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Abstract The rapid and abrupt transmission pattern of the SARS-CoV-2 unleashed the current COVID-19 pandemic, as recognized by the World Health Organization in March 2020. Considering the high risk of transmission of the virus in dental environments and the specificities in clinical practice, COVID-19 posed immediate challenges for dental care and education. Due to the need to establish infection prevention and control guidance in dental health settings to enable a safe clinical practice, this review aims to list the challenges and perspectives in managing dental care in services and schools. This review employed materials collected from PubMed and the main guidelines and studies on the novel coronavirus to provide an overview of the clinical procedures and decisions made by health care personnel in dental offices and dental schools. We expect the COVID-19 scenario to promote significant changes in clinical practice and dental education; dentists should seek specific and particular regulations for dental practice established by their state or country. Biosafety checklists are strongly recommended for appointments at dental services and face-to-face activities in dental schools.

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Abstract Objective: This study sought to identify the differences between the oral changes presented by patients with solid and hematologic tumors during chemotherapeutic treatment. Methodology: This is an observational, prospective and quantitative study using direct documentation by follow-up of 105 patients from 0 to 18 years using the modified Oral Assessment Guide (OAG). Of the 105 patients analyzed, 57 (54.3%) were boys with 7.3 years (±5.2) mean age. Hematologic neoplasms accounted for 51.4% of all cases. Results: Voice, lips, tongue, and saliva changes were not significantly different (p>0.05) between patients with solid or hematologic tumors and during the follow-up. From the 6th until the 10th week of chemotherapeutic treatment alterations in swallowing function, in the mucous membrane (buccal mucosa and palate), in the labial mucosa, and in the gingiva occurred and were distributed differently between the two tumors groups (p<0.05). The main alterations were observed in patients with hematologic tumors. Conclusion: It was concluded that the oral changes during the chemotherapeutic treatment occurred especially in swallowing function, in the mucous membrane, in the labial mucosa and in the gingiva, and these alterations were found mainly in patients with hematologic tumors.

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Abstract Periodontal therapy usually requires local anesthesia. If effective, a non-invasive, liposomal anesthetic gel could increase the levels of acceptance of patients in relation to periodontal therapy. Objective: This study investigated the efficacy of liposomal anesthetic gel for pain control during periodontal therapy. Methodology: Forty volunteers with moderate to severe chronic periodontitis were recruited, of which at least three sextants required periodontal therapy. At least one of the selected teeth had one site with a probing depth of ≥4 mm. The volunteers received the following three gels: a placebo, lidocaine/prilocaine (Oraqix®), or a liposomal lidocaine/prilocaine, which were applied to different sextants. Pain frequency was registered during treatment and the volunteers received a digital counter to register any painful or uncomfortable experiences. At the end of each session, the volunteers indicated their pain intensity using rating scales (NRS-101 and VRS-4). The volunteers had their hemodynamic parameters measured by a non-invasive digital monitor. Results: Pain frequency/intensity did not show statistical difference between intervention groups. The tested gels did not interfere with the hemodynamic indices. Dental anxiety, suppuration and probing depth could influence pain during periodontal therapy. Conclusion: Our results suggest limited indications for the use of non-invasive anesthesia when used for scaling and root planing. Intra-pocket anesthetic gel could be a good option for anxious patients, or those who have a fear of needles.

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Abstract Gap formation of composite resin restorations is a serious shortcoming in clinical practice. Polymerization shrinkage stress exceeds the tooth-restoration bond strength, and it causes bacterial infiltration within gaps between cavity walls and the restorative material. Thus, an intermediate liner application with a low elastic modulus has been advised to minimize polymerization shrinkage as well as gap formation. Objective: The purpose of this in vitro study was to assess gap formation volume in premolars restored with different bulk-fill composites, with and without a resin-modified glass-ionomer cement (RMGIC) liner, using x-ray micro-computed tomography (micro-CT). Methodology: Sixty extracted human maxillary premolars were divided into six groups according to bucco-palatal dimensions (n=10). Standardized Class II mesio-occluso-distal cavities were prepared. G-Premio Bond (GC Corp., Japan) was applied in the selective-etch mode. Teeth were restored with high-viscosity (Filtek Bulk Fill, 3M ESPE, USA)-FB, sonic-activated (SonicFill 2, Kerr, USA)-SF and low viscosity (Estelite Bulk Fill Flow, Tokuyama, Japan)-EB bulk-fill composites, with and without a liner (Ionoseal, Voco GmbH, Germany)-L. The specimens were subjected to 10,000 thermocycles (5-55°C) and 50,000 simulated chewing cycles (100 N). Gap formation based on the volume of black spaces at the tooth-restoration interface was quantified in mm3 using micro-computed tomography (SkyScan, Belgium), and analyses were performed. Data were analyzed using repeated-measures ANOVA and the Bonferroni correction test (p < 0.05). Results: The gap volume of all tested bulk-fill composites demonstrated that Group SF (1.581±0.773) had significantly higher values than Group EB (0.717±0.679). Regarding the use of a liner, a significant reduction in gap formation volume was observed only in Group SFL (0.927±0.630) compared with Group SF (1.581±0.773). Conclusion: It can be concluded that different types of bulk-fill composite resins affected gap formation volume. Low-viscosity bulk-fill composites exhibited better adaptation to cavity walls and less gap formation than did sonic-activated bulk-fill composites. The use of an RMGIC liner produced a significant reduction in gap formation volume for sonic-activated bulk-fill composites.

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Abstract Objectives: This study approaches the history of reclassifications and redefinitions around the odontogenic keratocyst (OK), as proposed by the World Health Organization (WHO), and aims to understand the impact of those changes on the prevalence and epidemiology of odontogenic tumors (OTs). Methodology: Cases of OTs diagnosed in an Oral Pathology service between January 1996 and December 2016 were reviewed. Demographic data of patients such as age, gender and site of lesions were retrieved from their respective records. Results: Within the studied period, 7,805 microscopic reports were elaborated and 200 (2.56%) of these were diagnosed as OTs. Out of these 200, between 1996 and 2005, prior to the 2005 WHO classification, there were 41 (20.5%) OTs cases, being odontoma the most frequent (23; 56.09%), followed by ameloblastoma (8; 19.51%) and myxoma (03; 7.31%). Between 2006 and 2016, after the previous 2005 WHO classification there were 159 (79.5%) OTs, being odontogenic keratocystic tumor (KCOT) the most frequent (68; 42.76%), followed by odontoma (39; 24.52%) and ameloblastoma (21; 13.20%). Conclusions: As of today, the most recent WHO classification to be followed brings KCOT back to the cyst category, which will impact on the prevalence and epidemiology of OTs; thus, this study was able to identify a considerable increase (287.80%) in the prevalence of OTs when the 2005 WHO classification was utilized. Despite being an important academic exercise, classifying odontogenic lesions and determining whether to place the odontogenic keratocyst in a cyst or tumor category is crucial to establish the correct diagnosis and treatment to follow, whether by oral medicine or oral surgery specialist, or by the general practitioner.

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Abstract Objective: The microbial composition of pericoronitis (Pc) is still controversial; it is not yet clear if the microbial profile of these lesions is similar to the profile observed in periodontitis (Pd). Therefore, the aim of the present study was to describe the microbial profile of Pc lesions and compare it directly with that of subjects with Pd. Methodology: Subjects with Pc and Pd were selected, and subgingival biofilm samples were collected from (i) third molars with symptomatic Pc (Pc-T), (ii) contralateral third molars without Pc (Pc-C) and (iii) teeth with a probing depth >3 mm from subjects with Pd. Counts and proportions of 40 bacterial species were evaluated using a checkerboard DNA-DNA hybridization technique. Results: Twenty-six patients with Pc and 18 with Pd were included in the study. In general, higher levels of microorganisms were observed in Pd. Only Actinomyces oris and Eubacterium nodatum were present in higher mean counts in the Pc-T group in comparison with the Pc-C and Pd-C groups (p<0.05). The microbiota associated with Pc-T was similar to that found in Pc-C. Sites with Pc lesions had lower proportions of red complex in comparison with the Pd sites. Conclusion: The microbiota of Pc is very diverse, but these lesions harbour lower levels of periodontal pathogens than Pd.

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Abstract Pathological parameters have been indicated as tumor prognostic factors in oral carcinoma. Objective: The objective of this study was to investigate the impact of pathological parameters on prognosis of patients affected only by tongue and/or floor of the mouth squamous cell carcinoma (SCC). Methodology: In total, 380 patients treated in the Brazilian National Cancer Institute (INCA) from 1999 to 2006 were included. These patients underwent radical resection followed by neck dissection. The clinical and pathological characteristics were recorded. The Kaplan-Meier method and Cox proportional hazards model were used in survival analysis. Overall survival (OS), cancer-specific survival (CSS) and disease-free interval (DFI) were estimated. Cox residuals were evaluated using the R software version 3.5.2. Worst OS, CSS and DFI were observed in patients with tumors in advanced pathological stages (p<0.001), with the presence of perineural invasion (p<0.001) and vascular invasion (p=0.005). Results: Advanced pathological stage and the presence of a poorly differentiated tumor were independent prognostic factors for OS and CSS. However, advanced pathological stage and perineural invasion were independent predictors of a shorter OS, DFI and CSS. Conclusion: Pathological stage and perineural invasion were the most significant pathological variables in survival analysis in tongue and/or floor of the mouth SCC.

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Abstract Oral and oropharyngeal cancer is considered a public health problem in several countries due to its high incidence and mortality rate. Objective: This study aimed to analyze oral and oropharyngeal cancer mortality in Uruguay from 1997 to 2014 by age, sex and country region. Methodology: A time series ecological study using secondary data was performed. Data on mortality due to oral and oropharyngeal cancers were obtained from the Vital Statistics Department of Uruguay's Ministry of Public Health. Results: The cumulative mortality rate due to oral and oropharyngeal cancer over the study period was of 19.26/100,000 persons in women and 83.61/100.000 in men, with a mean annual rate of 1.75/100,000 in women and 7.60/100,000 in men. Mortality rate from both sites during the study period was 4.34 times higher in men than in women. Malignant neoplasms of other parts of the tongue and base of tongue showed the highest mortality rate. The means of the annual coefficients of deaths were higher for the age groups between 50 and 69 years. Higher mortality rates of oral and oropharyngeal cancer were observed in Artigas (4.63) and Cerro Largo (3.75). Conclusions: Our study described a high mortality rate for oral and oropharyngeal cancer in Uruguay from 1997 to 2014. According to the country's health department, men, tongue cancer, and oral cavity had higher mortality rates, with some variation. Prevention strategies with control of risk factors and early diagnosis are necessary to improve survival in the Uruguayan population.

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Abstract Objective: The goal of the present study was to determine the effect of systemic and topical ozone application on alveolar bone loss (ABL) by evaluating the effect of Hypoxia-inducible factor −1 alpha (HIF-1-α) and receptor activator of NF-kB ligand (RANKL)-positive cells on histopathological and immunohistochemical changes in a rat periodontitis model. Methodology: Thirty male Wistar rats were divided into three groups: 1) Group C (control group); 2) Group SO (systemic ozone group) and 3) Group TO (topical ozone group). Experimental periodontitis was induced with a 3/0 silk suture placed at the mandibular left first molars of rats, and the suture was removed 14 days later. Ozone gas was injected intraperitoneally (0.7 mg/kg) in SO group. Topical ozone application protocol was performed using an ozone generator at 80% concentration (4th grade) 90- degree probe for the duration of 30 s. Both ozone applications were carried out for two weeks at intervals of two days. Histomorphometric and immunohistochemical analysis were performed. Results: ABL was significantly lower in Group SO compared to Group C (p: 0.0052). HIF-1α- positive cells were significantly lower in Group TO than in Group C (p: 0.0043). RANKL-positive cells were significantly lower in Group SO and in Group TO compared to the control group (p: 0.0033, p: 0.0075, respectively). Conclusion: Both ozone applications decreased RANKL-positive cell counts, TO application decreased HIF-1-α positive cells counts, and SO application was found to be more effective in reducing ABL compared to control group.

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Abstract Calcium aluminate cement (CAC) has been highlighted as a promising alternative for endodontic use aiming at periapical tissue repair. However, its effects on dental pulp cells have been poorly explored. Objective: This study assessed the impact of calcium chloride (CaCl2) and bismuth oxide (Bi2O3) or zinc oxide (ZnO) additives on odontoblast cell response to CAC. Methodology: MDPC-23 cells were exposed for up to 14 d: 1) CAC with 2.8% CaCl2 and 25% ZnO (CACz); 2) CAC with 2.8% CaCl2 and 25% Bi2O3 (CACb); 3) CAC with 10% CaCl2 and 25% Bi2O3 (CACb+); or 4) mineral trioxide aggregate (MTA), placed on inserts. Non-exposed cultures served as control. Cell morphology, cell viability, gene expression of alkaline phosphatase (ALP), bone sialoprotein (BSP), and dentin matrix protein 1 (DMP-1), ALP activity, and extracellular matrix mineralization were evaluated. Data were compared using ANOVA (α=5%). Results: Lower cell density was detected only for MTA and CACb+ compared with Control, with areas showing reduced cell spreading. Cell viability was similar among groups at days one and three (p>0.05). CACb+ and MTA showed the lowest cell viability values at day seven (p>0.05). CACb and CACb+ promoted higher ALP and BSP expression compared with CACz (p<0.05); despite that, all cements supported ALP activity. Matrix mineralization were enhanced in CACb+ and MTA. Conclusion: In conclusion, CAC with Bi2O3, but not with ZnO, supported the expression of odontoblastic phenotype, but only the composition with 10% CaCl2 promoted mineralized matrix formation, rendering it suitable for dentin-pulp complex repair.

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Abstract Objective: This clinical study sought to evaluate the effectiveness of passive ultrasonic activation (PUA) in eliminating microorganisms in primary endodontic infection (PEI) after instrumentation of root canals using microbiological culture and checkerboard DNA-DNA hybridization. Methodology: Twenty root canals with PEI and apical periodontitis were selected. The root canals were instrumented and then randomly divided into 2 groups, according to the irrigation method: PUA and conventional needle irrigation (CNI). Microbiological samples were collected before instrumentation (S1), after instrumentation (S2) and after irrigation with 17% EDTA (S3). The samples were subjected to anaerobic culture technique and checkerboard DNA-DNA hybridization analysis. Results: A statistically significant difference was found between CNI (23.56%) and PUA (98.37%) regarding the median percentage values for culturable bacteria reduction (p<0.05). In the initial samples, the most frequently detected species was S. constellatus (50%), and after root canal treatment was E. faecalis (50%). Conclusion: Both treatments significantly decreased the number of bacterial species compared with the initial sample. However, no statistical difference in the total microbial load between PUA and CNI groups was detected. The number of cultivable anaerobic bacteria reduced significantly using PUA, and the bacterial composition and number of bacterial species after using either CNI or PUA was similar.

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Abstract When exposure of the pulp to external environment occurs, reparative dentinogenesis can be induced by direct pulp capping to maintain pulp tissue vitality and function. These clinical situations require the use of materials that induce dentin repair and, subsequently, formation of a mineralized tissue. Objective: This work aims to assess the effect of tricalcium silicate cements and mineral trioxide aggregate cements, including repairing dentin formation and inflammatory reactions over time after pulp exposure in Wistar rats. Methodology: These two biomaterials were compared with positive control groups (open cavity with pulp tissue exposure) and negative control groups (no intervention). The evaluations were performed in three stages; three, seven and twenty-one days, and consisted of an imaging (nuclear medicine) and histological evaluation (H&E staining, immunohistochemistry and Alizarin Red S). Results: The therapeutic effect of these biomaterials was confirmed. Nuclear medicine evaluation demonstrated that the uptake of 99mTc-Hydroxymethylene diphosphonate (HMDP) showed no significant differences between the different experimental groups and the control, revealing the non-occurrence of differences in the phosphocalcium metabolism. The histological study demonstrated that in mineral trioxide aggregate therapies, the presence of moderate inflammatory infiltration was found after three days, decreasing during follow-ups. The formation of mineralized tissue was only verified at 21 days of follow-up. The tricalcium silicate therapies demonstrated the presence of a slight inflammatory infiltration on the third day, increasing throughout the follow-up. The formation of mineralized tissue was observed in the seventh follow-up day, increasing over time. Conclusions: The mineral trioxide aggregate (WhiteProRoot®MTA) and tricalcium silicate (Biodentine™) present slight and reversible inflammatory signs in the pulp tissue, with the formation of mineralized tissue. However, the exacerbated induction of mineralized tissue formation with the tricalcium silicate biomaterial may lead to the formation of pulp calcifications

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Abstract Chitosan is a natural, biocompatible chelating substance with potential for dental use. This study compared the effects of final canal irrigation with chitosan and EDTA on dentin microhardness, sealer dentin tubules penetration capacity, and push-out strength. Methodology: Fifty canine roots were distributed according to the final irrigation protocol (n=10): G1- 15% EDTA with conventional irrigation; G2- 15% EDTA with Endovac; G3- 0.2% chitosan with conventional irrigation; G4- 0.2% chitosan with Endovac; and G5- without irrigation. Specimens were obturated (AH Plus) and sectioned in 3 slices per root third. The first slice was used for microhardness and sealer penetration assessments under a laser confocal microscope. The second was utilized in a push-out strength test. The third slice was discarded. Data were analyzed using 2-way ANOVA and Tukey's post hoc test (α<0.05). Failure mode was determined at x40 magnification. Results: Microhardness reduction was more significant in groups G2 and G4 (p<0.05). Sealer penetration through dentin was significantly greater in group G2 (p<0.05). There was no significant difference between groups G1, G3, and G4 (p>0.05). In general, all experimental groups presented similar bond resistance (p>0.05) that significantly differed from the control (p<0.001). Mixed type failures were predominant. Conclusions: In general, 0.2% chitosan and 15% EDTA solutions act in a similar manner with regard to the variables studied. The use of Endovac potentiates the effect of these solutions.

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Abstract This study aimed to evaluate the physical properties and antifungal activities of polymethyl methacrylate (PMMA) acrylic resins after the incorporation of chlorhexidine diacetate salt (CDA). Methodology: First, acrylic resin specimens were fabricated with Vipi Cor® and DuraLay® resins with and without the incorporation of 0.5%, 1.0% or 2.0% CDA. The residual monomer and CDA release were measured at intervals ranging from 2 hours to 28 days using ultraviolet spectrometry combined with high-performance liquid chromatography. The antifungal activity against C. albicans was evaluated with the agar diffusion method. Fourier transform infrared spectroscopy was used to analyze the degree of resin conversion. Finally, the water sorption values of the resins were also measured. Results: The incorporated CDA concentration significantly changed the rate of CDA release (p<0.0001); however, the brand of the material appeared to have no significant influence on drug release. Subsequently, the inhibition zones were compared between the tested groups and within the same brand, and only the comparisons between the CDA 2% and CDA 1% groups and between the CDA 1% and CDA 0.5% groups failed to yield significant differences. Regarding the degrees of conversion, the differences were not significant and were lower only in the CDA 2% groups. Water sorption was significantly increased at the 1.0% and 2.0% concentrations. Conclusions: We concluded that the incorporation of CDA into PMMA-based resins enabled the inhibition of C. albicans growth rate, did not alter the degrees of conversion of the tested resins and did not change the release of residual monomers.

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Abstract Objective: This study evaluated the angiogenesis-enhancing potential of a tricalcium silicate-based mineral trioxide aggregate (ProRoot MTA), Biodentine, and a novel bioceramic root canal sealer (Well-Root ST) in human dental pulp stem cells (hDPSCs), human periodontal ligament stem cells (hPLSCs), and human tooth germ stem cells (hTGSCs). Methodology: Dulbecco's modified Eagle's medium was conditioned for 24 h by exposure to ProRoot MTA, Biodentine, or Well-Root ST specimens (prepared according to the manufacturers’ instructions). The cells were cultured in these conditioned media and their viability was assessed with 3-(4,5-dimethyl-thiazol-2-yl)-5-(3-carboxy-methoxy-phenyl)-2-(4-sulfo-phenyl)-2H tetrazolium (MTS) on days 1, 3, 7, 10, and 14. Angiogenic growth factors [platelet-derived growth factor (PDGF), basic fibroblast growth factor (FGF-2), and vascular endothelial growth factor (VEGF)] were assayed by sandwich enzyme-linked immunosorbent assay (ELISA) on days 1, 7, and 14. Human umbilical vein endothelial cell (HUVEC) migration assays were used to evaluate the vascular effects of the tested materials at 6–8 h. Statistical analyses included Kruskal–Wallis, Mann–Whitney U, and Friedman and Wilcoxon signed rank tests. Results: None of tricalcium silicate-based materials were cytotoxic and all induced a similar release of angiogenic growth factors (PDGF, FGF-2, and VEGF) (p>0.05). The best cell viability was observed for hDPSCs (p<0.05) with all tricalcium silicate-based materials at day 14. Tube formation by HUVECs showed a significant increase with all tested materials (p<0.05). Conclusion: The tricalcium silicate-based materials showed potential for angiogenic stimulation of all stem cell types and significantly enhanced tube formation by HUVECs.

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Abstract Objective This study aimed to assess the association between tooth size and root canal morphology by using CBCT analysis. Methodology In this retrospective study, tooth anatomic lengths (crown and root lengths, buccolingual and mesiodistal dimensions) of 384 patients were assessed and correlated with Vertucci’s root canal morphology classification. Data was analyzed for gender-related differences using the independent sample t-test, ANOVA, and the Pearson’s correlation for a possible relation between anatomic lengths and canal morphology. Results The maxillary first and second premolars showed a greater predilection for Type IV and II variants, respectively, while the mandibular first premolar showed a greater predilection for Type II canal system. The root canal system of the mandibular second premolar showed maximal diversity (47% Type I, 30% Type II, and 20% Type III). The dimensions were greater in men regardless of tooth type. The most significant relation (p<0.05) between the anatomic size and canal morphology was observed in the maxillary first premolars, followed by the mandibular canines (buccolingual dimension) and the lower second premolars (crown length). Negative correlations existed between the crown length and the patient’s age for the anterior teeth and mandibular second premolar (r=−0.2, p<0.01). Conclusions The most common canal formation for anterior teeth was the Type I. The anatomic lengths had the strongest influence on the canal configuration of the maxillary first premolar, with Type IV being the most common root canal system. The mandibular second premolars showed maximal diversity in the canal classification terms and had a significant correlation with their crown lengths. Clinical Relevance The complex relationship between the canal morphology and anatomic tooth sizes need meticulous awareness and recognition during endodontic procedures, in conjunction with the demographic variabilities.

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Abstract Ultrasonic wave technology is widely used during dental treatments. We previously demonstrated that this method protects the gingival tissue. However, the physiological change on the gingival microvasculature caused by this method remains unclear. Objective The aim of this study was to investigate the relationship between the morphological and physiological effects on gingival microcirculation when preparing teeth, using the conventional dental turbine or ultrasonic method. Methodology The lower premolar teeth of beagle dogs were prepared along the gingival margin by using a dental turbine or ultrasonic wave instrument. Gingival vasculature changes were investigated using scanning electron microscopy for corrosion resin casts. Gingival blood flow at the preparation site was determined simultaneously by laser Doppler flowmetry. These assessments were performed immediately (Day 0), at 7 days and 30 days after tooth preparation. Results At day 0, in the turbine group, blood vessels were destroyed and some resin leaked. Furthermore, gingival blood flow at the site was significantly increased. In contrast, the ultrasonic group demonstrated nearly normal vasculature and gingival blood flow similar to the non-prepared group for 30 days after preparation. No significant alterations occurred in gingival circulation 30 days after either preparation; however, the turbine group revealed obvious morphological changes. Conclusions Based on multiple approach analyses, this study demonstrated that ultrasonic waves are useful for microvascular protection in tooth preparation. Compared with a dental turbine, ultrasonic wave instruments caused minimal damage to gingival microcirculation. Tooth preparation using ultrasonic wave instruments could be valuable for protecting periodontal tissue.

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Abstract The site of the sinus tract depends on the rate of resistance against abscess exudate drainage, bone morphology, and distance from the root apex to the outer cortical bone. Objective To assess apical bone thickness in buccal and palatal/lingual aspects of maxillary and mandibular teeth, using a high-resolution cone-beam computed tomography (CBCT) system. Methodology In total, 422 CBCT examinations were included in the study, resulting in a sample of 1400 teeth. The scans were acquired by PreXion 3D, with a high-resolution protocol. The bone thickness was taken as the distance between the center of the apical foramen and the buccal and lingual/palatal cortical bone. The quantitative variables were expressed as mean values±standard deviation. The independent samples were analyzed using the t-test or the Mann-Whitney test (p<0.05). Results The lowest mean value of bone thickness was observed in the buccal cortical bone of the upper canines (1.49 mm±0.86) and in the upper central incisors (1.59 mm±0.67). In premolar teeth, the lowest values were found in the buccal cortical bone of upper first premolars (1.13 mm±0.68). In the posterior teeth, the lowest values were found in the buccal cortical bone of upper first molars (1.98 mm±1.33). In the lower second molar region, the buccal cortical bone (8.36 mm±1.84) was thicker than the lingual cortical bone (2.95 mm±1.16) (p<0.05). Conclusions The lowest mean values of bone thickness are in the buccal cortical bone of the maxillary teeth. In the mandible, bone thickness is thinner in the buccal bone around the anterior and premolar teeth, and in the lingual aspect of mandibular molars. All these anatomic characteristics could make the occurrence of the sinus tract more susceptible in these specific regions of the maxillary and mandibular alveolar bone.

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Abstract Objective The present study aimed to investigate the participation of focal adhesion kinases (FAK) in interactions between osteoblastic cells and titanium (Ti) surfaces with three different topographies, namely, untreated (US), microstructured (MS), and nanostructured (NS). Methodology Osteoblasts harvested from the calvarial bones of 3-day-old rats were cultured on US, MS and NS discs in the presence of PF-573228 (FAK inhibitor) to evaluate osteoblastic differentiation. After 24 h, we evaluated osteoblast morphology and vinculin expression, and on day 10, the following parameters: gene expression of osteoblastic markers and integrin signaling components, FAK protein expression and alkaline phosphatase (ALP) activity. A smooth surface, porosities at the microscale level, and nanocavities were observed in US, MS, and NS, respectively. Results FAK inhibition decreased the number of filopodia in cells grown on US and MS compared with that in NS. FAK inhibition decreased the gene expression of Alp, bone sialoprotein, osteocalcin, and ALP activity in cells grown on all evaluated surfaces. FAK inhibition did not affect the gene expression of Fak, integrin alpha 1 ( Itga1 ) and integrin beta 1 ( Itgb1 ) in cells grown on MS, increased the gene expression of Fak in cells grown on NS, and increased the gene expression of Itga1 and Itgb1 in cells grown on US and NS. Moreover, FAK protein expression decreased in cells cultured on US but increased in cells cultured on MS and NS after FAK inhibition; no difference in the expression of vinculin was observed among cells grown on all surfaces. Conclusions Our data demonstrate the relevance of FAK in the interactions between osteoblastic cells and Ti surfaces regardless of surface topography. Nanotopography positively regulated FAK expression and integrin signaling pathway components during osteoblast differentiation. In this context, the development of Ti surfaces with the ability to upregulate FAK activity could positively impact the process of implant osseointegration.

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Abstract Objective This study aims to evaluate the influence of different air-abrasion pressures and subsequent heat treatment on the flexural strength, surface roughness, and crystallographic phases of highly translucent partially stabilized zirconia (Y-PSZ), and on the tensile bond strength of resin cement to Y-PSZ. Methodology Fully sintered zirconia specimens were ground with SiC paper (control) and/or air-abraded with 50 µm particles of alumina at 0.1, 0.15, 0.2, or 0.3 MPa or left as-sintered. After air-abrasion at 0.2 MPa (0.2AB), additional specimens were then heated to 1500°C, and held for one hour at this temperature (0.2AB+HT1h). Flexural strength and surface roughness were evaluated. Crystalline phase identification was also carried out using X-ray diffraction. Bonded zirconia specimens with self-adhesive resin cement were stored in distilled water at 37°C for 24 h, either with or without aging (thermal cycling 4-60°C/20000). Results were analyzed statistically by ANOVA and Tukey-Kramer tests. Results The flexural strength decreased with the increase in air-abrasion pressure, while in contrast, the surface roughness increased. The lowest flexural strength and the highest roughness value were found for the 0.2AB and 0.3AB groups, respectively. All groups contained cubic-, tetragonal ( t )-, and rhombohedral ( r )-ZrO2 phases with the exception of the as-sintered group. Upon increasing the air-abrasion pressure, the relative amount of the r -ZrO2 phase increased, with a significant amount of r -ZrO2 phase being detected for the 0.2AB and 0.3AB groups. The 0.2AB+HT1h group exhibited a similar flexural strength and t -ZrO2 phase content as the as-sintered group. However, the 0.2AB group showed a significantly higher tensile bond strength (p<0.05) than the 0.2AB+HT1h group before and after aging. Conclusion Micromechanical retention by alumina air-abrasion at 0.2 MPa, in combination with chemical bonding of a resin to highly translucent Y-PSZ using a MDP-containing resin cement may enable durable bonding.

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Abstract Objective This study aimed to investigate the effects of Maras powder (a type of smokeless tobacco obtained from Nicotiana rustica Linn and mixed with the ashes of wood, especially from oak, walnut or grapevine) on the microRNA (miRNA) deregulation of oral mucosa, and it compares these effects with those of smoking. Methodology Oral mucosal samples were collected from 74 patients, consisting of 16 nonusers, 26 smokers, and 32 Maras powder users. Genes associated with oral cancer were selected and 90 microRNAs targeting these genes were identified. MicroRNA were isolated and purified using the microRNA isolation kit. MicroRNA were expressed using Fluidigm RT-PCR. Results A positive correlation between the duration of Maras powder use with miR-31 expression levels, and a negative correlation between the Maras powder chewing time and miR-372 expression levels was found. In addition, there is a negative correlation between the amount of Maras powder consumed and expression levels of miR-375, miR-378a, miR-145, and miR-10b; moreover, another negative correlation is observed between the number of cigarettes consumed and the expression levels of miR-23a, miR-23b, miR-203a, miR-200b, and miR-375. However, miR-200b and miR-92a levels were downregulated significantly more in Maras powder users when compared with smokers and nonusers (p<0.05). Conclusion The results show both chewing Maras powder and smoking have an effect on deregulation of miR-200b and miR-92a expressions. This leads to the belief that assessing the expression of these two miRNAs is a promising noninvasive method of analysis, especially in mutagen exposures. Finally, large-scale and high-throughput studies may help to identify an extensive miRNA expression profile associated with tobacco use and improve the understanding of oral malignancies.

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Abstract Objectives This study assessed the incidence and variability features of root canals system (RCS) and their ramifications according to Pucci & Reig (PR) (1944) and the American Association of Endodontists (AAE) (2017) by micro-computed tomography (μCT). Methodology 500 representative extracted human teeth of each tooth group (n=50) (maxillary/mandibular central and lateral incisors, canines, first and second premolars and molars) were scanned by μCT with a resolution of 26.70 μm. The reconstructed cross-sections images and the visualization of the continuous slices in the transversal axis were performed using DataViewer software. RCS were classified according to Pucci & Reig (main canal, collateral canal, lateral canal, secondary canal, accessory canal, intercanal, recurrent canal) and AAE (main canal, accessory canal, lateral canal). The apical deltas were assessed for both classifications. The prevalence of apical deltas was evaluated using the Chi-squared test (p<0.05). Results According to PR, a higher incidence of lateral canals was observed in maxillary canines (10%), central incisors (8%) and first premolars (6%). Using AAE, the highest incidence of lateral canals was observed in the mandibular first premolars (85%), first and second molars (84%), lateral incisors (67%), canines (59%), and in maxillary first premolars (52%). Regarding accessory canals, the PR showed a frequency in 2% of the maxillary lateral incisors and maxillary and mandibular first premolars and 3% of mandibular first and second molars. On the other hand, the AAE showed the highest incidence of accessory canals in 86% of the maxillary first premolars, 71% in mandibular lateral incisors, 69% in mandibular first premolars, 65% in mandibular canines, and 56% in maxillary canines. The PR showed the lowest incidence of apical deltas for all dental groups when compared with AAE (p=0.004). Interestingly, distal canals in maxillary molars showed a significant discrepancy between classifications (p=0.027). Conclusions μCT enabled accurately describing the RC system and related ramifications, adding to the PR and AAE classifications, with some discrepancies reported for maxillary molars. Clinical Relevance This μCT study enabled a thorough description of the variability among root canals and their ramifications, including clinically relevant details on the presence and location of lateral canals and accessories in all human tooth groups, beyond the currently existing classification systems.

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Abstract Objective To quantify the bone volume that can be safely withdrawn from 3 donor sites: (1) the mandibular symphysis, (2) the oblique mandibular line and (3) the skullcap. Methodology For the symphysis, 200 tomographic exams were evaluated by the extension of the anterior loop of mental foramen, by the nerve, by the distance of the foramens, by the distance between the vestibular cortical and the lingual plates and by the distance between the apexes, or lower anterior teeth, and the mandibular base, using the “distance” tool of the I-CAT Vision, in the panoramic and parasagittal reformations. For the oblique line, 70 TCFC exams were analyzed retrospectively in panoramic and parasagittal reformations, evaluating the thickness of the vestibular cortical and the distance between the cortical and the mandibular canal. For the cranial bone, a hexagonal donor site located in parietal area was considered. Results The average dimensions of the bone blocks that can be safely removed from the region of the mandibular symphysis are: 32.27 mm in length, 4.87 mm in height and 4 mm in thickness, providing a volume of 628.61 mm3 available for grafting. In the oblique line, the available bone volume for grafting was 859.61 mm3. In the region of the cranial vault, multiplying the average bone thickness by the area of the hexagon, an average volume of 2,499 mm3 was obtained. Conclusions Comparing the donor sites, the bone availability in the cranial vault is 3 times greater than in the mandibular posterior region, and at least 2 times greater than in the mandibular symphysis.

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Abstract Objective This in vitro study evaluated the effect of commercial whitening dentifrices on erosive tooth wear (ETW) of bovine enamel samples, in comparison with commercial regular dentifrices. Methodology Sixty bovine crowns were embedded in acrylic resin, polished and then had their baseline profile determined. They were randomly assigned to 5 groups (n=12/group), according to the type of commercial dentifrice to be tested: GI – Crest Anti-cavity Regular; GII – Crest 3D White; GIII – Colgate Total 12 Clean Mint; GIV – Colgate Optic White; GV – Placebo (negative control, fluoride-free dentifrice). The samples were submitted to daily erosive and abrasive challenges for 3 days. The erosive challenges were performed 3 times a day by immersing the specimens in 0.1% citric acid solution (pH 2.5) for 90 s. Each day after the first and last erosive challenges, the specimens were subjected to the abrasive challenge for 15 s, using a toothbrushing machine (Biopdi, São Carlos, SP, Brazil), soft toothbrushes and slurry (1:3 g/ml) of the tested toothpastes (1.5 N). The specimens were kept in artificial saliva between the challenges. The final profile was obtained and the ETW (µm) was calculated. Data were analyzed by Kruskal-Wallis and Dunn’s tests (p<0.05). Results All dentifrices tested significantly reduced the enamel wear in comparison with the Placebo, except GIII. The median (95% CI) ETW was 1.35 (1.25-1.46)bc for GI, 1.17 (1.01-1.34)cd for GII, 1.36 (1.28-1.45)ab for GIII, 1.08 (1.04-1.14)d for GIV and 2.28 (2.18-2.39)a for GV. Conclusion When dentifrices from the same manufacturer were compared, the whitening dentifrices led to similar or less wear than the regular ones.

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Abstract Isthmuses are reported as common anatomic complexities in teeth often associated with failures in endodontic treatment. They should be considered before starting treatment and a preoperative computed tomography scan (CT) may demonstrate these complexities. Objective To assess the diagnostic value of the highest resolution settings of a cone-beam CT (CBCT) system in identifying and measuring apical isthmuses, using micro-CT as reference. Methodology After micro-CT scanning, 40 humans’ lower first molars with isthmuses in the apical-3 mm of mesial roots were scanned by the highest resolution settings of the New Generation i-Cat ® CBCT equipment. Two blinded observers recorded the detection of isthmuses in CBCT scans. The lengths of isthmuses were compared between micro-CT and CBCT to assess the diagnostic value of CBCT. Quantitative data for sensitivity were represented as percentages (95% confidence interval). The Bland-Altman method was used to assess differences between gold standard lengths (micro-CT) and CBCT lengths. Results BCT demonstrated 30 positive findings, representing sensitivity for isthmus identification of 75% (95% CI=0.4114–1.1364). Differences between the lengths in micro-CT (1.99±0.40 mm) and CBCT (1.53±0.41 mm) were significant (p<0.0001). Conclusion The CBCT device used presented limited diagnostic value in the identification and measurement of apical isthmuses in the mesial roots of lower molars. In some cases, the actual anatomy of the apical root canal may not be completely delineated in this type of CBCT system, even using the highest resolution settings.

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Abstract Objective This clinical trial sought to evaluate the clinical effectiveness of concentrated growth factor (CGF) and compare it with connective tissue graft (CTG) with coronally advanced flap (CAF) in the treatment of Miller Class I gingival recessions (GR). Methodology This split-mouth study included 74 Miller Class I isolated (24 teeth) or multiple (50 teeth) GRs in 23 jaws of 19 patients. GRs were randomly treated using CGF (test group: 37 teeth; 12 teeth in isolated GRs, 25 teeth in multiple GRs) or CTG with CAF (control group: 37 teeth;12 teeth isolated GRs, 25 teeth in multiple GRs). Clinical variables, plaque index (PI), gingival index (GI), probing depth (PD), recession depth (RD), recession width (RW), clinical attachment level (CAL), keratinized tissue thickness (KTT), keratinized tissue width (KTW), and root coverage (RC) were assessed at the baseline as well as at three and six months post-surgery. Healing index (HI) were obtained in the second and third weeks post-surgery. Postoperative pain was assessed for the first seven days using a horizontal visual analog scale (VAS). Results No significant change was observed in PI, GI, or PD values in either the intergroup or the intragroup comparisons. A statistically significant decrease was observed in CAL, RD, and RW, and KTT increased in all groups at three and six months compared with the baseline. The control group had greater increases in KTW, KTT, and RC at three and six months. No significant difference was found in CAL or RD at the third and sixth months between the two groups. Healing was found to be similar for both groups in the second and third weeks post-surgery. The VAS values in the control group were higher than in the test group, especially at the second, fourth, fifth, and seventh days postoperatively. Conclusions CTG is superior to CGF with CAF for increasing KTT, KTW, and RC. CGF may be preferable due to decreased postoperative pain.

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Abstract Excessive weight is associated with periodontitis because of inflammatory mediators secreted by the adipose tissue. Periodontal impairments can occur during pregnancy due to association between high hormonal levels and inadequate oral hygiene. Moreover, periodontitis and excessive weight during pregnancy can negatively affect an infant’s weight at birth. Objective This observational, cross-sectional study aimed to evaluate the association between pre-pregnancy overweight/obesity, periodontitis during the third trimester of pregnancy, and the infants' birth weight. Methodology The sample set was divided into 2 groups according to the preconception body mass index: obesity/overweight (G1=50) and normal weight (G2=50). Educational level, monthly household income, and systemic impairments during pregnancy were assessed. Pocket probing depth (PPD) and clinical attachment level (CAL) were obtained to analyze periodontitis. The children’s birth weight was classified as low (<2.5 kg), insufficient (2.5–2.999 kg), normal (3–3.999 kg), or excessive (≥4 kg). Bivariate analysis (Mann-Whitney U test, t-test, chi-squared test) and logistic regression (stepwise backward likelihood ratio) were performed (p<0.05). Results G1 showed lower socioeconomic levels and higher prevalence of arterial hypertension and gestational diabetes mellitus during pregnancy than G2 (p=0.002). G1 showed higher means of PPD and CAL (p=0.041 and p=0.039, respectively) and therefore a higher prevalence of periodontitis than G2 (p=0.0003). G1 showed lower infants’ birth weight than G2 (p=0.0004). Excessive maternal weight and educational levels were independent variables associated with periodontitis during the third trimester of pregnancy (X2[2]=23.21; p<0.0001). Maternal overweight/obesity was also associated with low/insufficient birth weight (X2[1]=7.01; p=0.008). Conclusion The present findings suggest an association between excessive pre-pregnancy weight, maternal periodontitis, and low/insufficient birth weight.

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Abstract This study aimed to evaluate whether the presence of awake bruxism was associated with temporomandibular dysfunction symptoms, pain threshold at pressure, pain vigilance, oral health-related quality of life (OHRQoL), and anxiety and depression symptoms in patients undergoing orthodontic treatment. Methodology This observational study followed patients who had started receiving orthodontic treatment for six months. The following variables were measured three times (at baseline, one month, and six months): pressure pain threshold (PPT) in the right and left masseter, anterior temporalis, and temporomandibular joint (TMJ), and right forearm; pain vigilance and awareness questionnaire; and shortened form of the oral health impact profile (OHIP-14). Anxiety and depression symptoms were measured using the Beck anxiety inventory and the Beck depression inventory, respectively. The patients were divided into two main groups according to the presence (n=56) and absence (n=58) of possible awake bruxism. The multi-way analysis of variance (ANOVA) was applied on the date (p=0.050). Results TMJ and/or muscle pain were not observed in both groups. Time, sex, age group, and awake bruxism did not affect the PPT in the masticatory muscles and pain vigilance (p>0.050). However, the primary effect of awake bruxism was observed when anxiety (ANOVA: F=8.61, p=0.004) and depression (ANOVA: F=6.48, p=0.012) levels were higher and the OHRQoL was lower (ANOVA: F=8.61, p=0.004). Conclusion The patients with self-reported awake bruxism undergoing an orthodontic treatment did not develop TMJ/masticatory muscle pain. The self-reported awake bruxism is associated with higher anxiety and depression levels and a poorer OHRQoL in patients during the orthodontic treatment.

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Abstract The acquired pellicle formation is the first step in dental biofilm formation. It distinguishes dental biofilms from other biofilm types. Objective To explore the influence of salivary pellicle formation before biofilm formation on enamel demineralization. Methodology Saliva collection was approved by Indiana University IRB. Three donors provided wax–stimulated saliva as the microcosm bacterial inoculum source. Acquired pellicle was formed on bovine enamel samples. Two groups (0.5% and 1% sucrose–supplemented growth media) with three subgroups (surface conditioning using filtered/pasteurized saliva; filtered saliva; and deionized water (DIW)) were included (n=9/subgroup). Biofilm was then allowed to grow for 48 h using Brain Heart Infusion media supplemented with 5 g/l yeast extract, 1 mM CaCl2.2H2O, 5% vitamin K and hemin (v/v), and sucrose. Enamel samples were analyzed for Vickers surface microhardness change (VHNchange), and transverse microradiography measuring lesion depth (L) and mineral loss (∆Z). Data were analyzed using two-way ANOVA. Results The two-way interaction of sucrose concentration × surface conditioning was not significant for VHNchange (p=0.872), ∆Z (p=0.662) or L (p=0.436). Surface conditioning affected VHNchange (p=0.0079), while sucrose concentration impacted ∆Z (p<0.0001) and L (p<0.0001). Surface conditioning with filtered/pasteurized saliva resulted in the lowest VHNchange values for both sucrose concentrations. The differences between filtered/pasteurized subgroups and the two other surface conditionings were significant (filtered saliva p=0.006; DIW p=0.0075). Growing the biofilm in 1% sucrose resulted in lesions with higher ∆Z and L values when compared with 0.5% sucrose. The differences in ∆Z and L between sucrose concentration subgroups was significant, regardless of surface conditioning (both p<0.0001). Conclusion Within the study limitations, surface conditioning using human saliva does not influence biofilm–mediated enamel caries lesion formation as measured by transverse microradiography, while differences were observed using surface microhardness, indicating a complex interaction between pellicle proteins and biofilm–mediated demineralization of the enamel surface.

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Abstract This study investigated the effect of a calcium hydroxide (CH) paste (CleaniCal®) containing N-2-methyl pyrrolidone (NMP) as a vehicle on Enterococcus faecalis (E. faecalis) biofilms compared with other products containing saline (Calasept Plus™) or propylene glycol (PG) (Calcipex II®). Methodology Standardized bovine root canal specimens were used. The antibacterial effects were measured by colony-forming unit counting. The thickness of bacterial microcolonies and exopolysaccharides was assessed using confocal laser scanning microscopy. Morphological features of the biofilms were observed using field-emission scanning electron microscopy (FE-SEM). Bovine tooth blocks covered with nail polish were immersed into the vehicles and dispelling was observed. The data were analyzed using one-way analysis of variance and Tukey tests (p<0.05). Results CleaniCal® showed the highest antibacterial activity, followed by Calcipex II® (p<0.05). Moreover, NMP showed a higher antibacterial effect compared with PG (p<0.05). The thickness of bacteria and EPS in the CleaniCal® group was significantly lower than that of other materials tested (p<0.05). FE-SEM images showed the specimens treated with Calasept Plus™ were covered with biofilms, whereas the specimens treated with other medicaments were not. Notably, the specimen treated with CleaniCal® was cleaner than the one treated with Calcipex II®. Furthermore, the nail polish on the bovine tooth block immersed in NMP was completely dispelled. Conclusions CleaniCal® performed better than Calasept Plus™ and Calcipex II® in the removal efficacy of E. faecalis biofilms. The results suggest the effect might be due to the potent dissolving effect of NMP on organic substances.

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Abstract The tongue participates in the oral phase of swallowing by pushing the food bolus toward the oropharynx. This relationship between tongue function and swallowing is little addressed addressed in individuals with temporomandibular dysfunction (TMD). Objective: To analyze the association of functional tongue conditions on swallowing in individuals with TMD. Methodology: After approval by the Institutional Review Board, the study was conducted on 30 individuals of both sexes, aged 18 to 28 years, with TMD, and not treated for the disorder. Tongue function was assessed as to the mobility, pressure, and oral motor control. Swallowing was analyzed by clinical assessment during ingestion of solid (wafer biscuit) and liquid (water). Data regarding mobility and swallowing were collected using the orofacial myofunctional evaluation protocol. Tongue pressure was measured by the Iowa Oral Performance Instrument, during elevation, protrusion, swallowing, and resistance test. The oral motor control was assessed by the oral diadochokinesis (DDK) test by rapid and repeated emissions of syllables “ta” and “ka”. Data were statistically analyzed by the Spearman correlation coefficient, at a significance level of 5%. Results: Relationships were found between tongue function and swallowing for the following aspects: mobility (r=0.741), pressure in protrusion (r=-0.366), swallowing of saliva (r=-0.499), mean DDK rate in emissions “ta” (r=-0.424) and “ka” (r=-0.446), and mean DDK period in emissions “ta” (r=0.424) and “ka” (r=0.446). Thus, the greater the change in tongue mobility, the lower the tongue pressure in protrusion and swallowing of saliva, the lower the emissions per second, the longer the mean time between vocalizations, and the worse the swallowing of individuals with TMD. Conclusion: The functional conditions of the tongue regarding mobility, pressure, and oral DDK were associated with swallowing in individuals with TMD.

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Abstract Menopause induces oral bone loss, leading to various oral diseases. Mastication importantly affects bone metabolism in the jawbone. Objective: To analyze the effect of enhanced masticatory force on osteoprotegerin (OPG), receptor activator of nuclear factor kappa B ligand (RANKL), and mechano–growth factor (MGF) in alveolar bone of ovariectomized rats and to study the mechanics mechanism of the alveolar bone of ovariectomized rats response to enhanced masticatory force. Methodology: Thirty Sprague Dawley rats were randomly divided into three groups: sham–operation group (fat around the removed ovary + normal hard diet), model group (ovariectomy + normal hard diet), and experimental group (ovariectomy + high hard diet). It was a 2–month experiment. Enzyme–linked immunosorbent assay (ELISA) detected serum estradiol (E2), osteocalcin (BGP) and alkaline phosphatase (ALP) in rats. Bone histomorphometric indices in the third molar region of maxilla were detected by micro-CT; protein expressions of OPG, RANKL, and MGF in the third molar region of maxilla was detected by Western blot; and gene expression of OPG, RANKL, and MGF in the third molar region of maxilla was detected by Quantitative Real–Time PCR. Results: Comparing with model group, serum E2 in experimental group increased but not significantly, serum BGP and serum ALP in experimental group decreased but not significantly, OPG in experimental group in alveolar bone increased significantly, RANKL in experimental group in alveolar bone decreased significantly, RANKL/OPG ratio in experimental group decreased significantly, MGF in experimental group in alveolar bone increased significantly, bone volume to total volume fraction increased significantly in experimental group, trabecular thickness increased significantly in experimental group, and trabecular separation decreased significantly in experimental group. Conclusion: Enhanced masticatory force affected the expression of OPG, RANKL, and MGF in alveolar bone of ovariectomized rats, improved the quality of jaw bone of ovariectomized rats, and delayed oral bone loss by ovariectomy.

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Abstract The relationship between periodontitis and the pathogenesis of other inflammatory diseases, such as diabetes, rheumatoid arthritis and obesity has been an important topic of study in recent decades. The Th17 pathway plays a significant role in how local inflammation can influence systemic inflammation in the absence of systemic pathology. Objective: To determine Th17 biased-cells in systemically healthy patients in the presence of generalized chronic periodontitis. Methodology: A total of 28 patients were recruited without systemic inflammatory pathology, which was determined by clinical history, the Health Assessment Questionnaire (HAQ) and rheumatoid factor detection. Of these patients, 13 were diagnosed as healthy/gingivitis (H/G) and 15 as generalized chronic periodontitis (GCP). Th17 (CD4+CD161+) cells and Th17IL23R+ (CD4+CD161+IL-23R+) cells were quantified by flow cytometry, based on the total cells and on the lymphocyte region, termed the “enriched population” (50,000 events for each). Results: The percentages of Th17 cells of the H/G and periodontitis groups were similar on total cells and enriched population (19 vs 21.8; p=4.134 and 19.6 vs 21.8; p=0.55). However, Th17IL23R+ cells differ significantly between periodontally healthy patients and generalized chronic periodontitis patients in both total cell (0.22% vs 0.65%; p=0.0004) and enriched populations (0.2% vs 0.75%; p=0.0266). Conclusions: GCP patients (otherwise systemically healthy) were characterized by increased Th17-proinflammatory cell phenotype positive for the IL-23 receptor in peripheral blood. The proportion of Th17 cells that are negative for the IL-23 receptor in the peripheral blood of systemically healthy patients seemed to be unaffected by the presence or absence of chronic periodontitis.

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Abstract Genetic and epigenetic changes have been associated with periodontitis in various genes; however, little is known about genes involved in epigenetic mechanisms and in oxidative stress. Objective: This study aims to investigate the association of polymorphisms C677T in MTHFR (rs1801133) and −149C→T in DNMT3B (rs2424913), as well as the methylation profiles of MTHFR, miR-9-1, miR-9-3, SOD1, and CAT with periodontitis. The association between polymorphisms and DNA methylation profiles was also analyzed. Methodology: The population studied was composed of 100 nonsmokers of both sexes, divided into healthy and periodontitis groups. Genomic DNA was extracted from the epithelial buccal cells, which were collected through a mouthwash. Polymorphism analysis was performed through polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), while methylation-specific PCR (MSP) or combined bisulfite restriction analysis techniques were applied for methylation analysis. Results: For DNMT3B, the T allele and the TT genotype were detected more frequently in the periodontitis group, as well as the methylated profile on the miR-9-1 promoter region. There was also a tendency towards promoter region methylation on the CAT sequence of individuals with periodontal disease. Conclusion: The polymorphism −149C→T in DNMT3B (rs2424913) and the methylated profile of the miR-9-1 promoter region are associated with periodontitis.

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Abstract Objective Maxillary molar distalization with intraoral distalizer appliances is a non-extraction orthodontic treatment used to correct molar relationship in patients with Class II malocclusion presenting maxillary dentoalveolar protrusion and minor skeletal discrepancies. This study compares the changes caused by three distalizers with different force systems. Methodology 71 patients, divided into three groups, were included. The Jones jig group (JJG, n=30; 16 male, 14 female, 13.17 years mean age) was treated with the Jones jig for 0.8 years. The Distal jet group (DJG, n=25; 8 male, 17 female, 12.57 years mean age) was treated with the Distal jet for 1.06 years. The First Class group (FCG, n=16; 6 male, 10 female, 12.84 years mean age) was treated with the First Class for 0.69 years. Intergroup treatment changes were compared using one-way ANOVA, followed by post-hoc Tukey’s tests. Results Intergroup comparisons showed significantly greater maxillary incisor protrusion in DJG than in FCG (2.56±2.24 mm vs. 0.74±1.39mm, p=0.015). The maxillary first premolars showed progressive and significantly smaller mesial angulation in JJG, FCG and DJG, respectively (14.65±6.31º, 8.43±3.99º, 0.97±3.16º; p<0.001). They also showed greater mesialization in JJG than FCG (3.76±1.46 mm vs. 2.27±1.47 mm, p=0.010), and greater extrusion in DJG compared to JJG (0.90±0.77 mm vs 0.11±0.60 mm, p=0.004). The maxillary second premolars showed progressive and significantly smaller mesial angulation and mesialization in JJG, FCG and DJG, respectively (12.77±5.78º, 3.20±3.94º, -2.12±3.71º and 3.87±1.34 mm, 2.25±1.40 mm, 1.24±1.26 mm, respectively; p<0.001). DJG showed smaller distal angulation of maxillary first molars (-2.14±5.09º vs. -7.73±4.28º and -6.05±3.76º, for the JJG and FCG, respectively; p<0.001) and greater maxillary second molars extrusion (1.17±1.41 mm vs -0.02±1.16 mm and 0.16±1.40 mm, for the JJG and FCG, respectively; p=0.003). Overjet change was significantly larger in DJG compared to FCG (1.79±1.67 mm vs 0.68±0.84; p=0.046). Treatment time was smaller in FCG (0.69±0.22 years vs 0.81±0.33 years and 1.06±0.42 years, comparing it with the JJG and DJG, respectively; p=0.005). Conclusion The three appliances corrected the Class II molar relationship by dentoalveolar changes. The Distal jet produced smaller molar distal angulation than the Jones jig and First Class. The First Class appliance showed less anchorage loss, greater percentage of distalization and shorter treatment time than the Jones jig and Distal jet.

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Abstract Objectives To evaluate the acoustic properties of the /s/ sound in individuals with different occlusion types and to investigate relationships between these properties and cephalometric measurements. Methodology Sixty patients were divided into three groups based on malocclusion. Group 1 included 20 patients (mean age: 14.85±2.01 years) with Class I skeletal and dental relationships. Group 2 included 20 patients (mean age: 13.49±1.78 years) with Class II skeletal and dental relationships. Group 3 included 20 patients (mean age: 12.46±2.62 years) with Class III skeletal and dental relationships. Cephalometric tracings were obtained from cephalometric radiographs. All included patients were native speakers of Turkish. The /s/ sound was selected for center of gravity analysis. Correlations between cephalometric values and acoustic parameters were also investigated. Results The center of gravity of the /s/ sound had the lowest value in Group 2 (p<0.05). For the /s/ sound in Group 3, moderate positive correlations were found between center of gravity and Sella-Nasion to Gonion-Gnathion angle (p<0.05, r=0.444) Lower incisor to Nasion-B point (p<0.023, r=0.505), and Lower incisor to Nasion-B point angle (p<0.034; r=0.476). No correlation was found in other cephalometric measurements. Conclusions The /s/ sound was affected by malocclusion due to the changing place of articulation. Therefore, referral to an orthodontist for malocclusion treatment especially patients with class III in the early period is suggested for producing acoustically ideal sound.

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Abstract Enzymatic degradation of the hybrid layer can be accelerated by the activation of dentin metalloproteinases (MMP) during the bonding procedure. MMP inhibitors may be used to contain this process. Objective To evaluate the degree of conversion (DC%), dentin bond strength (µTBS) (immediate and after 1 year of storage in water), and nanoleakage of an experimental (EXP) and a commercial (SB) adhesive system, containing different concentrations of the MMP inhibitor GM1489: 0, 1 µM, 5 µM and 10 µM. Methodology DC% was evaluated by FT-IR spectroscopy. Dentin bond strength was evaluated by µTBS test. Half of beams were submitted to the µTBS test after 24 h and the other half, after storage for 1 year. From each tooth and storage time, 2 beams were reserved for nanoleakage testing. Data were analyzed using ANOVA and Tukey’s test to compare means (α=0.05). Results All adhesive systems maintained the µTBS after 1 year of storage. Groups with higher concentrations of inhibitor (5 µM and 10 µM) showed higher µTBS values than groups without inhibitor or with 1 µM. The nanoleakage values of all groups showed no increase after 1 year of storage and values were similar for SB and EXP groups, in both storage periods. The inhibitor did not affect the DC% of the EXP groups, but the SB5 and SB10 groups showed higher DC% values than those of SB0 and SB1. Conclusions The incorporation of GM1489 in the adhesive systems had no detrimental effect on DC%. The concentrations of 5 µM GM1489 for SB and 5 µM or 10 µM for EXP provided higher μTBS than groups without GM1489, in the evaluation after 1 year of storage; whereas the concentration of inhibitor did not affect adhesive systems nanoleakage.

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Abstract Natural products have emerged as a rich source of bioactive compounds for adjunctive treatments of many infectious and inflammatory conditions, including periodontitis. Among the monoterpenes with significant biological properties, there is the perillyl alcohol (POH), which can be found in several essential oils and has shown immunomodulatory properties in recent studies, which may be interesting in the treatment of non-neoplastic inflammatory disorders. Objective To determine the antibacterial and immune modulatory activities of the POH. Methodology The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the POH for two significant Gram-negative periodontal pathogens were determined by macrodilution and subculture, respectively. Cell proliferation and cytotoxicity in RAW 264.7 macrophages were determined by Trypan Blue and mitochondrial enzymatic activity assay. The modulation of reactive oxygen species (ROS) was analyzed by flow cytometry and expression of TNF and arginase-1 by real-time PCR. Results The POH was effective against P. gingivalis (ATCC 33277) and F. nucleatum (ATCC 25586) with MIC= MBC=1600 μM. No cytotoxicity up to 100 µM was observed on macrophages. The cell proliferation was inhibited from 48 hours at 100 μM (p<0.05) and 250 μM (p<0.01). The POH increased ROS production at both 10 μM and 100 μM (p<0.05) in unstimulated cells. The PMA-induced ROS production was not affected by POH, whereas 100 μM significantly reduced lipopolysaccharide-induced (LPS-induced) ROS. The expression of TNF was not affected by POH in unstimulated cells or in cells polarized to M1 phenotype, whereas both concentrations of POH reduced (p<0.05) the expression of arginase-1 in M2-polarized macrophages. Conclusion The POH has antibacterial activity against periodontal pathogens and reduced proliferation of murine macrophages without significant cytotoxicity at concentrations up to 100 μM. In addition, the POH reduced the LPS-induced ROS and the expression of arginase-1 in M2-polarized macrophages.

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Abstract Oral leukoplakia (OL) is a white lesion of an indeterminate risk not related to any excluded (other) known diseases or disorders that carry no increased risk for cancer. Many biological markers have been used in an attempt to predict malignant transformation; however, no reliable markers have been established so far. Objective To evaluate cell proliferation and immortalization in OL, comparing non-dysplastic (Non-dys OL) and dysplastic OL (Dys OL). Methodology This is a cross-sectional observational study. Paraffin-embedded tissue blocks of 28 specimens of Non-dys OL, 33 of Dys OL, 9 of normal oral mucosa (NOM), 17 of inflammatory hyperplasia (IH), and 19 of oral squamous cell carcinomas (OSCC) were stained for Ki-67 and BMI-1 using immunohistochemistry. Results A gradual increase in BMI-1 and K-i67 expression was found in oral carcinogenesis. The immunolabeling for those markers was higher in OSCC when compared with the other groups (Kruskal-Wallis, p<0.05). Ki-67 expression percentage was higher in OL and in IH when compared with NOM (Kruskal-Wallis/Dunn, p<0.05). Increased expression of BMI-1 was also observed in OL when compared with NOM (Kruskal-Wallis/Dunn, p<0.05). No differences were observed in expression of both markers when non-dysplastic and dysplastic leukoplakias were compared. A significant positive correlation between Ki-67 and BMI-1 was found (Spearman correlation coefficient, R=0.26, p=0.01). High-grade epithelial dysplasia was associated with malignant transformation (Chi-squared, p=0.03). Conclusions These findings indicate that BMI-1 expression increases in early oral carcinogenesis and is possibly associated with the occurrence of dysplastic changes. Furthermore, our findings indicate that both Ki-67 and BMI-1 are directly correlated and play a role in initiation and progression of OSCC.

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Abstract Objective To evaluate the influence of three levels of dental structure loss on stress distribution and bite load in root canal-treated young molar teeth that were filled with bulk-fill resin composite, using finite element analysis (FEA) to predict clinical failure. Methodology Three first mandibular molars with extensive caries lesions were selected in teenager patients. The habitual occlusion bite force was measured using gnathodynamometer before and after endodontic/restoration procedures. The recorded bite forces were used as input for patient-specific FEA models, generated from cone-beam computed tomographic (CT) scans of the teeth before and after treatment. Loads were simulated using the contact loading of the antagonist molars selected based on the CT scans and clinical evaluation. Pre and post treatment bite forces (N) in the 3 patients were 30.1/136.6, 34.3/133.4, and 47.9/124.1. Results Bite force increased 260% (from 36.7±11.6 to 131.9±17.8 N) after endodontic and direct restoration. Before endodontic intervention, the stress concentration was located in coronal tooth structure; after rehabilitation, the stresses were located in root dentin, regardless of the level of tooth structure loss. The bite force used on molar teeth after pulp removal during endodontic treatment resulted in high stress concentrations in weakened tooth areas and at the furcation. Conclusion Extensive caries negatively affected the bite force. After pulp removal and endodontic treatment, stress and strain concentrations were higher in the weakened dental structure. Root canal treatment associated with direct resin composite restorative procedure could restore the stress-strain conditions in permanent young molar teeth.

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Abstract Objective Ameloblastoma is a representative odontogenic tumor comprising several characteristic invasive forms, and its pathophysiology has not been sufficiently elucidated. A stable animal experimental model using immortalized cell lines is crucial to explain the factors causing differences among the subtypes of ameloblastoma, but this model has not yet been disclosed. In this study, a novel animal experimental model has been established, using immortalized human ameloblastoma-derived cell lines. Methodology Ameloblastoma cells suspended in Matrigel were subcutaneously transplanted into the heads of immunodeficient mice. Two immortalized human ameloblastoma cell lines were used: AM-1 cells derived from the plexiform type and AM-3 cells derived from the follicular type. The tissues were evaluated histologically 30, 60, and 90 days after transplantation. Results Tumor masses formed in all transplanted mice. In addition, the tumors formed in each group transplanted with different ameloblastoma cells were histologically distinct: the tumors in the group transplanted with AM-1 cells were similar to the plexiform type, and those in the group transplanted with AM-3-cells were similar to the follicular type. Conclusions A novel, stable animal experimental model of ameloblastoma was established using two cell lines derived from different subtypes of the tumor. This model can help clarify its pathophysiology and hasten the development of new ameloblastoma treatment strategies.

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Abstract Objective This study sought to analyze the gene expression of Candida albicans in sound root surface and root caries lesions, exploring its role in root caries pathogenesis. Methodology The differential gene expression of C. albicans and the specific genes related to cariogenic traits were studied in association with samples of biofilm collected from exposed sound root surface (SRS, n=10) and from biofilm and carious dentin of active root carious lesions (RC, n=9). The total microbial RNA was extracted, and the cDNA libraries were prepared and sequenced on the Illumina Hi-Seq2500. Unique reads were mapped to 163 oral microbial reference genomes including two chromosomes of C. albicans SC5314 (14,217 genes). The putative presence of C. albicans was estimated (sum of reads/total number of genes≥1) in each sample. Count data were normalized (using the DESeq method package) to analyze differential gene expression (using the DESeq2R package) applying the Benjamini-Hochberg correction (FDR<0.05). Results Two genes (CaO19.610, FDR=0.009; CaO19.2506, FDR=0.018) were up-regulated on SRS, and their functions are related to biofilm formation. Seven genes ( UTP20 , FDR=0.018; ITR1 , FDR=0.036; DHN6 , FDR=0.046; CaO19.7197 , FDR=0.046; CaO19.7838 , FDR=0.046; STT4 , FDR=0.046; GUT1 , FDR=0.046) were up-regulated on RC and their functions are related to metabolic activity, sugar transport, stress tolerance, invasion and pH regulation. The use of alternative carbon sources, including lactate, and the ability to form hypha may be a unique trait of C. albicans influencing biofilm virulence. Conclusions C. albicans is metabolically active in SRS and RC biofilm, with different roles in health and disease.

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Abstract Objective To investigate the effects of intro-oral injection of parathyroid hormone (PTH) on tooth extraction wound healing in hyperglycemic rats. Methodology 60 male Sprague-Dawley rats were randomly divided into the normal group (n=30) and DM group (n=30). Type 1 diabetes mellitus (DM) was induced by streptozotocin. After extracting the left first molar of all rats, each group was further divided into 3 subgroups (n=10 per subgroup), receiving the administration of intermittent PTH, continuous PTH and saline (control), respectively. The intermittent-PTH group received intra-oral injection of PTH three times per week for two weeks. A thermosensitive controlled-release hydrogel was synthesized for continuous-PTH administration. The serum chemistry was determined to evaluate the systemic condition. All animals were sacrificed after 14 days. Micro-computed tomography (Micro-CT) and histological analyses were used to evaluate the healing of extraction sockets. Results The level of serum glucose in the DM groups was significantly higher than that in the non-DM groups (p<0.05); the level of serum calcium was similar in all groups (p>0.05). Micro-CT analysis showed that the DM group had a significantly lower alveolar bone trabecular number (Tb.N) and higher trabecular separation (Tb.Sp) than the normal group (p<0.05). The histological analyses showed that no significant difference in the amount of new bone (hard tissue) formation was found between the PTH and non-PTH groups (p>0.05). Conclusions Bone formation in the extraction socket of the type 1 diabetic rats was reduced. PTH did not improve the healing of hard and soft tissues. The different PTH administration regimes (continuous vs. intermittent) had similar effect on tissue healing. These results demonstrated that the metabolic characteristics of the hyperglycemic rats produced a condition that was unable to respond to PTH treatment.

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Abstract Some conditions consolidated as risk factors for oropharyngeal dysphagia have already been identified in other diseases, such as neurological. Studies on cardiovascular diseases concentrate in individuals in the postoperative period; thus, it is unknown if these same factors occur in individuals hospitalized for clinical or surgical treatment of these diseases. Objective to correlate predictive risk factors for oropharyngeal dysphagia in individuals with cardiovascular disease admitted at a reference cardiology hospital. Methodology This is a retrospective clinical study. Medical records of 175 individuals hospitalized for clinical and/or surgical treatment at a reference cardiology hospital from January to June 2017, attendants of the Speech-Language Pathology and Nutrition team, were analyzed. Of these, 100 records were included in the study: 41 females and 59 males (mean age 67.56 years). Deaths and individuals from 0 to 18 years were excluded. Stroke, malnutrition, age and prolonged orotracheal intubation were considered predictive risk factors for oropharyngeal dysphagia. Mann-Whitney test and Fisher's test were used for statistical analysis. Results Stroke (OR=2.93 p=0.02), malnutrition (OR=2.89 p=0.02) and prolonged orotracheal intubation (OR=3.94 p=0.02) were statistically significant predictors for oropharyngeal dysphagia within this population. Age below 80 years was not significant (p=0.06), but within octogenarians, significance was found (p=0.033). Conclusion Stroke, malnutrition, prolonged orotracheal intubation and age > 80 years are predictive risk factors for oropharyngeal dysphagia in adult population with cardiovascular diseases.

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Abstract Objective The aim of this study was to compare the effects of music at 432 Hz, 440 Hz, and no music on the clinical perception of anxiety and salivary cortisol levels in patients undergoing tooth extraction. Methodology A parallel-group randomized clinical trial was conducted. Forty-two patients (average age: 23.8±7.8 years, 27 women) with a moderate level of anxiety were distributed in three groups: use of music for 15 minutes at a frequency of 432 Hz (n=15), at 440 Hz (n=15) and a control group without music (n=12). The CORAH Dental Anxiety Scale and salivary cortisol levels, estimated by the solid phase enzyme-linked immunosorbent assay (ELISA), were measured and compared before and after the music intervention between groups (two-way ANOVA-Tukey p<0.05, RStudio). Results Significantly lower anxiety level values were observed at 432 Hz (8.7±2.67) and 440 Hz (8.4±2.84) compared to the control group (17.2±4.60; p<0.05). The salivary cortisol level at 432 Hz (0.49±0.37 μg/dL) was significantly lower than 440 Hz (1.35±0.69 μg/dL) and the control group (1.59±0.7 μg/dL; p<0.05). Conclusion The use of music significantly decreased clinical anxiety levels, and the frequency of 432 Hz was effective in decreasing salivary cortisol levels before tooth extraction.

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Abstract Debonding, staining and wear are usually the reasons for denture teeth replacement by new ones from same or different brands. Objective This study investigates the possible differences in color of denture teeth of the same or different brands under different illuminations, since their metameric behavior in color under specific illumination may become unacceptable. Methodology For the purpose of this study, 10 denture teeth (#11), shade A3, of 4 different brands were selected (Creopal/KlemaDental Pro, Executive/DeguDent, Cosmo HXL/DeguDent, Ivostar/Ivoclar-Vivadent). Teeth stabilized in white silicone mold and the CIELAB color coordinates of their labial surface under 3 different illumination lights (D65, F2, A) were recorded, using a portable colorimeter (FRU/WR-18, Wave Inc). ΔE*ab values of all possible pairs of teeth of the same brand (n=45) or pair combinations of different brands (n=100) under each illumination light, in a dry and wet state were calculated. Data were analyzed statistically using 3-way ANOVA, Friedman’s and Wilcoxon’s tests at a significance level of α=0.05. Results The results showed that brand type affected significantly L*, a* and b* coordinates (p<0.0001), illumination a* and b* coordinates (p<0.0001), but none of them was affected by the hydration state of teeth (p>0.05). Intra-brand color differences ranged between 0.21-0.78ΔΕ* units with significant differences among brands (p<0.0001), among illumination lights (p<0.0001) and between hydration states (p=0.0001). Inter-brand differences ranged between 2.29-6.29ΔΕ* units with significant differences among pairs of brands (p<0.0001), illumination lights (p<0.0001) and hydration states (p<0.0001). Conclusions Differences were found between and within brands under D65 illumination which increased under F2 or A illumination affected by brand type and hydration status. Executive was the most stable brand than the others under different illuminations or wet states and for this reason its difference from other brands is the lowest. In clinical practice, there should be no blending of teeth of different brands but if we must, we should select those that are more stable under different illuminations

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Abstract Purpose To evaluate the kinetics of apical periodontitis development in vivo , induced either by contamination of the root canals by microorganisms from the oral cavity or by inoculation of bacterial lipopolysaccharide (LPS) and the regulation of major enzymes and receptors involved in the arachidonic acid metabolism. Methodology Apical periodontitis was induced in C57BL6 mice (n=96), by root canal exposure to oral cavity (n=48 teeth) or inoculation of LPS (10 µL of a suspension of 0.1 µg/µL) from E. coli into the root canals (n= 48 teeth). Healthy teeth were used as control (n=48 teeth). After 7, 14, 21 and 28 days the animals were euthanized and tissues removed for histopathological and qRT-PCR analyses. Histological analysis data were analyzed using two-way ANOVA followed by Sidak’s test, and qRT-PCR data using two-way ANOVA followed by Tukey’s test (α=0.05). Results Contamination by microorganisms led to the development of apical periodontitis, characterized by the recruitment of inflammatory cells and bone tissue resorption, whereas inoculation of LPS induced inflammatory cells recruitment without bone resorption. Both stimuli induced mRNA expression for cyclooxygenase-2 and 5-lipoxygenase enzymes. Expression of prostaglandin E 2 and leukotriene B 4 cell surface receptors were more stimulated by LPS. Regarding nuclear peroxisome proliferator-activated receptors (PPAR), oral contamination induced the synthesis of mRNA for PPARδ, differently from inoculation of LPS, that induced PPARα and PPARγ expression. Conclusions Contamination of the root canals by microorganisms from oral cavity induced the development of apical periodontitis differently than by inoculation with LPS, characterized by less bone loss than the first model. Regardless of the model used, it was found a local increase in the synthesis of mRNA for the enzymes 5-lipoxygenase and cyclooxygenase-2 of the arachidonic acid metabolism, as well as in the surface and nuclear receptors for the lipid mediators prostaglandin E2 and leukotriene B4.

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Abstract Objective Obesity is a chronic disease that negatively affects an individual’s general and oral health. The present study aimed to compare the clinical and microbiological effects of non-surgical periodontal therapy with the full mouth disinfection (FMD) protocol on obese and non-obese individuals at 9 months post-therapy. Methodology This clinical study was first submitted and approved by the Ethics Committee. Fifty-five obese patients and 39 non-obese patients with periodontitis were evaluated. The full-mouth periodontal clinical parameters, clinical attachment level (CAL), probing depth (PD), gingival index (GI), and plaque index (PI), were monitored at baseline, 3, 6, and 9 months after periodontal treatment with full mouth disinfection (FMD) protocol. The mean count of Tannerella forsythia , Porphyromonas gingivalis , Treponema Denticola , and Aggregatibacter actinomycetemcomitans was determined by quantitative polymerase chain reaction on subgingival biofilm samples. Demographic data were assessed by Chi-square test. For clinical and microbiological parameters, two-factor repeated-measures ANOVA was used. Results In both groups, periodontal therapy using the one-stage full-mouth disinfection protocol significantly improved CAL, PD, GI, and PI (p<0.05). Obese and non-obese patients equally responded to non-surgical periodontal therapy (p>0.05). Microbial count found no major differences (p>0.05) between obese and non-obese individuals who had undergone non-surgical periodontal therapy. Conclusions Obesity did not affect the clinical and microbiological outcomes of non-surgical periodontal therapy.

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Abstract Objective This study evaluated the clinical effect of violet LED light on in-office bleaching used alone or combined with 37% carbamide peroxide (CP) or 35% hydrogen peroxide (HP). Methodology A total of 100 patients were divided into five groups (n=20): LED, LED/CP, CP, LED/HP and HP. Colorimetric evaluation was performed using a spectrophotometer (ΔE, ΔL, Δa, Δb) and a visual shade guide (ΔSGU). Calcium (Ca)/phosphorous (P) ratio was quantified in the enamel microbiopsies. Measurements were performed at baseline (T 0 ), after bleaching (T B ) and in the 14-day follow-up (T 14 ). At each bleaching session, a visual scale determined the absolute risk (AR) and intensity of tooth sensitivity (TS). Data were evaluated by one-way (ΔE, Δa, ΔL, Δb), two-way repeated measures ANOVA (Ca/P ratio), and Tukey post-hoc tests. ΔSGU and TS were evaluated by Kruskal-Wallis and Mann-Whitney, and AR by Chi-Squared tests (a=5%). Results LED produced the lowest ΔE (p<0.05), but LED/HP promoted greater ΔE, ΔSGU and Δb (T 14 ) than HP (p<0.05). No differences were observed in ΔE and ΔSGU for LED/CP and HP groups (p>0.05). ΔL and Δa were not influenced by LED activation. After bleaching, LED/CP exhibited greater Δb than CP (p>0.05), but no differences were found between these groups at T 14 (p>0.05). LED treatment promoted the lowest risk of TS (16%), while HP promoted the highest (94.4%) (p<0.05). No statistical differences of risk of TS were found for CP (44%), LED/CP (61%) and LED/HP (88%) groups (p>0.05). No differences were found in enamel Ca/P ratio among treatments, regardless of evaluation times. Conclusions Violet LED alone produced the lowest bleaching effect, but enhanced HP bleaching results. Patients treated with LED/CP reached the same efficacy of HP, with reduced risk and intensity of tooth sensitivity and none of the bleaching protocols adversely affected enamel mineral content.

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Abstract Objectives This study evaluated if the use of a bioactive glass-ceramic-based gel, named Biosilicate (BS), before, after or mixed with bleaching gel, could influence the inflammation of the dental pulp tissue of rats’ molars undergoing dental bleaching with hydrogen peroxide (H2O2). Methodology The upper molars of Wistar rats (Rattus norvegicus, albinus) were divided into Ble: bleached (35% H2O2, 30-min); Ble-BS: bleached and followed by BS-based gel application (20 min); BS-Ble: BS-based gel application and then bleaching; BS/7d-Ble: BS-based gel applications for 7 days and then bleaching; Ble+BS: blend of H2O2 with BS-based gel (1:1, 30-min); and control: placebo gel. After 2 and 30 days (n=10), the rats were euthanized for histological evaluation. The Kruskal-Wallis and Dunn statistical tests were performed (P<0.05). Results At 2 days, the Ble and Ble-BS groups had significant alterations in the pulp tissue, with an area of necrosis. The groups with the application of BS-based gel before H2O2 had moderate inflammation and partial disorganization in the occlusal third of the coronary pulp and were significantly different from the Ble in the middle and cervical thirds (P<0.05). The most favorable results were observed in the Ble+BS, which was similar to the control in all thirds of the coronary pulp (P>0.05). At 30 days, the pulp tissue was organized and the bleached groups presented tertiary dentin deposition. The Ble group had the highest deposition of tertiary dentin, followed by the Ble-BS, and both were different from control (P<0.05). Conclusion A single BS-based gel application beforehand or BS-based gel blended with a bleaching gel minimize the pulp damage induced by dental bleaching.

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Abstract The study of dental development in individuals born with cleft lip and palate (CLP) serves to determine when orthodontic intervention should start. Objective To evaluate the permanent second molar development in children born with cleft lip and palate according to Demirjian’s and Nolla’s methods. Methodology Out of a total of 513 digital panoramic radiographs, 113 pairs of children aged 3 to 16 years were selected. The exams were from children born with or without cleft lip and palate, of the same sex, with an age difference of up to 30 days. The images were analyzed by three examiners and reliability was checked through intra-examiner agreement by the Kappa test. The data were analyzed by Wilcoxon's and Mann-Whitney tests according to each dataset. Results The findings indicated delayed development of the permanent second molars in children with CLP (P<0.001). The development of the right permanent second molar was delayed compared to the left molar in children with CLP. Moreover, mandibular teeth showed significantly earlier development than maxillary teeth in both the case and control groups. There was no significant difference in the development of permanent second molars between sexes. Conclusion Children with CLP presented delay in the development of permanent second molars.

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Abstract Laboratory tests are routinely used to test bonding properties of dental adhesives. Various aging methods that simulate the oral environment are used to complement these tests for assessment of adhesive bond durability. However, most of these methods challenge hydrolytic and mechanical stability of the adhesive- enamel/dentin interface, and not the biostability of dental adhesives. Objective To compare resin-dentin microtensile bond strength (μTBS) after a 15-day Streptococcus mutans (SM) or Streptococcus sobrinus (SS) bacterial exposure to the 6-month water storage (WS) ISO 11405 type 3 test. Methodology A total of 31 molars were flattened and their exposed dentin was restored with Optibond-FL adhesive system and Z-100 dental composite. Each restored molar was sectioned and trimmed into four dumbbell-shaped specimens, and randomly distributed based on the following aging conditions: A) 6 months of WS (n=31), B) 5.5 months of WS + 15 days of a SM-biofilm challenge (n=31), C) 15 days of a SM-biofilm challenge (n=31) and D) 15 days of a SS-biofilm challenge (n=31). μTBS were determined and the failure modes were classified using light microscopy. Results Statistical analyses showed that each type of aging condition affected μTBS (p<0.0001). For Group A (49.7±15.5MPa), the mean μTBS was significantly greater than in Groups B (19.3±6.3MPa), C (19.9±5.9MPa) and D (23.6±7.9MPa). For Group D, the mean μTBS was also significantly greater than for Groups B and C, but no difference was observed between Groups B and C. Conclusion A Streptococcus mutans- or Streptococcus sobrinus-based biofilm challenge for 15 days resulted in a significantly lower μTBS than did the ISO 11405 recommended 6 months of water storage. This type of biofilm-based aging model seems to be a practical method for testing biostability of resin-dentin bonding.

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Abstract Objective To clinically assess the effect of desensitizing gels and dentifrices on the reduction in pain sensitivity and color variation during tooth bleaching. Methodology A total of 108 volunteers were randomly separated into the following groups of n=12: GT/S-glycerine and thickener/sucralose; NF/S-potassium nitrate and sodium fluoride/sucralose; NA/S-potassium nitrate and arginine/sucralose; GT/AC-glycerine and thickener/arginine and calcium carbonate; NF/AC-potassium nitrate and sodium fluoride/arginine and calcium carbonate; NA/AC-potassium nitrate and arginine/arginine and calcium carbonate; GT/PN-glycerine and thickener/potassium nitrate; NF/PN-potassium nitrate and sodium fluoride/potassium nitrate; and NA/PN-potassium nitrate and arginine/potassium nitrate. Sensitivity was assessed with the numerical analogue scale, and color variation (ΔE) was measured with a spectrophotometer. The sensitivity values obtained were subjected to a multivariate analysis of variance (MANOVA) and color variation values were subjected to a randomized analysis of variance (p<0.05). Results The NF/AC, NA/AC, NF/PN, and NA/PN groups presented lower sensitivity values and reduced sensitivity compared to those of the other groups throughout the clinical sessions. None of the groups showed sensitivity at the 24-week assessment. Statistically, no significant difference were observed in the color values among the groups four weeks after the beginning of bleaching (p=0.074). Additionally, the color assessment of all groups was statistically similar four weeks (p=0.084) and 24 weeks (p=0.118) after the beginning. Conclusion Our results indicate that adding NF/S, NA/S, NF/AC, and NA/AC desensitizers to tooth bleaching protocols reduces pain sensitivity without affecting its effectiveness.

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Abstract Titanium dioxide nanotubes are nanostructures that can accelerate the oxidation reaction of bleaching procedures and promote a more effective whitening effect. Objective This study evaluated physicochemical properties of bleaching agents incorporated with titanium dioxide (TiO2) nanotubes, and the effects on tooth color change at different periods. Methodology 40 premolars were treated according to the following groups (n=10): CP - 10% carbamide peroxide (1 hour daily/21 days); CPN - CP incorporated into TiO2; HP - 40% hydrogen peroxide (three 40-minute sessions/7 days apart); HPN - HP incorporated into TiO2. Color shade was evaluated at five different periods (baseline, after 7, 14 and 21 days of bleaching, and 7 days after end of treatment) according to Vita Classical, CIELab and CIEDE2000 scales. Mean particle size (P), polydispersity (PO) and zeta potential (ZP) were evaluated using dynamic light scattering. Data on the different variables were analyzed by mixed model tests for measures repeated in time (ZP e L*), generalized linear models for measures repeated in time (P, PO, Vita Classical and b*), and Friedman and Mann-Whitney tests (a* and color change/ΔE and ΔE00). Results CP and CPN presented higher P, higher PO and lower ZP than HP and HPN (p≤0.05). All groups showed a significant decrease in Vita Classical color scores after 7 days of bleaching (p<0.05), and HPN presented a greater significant reduction than the other groups. L* increased in TiO2 presence, in all groups, without any differences (p>0.05) in bleaching time. A significant reduction occurred in the a* and b* values for all the groups, and HPN presented lower a* and b* values (p<0.05) than CPN. ΔE was clinically noticeable after 7 days, in all groups, and all groups resulted in a perceptible color change according to ΔE00. Conclusion TiO2 did not influence physicochemical properties of the bleaching agents. HPN presented more effective tooth bleaching than CPN.

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Abstract Objectives To compare the microtensile bond strength (µTBS) and depth of cure (DOC) of bulk-fill composites cured by monowave (MW) and polywave (PW) LED units using different curing times. Methodology Three composites were tested: Tetric EvoCeram Bulk Fill (TBF), Filtek Bulk Fill (FBF), and Tetric EvoCeram (T; control). Flat dentin surfaces treated with adhesive (AdheSE Universal®, Ivoclar Vivadent) were bonded with 4 mm cylindrical samples of each bulk-fill composite material (n=6) and cured with monowave (Satelec) or polywave (Bluephase Style) curing units for 10 or 20 seconds. After 24 hours, teeth were sectioned into individual 0.9 mm2 beams and tested for µTBS. Failure modes were analysed. Moreover, the DOC scrape test (IOS 4090) was completed (n=5) following the same curing protocols. Two-way ANOVA (a=0.05) was performed, isolating light-curing units. Results For samples cured with the MW light-curing unit, no significant effects were observed in the µTBS results between any of the resin composite brands and the curing times. Conversely, when resins were cured with a PW light unit, a significant effect was observed for TBF resin. In general, bulk-fill composites presented greater DOC and longer curing time resulted in higher DOC for all composites. Conclusion The µTBS of the composites to dentin was not affected by the curing mode of the resins, except for TBF cured with PW light unit. Bulk-fill composites exhibit greater DOC than conventional resin-based composites.

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Abstract Objective To compare two corticotomy surgical protocols in rats to verify whether they alter conventional orthodontic movement. Methodology Sixty Wistar rats were divided into three groups – orthodontic movement (CG), orthodontic movement and corticotomy (G1) and orthodontic movement with corticotomy and decortication (G2) – and euthanized after 7 and 14 days. Tooth movement (mm), bone volume fraction and bone volume ratio to total volume (BV/TV), and bone mineral density (BMD) were evaluated by micro-CT. The total amount of bone was measured in square millimeters and expressed as the percentage of bone area in the histomorphometry. The number of positive TRAP cells and RANK/RANKL/OPG interaction were also investigated. Results Day 14 showed a statistically significant difference in orthodontic tooth movement in CG compared with G1 (7.52 mm; p=0.009) and G2 (7.36 mm; p=0.016). A micro-CT analysis revealed a difference between CG, G1 and G2 regarding BV/TV, with G1 and G2 presenting a lower BV/TV ratio at 14 days (0.77 and 0.73 respectively); we found no statistically significant differences regarding BMD. There was a difference in the total amount of bone in the CG group between 7 and 14 days. At 14 days, CG presented a significantly higher bone percentage than G1 and G2. Regarding TRAP, G2 had more positive cells at 7 and 14 days compared with CG and G1. Conclusion Corticotomy accelerates orthodontic movement. Decortication does not improve corticotomy efficiency.

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Abstract Adding a biological apatite layer to the implant surface enhances bone healing around the implant. Objective This study aimed to characterize the mechanical properties and test human gingival fibroblasts behavior in contact with Zirconia and Titanium bioactive-modified implant materials. Methodology 6 groups were considered: Titanium (Ti6Al4V), Ti6Al4V with 5% HA and 5% ßTCP, Zirconia (YTZP), YTZP with 5% HA and 5% ßTCP. For each group, we produced discs using a novel fabrication method for functionally graded materials, under adequate conditions for etching and grit-blasting to achieve equivalent surface microroughness among the samples. Surface roughness (Ra, Rz), water contact angle, shear bond strength, and Vickers hardness were performed. Human gingival fibroblasts immortalized by hTERT gene from the fourth passage, were seeded on discs for 14 days. Cell viability and proliferation were assessed using a resazurin-based method, and cellular adhesion and morphology using field emission gun scanning electron microscopy (FEG-SEM). After 3 days of culture, images of fluorescent nucleic acid stain were collected by confocal laser scanning microscopy (CLSM). Results Results were presented as mean ± standard deviation (SD). We compared groups using one-way ANOVA with Tukey’s post-hoc test, and significance level was set at p<0.05. After 14 days of culture, cell viability and proliferation were significantly higher in YTZP group than in other groups (p<0.05). Samples of YTZP-ßTCP presented significantly higher wettability (p<0.05); yet, we observed no improvement in cell behavior on this group. Fibroblast spreading and surface density were more evident on YTZP specimens. Adding calcium-phosphate bioactive did not alter the tested mechanical properties; however, Ti6Al4V material shear bond strength was statistically higher than other groups (p<0.05). Conclusion Adding bioactive materials did not improve soft-tissue cell behavior. When compared to other zirconia and titanium groups, pure zirconia surface improved adhesion, viability and proliferation of fibroblasts. Cell behavior seems to depend on surface chemical composition rather than on surface roughness.

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Abstract Objective Our study seeks to investigate the effectiveness of kinesio taping (KT) on postoperative morbidity compared to placebo and control groups after impacted third molar surgery. Methodology Sixty patients with impacted mandibular third molar were included in this prospective, randomized, placebo-controlled clinical study. After surgical extraction of the impacted tooth, patients were allocated into three groups (20 patients each): group 1 received KT (kinesio), group 2 received placebo taping (placebo), and group 3 received no taping (control). The groups were compared regarding facial swelling, pain and trismus. Swelling was evaluated using a tape measuring method. Pain was assessed by a visual analog scale and the number of analgesic tablets taken. Trismus was determined by measuring maximum mouth opening. Results In the KT group, all parameters reduced significantly on 2nd and 4th postoperative days compared to other groups; however, placebo and control groups revealed comparable outcomes. On 7th day, all groups showed comparable results. Conclusions The KT application is an effective method for reducing morbidity after impacted mandibular third molar surgery. However, placebo taping is not as effective as proper taping. Placebo taping shows similar results compared to no taping regarding facial swelling percentage, pain and trismus.

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Abstract Objective There is increasingly common the consumption more times a day of foods and acidic drinks in the diet of the population. The present study aimed to evaluate and compare the effects of a calcium mesoporous silica nanoparticle single application of other calcium and/or fluoride products in reducing the progression of dental erosion. Methodology Half of the eroded area was covered of 60 blocks of enamel, after which the block was submitted to the following treatments: (Ca2+-MSN), casein phosphopeptide–amorphous calcium phosphate (CPP-ACP); CPP-ACP/F-(900 ppm F−); titanium tetrafluoride (TiF4 1%) (positive control); sodium fluoride (NaF 1.36%) (positive control); and Milli-Q® water (negative control) before being submitted to a second erosive challenge. A surface analysis was performed via a three-dimensional (3D) noncontact optical profilometry to assess the volumetric roughness (Sa) and tooth structure loss (TSL) and and through scanning electron microscopy (MEV). An analysis of variance (ANOVA) and Tukey’s test were performed. Results Regarding Sa, all experimental groups exhibited less roughness than the control (p<0.05). The TSL analysis revealed that the Ca2+-MSN and NaF groups were similar (p>0.05) and more effective in minimizing tooth loss compared with the other groups (p<0.05). Conclusions The Ca2+-MSN and NaF treatments were superior compared with the others and the negative control.

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Abstract Treatment of temporomandibular disorders (TMD) is a challenge for health care professionals. Therefore, new approaches have been investigated, such as the use of natural products. Objective This systematic review aims to summarize the natural products used in treatment of experimental models of TMD. Methodology A systematic search was performed in the databases Medline, Web of Science, Scopus, Embase, SciELO, LILACS, and Scholar Google databases in January 2020, dating from their inception. Pre-clinical studies with natural products for intervention in experimental TMD were included. Two reviewers independently selected the studies, extracted the data, and evaluated the risk of bias. Results 17 records were selected, and 17 different natural products were found, including three lectins, three plants or algae extracts, three sulfated polysaccharides, three cocoa preparations, and five isolated compounds. Concerning the risk of bias, most studies lacked on randomization and blinding. Nociception induced by phlogistic agents was evaluated in most articles, and in five studies it was associated with analysis of inflammatory parameters. In order to investigate the mechanism of action of the natural products used, eight studies evaluated expression of neural or glial molecular markers. Conclusions 16 of 17 natural products found in this review presented positive results, showing their potential for treatment of TMD. However, the lack of methodological clarity can influence these results.

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Abstract Aim To evaluate the cytotoxicity, biocompatibility and mineralization capacity of BIO-C PULPO, and MTA. Methodology L929 fibroblasts were cultured and MTT assay was used to determine the material cytotoxicity on 6, 24, and 48 h. A total of 30 male rats (Wistar) aged between 4 and 6 months, weighing between 250 and 300 g were used. Polyethylene tubes containing BIO-C PULPO, MTA, and empty tubes were implanted into dorsal connective tissue. After the experimental periods (7, 15, 30, 60, and 90 days) the tubes were histologically analyzed using hematoxylin-eosin (H&E), immunolabeling of IL-1β and TNF-α, and von Kossa staining, or without staining for polarized light analysis. The average number of inflammatory cells was quantified; the mineralization assessment was determined by the area marked in μm2 and semiquantitative immunolabeling analyses of IL-1β and TNF-α were performed. Then, data underwent statistical analysis with a 5% significance level. Results It was observed that BIO-C PULPO and MTA presented cytocompatibility at 6, 24, and 48 similar or higher than control for all evaluated period. On periods 7 and 15 days, BIO-C PULPO was the material with the highest number of inflammatory cells (p<0.05). On periods 30, 60, and 90 days, BIO-C PULPO and MTA presented similar inflammatory reactions (p>0.05). No statistical differences were found between Control, BIO-C PULPO, and MTA for immunolabeling of IL-1β and TNF-α in the different periods of analysis (p<0.05). Positive von Kossa staining and birefringent structures under polarized light were observed in all analyzed periods in contact with both materials, but larger mineralization area was found with BIO-C PULPO on day 90 (p<0.05). Conclusion BIO-C PULPO was biocompatible and induced mineralization similar to MTA.

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Abstract Saliva is the major contributor for the protein composition of the acquired enamel pellicle (AEP), a bacteria-free organic layer formed by the selective adsorption of salivary proteins on the surface of the enamel. However, the amount of proteins that can be recovered is even smaller under in vitro condition, due to the absence of continuous salivary flow. Objective This study developed an in vitro AEP protocol for proteomics analysis using a new formation technique with different collection solutions. Methodology 432 bovine enamel specimens were prepared (4x4 mm) and divided into four groups (n=108). Unstimulated saliva was provided by nine subjects. The new AEP formation technique was based on saliva resupply by a new one every 30 min within 120 minutes at 37ºC under agitation. AEP was collected using an electrode filter paper soaked in the collection solutions according with the group: 1) 3% citric acid (CA); 2) 0.5% sodium dodecyl sulfate (SDS); 3) CA followed by SDS (CA+SDS); 4) SDS followed by CA (SDS+CA). The pellicles collected were processed for analysis through LC-ESI-MS/MS technique. Results A total of 55 proteins were identified. The total numbers of proteins identified in each group were 40, 21, 28 and 41 for the groups CA, SDS, CA+SDS and SDS+CA, respectively. Twenty-three typical AEP proteins were identified in all groups, but Mucin was only found in CA and CA+SDS, while three types of PRP were not found in the SDS group. Moreover, a typical enamel protein, Enamelin, was identified in the CA+SDS group only. Conclusion The new technique of the in vitro AEP formation through saliva replacement was essential for a higher number of the proteins identified. In addition, considering practicality, quantity and quality of identified proteins, citric acid seems to be the best solution to be used for collection of AEP proteins.

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Abstract Tricalcium silicate-based cement are materials used in reparative and regenerative procedures in endodontics. A recently proposed formulation aimed to enhance handling during clinical use with a versatile material applicable by syringe. Although, the use of bismuth oxide as radiopacifier and grey raw powder are drawbacks considering aesthetics. Objectives Evaluate physicochemical, biological, and antimicrobial properties of Grey-MTAFlow (Ultradent) and assess whether the addition of zinc oxide (ZnO) prevents dentinal discoloration caused by bismuth oxide. Methodology Grey-MTAFlow was manipulated in 'thin' consistency for all tests. Luminosity, color change, ion migration to dentine, radiopacity, setting time, ISO 6876:2012 linear flow, volumetric lateral flow and central filling of simulated grooves scanned using micro-computed tomography (μCT), pH, calcium release, volumetric change using μCT, chemical characterisation, cytotoxicity, and antimicrobial activity were assessed. Addition of 5% ZnO to Grey-MTAFlow and a bismuth-containing experimental composition were comparatively tested. Statistical analyses used Shapiro-Wilk, T-test, ANOVA, and Kolmogorov-Smirnov (p<0.05). Results The addition of ZnO to Grey-MTAFlow prevented dentine darkening after 90 days due to bismuth migration reduction, although no statistical difference was found (p=0.863). ZnO addition significantly enhanced Grey-MTAFlow radiopacity without differences in initial setting time. Grey-MTAFlow presented an ISO linear flow of 10.9 mm and a balanced volumetric lateral flow with central filling in μCT evaluation. All compositions presented an alkaline pH after immersion. Grey-MTAFlow had a significantly higher calcium ion release after 28 days in comparison to 24 hours (p=0.011) and volumetric expansion of 0.4±1.8% after immersion. ZnO addition altered the hydrated cement matrix once calcium hydroxide (portlandite) could not be detected in characterisation. Neither of the materials produced inhibition halos nor reduced bacterial turbidity, but all presented cytocompatibility above 100%. Conclusion Grey-MTAFlow expanded after immersion and exhibited higher luminosity values after the evaluation period when ZnO was added, but chemical modifications after this addition occurred.

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Abstract The pursuit for quality of life urged a better understanding of aspects involved in ageing to minimize its consequences. Although many studies investigated older adults’ voice, aspects affecting this population voice-related quality of life have not yet been explored. Objective To investigate how aerodynamics and vocal aspects are associated with voice-related quality of life in older adults. Methodology fifty-six older adults aged 60 years or above – 39 women and 17 men – were evaluated. The following procedures were performed: application of the Voice-Related Quality of Life (V-RQOL) protocol; vocal assessment, including auditory-perceptual and acoustic analysis, from which we obtained fundamental frequency (F 0 ), standard deviation of fundamental frequency (SDF 0 ), shimmer, amplitude perturbation quotient (APQ), jitter, pitch period perturbation quotient (PPQ), and harmonics to noise ratio (HNR); aerodynamic assessment using a spirometer; and maximum phonation time (MPT) for /a/, /s/, /z/ and number counting. Results older adults tend to present high V-RQOL scores. Among women, roughness, APQ, and HNR parameters were negatively correlated with V-RQOL, whereas F 0 was positively. We found no correlation between spirometry measurements and V-RQOL. MPT for /a/, /z/, and number counting was positively correlated with V-RQOL solely among men. Conclusion Vocal roughness and acoustic parameters have a negative impact on the quality of life of older women. Respiratory aspects related to the available air support for speaking affected the most the voice-related quality of life of older men.

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Abstract The increased consumption of citrus sweets can contribute to the development of erosive tooth wear (ETW). Objective This in vitro study evaluated the erosive potential of citrus sweets on bovine enamel samples regarding the quantification of wear. Methodology Ninety bovine crowns were prepared and samples were randomly distributed into 6 groups (n=15): 0.1% citric acid solution (pH 2.5); Coca-Cola ® Soft Drink (pH 2.6); Fini ® Diet (lactic and citric acid, pH 3.3); Fini ® Jelly Kisses (lactic and citric acid, pH 3.5); Fini ® Fruit Salad Bubblegum (maleic acid, pH 2.6); Fini ® Regaliz Acid Tubes (maleic and citric acid, pH 3.1). Sweets were dissolved in the proportion of 40 g/250 mL of deionized water. Enamel samples were submitted to erosive challenges for 7 days (4 daily acid immersion cycles for 90 s each). Enamel wear was measured using contact profilometry (μm), and data (median values [interquartile range]) were submitted to Kruskal-Wallis/Dunn’s test (p<0.0001). Results All citrus sweets tested present a high erosive potential, Fini Diet ® (2.4 [1.2]) and Fini Regaliz Tubs ® (2.2 [0.5]) show the highest erosive potential, similar to 0.1% citric acid (2.3 [0.7]); Fini Regaliz Tubs ® is more erosive than Coca-Cola ® (1.4 [0.9]). Conclusion The evaluated citrus sweets have great erosive potential and play a key role in the development of ETW.

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Abstract Objectives This randomized, split-mouth, single-blinded trial assessed whether the use of reservoirs in at-home bleaching trays is equivalent to non-reservoir trays. Our choice of an equivalence trial was based on the expectation that a non-reservoir tray is sufficient to produce a color change. Secondary outcomes such as tooth sensitivity (TS) and gingival irritation (GI) were also assessed. Methodology Forty-six patients were selected with canines shade A2 or darker. In half of the patient’s arch, bleaching trays were made with reservoirs and the other half, without reservoirs. At-home bleaching was performed with carbamide peroxide (CP) 10% (3 h daily; 21 days). Color change was evaluated with a digital spectrophotometer (ΔE, ΔE00, and Whiteness Index) and shade guide units (ΔSGU) at baseline, during and one-month post-bleaching. TS and GI were assessed with a numeric scale (NRS) and a visual analog scale (VAS). Results After one month, the equivalence of reservoir and non-reservoir groups were observed in all color instruments (p>0.05). Fifteen and sixteen patients presented pain (absolute risk: 33% and 35%, 95%, confidence interval (CI) 21-46% and 23-49%) in the reservoir and non-reservoir side, respectively. The odds ratio for pain was 0.8 (95%CI 0.2-3.0) and the p-value was non-significant (p=1.0). TS intensity was similar between both groups in any of the pain scales (p>0.05). No difference in the GI was observed (p>0.05). Conclusions The protocol with reservoirs is equivalent in color change to the non-reservoir, although no superiority of the latter was observed in terms of reduced TS and GI with at-home 10% carbamide peroxide bleaching. Clinical Relevance The presence of reservoirs in a bleaching tray did not improve color change or affect tooth sensitivity and gingival irritation.

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Abstract Painful temporomandibular disorders (TMD) in children and adolescents may impact negatively the individual´s life. The presence of comorbidities associated with TMD tends to increase the persistence of pain and to facilitate its chronification. Objective To investigate the presence of other painful conditions and systemic diseases and their association with painful TMD. Methodology In this cross-sectional study, 690 adolescents aged between 12-14 years old were evaluated through questionnaires and clinical examinations. Results Painful TMD was found in 16.2% of the sample, with a significant association with bronchitis (OR= 2.5; p=0.003) and asthma (OR=3.1; p=0.013), reported by the parents/legal guardians of the participants. Adolescents with regional and widespread pain were 2.7 (95% CI: 1.65-4.55) and 3.6 (95% CI: 1.29-10.14) more likely to also present painful TMD. Painful TMD was associated with a higher number of body pain sites in the last 12 months (4.26 vs. 2.90; p<0.001), as well as a higher number of systemic diseases (1.48 vs. 1.18; p=0.048), when compared to adolescents without painful TMD. Conclusion The findings of this study point out the importance of considering the presence of comorbid conditions in the diagnosis and management of painful TMD in adolescents. A multidisciplinary approach would contribute to better control of painful TMD and decrease its chronification risk.

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Abstract Objectives To evaluate apoptotic levels of peripheral blood mononuclear cells (PBMCs) and apoptotic regulatory proteins (Bax and Bcl-2) in lymphocyte subsets of oral cancer (OC) patients and healthy controls (HC). Methodology The percentage of apoptotic cells and lymphocyte counts were measured in the first cohort using PBMCs obtained from 23 OC patients and 6 HC. In the second cohort, (OC, 33; HC, 13), the mean fluorescence intensity (MFI) of Bax and Bcl-2 in CD19+ B, CD4+ T, CD8+ T, and CD16+56+ natural killer (NK) cells was determined via flow cytometry. Results The percentage of apoptotic cells was higher in the PBMCs of OC patients than in HC patients, particularly in patients with stage IV cancer (p<0.05). However, lymphocyte counts were significantly lower in stage IV patients (p<0.05). NK CD19+ B and CD16+56+ cell counts were significantly lower in OC patients compared with HC patients (p<0.001 and p<0.01, respectively), but CD4+ T cells were interestingly significantly higher in OC patients (p<0.001). While Bax MFI was slightly higher, Bcl-2 MFI was significantly lower for all four lymphocyte subsets in OC samples, particularly in stage IV patients, when compared with HC. Consequently, Bax/Bcl-2 ratios showed an upward trend from HC to OC patients, particularly those in stage IV. We found similar trends in Bax and Bcl-2 MFI for tumor stage, tumor size, and lymph node involvement. Conclusions The increased lymphocyte apoptosis in stage IV OC patients may be related to higher Bax levels and lower Bcl-2 levels. The Bax/Bcl-2 ratio in lymphocytes may be useful to determine the prognosis of OC patients, and could be considered a mean for supportive treatment in the future.

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Abstract Mineralization-promoting peptides are attractive candidates for new remineralization systems. In previous studies, peptides have been applied as aqueous solutions, which is not a clinically relevant form. Objective This study aims to investigate the efficiency of a mineralization-promoting peptide, applied in varnish, on remineralizing artificial caries on primary teeth. Methodology 55 primary molars were collected. Specimens were immersed in a demineralizing solution for 7 days and then, divided into 7 groups: Baseline: No-remineralization, Placebo: Blank colophony, F: Colophony 5% fluoride, P: Colophony 10% peptide, P+F: Colophony 5% fluoride and 10% peptide, Embrace: Embrace™ varnish, Durashield: Durashield™ varnish. A mixture of 35% w/v colophony varnishes were prepared in ethanol and applied accordingly. Specimens were immersed in a remineralization solution for 4 weeks and it was evaluated using PLM and SEM. Lesion depth reduction was examined by one-way ANOVA. Results There was no significant difference in mean lesion depths between baseline (147.04 ± 10.18 µm) and placebo groups (139.73 ± 14.92 µm), between F (120.95 ± 12.23 µm) and Durashield (113.47 ± 14.36 µm) groups and between P (81.79 ± 23.15 µm) and Embrace (90.26 ± 17.72 µm) groups. Lesion depth for the P+F group (66.95±10.59 µm) was significantly higher compared to all other groups. All groups contained samples with subsurface demineralized regions. Number of subsurface demineralized regions were higher in fluoride-containing groups. Conclusions We conclude that the mineralization-promoting peptide (MPP3) is effective in this in vitro study and the peptide shows benefits over fluoride as it yields less subsurface demineralized regions.

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Abstract Objective This study aims to evaluate bone repair and the development of the medication related osteonecrosis of the jaw (MRONJ) associated with the use of zoledronic acid in Wistar rats. Methodology 48 male Wistar rats were divided into four groups: ZA, treated with intraperitoneal zoledronic acid, 0.6 mg/kg every 28 days, totaling five doses; control (C), treated with 0.9% sodium chloride; ZA-surgical (SZA) and C-surgical (SC), submitted to extraction of the right upper molars 45 days after the first application. Alveolar bone repair was evaluated by macroscopic and histological analysis. Protein expression evaluations were performed by qPCR. Results Macroscopic evaluation showed that 91.66% (11) of the animals in the SZA group and 41.66% (5) from the SC group presented solution of epithelium continuity (P<0.05). All animals in the SZA group and none in the SC group had bone sequestration. The area of osteonecrosis was higher in the SZA group than in the SC group (P<0.05). In molecular evaluation, the SZA group presented changes in the expression of markers for osteoclasts, with increased RANK and RANKL, and a decrease in OPG. Conclusion The results highlighted strong and evident interference of zoledronic acid in bone repair of the socket, causing osteonecrosis and delayed bone remodeling.

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Abstract Objective Pulp revascularization is an effective treatment for immature necrotic teeth. Calcium hydroxide has been used in pulp revascularization as an intracanal medication due to its antimicrobial action and the non-exhibition of crown discoloration and cytotoxicity for stem cells from the apical papilla. Our study aimed to investigate the clinical success and quantitative radiographic changes of root development in immature traumatized teeth using calcium hydroxide plus 2% chlorhexidine gel as intracanal medication. Methodology In this retrospective study, 16 patients were treated with a standardized pulp revascularization protocol. Calcium hydroxide and 2% chlorhexidine gel were manipulated in a 1:1 (v/v) ratio and inserted into root canals with Lentulo spirals (Dentsply Maillefer, Baillaigues, Switzerland). Patients were followed up for a period from 9 to 36 months for the evaluation of clinical and radiological data. Radiographic measurements of root length, root width, apical diameter, and MTA placement from the apex were quantified using software ImageJ. Wilcoxon test and t-test were used, according to nonparametric or parametric data, respectively, for changes over time in root length, root width, and apical diameter. Results Fifteen teeth survived during the follow-up period (93.75%) and met the criteria for clinical success. Although the changes seem to be very small in many cases, significant increases in the average root length (14.28%, p<0.0001), root width (8.12%, p=0.0196), and decrease in apical diameter (48.37%, p=0.0007) were observed. MTA placement from the apex and age at the time of treatment was not significantly associated with the quantitative radiographic outcomes. Conclusions Pulp revascularization in traumatized immature teeth treated with calcium hydroxide plus 2% chlorhexidine gel as intracanal medication had high success and survival rates, showing periodontal healing and resolution of signs and symptoms. However, concerning the continued root development, the outcomes can still be considered unpredictable.

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Abstract Objective: This study analyzed the effect of ionizing radiation on bone microarchitecture and biomechanical properties in the bone tissue surrounding a dental implant. Methodology: Twenty rabbits received three dental morse taper junction implants: one in the left tibia and two in the right tibia. The animals were randomized into two groups: the nonirradiated group (control group) and the irradiated group, which received 30 Gy in a single dose 2 weeks after the implant procedure. Four weeks after the implant procedure, the animals were sacrificed, and the implant/bone specimens were used for each experiment. The specimens (n=10) of the right tibia were examined by microcomputed tomography to measure the cortical volume (CtV, mm3), cortical thickness (CtTh, mm) and porosity (CtPo, %). The other specimens (n=10) were examined by dynamic indentation to measure the elastic modulus (E, GPa) and Vickers hardness (VHN, N/mm2) in the bone. The specimens of the left tibia (n=10) were subjected to pull-out tests to calculate the failure load (N), displacement (mm) up to the failure point and interface stiffness (N/mm). In the irradiated group, two measurements were performed: close, at 1 mm surrounding the implant surface, and distant, at 2.5 mm from the external limit of the first measurement. Data were analyzed using one-way ANOVA, Tukey’s test and Student’s t-test (α=0.05). Results: The irradiated bone closer to the implant surface had lower elastic modulus (E), Vickers hardness (VHN), Ct.Th, and Ct.V values and a higher Ct.Po value than the bone distant to the implant (P<0.04). The irradiated bone that was distant from the implant surface had lower E, VHN, and Ct.Th values and a higher Ct.Po value than the nonirradiated bone (P<0.04). The nonirradiated bone had higher failure loads, displacements and stiffness values than the irradiated bone (P<0.02). Conclusion: Ionizing radiation in dental implants resulted in negative effects on the microarchitecture and biomechanical properties of bone tissue, mainly near the surface of the implant.

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Abstract Objective: This study aimed to determine serum and salivary levels of neutrophil gelatinase-associated lipocalin (NGAL) and evaluate NGAL correlation with key anti-interleukin 10 (IL-10) and pro-inflammatory (IL-1β) cytokines in different severities of periodontal diseases. We also calculated the systemic inflammation using the periodontal inflamed surface area (PISA) to evaluate its correlation with NGAL in the study groups. Methodology: Eighty systemically healthy and non-smoking individuals were separated into four groups of 20: clinically healthy (Group 1), gingivitis (Group 2), stage I generalized periodontitis (Group 3, Grade A), and stage III generalized periodontitis (Group 4, Grade A). Sociodemographic characteristics and periodontal parameters were recorded, and PISA was calculated. The serum and salivary levels of interleukin (IL)-1β, IL-10, and NGAL were determined using the enzyme-linked immunosorbent assay (ELISA). Results: We observed a significant increase in serum and salivary NGAL levels from healthy to periodontitis groups (p=0.000). Group 2 presented significantly higher serum and salivary IL-10 levels and salivary IL-1β levels than Group 3 (p=0.000). Serum and salivary parameters (IL-1β, IL-10, and NGAL levels) were strongly positively correlated to periodontal parameters and PISA values (p=0.000). Groups 2 and 3 showed overlapping PISA values. Conclusion: The overlapping PISA values found in Groups 2 and 3 suggest that gingivitis might progress to a systemic inflammatory burden somewhat comparable to stage I periodontitis. This finding is supported by the higher serum and salivary cytokines/mediators levels in the gingivitis group than in stage I periodontitis group. Serum and salivary NGAL levels increased proportionally to disease severity and PISA. NGAL seems to play a role in the pathogenesis of periodontal disease, within the limitation of our study.

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Abstract Gingival conditions and tooth sensitivity of young patients with amelogenesis imperfecta lack in depth studies. This case-control study aimed to compare (1) the gingival inflammation, the presence of enamel defects, and tooth sensitivity in young patients with and without amelogenesis imperfecta and (2) to investigate if any difference exists between subtypes of amelogenesis imperfecta. Methodology We compared forty-two participants with amelogenesis imperfecta with forty-two controls matched for age, gender, and the number of examined sites. Based on interview, clinical examination, and intraoral photography, we collected data on periodontal conditions, enamel defects and the presence of tooth sensitivity. Comparison tests were performed to investigate if any difference existed between cases and controls; and among cases, between the different subtypes of amelogenesis imperfecta. We performed a post-hoc analysis for any significant difference observed. Results We observed more gingival inflammation, enamel defects and tooth sensitivity among cases (all p<0.05). Participants with hypocalcified amelogenesis imperfecta had more gingival inflammation, enamel defects, and tooth sensitivity than patients with the hypoplastic and hypomature subtypes (all p<0.05). After adjustment for dental plaque, gingival inflammation was associated with the presence of amelogenesis imperfecta (OR (95%CI) = 1.14 (1.05; 1.24). p<0.01). Conclusion Gingival inflammation, enamel defect and tooth sensitivity are more frequently observed among young patients with amelogenesis imperfecta, and more specifically among children with the hypocalcified subtype.

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Abstract Glaze application on monolithic zirconia (Y-TZP) can be a practical approach to improve the mechanical properties of this material. Objective Our study evaluated the effect of glazing side and mechanical cycling on the biaxial flexure strength (BFS) of a Y-TZP. Methodology Eighty sintered Y-TZP discs (Ø:12 mm; thickness: 1.2 mm - ISO 6872) were produced and randomly assigned into eight groups (n=10), according to the factors “glazing side” (control – no glazing; GT – glaze on tensile side; GC – glaze on compression side; GTC – glaze on both sides) and “mechanical aging” (non-aged and aged, A – mechanical cycling: 1.2×106, 84 N, 3 Hz, under water at 37°C). Specimens were subjected to BFS test (1 mm/min; 1,000 Kgf load cell) and fractured surfaces were analyzed by stereomicroscopy and SEM. Hsueh’s rigorous solutions were used to estimate the stress at failure of glazed specimens. Two-way ANOVA, Tukey’s test (5%), and Weibull analysis were performed. Results The “glazing side”, “mechanical aging” and the interaction of the factors were significant (p<0.05). Groups GC (1157.9±146.9 MPa), GT (1156.1±195.3 MPa), GTC (986.0±187.4 MPa) and GTC-A (1131.9±128.9 MPa) presented higher BFS than control groups (Tukey, 5%). Hsueh’s rigorous solutions showed that the maximum tensile stress was presented in the bottom of zirconia layer, at the zirconia/glaze interface. Weibull characteristic strength (σo) of the GC was higher than all groups (p<0.05), except to GT, GTC-A and GTC, which were similar among them. The fractography showed initiation of failures from zirconia the tensile side regardless of the side of glaze application and fatigue. Conclusion Glazing zirconia applied on both tensile and compression sides improves the flexural strength of Y-TZP, regardless the mechanical aging.

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Abstract Objective This study aimed to evaluate the effects of different toothpastes on the surface wear of enamel, dentin, composite resin (CR), and resin-modified glass ionomer cement (RMGIC), and to perform a topographic analysis of the surfaces, based on representative images generated by atomic force microscopy (AFM) after erosion-abrasion cycles. Methodology One hundred and forty bovine incisors were collected and divided into two groups: 72 enamel and 72 dentin blocks (4×4 mm). Half of the specimens were restored with CR (Filtek Z350 XT) and the other half with RMGIC (Fuji II LC). Then, samples were submitted to a demineralization cycle (5 days, 4×2 min/day, 1% citric acid, pH 3.2) and exposed to three different toothpastes (2×15 s/day): without fluoride (WF, n=12), sodium fluoride-based (NaF, n=12), and stannous fluoride-based (SnF2, n=12). Surface wear, as well as restoration interfaces wear, were investigated by profilometry of the dental substrates and restorative materials. All representative surfaces underwent AFM analysis. Data were analyzed by two-way analysis of variance and Tukey’s tests (α=0.05). Results NaF-based toothpaste caused the greater dentin surface wear (p<0.05). Toothpastes affected only enamel-restoration interfaces. AFM analysis showed precipitate formation in dentinal tubules caused by the use of fluoride toothpastes. Conclusions NaF-based toothpastes had no protective effect on enamel adjacent to CR and RMGIC against erosion-abrasion challenges, nor on dentin adjacent to RMGIC material. SnF2-based toothpastes caused more damage to interfaces between enamel and RMGIC.

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Abstract Proanthocyanidin has been shown to be efficient in inhibiting matrix metalloproteinases. Objective The aim of this in situ study was to evaluate the protective effect of Proanthocyanidin-based mouthrinses either with naturally acidic or with a neutral pH applied on dentin subjected to erosion. Methodology Eight volunteers wore one palatal device in two phases (7 days washout) with 16 samples per group (n=8). The groups under study were: First Phase/ G1 – 10% proanthocyanidin mouthrinse (pH 7.0, Experimental group 1 – Purified Grape Seeds Oligomeric Proanthocyanidins), G2 – 10% proanthocyanidin mouthrinse (pH 3.0, Experimental group 2 – Purified Grape Seeds Oligomeric Proanthocyanidins). Second Phase/ G3 – 0.12% chlorhexidine mouthrinse (pH 7.0, Positive control group), G4 – no previous treatment (Negative control group). Each device was subjected to 3 erosive cycles (5 minutes) per day for 5 days. Treatments with different mouthrinses were applied once after the second erosive challenge (5 minutes). Profilometry was used to quantify dentin loss (µm). Results Data were analyzed by repeated measures of ANOVA followed by Fisher’s test (p<0.05). G1 (1.17±0.69) and G3 (1.22±0.25) showed significantly lower wear values with no statistical difference between them. G2 (2.99±1.15) and G4 (2.29±1.13) presented higher wear values with no significant differences between them. Conclusion The 10% proanthocyanidin mouthrinse (pH 7.0) could be a good strategy to reduce dentin wear progression.

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Abstract The period of functional adaptation to a new conventional complete denture embraces many transitory issues, and this period is directly related to the rehabilitation success. Objective This clinical trial evaluated the influence of the height of mandibular ridge on the masticatory function of complete denture (CD) wearers during the adaptation period. Methodology A total of 28 individuals wearing new CDs (NR, n=14, normal mandibular ridges, 64±12.5 years, 9 female; RR, n=14, resorbed mandibular ridges, 69±6.8 years, 9 female) were assessed at 24 hours, 30 days, three months and six months after the insertion of the CDs for masticatory performance (MP, sieves method), satisfaction with CDs (questionnaire) and maximum occlusal bite force (MOBF) (gnatodynamometer). The classification of the mandibular ridges followed the Kapur index. Data of MP and MOBF were analyzed by two-way ANOVA and satisfaction with CDs was analyzed by Generalized Estimating Equations (GEE), α=.05. Results Participants with NR presented better masticatory performance (p=.000 - NR 30.25±9.93%, RR 12.41±7.17%), general satisfaction (p=.047), retention of mandibular denture (p=.001), chewing ability (p=.037), and comfort of wearing a mandibular denture (p=.000). Regardless of the mandibular ridge, MP (p=.000) was higher at three (21.26±12.07%) and six months (24.25±12.26%) in comparison to 24 hours (18.09±10.89%), the MOBF (p=.000) was higher at three months (78.50±6.49 N) compared to 24 hours (57.34±5.55 N) and 30 days (62.72±5.97 N), and the comfort of wearing a mandibular denture (p=.002) at three months (1.61 ± 0.07) was greater than 24 hours (1.29±0.10) and 30 days (1.36±10). Conclusions The study suggests that the participants with NR have higher MP and satisfaction with their CD, regardless of the follow-up period after the insertion of the new CD. After subjects received the CD, a period of 3 months was necessary for achieving better achievement MOBF, MP, and self-perceived comfort with the mandibular denture, regardless of the height of the mandibular ridge.

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Abstract Heterogeneous cell populations of osteo/cementoblastic (O/C) or fibroblastic phenotypes constitute the periodontal dental ligament (PDL). A better understanding of these PDL cell subpopulations is essential to propose regenerative approaches based on a sound biological rationale. Objective Our study aimed to clarify the differential transcriptome profile of PDL cells poised to differentiate into the O/C cell lineage. Methodology To characterize periodontal-derived cells with distinct differentiation capacities, single-cell-derived clones were isolated from adult human PDL progenitor cells and their potential to differentiate into osteo/cementoblastic (O/C) phenotype (C-O clones) or fibroblastic phenotype (C-F clones) was assessed in vitro. The transcriptome profile of the clonal cell lines in standard medium cultivation was evaluated using next-generation sequencing technology (RNA-seq). Over 230 differentially expressed genes (DEG) were identified, in which C-O clones showed a higher number of upregulated genes (193) and 42 downregulated genes. Results The upregulated genes were associated with the Cadherin and Wnt signaling pathways as well as annotated biological processes, including “anatomical structure development” and “cell adhesion.” Both transcriptome and RT-qPCR showed up-regulation of WNT2, WNT16, and WIF1 in C-O clones. Conclusions This comprehensive transcriptomic assessment of human PDL progenitor cells revealed that expression of transcripts related to the biological process “anatomical structure development,” Cadherin signaling, and Wnt signaling can identify PDL cells with a higher potential to commit to the O/C phenotype. A better understanding of these pathways and their function in O/C differentiation will help to improve protocols for periodontal regenerative therapies.

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Abstract Objective This randomized and clinical trial aimed to evaluate the performance of a new restorative Glass Ionomer Cement (GIC) for the restoration of non-carious cervical lesions (NCCLs) of patients with systemic diseases compared with a posterior resin composite after 12 months. Methodology 134 restorations were placed at 30 patients presenting systemic diseases by a single clinician. NCCLs were allocated to two groups according to restorative system used: a conventional restorative GIC [Fuji Bulk (GC, Tokyo Japan) (FB)] and a posterior resin composite [G-ænial Posterior (GC, Tokyo Japan) (GP)] used with a universal adhesive using etch&rinse mode. All restorative procedures were conducted according to manufacturer’s instructions. Restorations were scored regarding retention, marginal discoloration, marginal adaptation, secondary caries, surface texture, and post-operative sensitivity using modified United States Public Health Service (USPHS) criteria after 1 week (baseline), 6, and 12 months. Descriptive statistics were performed using chi-square tests. Cochran Q and Mc Nemar’s tests were used to detect differences over time. Results After 12 months, recall rate was 93% and the rates of cumulative retention failure for FB and GP were 4.9% and 1.6% respectively. Both groups presented similar alpha rates for marginal adaptation (FB 86.2%, GP 95.5%) and marginal discoloration (FB 93.8%, GP 97%) at 6-month recall, but FB restorations showed higher bravo scores than GP restorations for marginal adaptation and marginal discoloration after 12 months (p<0.05). Regarding surface texture, 2 FB restorations (3.1%) were scored as bravo after 6 months. All restorations were scored as alpha for secondary caries and postoperative sensitivity after 12 months. Conclusion Although the posterior resin composite demonstrated clinically higher alpha scores than the conventional GIC for marginal adaptation and discoloration, both materials successfully restored NCCLs at patients with systematic disease after a year. Clinical relevance Due to its acceptable clinical results, the tested conventional restorative GIC can be used for the restoration of NCCLs of patients with systemic diseases.

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Abstract This paper aims to evaluate the effect of different surface treatments on surface topography, wettability, and shear bond strength of resin cement to glass ceramic. Methodology: For SBS test, 32 blocks (7x7x2 mm) of lithium disilicate were obtained and randomly divided into eight groups (four blocks per group) according to each surface treatment (HF 20 s, 60 s, 120 s + silanization/S or Scotch Bond Universal/ SBU) and the Monobond Etch & Prime - MEP application followed or not by SBU. On each treated surface ceramic block, up to four dual-curing resin cement cylinders were prepared and light-cured for 40s (N=120/n=15). The specimens were thermocycled (10,000 cycles, 5-55°C, 30 s) and the SBS test (50KgF, 0.5 mm/min) was performed. Furthermore, failure analysis, wettability, AFM, and SEM were carried out. SBS data (MPa) were analyzed using Student's t-test, two-way ANOVA, Tukey's test (5%) and Weibull's analysis. Results: For HF experimental groups, two-way ANOVA presented the factors “etching time” and “bonding agent” as significant (p<0.05). After silane application, the HF groups presented similar bond strength. SBU application compromised the SBS, except for 120s etching time (HF120sS: 23.39ᵃ±6.48 MPa; HF120sSBU: 18.76ᵃ±8.81MPa). For MEP groups, SBU application did not significantly affect the results (p=0.41). The MEP group presented the highest Weibull modulus (4.08A) and they were statistically different exclusively from the HF20sSBU (0.58B). Conclusion: The HF 20s, 60s, 120 s followed by silane, promoted similar resin-bond strength to ceramic and the SBU application after HF or MEP did not increase the SBS.

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Abstract Objective: This study aims to replicate the phenotype of Ltbp1 knockout mice in zebrafish, and to address the function of LTBP1 in craniofacial development. Methods: Whole mount in situ hybridization (WISH) of ltbp1 was performed at critical periods of zebrafish craniofacial development to explore the spatial-temporal expression pattern. Furthermore, we generated morpholino based knockdown model of ltbp1 to study the craniofacial phenotype. Results: WISH of ltbp1 was mainly detected in the mandibular jaw region, brain trunk, and internal organs such as pancreas and gallbladder. And ltbp1 colocalized with both sox9a and ckma in mandibular region. Morpholino based knockdown of ltbp1 results in severe jaw malformation. Alcian blue staining revealed severe deformity of Meckel's cartilage along with the absence of ceratobranchial. Three-dimension measurements of ltbp1 morphants jaws showed decrease in both mandible length and width and increase in open mouth distance. Expression of cartilage marker sox9a and muscle marker ckma was decreased in ltbp1 morphants. Conclusions: Our experiments found that ltbp1 was expressed in zebrafish mandibular jaw cartilages and the surrounding muscles. The ltbp1 knockdown zebrafish exhibited phenotypes consistent with Ltbp1 knockout mice. And loss of ltbp1 function lead to significant mandibular jaw defects and affect both jaw cartilages and surrounding muscles.

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Abstract Objective: This study evaluated the effects of local vitamin C treatment on tissue advanced glycation end products (AGE), interleukin (IL)-6, 8-hydroxy-2-deoxyguanosine (8-OHdG), and matrix metalloproteinases (MMP)-8 in tissues; serum C-terminal telopeptide fragments (CTX); and alveolar bone loss (ABL) in rats. Methodology: 35 male Sprague Dawley rats were divided equally into five groups: 1) control (C), 2) experimental periodontitis (P), 3) experimental diabetes (D), 4) experimental diabetes and experimental periodontitis (D + P), and 5) experimental diabetes–experimental periodontitis–locally applied vitamin C (D + P + LvitC). Diabetes was induced in rats with alloxan monohydrate, after which periodontitis was induced by ligature placement in the right mandibular first molar teeth for 11 days. In the treatment group, vitamin C was administered locally three times with two-days interval after ligature removal. The animals were sacrificed, and the samples were analyzed histometrically and immunohistochemically. Results: CTX, 8-OHdG, and AGE values significantly decreased in the treatment group compared to the D + P group. IL-6 and MMP-8 values decreased in the treatment group compared to the D + P group, but this is not significant. ABL was significantly reduced by the local delivery of vitamin C. Conclusion: This study reveals that vitamin C treatment may be beneficial to reduce serum CTX and gingival MMP-8 levels, oxidative stress, inflammation, and AGE accumulation in periodontal tissue. Vitamin C may be an immunomodulator and antioxidant locally applied in the treatment of periodontitis to reduce the adverse effects of diabetes in periodontal tissues.

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Abstract Dual-cured buildup composites and simplified light-cured adhesive systems are mixed with a chemical activator to prevent the incompatibility between them. Objective: To evaluate microshear bond strength (μSBS) and nanoleakage (NL) of three universal adhesives used under buildup composites using different curing modes, at baseline and after 6-months (6m). Methodology: Dentin specimens of 55 molars were assigned to: Clearfil Universal Bond[CFU], Prime&Bond Elect[PBE] and One Coat 7 Universal[OCU]. All-Bond Universal[ABU] and Adper Scotchbond Multi-Purpose[SMP] were used as controls. CFU, PBE, and OCU were: light-cured [LC], dual-cured using a self-curing activator [DC], and self-cured, using a self-curing activator and waiting for 20 min [SC]. Upon the application of the adhesive, transparent matrices were filled with a dual-cured buildup composite and light cured, then tested in mSBS. For NL, the specimens were submersed in ammoniacal silver nitrate and sectioned to observe under the SEM. Three-way ANOVA and Tukey's test were applied (α=0.05). Results: OCU/LC-PBE/LC resulted in higher mean μSBS than ABU/LC. For SMP/DC higher mean μSBS were obtained than for both CFU/DC and OCU/DC (baseline). No universal adhesive was significantly affected by curing mode or storage time. CFU, PBE, and OCU did not undergo significant changes in any curing mode (p>0.05). NL (baseline) PBE/LC resulted in higher %NL compared to ABU/LC. SMP/DC resulted in higher %NL than CFU/DC-OCU/DC. CFU/LC/DC resulted in lower %NL than CFU/SC. PBE/SC resulted in lower %NL than PBE/DC. OCU/LC/SC showed lower %NL than OCU/DC. OCU showed significant lower %NL than CFU and PBE. All CFU groups, as well as OCU/SC, resulted in increased %NL at 6m when compared with baseline. Conclusion: For universal adhesives used in etch-and-rinse mode, self-cured activator and different curing modes did not influence μSBS. However, some interactions were observed for NL, but this influence was material-specific.

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Abstract An important factor affecting the biomechanical behavior of implant-supported reconstructions is the implant-abutment misfit. Objective: This study evaluated the misfit between Ti-Base abutments and implants by means of polyvinyl siloxane replica technique using microcomputed tomography (μCT). Methodology: Volumetric and linear (central and marginal) gaps of four Ti-base abutments (n=10/group): (i) Odontofix LTDA (OD), (ii) Singular Implants (SING), (iii) EFF Dental Components (EFF), and (iv) Control Group (S.I.N implants) compatible with an implant system (Strong SW, S.I.N Implants) were measured using μCT reconstructed polyvinyl siloxane replicas. Results: The results showed significantly lower volume gap for Control S.I.N (0.67±0.29 mm3) and SING (0.69±0.28 mm3) Ti-base abutments relative to OD (1.42±0.28 mm3) and EFF groups (1.04±0.28 mm3) (p<0.033), without significant difference between them (p=0.936). While gap values were homogenous in the central region, EFF presented a significantly higher marginal gap. Accordingly, the Control S.I.N and Singular Ti-base abutments showed improved volumetric and marginal fit relative to Odontofix and EFF. Conclusion: The method of manufacturing abutments influenced the misfit at the implant-abutment interface.

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Abstract Temporomandibular dysfunction (TMD), anxiety, and depression are disorders that, due to the current lifestyle, are affecting an increasing portion of the population. Investigating the prevalence of the symptoms of these disorders during the quarantine due to the coronavirus 2019 pandemic (COVID-19) is important to outline clinical strategies for patient care. Objective: This study assessed the prevalence of TMD symptoms, anxiety, depression, and oral behaviors and their associations during the social isolation due to COVID-19. Methodology: Questionnaires were used to assess TMD symptoms in accordance with the Diagnostic Criteria for Temporomandibular Disorders: clinical protocol and assessment instruments, a questionnaire to verify oral behaviors and Hospital Anxiety and Depression Scale to assess symptoms of anxiety and depression in students of dentistry at the Faculty of Health Sciences of the University of Brasília in May 2020. Qualitative data were subjected to descriptive statistics and chi-squared analysis (p<0.05). The relationship between quantitative and qualitative data was evaluated using Spearman's rho correlation (p<0.05). Results: There was a high prevalence of TMD symptoms, anxiety, and depression in the participants, resulting in association between gender and anxiety symptoms (p=0.029). There was a positive correlation between oral behaviors and TMD symptoms (r=0.364; p<0.001), between oral behaviors and anxiety symptoms (r=0.312; p=0.001), and between oral behaviors and symptoms of depression (r=0.216; p=0.021). Conclusion: Social isolation due to the COVID-19 pandemic has an impact on the prevalence of TMD symptoms, anxiety, and depression.

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Abstract Implant surface decontamination is a challenging procedure for therapy of peri-implant disease. Objective: This study aimed to compare the effectiveness of decontamination on oral biofilm-contaminated titanium surfaces in Er:YAG laser, Er, Cr:YSGG laser, and plastic curette. Methodology: For oral biofilms formation, six participants wore an acrylic splint with eight titanium discs in the maxillary arch for 72 hours. A total of 48 contaminated discs were distributed among four groups: untreated control; decontamination with plastic curettes; Er, Cr:YSGG laser; and Er:YAG laser irradiation. Complete plaque removal was estimated using naked-eye and the time taken was recorded; the residual plaque area was measured and the morphological alteration of the specimen surface was observed by scanning electron microscopy. The total bacterial load and the viability of adherent bacteria were quantified by live or dead cell labeling with fluorescence microscopy. Results: The mean treatment time significantly decreased based on the treatment used in the following order: Er:YAG, Er, Cr:YSGG laser, and plastic curettes (234.9±25.4 sec, 156.1±12.7 sec, and 126.4±18.6 sec, P=0.000). The mean RPA in the Er, Cr:YSGG laser group (7.0±2.5%) was lower than Er:YAG and plastic curettes groups (10.3±2.4%, 12.3±3.6%, p=0.023). The viable bacteria on the titanium surface after Er, Cr:YSGG laser irradiation was significantly lower compared to the decontamination with plastic curette (P=0.05) but it was not significantly different from the Er:YAG laser irradiation. Conclusion: We found that Er:YAG laser and Er, Cr:YSGG laser irradiation were effective methods for decontaminations without surface alterations.

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Abstract Objective This study aimed to clarify the association between oral human cytomegalovirus (HCMV) and periodontitis in Japanese adults. Methodology In total, 190 patients (75 men and 115 women; mean age, 70.2 years) who visited Hiroshima University Hospital between March 2018 and May 2020 were included. Oral rinse samples were taken to examine the presence of HCMV DNA using real-time polymerase chain reaction (PCR). P. gingivalis was detected by semi-quantitative PCR analysis. Results HCMV DNA was present in nine of 190 patients (4.7%). There were significant associations between HCMV presence and the presence of ≥4-mm-deep periodontal pockets with bleeding on probing (BOP) (P<0.01) and ≥6-mm-deep periodontal pockets with BOP (P=0.01). However, no significant relationship was observed between HCMV presence and periodontal epithelial surface area scores. Logistic regression analysis revealed that the presence of ≥4-mm-deep periodontal pockets with BOP was significantly associated with HCMV (odds ratio, 14.4; P=0.01). Propensity score matching was performed between patients presenting ≥4-mm-deep periodontal pockets with BOP (i.e., active periodontitis) and patients without ≥4-mm-deep periodontal pockets with BOP; 62 matched pairs were generated. Patients who had ≥4-mm-deep periodontal pockets with BOP showed a higher rate of HCMV presence (9.7%) than those who lacked ≥4-mm-deep periodontal pockets with BOP (0.0%). There was a significant relationship between HCMV presence and ≥4-mm-deep periodontal pockets with BOP (P=0.03). A significant relationship was found between HCMV/P. gingivalis DNA presence and ≥4-mm-deep periodontal pockets with BOP (P=0.03). Conclusions Coinfection of oral HCMV and P. gingivalis was significantly associated with active periodontitis. Moreover, interactions between oral HCMV and P. gingivalis may be related to the severity of periodontal disease.

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Abstract Objective To evaluate the surface morphology and in vitro leachability of temporary soft linings modified by the incorporation of antifungals in minimum inhibitory concentrations (MIC) for Candida albicans biofilm. Methodology Specimens of soft lining materials Softone and Trusoft were made without (control) or with the addition of nystatin (Ny), miconazole (Mc), ketoconazole (Ke), chlorhexidine diacetate (Chx), or itraconazole (It) at their MIC for C. albicans biofilm. The surface analyses were performed using Confocal laser scanning microscopy after 24 h, 7 days, or 14 days of immersion in distilled water at 37ºC. In vitro leachability of Chx or Ny from the modified materials was also measured using Ultraviolet visible spectroscopy for up to 14 days of immersion in distilled water at 37ºC. Data (μg/mL) were submitted to ANOVA 1-factor/Bonferroni (α=0.05). Results Softone had a more irregular surface than Trusoft. Morphological changes were noted in both materials with increasing immersion time, particularly, in those containing drugs. Groups containing Chx and It presented extremely porous and irregular surfaces. Both materials had biexponential release kinetics. Softone leached a higher concentration of the antifungals than Trusoft (p=0.004), and chlorhexidine was released at a higher concentration than nystatin (p<0.001). Conclusions The surface of the soft lining materials changed more significantly with the addition of Chx or It. Softone released a higher concentration of drugs than Trusoft did, guiding the future treatment of denture stomatitis.

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Abstract The evidence is inconclusive regarding the effect of periodontal treatment on glycemic control and systemic inflammation in patients with type 2 diabetes (T2D) and periodontitis Objective: To evaluate the effect of scaling and root planing (SRP) on the metabolic control and systemic inflammation of patients with type 2 diabetes (T2D). Methodology: A literature search was conducted using the MEDLINE database via PubMed and the Cochrane Central Register of Controlled Trials, from their oldest records up to July 2018. Only randomized clinical trials (RCT) were considered eligible for evaluating the effect of periodontal treatment on markers of metabolic control [glycated hemoglobin (HbA1C)] and systemic inflammation [C-reactive protein (CRP)] in patients with T2D. The quality of the studies was evaluated using the Cochrane Collaboration risk assessment tool. Meta-analyses were performed for HbA1c and CRP using random effects models. The size of the overall intervention effect was estimated by calculating the weighted average of the differences in means (DM) between the groups in each study. Heterogeneity was assessed using the Q-statistic method (x2 and I²). The level of significance was established at p<0.05. Results: Nine RCT were included. SRP was effective in reducing HbA1c [DM=0.56 (0.36-0.75); p<0.01] and CRP [DM=1.89 (1.70-2.08); p<0.01]. No heterogeneity was detected (I2=0%, p>0.05). Conclusions: SRP has an impact on metabolic control and reduction of systemic inflammation of patients with T2D.