing gum was not able to enhance the anti-erosive effect of conventional chewing gum against enamel loss.

ABSTRACT Objective This in situ/ex vivo study investigated the effect of CO2 laser irradiation and acidulated phosphate fluoride gel (APF) application, separately and in combination, on enamel resistance to erosion. Material and Methods During 2 experimental 5-day crossover phases, 8 volunteers wore intraoral appliances containing bovine enamel blocks which were submitted to four groups: 1st phase - control, untreated and CO2 laser irradiation, 2nd phase - fluoride application and fluoride application before CO2 laser irradiation. Laser irradiation was performed at 10.6 µm wavelength, 5 µs pulse duration and 50 Hz frequency, with average power input and output of 2.3 W and 2.0 W, respectively (28.6 J/cm2). APF gel (1.23%F, pH 3.5) was applied on enamel surface with a microbrush and left on for 4 minutes. Then, the enamel blocks were fixed at the intraoral appliance level. The erosion was performed extraorally 4 times daily for 5 min in 150 mL of cola drink. Enamel loss was measured profilometrically after treatment and after the in situ phase. The data were tested using one-way Repeated Measures Anova and Tukey's test (p<0.05). Results CO2 laser alone (2.00±0.39 µm) did not show any significantly preventive effect against enamel erosion when compared with the control group (2.41±1.20 µm). Fluoride treated enamel, associated (1.50±0.30 µm) or not (1.47±0.63 µm) with laser irradiation, significantly differed from the control. Conclusion The APF application decreased enamel wear; however, CO2 laser irradiation did not enhance fluoride ability to reduce enamel wear.