Abstract Objective This study aimed to evaluate the cytotoxicity and synergistic effect of epigallocatechin gallate (EGCG) and fosfomycin (FOSFO) on biofilms of oral bacteria associated with endodontic infections. Methodology This study determined minimum inhibitory and bactericidal concentration (MIC/MBC) and fractionated inhibitory concentration (FIC) of EGCG and FOSFO against Enterococcus faecalis, Actinomyces israelii, Streptococcus mutans, and Fusobacterium nucleatum. Monospecies and multispecies biofilms with those bacteria formed in polystyrene microplates and in radicular dentin blocks of bovine teeth were treated with the compounds and control chlorhexidine (CHX) and evaluated by bacterial counts and microscopy analysis. Toxicity effect of the compounds was determined on fibroblasts culture by methyl tetrazolium assays. Results The combination of EGCG + FOSFO demonstrated synergism against all bacterial species, with an FIC index ranging from 0.35 to 0.5. At the MIC/FIC concentrations, EGCG, FOSFO, and EGCG+FOSFO were not toxic to fibroblasts. EGCG+FOSFO significantly reduced monospecies biofilms of E. faecalis and A. israelli, whereas S. mutans and F. nucleatum biofilms were eliminated by all compounds. Scanning electron microscopy of multispecies biofilms treated with EGCG, EGCG+FOSFO, and CHX at 100x MIC showed evident biofilm disorganization and substantial reduction of extracellular matrix. Confocal microscopy observed a significant reduction of multispecies biofilms formed in dentin tubules with 84.85%, 78.49%, and 50.6% of dead cells for EGCG+FOSFO, EGCG, and CHX at 100x MIC, respectively. Conclusion EGCG and fosfomycin showed a synergistic effect against biofilms of oral pathogens related to root canal infections without causing cytotoxicity.

ABSTRACT Objective: To assess the efficacy and safety of the use of midazolam as monotherapy, compared to the associated use of midazolam and hydroxyzine for minimum and moderate sedation of children in dental offices, using data obtained from clinical trials. Material and Methods: A systematic review protocol was developed and registered on PROSPERO (CR42020208633). An electronic search was carried out in Pubmed, Lilacs, Science Direct, Open Gray, Web of Science, and central Cochrane Library. No language restrictions were included. Clinical trials were carried out with children aged 0-12 years, using midazolam as monotherapy compared to the use of midazolam associated with hydroxyzine to verify the effectiveness and safety of oral sedation. The quality of the studies was individually assessed and grouped using the RoB 2 (Revised Cochrane risk-of-bias tool for randomized trials) and GRADE (Grading of Recommendations Assessment, Development and Evaluation) systems, respectively. Results: A total of 749 studies were found. After analyzing the inclusion and removal of duplicates, two studies were analyzed for the quality of evidence. Through this analysis, it was possible to verify the very low level of scientific evidence on the superiority of the efficacy and safety of the combined use of midazolam and hydroxyzine for oral sedation in children in dental offices. Conclusion: The conflicting results and limitations of the studies enabled to establish that there is insufficient evidence to support the use of these drugs combined. There is only evidence for the use of midazolam as monotherapy.

Abstract Objectives: To evaluate the influence of photobiomodulation with infrared laser (IRL) in the rat pulp tissue after bleaching, considering the immunolabeling of interleukin (IL)-23 and hypoxia-inducible factor (HIF)-1α. Methodology: The right and left molars of forty rats were divided into groups: Control – with placebo gel and Bleached – with 35% hydrogen peroxide (H2O2). Half of the rats received one IRL application on both sides, establishing a split-mouth design, which resulted in 4 groups with 20 hemi-maxillae each: Control, Bleach, IRL, and Bleached-IRL. Rats (n=10) from each group were euthanized, at 2- and 30-days mark, and the pulp tissue was evaluated using inflammation and immunolabeling scores. Wilcoxon and Mann-Whitney statistical tests were performed (p<0.05). Results: At the 2-days mark, the Bleached group had severe inflammation and necrosis in the occlusal thirds of the pulp, and moderate to severe inflammation in cervical third, whereas the Bleached-IRL had mild to moderate inflammation (p<0.05). At the 30-days mark, there was no inflammation, but tertiary dentine formation in the bleached groups. Regarding IL-23, severe immunolabeling was observed in the Bleached group (p<0.05) at the 2-days mark; at the 30-days mark, there was a reduction in immunolabeling, in which the Bleached group had moderate and the Bleached-IRL group had mild immunolabeling (p>0.05). HIF-1α was more evident at the 2-days mark in the Bleached group, without significant difference with the Bleached-IRL (p>0.05). The difference was observed between the bleached and control groups, without immunolabeling (p<0.05); at the 30-days mark, the Bleached group had reduction in HIF-1α immunolabeling, while the Bleached-IRL had an increase; the difference remained between the bleached and the controls groups (p<0.05) Conclusion: Photobiomodulation using IRL minimized the inflammation and IL-23 immunolabeling in the pulp tissue of rats after dental bleaching, but did not influence significantly the HIF-1α immunolabeling.

Abstract Objective The aim of this study was to evaluate whether probiotics multi-strain formula affects the development of apical periodontitis (AP) induced in rats. Methodology 16 Wistar rats were divided in two groups (n=8): rats with AP fed with regular diet (Control-C (CG)); rats with AP, fed with regular diet and supplemented with multi-strain formula (one billion colony-forming units (CFU)): GNC Probiotic Complex (PCG) ( Lactobacillus acidophilus, Lactobacillus salivaris, Lactobacillus plantarum, Lactobacillus rhamnosus, Bifidobacterium bifidum, Bifidobacterium animalis subs. lactis and Streptococcus thermofilus ). AP was induced in the upper and lower first molars by dental pulp exposure to the oral environment. PCG was administered orally through gavage for 30 days during the AP development. After this period the animals were euthanized and the mandibles were removed and processed for histologic analysis, and immunochemical assays for interleukin (IL)-6, IL-10, IL-1β, RANKL, OPG, and TRAP. The Mann–Whitney U test and Student’s t test were performed (P<.05). Results The CG showed more intense inflammatory infiltrate than the PCG group (P<.05). IL-1β, IL 6 and RANKL decreased in the PCG group compared with CG (P<.05). The IL-10 level increased in the PCG group (P<.05). The OPG level was similar in both groups (P>.05). The number of mature osteoclasts (TRAP-positive multinucleated cells) was lower in PCG group when compared to the CG (P<.05). Conclusion Probiotic Complex modulates inflammation and bone resorption in apical periodontitis.

ABSTRACT Objective: To investigate the tissue response and the biomineralization ability of CER prepared with epoxy resin or water compared to Mineral Trioxide Aggregate (MTA). Material and Methods: Polyethylene tubes containing materials or empty tubes for control were inserted into the subcutaneous tissues of 30 rats. After 7, 15, 30, 60, and 90 days, the rats were killed and the tubes were removed for analysis using hematoxylin-eosin staining, von Kossa staining, and under polarized light. Inflammation was graded through a score system; the thickness of the fibrous capsule was classified as thin or thick; the biomineralization ability was recorded as present or absent. The results were statistically analyzed using the Kruskal-Wallis test (p<0.05). Results: Histologic analysis performed after 7 and 15 days for CER prepared with epoxy resin or water and for MTA showed moderate inflammation and a thick fibrous capsule (p>0.05). After 30, 60, and 90 days, mild inflammation, and a thin fibrous capsule were observed in all groups (p>0.05). Conclusion: All materials had structures positive for von Kossa and birefringent to polarized light. CER epoxy resin showed biocompatibility and biomineralization similar to CER water and MTA.

ABSTRACT Objective: To evaluate the pulp tissue of rat molars after pulpotomies with mineral trioxide aggregate (MTA), BiodentineTM (BDT) and calcium hydroxide (CH) mixed with sterile saline solution (24 hours, 72 hours, 7 days and 15 days), through correlating MPO activity with active neutrophils and MMP8 activity with tissue remodeling. Material and Methods: Thirty-eight Wistar rats were randomly distributed into groups (control, I (MTA gray), II (BDT), and III (CH)) and subdivided according to the study period of 24 hours, 72 hours, 7 days or 15 days after pulpotomy. MMP8 activity was assessed through fluorescence technique, and MPO activity was determined using the MPO assay. Results: A gradual decrease of MPO and MM8 activity occurred in the group MTA over the experimental periods (p<0.05). Groups BDT and CH exhibited an increase in the activity at 7 and 15 days (p<0.05). Conclusion: MTA demonstrated a decrease in the values of MPO e MMP8. BDT and CH showed high neutrophilic and collagenase activity over the experimental periods.

ABSTRACT This paper discusses the potential risk that COVID-19 generates for the development of enamel defects. This hypothesis was built based on the etiopathogenesis of enamel defects and the relationship with the symptom's characteristic of COVID-19. Pregnancy is a critical period for the child's development; exposure to pathological agents can cause systemic imbalances and risks of adverse perinatal and prenatal outcomes. The main clinical symptoms of this disease and its association with that dental outcome were considered. Fever, breathing, cardiovascular disorders, and diarrhea were related as potential etiological factors of ameloblast metabolism imbalance, which can interfere qualitatively and quantitatively in the development, maturation and mineralization of the tooth enamel. Molecular disorders derived from COVID-19, as well as their clinical symptoms, can be considered potential risk factors for the development of enamel defects. Individuals with enamel defects experienced high stress levels during pregnancy or early childhood. The approach adopted may help build new research to ensure understanding of the etiology of the development of dental enamel defects and its relationship with COVID-19. However, longitudinal studies need to be conducted to confirm the association between COVID-19 and adverse events during pregnancy.

ABSTRACT Objective: To evaluate both knowledge and attitude of public elementary school teachers in Alfenas, Minas Gerais, Brazil, towards Traumatic Dental Injuries (TDIs). Material and Methods: Four hundred and seventy-one teachers from 10 schools were invited to participate in the research. Two hundred and twelve were accepted, being randomly selected to answer semi-structured questionnaires. Statistical tests Chi-Square and Fisher’s Exact Test were also used for the other variables at a significance level of 5% (p<0.05). Results: 212 teachers answered; 34% had first-aid training in college or even voluntarily (22.6%), but only 7.1% had taken training about TDIs. Only 7.5% were satisfied with the knowledge about TDIs, and 12.3% felt prepared to face it. About 10.8% had seen any type of TDI; 53.3% would report to the headmaster and 8% to the school dentist in a TDI event. The teachers’ first attitude towards TDIs would be contacting the child’s parents (63.2% and 58.5%, respectively). About 47.2% would handle the tooth properly (by the crown). Nearly 90.1% would carry out a permanent tooth dental reimplant, which would be conducted immediately (71.2%). About 36.3% of the teachers storage the avulsed tooth in a liquid: milk (46.7%) and saline solution (24.7%). The dry media mentioned were paper (18.9%) and gauze or cotton (15.1%). Most of the teachers (96.2%) stated that they would like to receive information on TDI. Conclusion: Elementary school teachers in Alfenas have partial knowledge about traumatic dental injuries.

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.

Abstract Objective: To evaluate sleep quality and excessive daytime sleepiness in undergraduate dentistry students. Material and Methods: This research is characterized as an observational study of transversal type, having analysed undergraduate students in dentistry from a public university in the State of Pernambuco, Brazil. The sample consisted of 325 students enrolled between the 1st and 10th academic semester. Sleep quality was evaluated using the Pittsburgh Sleep Quality Index (PSQI), while excessive daytime sleepiness was analysed through the Epworth Sleepiness Scale (ESS). The t-Student and Mann-Whitney tests were used for the numerical values, with the F-test (ANOVA) or the Kruskal-Wallis test being used to compare more than two categories. Results: It was observed that 71.1% of students presented a poor sleep quality, with more than half of the students exhibiting excessive daytime sleepiness (58.1%). A relationship between sleep quality and the academic semester was verified. However, gender and age were not associated with sleep quality or with excessive daytime sleepiness. Conclusion: A high prevalence of poor sleep quality and excessive daytime sleepiness was observed among undergraduate students in dentistry. The data suggest that the undergraduate degree in Dentistry can influence the students’ quality of sleep, regardless of age or gender.

Resumo Novas formulações de agregado de trióxido mineral (MTA) são constantemente introduzidas no mercado, geralmente em forma de pó e líquido. O Biocerâmico (Bio-C) Reparador (Repair) é um material pronto para uso sugerido como substituto do MTA, mas suas propriedades precisam ser estudadas. Este estudo avaliou a citotoxicidade, biocompatibilidade e biomineralização do Bio-C Repair comparado ao MTA-High Plasticity (MTA-HP) e MTA branco da Angelus (MTA-Ang). Fibroblastos L929 foram expostos a extratos dos materiais (não diluído, ½ e ¼ diluições; 6, 24 e 48 h). Tubos de polietileno contendo os materiais ou vazios (controle) foram implantados no tecido subcutâneo de ratos. Após 7 e 30 dias (n=8), os espécimes foram removidos para análises (hematoxilina-eosina, von Kossa e luz polarizada). Os dados da citotoxicidade foram analisados estatisticamente pelo teste de two-way ANOVA, e os dados da biocompatibilidade pelos testes de Kruskal-Wallis e Dunn (p<0,05). As células expostas aos materiais apresentaram maior viabilidade celular na maior parte dos períodos, comparados com o controle (p<0,05). O extrato não diluído e ½ diluição do MTA-HP apresentaram maior citocompatibilidade do que Bio-C Repair às 6h, e com ¼ diluição às 24h (p<0,05); o MTA-Ang branco apresentou maior citocompatibilidade do que o Bio-C Repair na maior parte dos períodos (p<0,05). O extrato não diluído do MTA-Ang branco apresentou maior citocompatibilidade às 6 e 24 h comparado ao MTA-HP, e com ½ diluição às 24h (p<0,05). A citocompatibilidade dos materiais foi semelhante às 48 h para a maior parte das diluições (p>0,05). Aos 7 e 30 dias, os grupos apresentaram inflamação moderada e leve, respectivamente (p>0,05). Todos os materiais mostraram estruturas positivas para von Kossa e luz polarizada. Em conclusão, o Bio-C Repair teve citocompatibilidade semelhante aos materiais à base de MTA, é biocompatível e induz à biomineralização.

Abstract New mineral trioxide aggregate (MTA) formulations are constantly introduced in the market, usually in a powder-and-liquid form. Bioceramic (Bio-C) Repair is a ready-for-use material suggested as substitute for MTA, but its properties need to be studied. This study evaluated the cytotoxicity, biocompatibility and biomineralization of Bio-C Repair compared to MTA Repair High-Plasticity (MTA-HP) and white MTA-Angelus (MTA-Ang). L929 fibroblasts were exposed to material-extracted (undiluted, ½ and ¼ dilutions; 6, 24 and 48h). Polyethylene tubes with material or empty (control) were implanted in the subcutaneous tissue of rats. After 7 and 30 days (n=8), the specimens were removed for analysis (hematoxylin-eosin, von Kossa and polarized light). Cytotoxicity data were statistically analyzed by two-way ANOVA, and biocompatibility data by Kruskal-Wallis and Dunn tests (p<0.05). The cells exposed to the materials had greater viability at most of the periods compared with control (p<0.05). The undiluted and ½ dilutions of MTA-HP extract showed higher cytocompatibility than Bio-C Repair at 6 h and with the ¼ dilution at 24 h (p<0.05); the white MTA-Ang showed higher cytocompatibility than Bio-C Repair at most of periods (p<0.05). The undiluted white MTA-Ang extract had higher cytocompatibility at 6 and 24h than MTA-HP, and with ½ dilution at 24h (p<0.05). The materials’ cytocompatibility was similar at 48h for most dilutions (p>0.05). At 7 and 30 days, the groups had moderate and mild inflammation, respectively (p>0.05). All materials showed positive structures for von Kossa and polarized light. In conclusion, Bio-C Repair had similar cytocompatibility to MTA-based materials is biocompatible and induces biomineralization.

Abstract Objectives: To evaluate the effect of calcium hydroxide (CH) associated with two different vehicles as a capping material for pulp tissue in primary molars, compared with mineral trioxide aggregate (MTA). Methodology: Forty-five primary mandibular molars with dental caries were treated by conventional pulpotomy using one of the following materials: MTA only (MTA group), CH with saline (CH+saline group) and CH with polyethylene glycol (CH+PEG group) (15 teeth/group). Clinical and periapical radiographic examinations of the pulpotomized teeth were performed 3, 6, and 12 months after treatment. Data were tested by chi-squared analysis and a multiple comparison post-test. Results: The MTA group showed both clinical and radiographic treatment success in 14/14 teeth (100%), at all follow-up appointments. By clinical evaluation, no teeth in the CH+saline and CH+PEG groups had signs of mobility, fistula, swelling or inflammation of the surrounding gingival tissue. However, in the CH+saline group, radiographic analysis detected internal resorption in up to 9/15 teeth (67%), and inter-radicular bone resorption and furcation radiolucency in up to 5/15 teeth (36%), from 3 to 12 months of follow-up. In the CH+PEG group, 2/11 teeth (18%) had internal resorption and 1/11 teeth (9%) presented bone resorption and furcation radiolucency at all follow-up appointments. Conclusion: CH with PEG performed better than CH with saline as capping material for pulpotomy of primary teeth. However, both combinations yielded clinical and radiographic results inferior to those of MTA alone.

Abstract This study evaluated the effect of hypertension on tissue response and biomineralization capacity of white Mineral Trioxide Aggregate (MTA), High-plasticity MTA (MTA HP), and Biodentine® (BDT) in rats. Polyethylene tubes filled with MTA, MTA HP, BDT, and the control group (empty tubes) were placed into the dorsal subcutaneous tissue of 32 male rats (16 normotensive (NT) and 16 hypertensive rats - 8 per group). After 7 and 30 days, the polyethylene tubes surrounded by connective tissue were removed, fixed, and embedded in histological resin. The mean number of inflammatory cells was estimated in HE-stained sections, biomineralization was quantified as area (µm2) by Kossa (VK) staining, and examination by polarized light (LP) microscopy was performed. The differences amongst the groups were analyzed statistically by the Mann-Whitney or Student’s t test, according to Shapiro-Wilk test of normality (p < 0.05). The inflammatory responses to all materials were greater in hypertensive rats than in NT rats (p < 0.05). Positive VK staining in MTA and BDT were more pronounced in NT rats at 7 and 30 days (p < 0.05). Birefringent structures in LP for MTA, MTA HP, and BDT were more pronounced in NT rats at 7 days (p<0.05). In rats, hypertension was able to increase inflammatory infiltrate and decrease biomineralization of the tested materials.

Abstract Purpose To compare, both qualitatively and quantitatively, the inflammatory cells, vascular density and IL-6 immunolabeled cells present in the pulp after pulpotomy with white MTA versus 15.5% ferric sulfate (FS). Methodology Forty-eight mandibular first molars from 24 Wistar rats were divided into MTA or FS groups and subdivided according to the period after pulpotomy procedure (24, 48 and 72 hours). Four teeth (sound and untreated) were used as controls. Histological sections were obtained and assessed through the descriptive analysis of morphological aspects of pulp tissue and the quantification of inflammatory cells, vascular density and interleukin-6 (IL-6) expression. Data were statistically analyzed (p<0.05). Results The number of inflammatory cells was similar in both groups, being predominantly localized at the cervical radicular third. In the MTA group, increased inflammation was observed at 48 hours. Vascular density was similar in both groups and over time, being predominant in the medium radicular third. No correlation was found between the number of inflammatory cells and the vascular density. Pulp tissue was more organized in MTA-treated teeth. In both groups, a weak to moderate IL-6 expression was detected in odontoblasts and inflammatory cells. Comparing both groups, there was a greater IL-6 expression in the cervical radicular third of teeth treated with MTA at 24 hours and in the medium and apical thirds at 72 hours, while in the FS group a greater IL-6 expression was found in the apical third at 24 hours. Conclusion The MTA group presented better histological features and greater IL-6 expression than the FS group. However, no difference was observed between the groups regarding the inflammatory status and vascularization, suggesting the usefulness of FS as a low-cost alternative to MTA.

Resumo O objetivo deste estudo foi avaliar os resultados clínicos, radiográficos e histológicos do complexo dentino-pulpar de molares decíduos após pulpotomia com agregado trióxido mineral (MTA) e sulfato férrico (SF) 15,5%. Trinta e um molares decíduos foram alocados aleatoriamente nos grupos MTA ou SF. As avaliações clínicas e radiográficas foram realizadas aos 3, 6, 12 e 18 meses de acompanhamento. Os dentes no período regular de esfoliação foram extraídos e processados para análise histológica. Os dados clínicos e radiográficos foram analisados estatisticamente (p≤0,01). O resultados histológicos foram analisados descritivamente. Todos os dentes tratados apresentaram sucesso clínico ao longo dos períodos experimentais. Ambos os grupos exibiram 100% de sucesso radiográfico aos 3, 6 e 12 meses. Aos 18 meses de acompanhamento, um dente do grupo SF apresentou radiolucidez interradicular (p>0,01). Histologicamente, os dentes tratados apresentaram vitalidade pulpar e ausência de infiltrado inflamatório no tecido conjuntivo. Somente o grupo MTA mostrou barreira de tecido duro rodeada por odontoblastos sobre os cotos pulpares. MTA e SF 15,5% são eficazes para pulpotomias de dentes decíduos. Embora o MTA seja considerado o material de primeira escolha, o SF pode ser uma alternativa adequada quando o custo do tratamento é um problema.

Abstract The aim of this study was to evaluate the clinical, radiographic and histological outcomes of the dentin-pulp complex from primary molars after pulpotomy with mineral trioxide aggregate (MTA) and 15.5% ferric sulfate (FS). Thirty-one primary molars were randomly allocated into MTA or FS groups. Clinical and radiographic evaluations were recorded at 3-, 6-, 12- and 18-month follow-up. Teeth at the regular exfoliation period were extracted and processed for histological analysis. Clinical and radiographic data were tested by statistical analysis (p≤0.01). Histological outcomes were analyzed descriptively. All of the treated teeth presented clinical success over the experimental periods. Both groups exhibited 100% of radiographic success at 3, 6 and 12 months. At the 18-month follow-up, one tooth from FS group presented inter-radicular radiolucency (p>0.01). Histologically, the treated teeth presented pulp vitality and absence of inflammatory infiltrate into the connective tissue. Only MTA group showed hard tissue barrier surrounded by odontoblasts over the pulp stumps. Both MTA and 15.5% FS are effective for pulpotomies of primary teeth. Although MTA is considered the first-choice material, FS may be a suitable alternative when treatment cost is an issue.