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Aim: To investigate the effect of chronic alcohol consumption on apical periodontitis in rats. Methodology: Thirty‐two male Wistar rats were arranged into four groups: Control (C): without apical periodontitis and nonalcoholic diet; (AL): without apical periodontitis and alcoholic diet; (AP): with apical periodontitis and nonalcoholic diet; and (AP + AL): with apical periodontitis and alcoholic diet. The alcoholic solution at 20% was given to the AL and AP + AL groups as the sole source of hydration throughout the experiment. AP was induced in the mandibular left first molars at the end of the 4th week. Weight changes and the amount of solid and liquid foods were recorded for 8 weeks. At the end, the animals were euthanized and the jaws removed followed by histological processing for histopathological and RANKL, OPG, TRAP and HIF‐1α analyses. The Mann–Whitney test was used for nonparametric data, and anova followed by the Tukey test was performed for parametric data, with P < 0.05. Results: Animals that received the alcoholic diet had a lower weight gain than the other groups (P < 0.05). Control and AL groups did not have an inflammatory response in the periapical tissues. The median score of inflammatory infiltrate was significantly higher in the AP + AL group (2.5) compared to the AP group (1.5; P < 0.05). In the same comparison, AP + AL was associated with score 3 for RANKL and HIF‐1α versus score 2 for AP group (P < 0.05). Moreover, the values for TRAP were 3.88 ± 0.70 cells mm−1 for the AP + AL group and 2.43 ± 0.94 cells mm−1 for the AP group (P < 0.05). Conclusion: In rats, an alcoholic diet had a significant effect on the severity of apical periodontitis, exacerbating the inflammatory response and osteoclastogenesis. [ABSTRACT FROM AUTHOR]

Objective: Evaluate whether a multi-strain probiotic formula affects blood parameters (hematologic, calcium, and phosphorus levels) and alters the expression of β-defensin-2, β-defensin-3, and toll-like receptor 4 in male rats with induced apical periodontitis (AP)., Design: Wistar rats were divided into two groups (n = 8 each): (1) rats with AP on a regular diet (Control) and (2) rats with AP on a regular diet supplemented with the multi-strain probiotic GNC Probiotic Complex (GCP) at one billion CFU. AP was induced by exposing the dental pulp of the first molars to the oral environment. GCP was administered daily via gavage for 30 days during AP development. After 30 days, animals were anesthetized, a cardiac puncture was performed, and 5 mL of blood was collected for hematologic, calcium, and phosphorus analysis. Animals were then euthanized, and mandibles were removed for histological and immunochemical analysis of β-defensin-2, β-defensin-3, and toll-like receptor 4. Statistical analyses used Mann-Whitney U and Student's t-tests, with significance at P < 0.05., Results: No significant differences were observed in blood parameters between the Control and GCP groups (P > 0.05). In AP, the Control group showed more intense inflammatory infiltrates and higher median severity scores than the GCP group (P < 0.05). Immunoreactivity levels for β-defensin-2, β-defensin-3, and toll-like receptor 4 were significantly increased in the GCP group (P < 0.05)., Conclusion: Probiotic complex reduces inflammation and enhances immunolabeling of β-defensin-2, β-defensin-3, and toll-like receptor 4 in AP., Competing Interests: Declaration of Competing Interest Nothing to declare, (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Objective: This study evaluated the influence of melatonin supplementation on tissue's response of endodontic sealers in Wistar rats., Methodology: Forty-eight rats received subcutaneous implants of four polyethylene tubes: one empty (control) and three filled with endodontic sealers (AH Plus, Endofill and Sealapex). Half of the animals were supplemented with melatonin (ME) and the remaining treated with water (WA) for 15 days before the implantation until euthanasia, forming the groups: control-WA, AH Plus-WA, Endofill-WA, Sealapex-WA, Control-ME, AH Plus-ME, Endofill-ME and Sealapex-ME. After 5, 15 and 30 days, (n = 8) tubes were removed and evaluated in H&E., immunohistochemistry, PSR, VK and POL. The results were statistically analyzed (p < 0.05)., Results: In animals treated with water, Endofill-WA evoked more intense inflammatory infiltrate compared to AH Plus-WA and Control-WA in a 30-day period (p < 0.05). In animals supplemented with melatonin, there was any difference among endodontic sealers' response in any period of analysis (p > 0.05). Comparing the individual response of each sealer, over a 30-day period, Endofill-ME and Sealapex-ME showed less inflammatory infiltrate compared to Endofill-WA and Sealapex-WA, respectively (p < 0.05). Immunostaining for IL-6 and TNF-α was less intense for all groups in animals supplemented with melatonin, in most periods, except for the Endofill sealer (p < 0.05). Furthermore, Endofill-ME at 5 days and AH-Plus-ME at 30 days showed a higher percentage of mature collagen fibers compared to the Endofill-WA and AH Plus-WA, respectively (p < 0.05). Positive structures for von Kossa staining and birefringent to polarized light were observed only for Sealapex-WA and Sealapex-ME in all periods., Conclusions: It can be concluded that melatonin influences the tissue response to endodontic sealers, modulating the inflammatory and reparative process., Competing Interests: Declarations. Conflict of interest: The authors have no conflict of interest to declare that is relevant to the content of this article., (© 2024. The Author(s), under exclusive licence to The Society of The Nippon Dental University.)

Objectives: This systematic review (PROSPERO CRD42021227711) evaluated the influence of diabetes mellitus (DM) on the response of the pulp tissue and in the pulp cells behaviour., Materials and Methods: Searches in PubMed/MEDLINE, Embase, Web of Science and OpenGrey were performed until March 2022. Studies evaluating the effects of DM in the pulp tissue inflammation and in the cell behaviour were included, followed by risk of bias assessment (Methodological Index for Non-Randomized Studies and SYRCLE's RoB tools). The meta-analysis was unfeasible, and a narrative synthesis for each outcome was provided., Results: Of the 615 studies, 21 were eligible, mainly with in vivo analysis (16 studies). The pulp inflammation (10 studies) was analysed mainly by haematoxylin-eosin stain; DM increased pulp inflammation/degeneration in 9 studies, especially after dental procedures. The cell viability (5 studies) was analysed mostly using MTT assay; DM and glycating agents decreased cellular viability in 3 studies. DM reduced collagen in all of three studies. There were controversial results regarding mineralization; however, increased alkaline phosphatase was reported in three of four studies., Conclusions: DM seems to increase inflammation/degeneration and mineralization in the pulp tissue while reducing cell proliferation. Further analyses in human pulp are important to provide stronger evidence., (© 2022 Wiley Periodicals LLC.)

The purpose of this study was to evaluate, in vivo, the biocompatibility, biomineralization, collagen maturation and the in vitro antibacterial and cytotoxicity of resinous endodontic sealers containing calcium hydroxide. Forty rats were implanted with polyethylene tubes containing Sealer 26, Sealer Plus, Dia-ProSeal and an empty tube, examined after 7, 15, 30 and 60 days. Antimicrobial activity was evaluated against Enterococcus faecalis by Agar Diffusion Test (ADT) through inhibition zones. For cytotoxicity, undifferentiated pulp cells (OD-21) were cultured and assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, exposed to dilution of serial extracts at 6, 24, 48h. Cytotoxicity was analyzed by two-way ANOVA and Bonferroni correction. Kruskal-Wallis test followed by Dunn test was performed for nonparametric data (p<0.05). MTT assay revealed cell proliferation affected by sealers extract in all periods (p<0.0001), except for Dia-Proseal and Sealer Plus ⅛ dilution. Subcutaneous analysis showed at day 7th moderate inflammatory infiltration. After 30 days, Sealer 26 still showed moderate inflammatory infiltrate compared to mild inflammation from control and Dia-ProSeal (p = 0.006). At day 60th, all groups showed similar mild inflammatory infiltrate (p>0.05). Sealer 26 induced more biomineralization than other sealers in all periods. At 7 and 15 days, all sealers had significant percentage of immature collagen fibers. After 60 days Sealer 26 showed more mature fibers compared to other sealers (p<0.001). All sealers had a smaller zone of inhibition than chlorhexidine, but with no significant difference among any group (p>0.05). All sealers showed satisfactory biological responses with in vitro/in vivo biocompatibility and antimicrobial activity against planktonic bacteria. Sealer 26 induced more biomineralization than Sealer Plus and Dia-ProSeal., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Bueno et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

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.

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.

Aim: To evaluate the effect of red wine consumption or its polyphenols on the inflammation/resorption processes associated with apical periodontitis in rats., Methodology: Thirty-two three-month-old Wistar rats had apical periodontitis induced in four first molars and were then arranged into four groups: control (C)-rats with apical periodontitis; wine (W)-rats with apical periodontitis receiving 4.28 ml/kg of red wine; resveratrol+quercetin (R+Q)-rats with apical periodontitis receiving 4.28 ml/kg of a solution containing 1.00 mg/L of quercetin and 0.86 mg/L of resveratrol and alcohol (ALC)-rats with apical periodontitis receiving the alcoholic dose contained in the wine. The oral gavage treatments were administered daily, from day 0 to day 45. On the 15th day, apical periodontitis was induced, and on the 45th day, the animals were euthanized. Histological, immunohistochemical (RANKL, OPG, TRAP, IL-10, TNF-⍺ and IL-1β) and micro-computed tomography for bone resorption analysis were performed in the jaws. The Kruskal-Wallis with Dunn's test was performed for nonparametric data, and the anova with Tukey's test for parametric data, p < .05., Results: The median score of the inflammatory process was significantly lower in the R+Q group (1) compared to the C (2) (p = .0305) and ALC (3) (p = .0003) groups, and not different from the W (1.5) group. The immunolabeling for OPG was significantly higher in the R+Q group (p = .0054) compared to all groups; the same was observed for IL-10 (p = .0185), different from groups C and ALC. The R+Q group had the lowest TRAP cell count (p < .0001), followed by the W group, both inferior to C and ALC groups. The lowest bone resorption value was in the R+Q group (0.50mm 3  ± 0.21mm 3 ), significantly lower (p = .0292) than the C group (0.88mm 3  ± 0.10mm 3 ). The W group (0.60 mm 3  ± 0.25 mm 3 ) and R+Q group had less bone resorption compared to the ALC group (0.97 mm 3  ± 0.22 mm 3 ), p = .0297 and p = .0042, respectively., Conclusion: Red wine administration to rats for 15 days before induction of apical periodontitis decreased inflammation, TRAP marking and periapical bone resorption compared to alcohol. Resveratrol-quercetin administration reduced the inflammatory process in apical periodontitis, periapical bone resorption, and altered the OPG, IL-10 and TRAP expression compared to C and ALC groups., (© 2021 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Endodontics has gained emphasis in the scientific community in recent years due to the increase in clinical and in animal models studies focused on endodontic medicine, which aims to evaluate the interrelationship between systemic and periapical tissues pathological conditions. These studies have shown that systemic changes can boost the pathogenesis of endodontic infection, favoring its development and progression. A contrary relationship is reported in numerous studies that affirm the potential of endodontic infection to trigger systemic damage and may lead to the worsening of pre-existing pathologies. Recently, the potential of filling materials to develop systemic changes such as neurological alterations had been evaluated, also showing that systemic diseases can negatively influence tissue responses to filling materials after endodontic treatment. Despite advances in endodontic medicine studies, there are still gaps in knowledge on the mechanisms of interactions between apical periodontitis (AP) and systemic diseases and much research to be done. In this sense, this critical narrative literature review aimed to show the evolution of studies in endodontic medicine to help the endodontist to know the role of systemic diseases in the pathogenesis of AP and the possible interference in the repair of periapical tissues after endodontic treatment, as well as to evidence the systemic complications that can be triggered or aggravated in the presence of endodontic infection., (© 2021. The Society of The Nippon Dental University.)

Aim: To evaluate the effect of excessive caffeine intake on the inflammation/resorption processes associated with periapical periodontitis (PP) in rats., Methodology: Sixteen Wistar rats were used. Periapical periodontitis was induced in the four first molars in each animal. The animals were arranged into two groups: control (C)-rats with periapical periodontitis; and caffeine (CAF)-rats with periapical periodontitis under caffeine administration protocol. The CAF animals received 10 mg/100 g of body weight/day of caffeine via gavage starting fifteen days before PP induction and continuing for thirty more days until euthanasia. On the 30th day, the animals were euthanized and the jaws removed for microcomputed tomography, histological and immunohistochemical analysis for RANKL, OPG, TRAP, IL-10, TNF-⍺ and IL-1β. The Mann-Whitney test was performed for nonparametric data, and Student's t test was performed for parametric data, using p < .05., Results: There was no significant difference in the weight change between the groups. The median score of the inflammatory process was significantly greater in the CAF group (3) compared with the C group (2), p = .0256. Bone resorption was greater in the group consuming caffeine (1.08 ± 0.15 mm 3 ) compared with the C group (0.88 ± 0.10 mm 3 ), p = .0346. The immunolabelling for RANKL, TRAP and IL-1β was significantly higher in the CAF group when compared to the control, p < .05. No differences were found for the OPG, IL-10 and TNF-⍺ immunolabelling., Conclusion: Excessive caffeine exposure via gavage in rats was able to exacerbate the volume of periapical bone destruction, and the inflammatory pattern deriving from periapical periodontitis altering the expression of RANKL, IL-1β and TRAP., (© 2021 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

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.

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.

Aim: To investigate the effect of different alcohol concentrations on the development of apical periodontitis (AP) in rats., Methods: Forty Wistar rats were arranged into five groups: (C) - control rats receiving sterile water as the only liquid; (G5) - animals receiving an alcohol solution at 5%, (G10) - alcohol solution at 10%, (G15) - alcohol solution at 15%, and (G20) - alcohol solution at 20%. The alcoholic solution or water was given to the groups as the sole source of hydration throughout the 30 days of the experiment. AP was induced in the mandibular molars on the first day. In the end, the animals were euthanized for histopathological and IL-1b, RANKL, OPG, and TRAP analyses. The Kruskal-Wallis test was used for nonparametric data, and ANOVA followed by the Tukey test were performed for parametric data, p < 0.05., Results: G15 and G20 had a greater chronic inflammatory infiltrate (Score 3) and AP size bigger (1.59 ± 0.41 and 1.83 ± 0.38, respectively) than the C, G5 and G10 (p < 0.05). No significant difference was found in the IL-1b analyses. The G15 and G20 showed the highest immunolabeling pattern for RANKL and the lowest for OPG. The G20 had greater TRAP cells per mm (4.70 ± 0.99) compared to the C, G5, and G10 (p < 0.05). Furthermore, G15 presented 3.92 ± 0.64 TRAP cells/mm, higher than C (p < 0.05)., Conclusions: G5 and G10 did not exert a protective or aggravating effect on the AP development. However, G15 and G20 had a significant effect on the AP severity, exacerbating the inflammation and osteoclast markers., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Aim: The aim of the present study was to evaluate apical periodontitis (AP) development in rats under a chronic alcohol diet by calcium, phosphorus, and alkaline phosphatase blood levels in addition to histological and radiographic analyses., Methods: Thirty-two rats were arranged into four groups: (a) group 1: without apical periodontitis and on a regular diet; (b) group 2: AP and on a regular diet; (c) group 3: alcoholic diet without apical periodontitis; and (d) group 4: alcoholic diet and apical periodontitis. Alcoholic solution at 20% was given throughout the 8-week experiment. AP was induced in the first molars at the end of the 7th week. At the end, the animals were anesthetized for blood collection, followed by euthanasia, and jaws were removed for digital radiography and histological processing. The level of significance was 5%., Results: Calcium levels remained constant in all groups (P > 0.05). Group 4 showed a higher phosphorous level than group 2 (P < 0.05). The alkaline phosphatase activity was higher in group 3 compared with group 1 (P < 0.05). Three animals in group 4 exhibited a severe inflammatory reaction, whereas the animals in group 2 did not demonstrate any reaction (P < 0.05). The lowest value of radiographic density was given by group 4 (P < 0.05)., Conclusions: Chronic alcohol consumption increased serum phosphorus and decreased bone density in the periapical region, favoring AP development., (© 2019 John Wiley & Sons Australia, Ltd.)

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.

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.

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.

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.

Aim: To evaluate the inflammatory response, biomineralization and production of osteocalcin (OCN), osteopontin (OPN), and bone sialoprotein (BSP) of a new root filling material for primary teeth (Bio-C Pulpecto) compared to MTA., Design: Polyethylene tubes containing Bio-C Pulpecto, MTA, and empty tubes were implanted into the dorsal connective tissue. After 7, 15, 30, 60, and 90 days, the tubes with surrounding tissue were removed and histologically processed to be analysed using haematoxylin and eosin, von Kossa staining, or no staining for observation under polarized light and immunohistochemistry for the detection of OCN, OPN, and BSP. The Kruskal-Wallis test was used followed by Dunn's test. The significance level was set at 5%., Results: The inflammatory response observed with MTA and Bio-C Pulpecto was more exacerbated until the 15th day and decreased from 30 days on. No significant difference was found between control, MTA, and Bio-C Pulpecto (P > 0.05). Bio-C Pulpecto and MTA showed positivity for von Kossa and birefringent to polarized light. The immunolabelling for OCN, OPN, and BSP was more intense for MTA and Bio-C Pulpecto on days 60 and 90 (P > 0.05)., Conclusion: Bio-C Pulpecto was biocompatible, induced biomineralization and was immunopositive for osteogenic markers such as OCN, OPN, and BSP, similarly to MTA., (© 2019 BSPD, IAPD and John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

The aim of this preliminary study was to compare the effects of different energy densities from red and infrared low-level laser (LLL) on viability and proliferation of stem cells from human exfoliated deciduous teeth (SHED). SHED were irradiated with red laser (R) or infrared laser (IR) set with the following dosimetry: 1.2 J/cm 2 (0.05 J), 2.5 J/cm 2 (0.1 J), 5.0 J/cm 2 (0.2 J), and 7.5 J/cm 2 (0.3 J). Positive (C+) and negative (C-) control groups comprised non-irradiated cells. Data were analyzed by two-way ANOVA followed by Tukey's test (P < 0.05). At 24- and 48-h period, group R5.0 showed significantly higher cell viability rates than R1.2 and R2.5. At 48 h, R2.5 also revealed lower proliferation than R5.0. Comparing to the C+ group, R2.5 exhibited lower viability at 72 h, and proliferation at 24 and 48 h. Groups R1.2, IR1.2, and IR5.0 were less viable at 24 h, while R1.2, IR2.5, and IR5.0 revealed lower proliferative capacity at 48 h. Overall, our results showed that LLL can favor viability and proliferation of SHED, especially when cells receive red laser irradiation at 5.0 J/cm 2 . Therefore, according to this preliminary investigation, 5 J/cm 2 applied by red LLL induced high rates of cell viability and proliferation, while the same irradiation dose using infrared laser led to negative effects. LLL irradiation with 1.2 and 2.5 J/cm 2 was deleterious to metabolic activity and proliferation of SHED regardless of the type of laser. Further studies are necessary to gain in-depth knowledge about the effects of different wavelengths of LLL on SHED viability and proliferation.

Objective: The aim of this retrospective clinical study was to evaluate the incidence of ProTaper Universal System instrument fractures, associated with observation of the arch, group of teeth, and root thirds in which these fractures occurred., Methods: From analysis of charts, clinical record cards and radiographs of endodontic treatments performed by postgraduate students using the ProTaper Universal System at a reference center, a total of 1031 teeth and 2355 canals were analyzed. The general incidence of instrument fractures and their frequency, considering the group of teeth, arch and root thirds, were cataloged and the data obtained were statistically analyzed (Exact Fischer test, with level of significance of 1%)., Results: The general percentage of fractures, considering the number of teeth and number of root canals evaluated was 4.4% and 1.9%, respectively. Instrument fractures occurred more frequently in the mandibular first (8.8%) and second (9.6%) molars, however, without statistically significant difference between them (P=0.81). In the first and second maxillary molars, the incidence of fracture was 4.7% and 5.1%, respectively, also without significant difference (P=0.81). Considering the dental arches (maxillary and mandibular), the fractures occurred with significantly higher frequency in the mandibular arch (66.7%), in comparison with the maxillary arch (33.3%) (P<0.01). A significantly higher percentage of fractures occurred in the apical third (84.4%) compared with the middle third (15.6%) (P<0.01)., Conclusion: The general percentage of fractures, considering the number of teeth and number of root canals evaluated was 4.4% and 1.9%, respectively. However, the arch (mandibular) and root third (apical) had a significant effect on the incidence of instrument fractures., Competing Interests: Disclosures Conflict of interest: The authors deny any conflict of interest., (Copyright: © 2018 European Endodontic Journal.)

Objective: The aim of this study was to evaluate the periodontal status of teeth indicated for undergoing endodontic treatment., Materials and Methods: Two hundred and nine teeth were evaluated using probing depth tests at three vestibular and three palatine/lingual sites before the patients underwent treatment. Teeth that presented up to 3-mm probing depth were considered as healthy. Those that presented at least one site with probing depth >3 mm were considered periodontally compromised. The data were statistically analyzed by means of applying the Chi-square test, with a level of significance of 5%., Results: Of the total of 209 teeth evaluated, 40 (19.10%) presented periodontal compromise. There was statistically significant difference related to the compromised teeth of patients of the female gender (22.80%) compared with teeth of patients of the male gender (12.30%) ( P < 0.05)., Conclusions: Fewer than 20% of the teeth evaluated presented a periodontal compromise. Patients of the female gender presented a higher number of periodontally compromised teeth than patients of the male gender., Competing Interests: There are no conflicts of interest.

Permanent maxillary central incisor impaction is a rare condition of multifactorial etiology that should be treated on diagnosis. This article describes multidisciplinary treatment of an 11-year-old patient presenting with a permanent maxillary right central incisor that was impacted due to the presence of 2 supernumerary teeth. The diagnosis was reached through clinical evaluation, radiography, and cone beam computed tomography. Treatment involved the surgical removal of the supernumerary teeth followed by traction of the impacted tooth through a combination of removable and fixed orthodontic appliances. Orthodontic treatment was completed after 18 months, resulting in adequate positioning of the right central incisor in the arch. Periodic follow-up visits were scheduled for prophylaxis and oral hygiene reinforcement. After 2 years, the teeth were adequately aligned, the gingiva was healthy, and the patient was esthetically satisfied. The multidisciplinary approach allowed functional and esthetic oral rehabilitation, thus helping the patient to avoid the psychological impact of a missing anterior tooth.

Background/aims: The prognosis of traumatized teeth can be optimized if appropriate procedures are following at the time of the accident. The aim of this study was to evaluate the level of knowledge of students' parents from a group of Brazilian schools regarding fracture and tooth avulsion in permanent teeth., Methods: Parents of 802 students were interviewed using a pre-tested questionnaire designed to explore how they would manage their child's dental avulsion or fracture., Results: Among the parents, 8.6% reported their child had already suffered tooth injuries. Concerning dental fracture, 49.5% considered the possibility of bonding the tooth fragment and, for dental avulsion, 65.5% considered tooth replantation. Approximately 33.5% of parents would take the child to the dental office in case of fracture or dental avulsion, and of those, 66.2% would do so immediately. Further, 69.5% (for fracture) and 70.4% (for avulsion) would do so regardless of symptoms; only 24.2% and 21.3%, respectively, would seek care only if there was pain. In case of avulsion, 22.3% of those interviewed would store the tooth in liquid, such as saline solution (17.8%), water (16.4%) and fresh milk (12.2%)., Conclusion: Parents had partial and insufficient knowledge as to the proper approach to be adopted in cases of fracture and dental avulsion. Previous experience of dental trauma was not related to more appropriate behavior regarding these injuries., (© 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

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.

The aim of the study was to evaluate the effects of the capping materials mineral trioxide aggregate (MTA), calcium hydroxide (CH) and BiodentineTM (BD) on stem cells from human exfoliated deciduous teeth (SHED) in vitro. SHED were cultured for 1 - 7 days in medium conditioned by incubation with MTA, BD or CH (1 mg/mL), and tested for viability (MTT assay) and proliferation (SRB assay). Also, the migration of serum-starved SHED towards conditioned media was assayed in companion plates, with 8 μm-pore-sized membranes, for 24 h. Gene expression of dentin matrix protein-1 (DMP-1) was evaluated by reverse-transcription polymerase chain reaction. Regular culture medium with 10% FBS (without conditioning) and culture medium supplemented with 20% FBS were used as controls. MTA, CH and BD conditioned media maintained cell viability and allowed continuous SHED proliferation, with CH conditioned medium causing the highest positive effect on proliferation at the end of the treatment period (compared with BD and MTA) (p<0.05). In contrast, we observed increased SHED migration towards BD and MTA conditioned media (compared with CH) (p<0.05). A greater amount of DMP-1 gene was expressed in MTA group compared with the other groups from day 7 up to day 21. Our results show that the three capping materials are biocompatible, maintain viability and stimulate proliferation, migration and differentiation in a key dental stem cell population.

Objectives: This study evaluated smear layer removal by different chemical solutions used with or without ultrasonic activation after post preparation., Materials and Methods: Forty-five extracted uniradicular human mandibular premolars with single canals were treated endodontically. The cervical and middle thirds of the fillings were then removed, and the specimens were divided into 9 groups: G1, saline solution (NaCl); G2, 2.5% sodium hypochlorite (NaOCl); G3, 2% chlorhexidine (CHX); G4, 11.5% polyacrylic acid (PAA); G5, 17% ethylenediaminetetraacetic acid (EDTA). For the groups 6, 7, 8, and 9, the same solutions used in the groups 2, 3, 4, and 5 were used, respectively, but activated with ultrasonic activation. Afterwards, the roots were analyzed by a score considering the images obtained from a scanning electron microscope., Results: EDTA achieved the best performance compared with the other solutions evaluated regardless of the irrigation method ( p < 0.05)., Conclusions: Ultrasonic activation did not significantly influence smear layer removal., Competing Interests: Conflict of Interest: No potential conflict of interest relevant to this article was reported.

This case report describes the diagnosis of a traumatic periapical lesion, caused by parafunctional habits (bruxism) and associated with bone rarefaction, that had not been identified by periapical radiography. Slight edema was observed clinically at the apical region of the maxillary left canine; however, the tooth demonstrated only physiologic mobility, and the results of a pulpal sensitivity test were inconclusive. A cone beam computed tomogram (CBCT) showed the presence of a periapical lesion. After preparation of the root canal, intracanal medication (calcium hydroxide and paramonochlorophenol) was placed and changed once a month for 2 months. After 60 days (at the time of the second medication change), the edema at the apical region was no longer visible. The root canal was filled, and CBCTs obtained 30 days and 24 months after completion of treatment showed that the lesion had regressed with bone repair. Although it is an expensive procedure, CBCT is important in the examination and diagnosis of periapical lesions that may not be seen in periapical radiographs.

Background: Iatrogenic complications such as accidental perforation of the root or the floor of the pulp chamber may occur., Case Report: Patient was referred for root canal retreatment of the mandibular left second molar with periapical lesion evidenced through radiographic examination. During post removal, iatrogenic perforation occurred at the mesial face of the distal root. After clinical localization of the perforation and bleeding control, MTA was applied. In a second appointment, the root canal filling was removed and the chemical-surgical retreatment of the canals was performed, followed by the obturation with gutta-percha and sealer. Patient returned after three days reporting no pain. After 6 months, 3, 7 and 10 years of follow-up., Conclusion: Absence of pain, normal periodontal probing and lack of radiolucent area at the region of perforation and the periapices were detected, which evidenced the successful repair of the tooth.

Bioactive molecules stored in dentin, such as transforming growth factor beta1 (TGF-b1), may be involved in the signaling events related to dental tissue repair. The authors conducted an in vitro evaluation of the amount of TGF-b1 released from dentin slices after treatment with 10% ethylenediaminetetraacetic acid (EDTA), 2.5% sodium hypochlorite (NaOCl) or phosphate-buffered saline (PBS), and the effect of this growth factor on stem cell migration from human exfoliated deciduous teeth (SHED). Sixty 1-mm-thick tooth slices were prepared with or without the predentin layer, and treated with either 10% EDTA for 1 minute, 2.5% NaOCl for 5 days or kept in PBS. Tooth slice conditioned media were prepared and used for TGF-b1 ELISA and migration assays. Culture medium with different concentrations of recombinant human TGF-b1 (0.5, 1.0, 5.0 or 10.0 ng/mL) was also tested by migration assay. The data were evaluated by ANOVA and Tukey's test. Optical density values corresponding to media conditioned by tooth slices either containing or not containing the predentin layer and treated with 10% EDTA were statistically greater than the other groups and close to 1 ng/mL. Increased rates of migration toward media conditioned by tooth slices containing the predentin layer and treated with PBS, 10% EDTA or 2.5% NaOCl were observed. Recombinant human TGF-b1 also stimulated migration of SHED, irrespective of the concentration used. EDTA may be considered an effective extractant of TGF-b1 from the dentin matrix. However, it does not impact SHED migration, suggesting that other components may account for the cell migration.

Impaction of permanent maxillary central incisors is a rare condition that can affect esthetics and oral function. The etiology of impaction is multifactorial and may be related to embryologic reasons, hereditary factors, mechanical obstacles, and pathologic elements, such as the presence of supernumerary teeth. The purpose of this report is to discuss the case of a 10-year-old patient who presented an impacted permanent maxillary right central incisor due to a supernumerary tooth. Surgical removal of the supernumerary tooth and orthodontic traction of the impacted central incisor by the closed eruption technique were performed. After 12 months, the permanent maxillary right central incisor was adequately positioned in the arch. The multidisciplinary treatment allowed the functional and esthetic rehabilitation to happen in a short period of time, thus reducing the patient's psychological impact caused by the lack of an anterior tooth.