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To evaluate the inflammatory tissue response to BioRoot™ RCS (BR) and AH Plus Jet (AHPJ) sealers implanted in mice subcutaneous tissue. It was hypothesized that the inflammatory tissue response to BR would be milder than to AHPJ. An in vivo study was carried out using isogenic mice. The sealers were implanted during standardized surgical procedures. The inflammatory response was evaluated by microscopic analysis and von Kossa reaction in the reactionary tissue around the specimens after 7, 21, and 63 days. For comparisons, a zinc oxide and eugenol sealer (ZOE) was used as a positive control, in addition to a negative control without a sealer (n = 10 per group/period). All statistical analyses considered a significance level of 5%. All endodontic sealers triggered an inflammatory tissue response after 7 days. BR had a higher inflammatory cell count and a thicker fibrous capsule when compared with AHPJ, but both were less inflammatory than ZOE (p < .001). After 21 days, BR continued to trigger an intense inflammatory tissue response, higher in both microscopic parameters compared to AHPJ, and a thicker fibrous capsule than ZOE (p < .001). After 63 days, the inflammatory tissue response decreased in BR, matching the fibrous capsule thickness with AHPJ and ZOE. BR promoted intense calcium precipitation in all study periods. After 63 days, AHPJ and BR sealers were more biocompatible to subcutaneous mice tissue, but AHPJ present better early inflammatory response, as well as BR showed potential bioactivity. RESEARCH HIGHLIGHTS: The inflammatory tissue response triggered by a bioceramic endodontic sealer (BR) was not milder than that triggered by an epoxy-resin based endodontic sealer (AHPJ) during the first 3 weeks, considering the microscopic analysis of the reactionary tissue., (© 2024 Wiley Periodicals LLC.)

This study evaluated territorial disparities in dental care for disabled persons in Brazil's public healthcare system from 2014 to 2023. The person-year incidence of outpatient dental procedures carried out by special care dentistry specialists and hospitalizations for dental procedures for disabled persons were compared across different regions and against the national estimate. In addition, productivity was correlated with oral health-related indicators. The significance level was set at 5%. The northern region exhibited the highest outpatient productivity, while the southern region showed lower productivity compared to the national estimate (both p -value < 0.05). This pattern was reversed in inpatient productivity (both p -value < 0.05), with the northeastern and central-western regions also below average (both p -value < 0.05). There were no significant correlations between the indicators and inpatient productivity, but outpatient productivity was positively correlated with the proportions of inhabitants who self-rated their general and oral health as "poor" or "very poor", who have never visited a dentist, and who visited a dentist for tooth extraction (all p -values < 0.05). Territorial disparities in dental care for disabled persons were observed within Brazil's public healthcare system, and they were correlated with unfavorable oral health-related indicators at the population level.

Objective: To evaluate the effects of NLRP3 inflammasome inhibition or knockout in experimental apical periodontitis (AP) induced in mice., Methods: The experimental AP was induced by pulpal exposure. To evaluate NLRP3-specific inhibitor medication (MCC950), WT mice received intraperitoneal injections, while the control received PBS (n = 10). In addition, to evaluate NLRP3 knockout, 35 wild-type (WT) and 35 NLRP3 -/- mice were divided into a control group (without pulpal exposure, n = 5) and three experimental groups: after 2, 14 and 42 days after pulpal exposure (n = 10). Microscopic and molecular analyzes were carried out using a significance level of 5%., Results: Exposure to MCC950 did not affect the periapical lesion size after 14 days (P = 0.584). However, exposed mice had a lower expression of IL-1β, IL-18 and caspase-1 (P = 0.010, 0.016 and 0.002, respectively). Moreover, NLRP3 -/- mice showed a smaller periapical lesion after 14 and 42 days (P = 0.023 and 0.031, respectively), as well as a lower expression of IL-1β after 42 days (P < 0.001), of IL-18 and caspase-1 after 14 (P < 0.001 and 0.035, respectively) and 42 days (P = 0.002 and 0.002, respectively). NLRP3 -/- mice also showed a lower mRNA for Il-1β, Il-18 and Casp1 after 2 (P = 0.002, 0.036 and 0.001, respectively) and 14 days (P = 0.002, 0.002 and 0.001, respectively)., Conclusions: NLRP3 inflammasome inhibition or knockout can attenuate the inflammatory events that result in the periapical lesion (AP) formation after pulpal exposure in mice., Clinical Relevance: The NLRP3 inflammasome may be a therapeutic target for AP, and new approaches may verify the impact of its inhibition (through intracanal medications or filling materials) on the bone repair process and treatment success., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: The conicity of the root canals of primary teeth is an important measure for endodontic therapies. However, determining this conicity depends on the methods employed, which requires further investigation., Aim: The aim of this study was to determine the conicity of the root canals of the upper and lower primary second molars using nanotomography (nCT)., Design: An in vitro study was performed using nine primary second molars, both upper and lower, subjected to nCT. Comparisons between the diameters of root canals were performed between the thirds (cervical-D0, middle-D5, and apical-D7). The conicity (%) was determined for each root canal from cervical to apical. Data were statistically analyzed with a significance level of 5%., Results: The conicity ranged from 2% to 8% for the upper primary second molars. Significant differences in root canal diameter between the thirds (D0, D5, and D7 points) were observed in the mesio- and distobuccal roots (p < .05), but not in the palatal roots (p > .05). For the lower primary second molars, the conicity ranged from 2% to 17%, as well as significant differences in root canal diameter between the thirds (D0, D5, and D7 points) were observed in all roots (distal, mesiobuccal, and mesiolingual; p < .05)., Conclusion: The conicity of the upper primary second molars was different from that of the lower ones, which showed a greater variability., (© 2024 BSPD, IAPD and John Wiley & Sons Ltd.)

Aim: To evaluate the role of regulatory T lymphocytes (Tregs) in the presence or absence of the synthetic ligand Pam3Cys during the progression of periapical lesion in wild-type (WT) and toll-like receptor 2 knockout (TLR2KO) mice., Methodology: A total of 130 C57BL/6 male WT and TLR2KO mice were allocated into control (n = 5) and experimental (periapical lesion induction) (n = 10) groups. In specific groups (WT+Pam3cys and TLR2KO+Pam3cys), the synthetic ligand Pam3cys was administered intraperitoneally every 7 days, according to the experimental period (14, 21 and 42 days). At the end of those periods, the animals were euthanized, and the mandible and the spleen were submitted to histotechnical processing. Mandible histological sections were analysed by haematoxylin and eosin, TRAP histoenzymology and immunohistochemistry (FOXP3, RANK, RANKL and OPG). Spleen sections were analysed by immunohistochemistry (FOXP3)., Results: The inflammatory infiltrate and bone resorption were more intense in the TLR2KO group compared to the WT group. The animals that received the Pam3cys had smaller periapical lesions when compared to the animals that did not receive the ligand (p < .05). TLR2KO animals showed a significant increase in the number of osteoclasts when compared to TLR2KO+Pam3cys group (p < .05). At 21 days, the WT+Pam3cys group had a lower number of osteoclasts when compared to the WT animals (p = .02). FOXP3 expression was more intense in the WT+Pam3cys groups when compared to the WT animals in the 42 days (p = .03). In the spleen analysis, the WT+Pam3cys group also had a higher expression of FOXP3 when compared to the WT animals at 14 and 42 days (p = .02). Concerning RANKL, there was a reduction in staining in the KOTLR2+Pam3cys groups at 21 and 42 days (p = .03) and a higher binding ratio between RANK/RANKL in animals that did not receive the ligand., Conclusion: Administration of the Pam3cys increased the proliferation of Tregs, showed by FOXP3 expression and prevented the progression of the periapical lesion in WT mice. On the other hand, in the TLR2KO animals, Treg expression was lower with larger areas of periapical lesions. Finally, systemic administration of the Pam3cys in KO animals was able to limit the deleterious effects of the absence of the TLR2 receptor., (© 2024 British Endodontic Society. Published by John Wiley & Sons Ltd.)

Microfluidic cotton thread-based electroanalytical devices (μTEDs) are analytical systems with attractive features such as spontaneous passive flow, low cost, minimal waste production, and good sensitivity. Currently, sample injection in µTEDs is performed by hand using manual micropipettes, which have drawbacks such as inconstant speed and position, dependence of skilled analysts, and need of physical effort of operator during prolonged times, leading to poor reproducibility and risk of strain injury. As an alternative to these inconveniences, we propose, for the first time, the use of electronic micropipettes to carry out automated injections in µTEDs. This new approach avoids all disadvantages of manual injections, while also improving the performance, experience, and versatility of µTEDs. The platform developed here is composed by three 3D-printed electrodes (detector) attached to a 3D-printed platform containing an adjustable holder that keeps the electronic pipette in the same x/y/z position. As a proof-of-concept, both injection modes (manual and electronic) were compared using three model analytes (nitrite, paracetamol, and 5-hydroxytryptophan) on µTED with amperometric detection. As result, improved analytical performance (limits of detection between 2.5- and 5-fold lower) was obtained when using electronic injections, as well as better repeatability/reproducibility and higher analytical frequencies. In addition, the determination of paracetamol in urine samples suggested better precision and accuracy for automated injection. Thus, electronic injection is a great advance and changes the state-of-art of µTEDs, mainly considering the use of more modern and versatile electronic pipettes (wider range of pre-programmed modes), which can lead to the development of even more automated systems., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

The aim of the present study was to evaluate, in vitro, the antimicrobial activity of the probiotic Bifidobacterium animalis subsp. lactis HN019, through the well technique, against 10 microorganisms can be found involved in endodontic infections. The antimicrobial activity of the probiotic was performed on Streptococcus mutans, Streptococcus sobrinus, Lacticaseibacillus casei, Enterococcus faecalis, Staphylococcus aureus, Candida albicans, Porphyromonas gingivalis, Porphyromonas endodontalis, Fusobacterium nucleatum and Prevotella intermedia. For the control group, it was used non-pathogenic bacteria Escherichia coli, Saccharomyces cerevisiae, and Kocuria rizhopilla. After 48 to 72 h of incubation of the petri dishes containing the culture medium, the microorganism strains, and the probiotic, the plates were examined to assess the uniformity of microbial growth, presence of contaminants, and the halo of inhibition. After visual inspection, the reading of the halo of inhibition was performed with the aid of a digital caliper using a reflected light source to illuminate the inverted plate on a black, opaque background after removing the cap. Thus, 3 values were obtained from each bacterial inoculum, which were added and divided by three to obtain the average of the values. The results of the in vitro study demonstrated that the probiotic B. animalis subsp. lactis HN019 promoted the inhibition of all strains of the pathogens evaluated, with the exception of Candida albicans, demonstrating antimicrobial activity on these microorganisms., (© 2023. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

The development of miniaturized, sustainable and eco-friendly analytical sensors with low production cost is a current trend worldwide. Within this idea, this work presents  the innovative use of masked stereolithography (MSLA) 3D-printed substrates for the easy fabrication of pencil-drawn electrochemical sensors (MSLA-3D-PDE). The use of a non-toxic material such as pencil (electrodes) together with a biodegradable 3D printing resin (substrate) allowed the production of devices that are quite cheap (ca. US$ 0.11 per sensor) and with low environmental impact. Compared to paper, which is the most used substrate for manufacturing pencil-drawn electrodes, the MSLA-3D-printed substrate has the advantages of not absorbing water (hydrophobicity) or becoming crinkled and weakened when in contact with solutions. These features provide more reproducible, reliable, stable, and long-lasting sensors. The MSLA-3D-PDE, in conjunction with the custom cell developed, showed excellent robustness and electrochemical performance similar to that observed of the glassy carbon electrode, without the need of any activation procedure. The analytical applicability of this platform was explored through the quantification of omeprazole in pharmaceuticals. A limit of detection (LOD) of 0.72 µmol L -1 was achieved, with a linear range of 10 to 200 µmol L -1 . Analysis of real samples provided results that were highly concordant with those obtained by UV-Vis spectrophotometry (relative error ≤ 1.50%). In addition, the greenness of this approach was evaluated and confirmed by a quantitative methodology (Eco-Scale index). Thus, the MSLA-3D-PDE appears as a new and sustainable tool with great potential of use in analytical electrochemistry., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Purpose: To estimate the taper of root canals of deciduous maxillary and mandibular canines by nano computed tomography (nano-CT)., Methods: This in vitro study involved CT scan analysis of nine maxillary and five mandibular primary canines. The images of each tooth were reconstructed using OnDemand3D software. Thereon, diameter and taper analyses were performed on the free FreeCAD 0.18 software for the three-dimensional (3D) computer-aided design model. Statistical analysis was conducted using Stata v14.0 software, adopting a significance level of 5%., Results: 3D image reconstruction was performed, considering the diameters obtained along the entire length of the tooth root, and the conical model was built with a height of 10 mm. The diameters of the maxillary canine at points D0 (0 mm), D5 (5 mm), D7 (7 mm), and D10 (10 mm) were 1.62, 1.07, 0.78, and 0.49 mm, respectively, with a significant difference between the four points (p = 0.0001). Regarding maxillary canine root taper values in the cervical, middle, and apical regions, the values were 12%, 14%, and 10%, respectively. For mandibular canines, the mean diameter values obtained at points D0, D5, D7, and D10 were 1.51, 0.83, 0.64, and 0.45 mm, respectively, with significant differences among the four points (p = 0.005). The inferior canine root tapers in the cervical, middle, and apical regions were 14%, 10%, and 6%, respectively., Conclusion: The detailed knowledge of the root morphology of maxillary and mandibular deciduous canines, as it has been shown in vitro using nano-CT, is critical to achieve accurate and efficient endodontic treatments., (© 2023. The Author(s), under exclusive licence to European Academy of Paediatric Dentistry.)

The aim of this study was to evaluate the sensitivity, specificity, and predictive values of the fluorescence microscopy method in the detection of apical dental reabsorption after induction of apical periodontitis in animal models. Forty-first molars of mice, aged 6 to 8 weeks, had their root canals exposed to the oral environment or were maintained healthy as controls ( n = 20). After 14 and 42 days, mice were euthanized and tissues were collected for histological evaluation by means of bright field and fluorescence microscopy. The accuracy of fluorescence microscopy in identifying apical external dental resorption was investigated using a diagnostic validation test based on the sensitivity (S) and specificity (E) properties. Bright-field microscopy revealed a higher number of specimens with scores of 1 to 3 - absence of apical dental resorption ( n = 29; 52%), while fluorescence microscopy revealed a higher number of specimens with scores of 4 to 6 - presence of apical dental resorption ( n = 37; 66%). Out of 56 specimens, 26 were TP, 11 were FP, and 19 were TN. No FN result was observed. Fluorescence microscopy presented a sensitivity value of 1, similar to the bright-field method, while specificity was lower (0.633). The accuracy of the fluorescent method to detect apical dental resorption was 0.804. Fluorescence microscopy revealed a higher number of false positive apical dental resorption than bright-field microscopy. The detection of apical dental resorption was not impacted by the sensitivity of the method but by its specificity., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2023 Marcio Santos de Carvalho et al.)

Purpose: We evaluated bacterial endotoxin adhesion, superficial micromorphology and mechanical properties of latex and non-latex intermaxillary orthodontic elastics., Methods: To quantify the adhered bacterial endotoxin, elastics were divided into 5 groups: experimental (n = 12) latex and non-latex elastics, previously contaminated by an endotoxin solution, negative control (n = 6) latex and non-latex elastics without contamination, and positive control (n = 6) stainless steel specimens (metallic replicas), contaminated by an endotoxin solution. In parallel, the structural micromorphology (n = 6) and surface roughness of latex and non-latex intermaxillary orthodontic elastics were assessed using confocal laser microscopy. Force degradation (g) and deformation of the internal diameter change (mm) were also evaluated. Structural micromorphology, surface roughness (µm), force degradation (g) and internal diameter (mm) change were evaluated at time 0 and after 24 and 72 h in a deformation test. Data were analyzed by the Shapiro-Wilk, Kruskal-Wallis, Dunn, ANOVA and Bonferroni tests (α = 5%)., Results: Endotoxin adhered similarly to both types of elastics with scores of 3 (> 1.0 EU/mL). The surface microstructure of both types of elastics showed irregularities and porosities at all times. Initially, the latex elastics had a higher surface roughness (p < 0.001) than the non-latex ones. After 24 h loading, surface roughness of the latex elastics was significantly reduced (p < 0.001), while after 72 h, the values were similar for both types (p > 0.05). The non-latex elastics had significantly higher force generation values (p < 0.05) at 0, 24 and 72 h compared with the latex elastics, although there was a significant reduction (p < 0.001) in force over time for both elastics. Despite similar initial values, non-latex elastics had a significantly larger internal diameter (p < 0.001) after the loading periods of 24 and 72 h compared with the latex elastics., Conclusion: Both elastics showed high affinity with endotoxin and microstructural irregularities of their surface. The non-latex elastics generated higher force values but demonstrated greater deformation of the internal diameter after loading., (© 2022. Springer Medizin Verlag GmbH, ein Teil von Springer Nature.)

Background: To investigate if 5-LO selective inhibitor (MK-886) could be used for systemic treatment of experimentally induced apical periodontitis in a mouse model., Methods: Twenty-four C57BL/6 mice were used. After coronal opening, a solution containing Escherichia coli LPS (1.0 µg/µL) was inoculated into the root canals of the lower and upper right first molars (n = 72 teeth). After 30 days apical periodontitis was established, and the animals were treated with MK-886 (5 mg/kg), a 5-LO inhibitor, for 7 and 14 days. The tissues were removed for histopathological and histometric analyses, evaluation of osteoclast number and gene expression for receptor activator of nuclear factor kappa-B (Tnfrsf11a), receptor activator of nuclear factor kappa-B ligand (Tnfsf11), osteoprotegerin (Tnfrsf11b), tartrate-resistant acid phosphatase (Acp5), matrix metalloproteinase-9 (Mmp9), cathepsin K (Ctsk) and calcitonin receptor (Calcr). Statistical data analysis was performed using Kruskal Wallis followed by Dunn's tests (α = 0.05)., Results: Administration of MK-886 for 7 days exerted no effect on apical periodontitis progression compared to LPS inoculation without treatment (p = 0.3549), while treatment for 14 days exacerbated bone loss (p < 0.0001). Administration of MK-886 enhanced osteoclastogenesis signaling and osteoclast formation within 7 days (p = 0.0005), but exerted no effect at 14 days (p > 0.9999). After 7 days of treatment, MK-886 induced mRNA expression for Acp5 (p = 0.0001), Calcr (p = 0.0003), Mmp9 (p = 0.0005) and Ctsk (p = 0.0008), however no effect in those gene expression was observed after 14 days (p > 0.05)., Conclusion: Systemic treatment with MK-886 exacerbated LPS-induced apical periodontitis in a mouse model., (© 2023. The Author(s).)

Aim: The aim of this study was to evaluate the IFI16 and IFN-α/β receptors expression during the genesis and development of experimental apical periodontitis (AP) in mice teeth., Methodology: Apical periodontitis was induced in the lower first molars of 40 C57BL/6 mice. They were divided according to the experimental periods 2, 7, 14, 21 and 42 days (n = 8 per group). Five animals were used as a control group (without AP). Specimens were submitted to histological processing for description of the inflammatory process, immunostaining for the presence/absence and localization of IFI16 and IFN-α/β receptors (qualitative and semi-quantitative analysis) and tartrate-resistant acid phosphatase (TRAP) histoenzimology., Results: The results showed a gradual development of AP over the experimental times. The expression of IFI16 was noticeably more exacerbated in the experimental early period (day 2) whilst the lowest expression was observed in the control group (p = .02). For IFN-α/β receptors, a higher intensity staining was observed 42 days after AP induction, that was statistically different from the control group (p = .02). In addition, the number of TRAP-positive cells was higher on the later periods (days 21 and 42; p < .001)., Conclusion: IFI16 protein expression was highest during the early periods after AP induction in mice teeth, whilst IFN-α/β receptor expression was highest after AP became established., (© 2022 British Endodontic Society. Published by John Wiley & Sons Ltd.)

Background: Periodontal destruction can be the result of different known and yet-to-be-discovered biological pathways. Recent human genetic association studies have implicated interferon-gamma inducible protein 16 (IFI16) and absent in melanoma 2 (AIM2) with high periodontal interleukin (IL)-1β levels and more destructive disease, but mechanistic evidence is lacking. Here, we sought to experimentally validate these observational associations and better understand IFI16 and AIM2's roles in periodontitis., Methods: Periodontitis was induced in Ifi204 -/- (IFI16 murine homolog) and Aim2 -/- mice using the ligature model. Chimeric mice were created to identify the main source cells of Ifi204 in the periodontium. IFI16-silenced human endothelial cells were treated with periodontal pathogens in vitro. Periodontal tissues from Ifi204 -/- mice were evaluated for alveolar bone (micro-CT), cell inflammatory infiltration (MPO+ staining), Il1b (qRT-PCR), and osteoclast numbers (cathepsin K+ staining)., Results: Ifi204-deficient mice> exhibited >20% higher alveolar bone loss than wild-type (WT) (P < 0.05), while no significant difference was found in Aim2 -/- mice. Ifi204's effect on bone loss was primarily mediated by a nonbone marrow source and was independent of Aim2. Ifi204-deficient mice had greater neutrophil/macrophage trafficking into gingival tissues regardless of periodontitis development compared to WT. In human endothelial cells, IFI16 decreased the chemokine response to periodontal pathogens. In murine periodontitis, Ifi204 depletion elevated gingival Il1b and increased osteoclast numbers at diseased sites (P < 0.05)., Conclusions: These findings support IFI16's role as a novel regulator of inflammatory cell trafficking to the periodontium that protects against bone loss and offers potential targets for the development of new periodontal disease biomarkers and therapeutics., (© 2022 American Academy of Periodontology.)

Different types of brackets seem to influence the disruption of the oral microbial environment. Therefore, the aim of this study was to evaluate the influence of self-ligating brackets on the gingival crevicular fluid levels of the putative periodontal pathogens Aggregatibacter actinomycetemcomitans sorotype a (Aaa), Tannerella forsythia, Fusobacterium nucleatum, and Porphyromonas gingivalis. Sixty samples of crevicular fluid of twenty patients (11 boys and 9 girls) were analysed at baseline (T0) and after 30 (T1) and 60 (T2) days of bonding of the self-ligating (In-Ovation ® R, Dentsply, GAC or SmartClip™, 3 M Unitek, Monrovia, CA, USA) and of one conventional bracket (Gemini™, 3 M Unitek, Monrovia, CA, USA) used with elastomeric ligatures. Total DNA from samples was extracted using CTAB-DNA precipitation method and Real-time PCR was performed to analyse bacterial level. Non-parametric Friedman and Wilcoxon tests were used for data analysis (p value of < 0.05). F. nucleatum presented a different level among the different brackets at T1 (p = 0.025), the highest level in the Gemini™ bracket when compared to the SmartClip™ bracket (p = 0.043). P. ginigvalis levels increased in the In-Ovation ® R (p = 0.028) at T1. The subgingival levels of bacterial species associated with periodontal disease P. ginigvalis increased in the self-ligating brackets In-Ovation ® R.Clinical Relevance: Some kinds of brackets could provide more retentive sites than others, and it seems to modulate the subgingival microbiota, since, in this study, we could observe the increase of the species associated with periodontal disease. Preventive protocols should be adopted in the use of self-ligating brackets., (© 2021. The Society of The Nippon Dental University.)

Apical periodontitis is an immune inflammatory response around periapical tissues as a result of pathogens invasion into the root canal. The host immunoinflammatory response could determine the progression of this disease, which involves the recruitment of immune cells, and the release of several cytokines in the lesion site. The 5-lipoxygenase pathway has been activated in some osteolytic diseases due to its capacity to interfere in the proliferation and differentiation of bone cells, including the osteoclasts. As mean to understand the inflammatory genes regulation in the apical periodontitis progression, we evaluated the network of 66 genes related to cytokines, chemokines and other inflammatory mediators and receptors in the wild-type (WT) and 5-lipoxygenase enzyme genetically deficient mice (KO). This article presents data not published but related to the research article "Effects of 5-lipoxygenase gene disruption on inflammation, osteoclastogenesis and bone resorption in polymicrobial apical periodontitis" ., Competing Interests: The authors declare no conflict of interest for this article., (© 2022 The Author(s). Published by Elsevier Inc.)

Objective: To verify the association between 25(OH)D level and polymorphisms in the vitamin D receptor gene ( VDR ) with the disturbance in the dental development and eruption., Design: A total of 183 children from two datasets were evaluated. The first dataset was a case-control (15:15) designed to assess if persistent primary tooth (PPT) is associate with serum 25(OH)D level and with genetic polymorphisms in VDR . The second dataset of genomic DNA samples from 54 children with delayed tooth eruption (DTE) and 99 controls were analysed to verify if genetic polymorphisms in VDR (rs2228570 and rs739837) are associated with DTE. The 25(OH)D and the genotyping/allele distribution were analysed using the T -test and chi-square test, respectively., Results: The level of 25(OH)D in the PPT group (24.9 ± 6.4 mg/mL) was significantly lower than the control (30.0 ± 7.0 mg/mL) ( p =.047). Our data show that children with 25(OH)D deficiency are more likely to present PPT (OR = 2.36; 95%CI: 1.51, 3.70). The rs739837 and rs2228570 polymorphisms were not associated with DTE (OR = 1.44; 95%CI: 0.87, 2.39 and OR = 0.80; 95%CI: 0.45, 1.44, respectively)., Conclusions: Vitamin D deficiency is a risk factor for PPT.

Objectives: The objective of this study was to evaluate the effect of non-steroidal anti-inflammatory drugs (NSAIDs) in controlling pulpal and periapical inflammation in vivo as a potential coadjutant systemic therapy for pulpitis., Materials and Methods: A suspension containing E. coli lipopolysaccharide (LPS; 1.0 μg/μL) was inoculated into the pulp chamber of the first molars of C57BL/6 mice (n = 72), and the animals were treated daily with indomethacin or celecoxib throughout the experimental periods. After 7, 14, 21, and 28 days, the tissues were removed for histopathological, histoenzymology, histometric, and immunohistochemical evaluation., Results: Inoculation of LPS into the pulp chamber induced the synthesis of the enzyme cyclooxygenase-2 (COX-2) in dental pulp and periapical region. Indomethacin and celecoxib treatment changed the profile of inflammatory cells recruited to dental pulp and to the periapex, which was characterized by a higher mononuclear cell infiltrate, compared to LPS inoculation alone which recruited a higher amount of polymorphonuclear neutrophils. Administration of indomethacin for 28 days resulted in the development of apical periodontitis and increased osteoclast recruitment, unlike celecoxib., Conclusions: NSAIDs indomethacin and celecoxib changed the recruitment of inflammatory cells to a mononuclear profile upon inoculation of LPS into the pup chamber, but indomethacin enhanced periapical bone loss whereas celecoxib did not., Clinical Relevance: Celecoxib, a selective COX-2 inhibitor, can change the profile of inflammatory cells recruited to the dental pulp challenged with LPS and might a be potential systemic coadjutant for treatment of pulpitis., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

3D printing is a hot topic in electroanalytical chemistry, allowing the construction of custom cells and sensors at affordable prices. In this work, we describe a novel small and practical 3D-printed electrochemical cell. The cell's body, manufactured in ABS on a 3D printer, is composed by three parts easily screwed: solution vessel, stick and cover with two embedded 3D-pen-printed carbon black-polylactic acid (CB-PLA) electrodes (counter and pseudo-reference). The cell is compatible with any planar working electrode, in which boron-doped diamond, graphite sheet (GS) and 3D-printed CB-PLA were shown as examples. A new alternative protocol to quickly produce 3D-printed sensors using a 3D pen and other low-cost apparatus is also proposed. The voltammetric performance of each evaluated sensor was carried out in the presence of redox probe ferricyanide and paracetamol as model analyte, and the surfaces were characterized by electrochemical impedance spectroscopy and scanning electrochemical microscopy. To present an analytical application of the 3D-printed cell, low-cost flexible sensors (GS and CB-PLA) were used as integrated platforms for sampling and detection of solid drugs. As a proof-of-concept, traces of drugs with a historic of counterfeit or adulteration (sildenafil citrate, tadalafil, losartan and 17α-ethinylestradiol) were abrasively sampled over the sensor and assembled on 3D-printed cell to perform a fast voltammetric scan in the presence of only 500 μL of electrolyte. This protocol is attractive for pharmaceutical and forensic sciences as a simple preliminary screening method which could identify the presence or absence of the suspicious drug as well as impurities or adulterants. The 3D-printed cell was also used for the determination of 17α-ethinylestradiol in a contraceptive pill to demonstrate a quantitative analysis. The cell is quickly printed (90 min), cheap (US$ 0.30) and requires low electrolyte volumes (0.5-3.0 mL), being suitable to be used in several other electroanalyses, especially for on-site applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Antimicrobial photodynamic therapy (aPDT) is a complementary therapeutic modality for periodontal and endodontic diseases, in which Gram-negative bacteria are directly involved. Currently, there are few evidences regarding the effects of aPDT on bacterial components such as lipopolysaccharide (LPS) and it would represent a major step forward in the clinical use of this therapy. In this context, this study aimed to evaluate the efficacy of different photosensitizers (PSs) used in aPDT in LPS inhibition. Four PSs were used in this study: methylene blue (MB), toluidine blue (TBO), new methylene blue (NMB), and curcumin (CUR). Different approaches to evaluate LPS interaction with PSs were used, such as spectrophotometry, Limulus amebocyte lysate (LAL) test, functional assays using mouse macrophages, and an in vivo model of LPS injection. Spectrophotometry showed that LPS decreased the absorbance of all PSs used, indicating interactions between the two species. LAL assay revealed significant differences in LPS concentrations upon pre-incubation with the different PSs. Interestingly, the inflammatory potential of LPS decreased after previous treatment with the four PSs, resulting in decreased secretion of inflammatory cytokines by macrophages. In vivo, pre-incubating curcumin with LPS prevented animals from undergoing septic shock within the established time. Using relevant models to study the inflammatory activity of LPS, we found that all PSs used in this work decreased LPS-induced inflammation, with a more striking effect observed for NMB and curcumin. These data advance the understanding of the mechanisms of LPS inhibition by PSs.

Objectives: To answer the questions: (1) Does reducing estrogen levels influence the microbial composition of the oral cavity? (2) Does the presence of periapical lesion (PL) cause changes in the oral microbiota? (3) Since estrogen deficiency alters the oral microbiota, can this be one of the factors that contribute to the increase of the PL?, Materials and Methods: Thirty-six rats were divided into four groups: sham (control), ovariectomy (OVX), control with PL (Sham + PL), and OVX + PL. After 9 weeks of OVX, the lower first molars were submitted to PL induction. After 21 days, the microbiological collection of the oral cavity was performed, and the animals were euthanized. The contents were evaluated by the checkerboard DNA-DNA hybridization method, to verify the prevalence of 40 bacterial species (divided into 7 microbial complexes). The blocks containing the lower first molars were submitted to histotechnical processing and staining with hematoxylin and eosin (HE), for the measurement of the periapical lesion area. The results were submitted to ANOVA and Kruskal-Wallis tests and Tukey and Dunn post-tests, with a significance level of 5%., Results: In conditions of estrogen deficiency, there was alteration of the oral microbiota. The OVX groups had a higher amount of bacteria compared to the SHAM group in most of the microbial complexes (p < 0.001). The animals in the control group (with or without lesion) did not present a statistically significant difference (p > 0.001) in any of the microbial complexes. The PLs in OVX animals were significantly higher compared to SHAM animals (p < 0.001)., Conclusions: Hypoestrogenicity conditions interfere in the oral microbiota by increasing the amount of bacteria in the saliva and influencing the progression of periapical lesions., Clinical Relevance: This inedited study shows that deficiency of estrogen leads to alteration of the oral microbiota.

Introduction: The objective of this study was to evaluate in vitro and in vivo bacterial endotoxin (LPS) adhesion in polyurethane and silicone esthetic elastomeric orthodontic ligatures. The null hypotheses tested were: (1) there is no LPS adhesion in esthetic elastomeric orthodontic ligatures; and (2) there is no difference in the LPS adhesion between different brands of these ligatures., Methods: For the in vitro study, 4 types of esthetic elastomeric ligatures were used (Sani-Ties and Sili-Ties [Dentsply GAC, Islandia, NY;] and Mini Single Case Ligature Stick and Synergy low-friction ligatures [Rocky Mountain Orthodontics, Denver, Colo]), contaminated or not with endotoxin solution. Replicas of twisted wire and cast stainless steel ligatures were used as control. For the in vivo study, 10 male and 10 female patients, aged 15-30 years, received the same 4 types of ligatures, 1 of each inserted in the maxillary and mandibular canines, randomly. Twenty-one days later, the ligatures were removed, and endotoxin quantification was performed using the Limulus amebocyte lysate test. Data were analyzed (α = 0.05) using the Kruskal-Wallis test and Dunn's posttest or analysis of variance and Tukey's posttest., Results: GAC silicone group had the lowest median contamination (1.15 endotoxin units/mL; P <0.0001) in vitro. In the in vivo study, the GAC silicone group had the lowest mean contamination (0.577 endotoxin units/mL; P <0.001). In both studies, the other groups did not present a significant difference when compared with each other (P >0.05)., Conclusions: LPS exhibited an affinity for all the tested polyurethane and silicone elastomeric ligatures. GAC silicone ligatures presented with lower amounts of LPS attached to their surfaces. Thus, both null hypotheses were rejected., (Copyright © 2021 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

The aim of this study was to evaluate the M1 and M2 macrophage modulation after stimuli with different materials used during endodontic treatment. In bone marrow-derived macrophage cell culture, from males C57BL/6 wild-type (WT) mice, gene expression analysis of markers to M1 and M2 macrophages was performed by qRT-PCR (Cxcl10, CxCL9, iNOS, Arg1, Chil3, Retnla and MRC1) and cytokine quantification by Luminex® (GM-CSF, IL-10, IL-6, IL-1β and TNF-α) after exposure to the five endodontic sealers: AH Plus, Sealapex Xpress, Endosequence BC Sealer, BioRoot RCS and a calcium hydroxide-based paste. For normal values, ANOVA test was used, followed by Tukey post-test. For non-normal values, the Kruskall-Wallis test was used. BioRootTM RCS and EndoSequence BC SealerTM stimulated the highest expression of markers for M1 macrophages, while calcium hydroxide-based paste stimulated the lowest expression of these gene markers. For M2 protein markers, BioRootTM RCS presented the highest stimulation while calcium hydroxide-based paste also presented the lowest stimulation. It was concluded that all the evaluated filling materials increased the genetic expression of pro- and anti-inflammatory markers: TNF-α and IL-10 respectively. The others proinflammatory mediators showed differences against the filling materials. However, this process did not induce the inflammatory response polarization, resulting in a hybrid macrophage.

Aim: To characterize plasma cell subsets in chronic periapical lesions affecting permanent and primary teeth., Methodology: Only chronic periapical lesions without root canal treatment were selected. Twenty-one radicular cysts and 7 periapical granulomas affecting permanent teeth and 19 radicular cysts and 4 periapical granulomas affecting primary teeth were assessed for immunoglobulin (Ig) light chain (kappa and lambda), Ig heavy chain (IgG, IgG4, IgA, IgM and IgD) and plasma cell immunohistochemical markers (MUM1/IRF4, EMA and CD138). The data acquired were analysed by Student's t test, Mann-Whitney U, Friedman test followed by Dunn's multiple comparison test and Spearman's rank correlation., Results: All cases were polyclonal (having similar kappa/lambda light chain ratios). IgG was most abundant compared to other Ig heavy chains (all, P < 0.001); like Ig light chains, but unlike IgA, there was greater expression of IgG in the primary compared to the permanent dentition, for both radicular cysts (P < 0.001) and periapical granulomas (P = 0.53). Notably, IgG4 expression was greater in the permanent than the primary dentition, for both radicular cyst (P < 0.05) and periapical granuloma (P = 0.65). IgM and IgD expression was scarce and variable, whereas plasma cell populations were detected efficiently through EMA, CD138 and MUM1/IRF4 markers, the latter being more sensitive in both dentitions., Conclusions: There were slight variations in the Ig light and heavy chain profiles in chronic periapical lesions when comparing the permanent and primary dentitions. The ability of IgG4+ plasma cell infiltration to modulate inflammatory responses in chronic periapical lesions arising from permanent as opposed to primary teeth should be considered in future studies., (© 2020 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Objectives: To evaluate denosumab, a human monoclonal antibody that mimics the effects of osteoprotegerin in bone metabolism, as a topical treatment of root surface to be used prior to delayed tooth replantation., Materials and Methods: Thirty-six rats' right incisors were used. Teeth were extracted and divided into: delayed replantation without root surface treatment (control); delayed replantation with root surface treatment with denosumab 60 mg/mL and 30 mg/mL, respectively, for 10 min both experimentals groups. After that, the root canals were filled with calcium hydroxide and replanted. After 15 and 60 days, the animals were euthanized, and the samples were collected and processed for microscopic analysis. Histological sections were performed, and stained with HE to describe the dental characteristics, measure ankylosis, replacement resorption, and dental resorption by conventional microscopy. Also, was performed Brown & Brenn staining and immunohistochemistry for RANKL, OPG, and periostin., Results: Denosumab 60 mg/mL reducted ankylosis (p < 0.0001), replacement resorption (p < 0.0001), and tooth resorption, 60 days after replantation, compared to untreated replanted teeth (p < 0.005). Lower bacterial contamination in root surface in the denosumab treatment groups was found, regardless of the concentration used (p < 0.001). Also, denosumab treatment inhibited the expression of RANKL without modulating OPG. Periostin was observed in periodontal ligament of replanted tooth, although this labelling was absent in the ankylosis areas, in both experimental periods., Conclusion: Treatment of the root surface with denosumab at 60 mg/mL of rat teeth before delayed replantation reduced dental root resorption compared with the untreated teeth after 60 days., Clinical Relevance: Survival of a replanted tooth has been a challenge in clinical practice. The use of a medication, such as denosumab, to limit dental root resorption represents an important therapeutical approach.

This work presents the association of cloud point extraction (CPE) and electroanalysis for the selective and sensitive determination of methyl parathion (MP) in honey. The CPE step provided the pre-concentration of MP from a complex sample, in which the optimized extraction parameters (Triton X-100 concentration of 0.75% w/v, NaCl concentration of 1.0% w/v and heating time of 30 min) were investigated using a factorial design (23). The detection of MP was performed using a cathodically pre-treated boron-doped diamond (BDD) working electrode and square wave voltammetry (SWV), after a suitable dilution of the CPE extract in Britton-Robinson buffer pH 6.0 as the supporting electrolyte. MP presented three electrochemical processes over the BDD surface, but only the reduction peak at around -0.7 V was monitored for the MP determination (higher detectability). Improved reproducibility was reached by applying an in situ cleaning step (+2.0 V for 15 s) followed by a re-activation process (-2.0 V for 15 s) between measurements. Using the optimized variables, a linear range between 0.1 and 2.0 μmol L-1 was obtained for MP with a limit of detection of 0.006 μmol L-1, a 6-fold lower value when compared with the value attained without the CPE step. The experimental enrichment factor of MP was 6.1. Also, the optimized CPE allowed the determination of MP in honey samples with good accuracy (recovery between 94 and 106%), which was not possible using direct detection (without CPE) due to the matrix interference. This is the first paper that demonstrates the combination of CPE and electroanalysis for the determination of an organic compound.

Aim: To investigate the presence, localization and the possible correlation of the fibroblast growth factor receptor-2 (FGFR2) with inflammatory resorption of cementum, periodontal ligament and alveolar bone during development of apical periodontitis in mice., Methodology: Apical periodontitis was experimentally induced in mandibular first molars of mice by pulp exposure to the oral environment. Healthy teeth without pulp exposure were used as controls. At 7, 21 and 42 days following pulp exposure, the animals were euthanized and the jaws were prepared for analysis under conventional and fluorescence microscopy, immunohistochemistry (FGFR2), RT-PCR (RNAm levels of RANK, RANKL, OPG, Runx2 and cathepsin K) and enzyme histochemistry (cementoclasts and osteoclasts). Statistical analysis was performed by Kruskal-Wallis tests and Dunn's post hoc tests for multiple comparisons (α = 0.05) using SAS 9.4 software., Results: FGFR2-positive cells were not observed in the tissues surrounding healthy teeth but were observed in teeth with periapical lesions from seven days after root canal contamination. At days 21 and 42 after endodontic infection, the increase in periapical lesion size was accompanied by significantly enhanced expression of FGFR2 (P < 0.0001), significantly increased intensity of inflammatory cells, number of osteoclasts (P < 0.0001) and cementoclasts (P < 0.0001), and significantly enhanced RNAm levels of RANK/RANKL/OPG, Runx2 and cathepsin K compared to day 0 (P < 0.0001). At 21 and 42 days, FGFR2 was also expressed on osteoblasts, fibroblasts and inside enlarged lacunae of cementocytes along with acute and chronic inflammatory cells (macrophages, plasma cells and neutrophils). At all periods and cells, FGFR2 expression was observed in the cell membrane and cytoplasm, but not in the nucleus., Conclusion: In mice, FGFR2 was not expressed in tissues surrounding healthy teeth but was expressed in apical periodontitis, specifically in the membrane and cytoplasm of osteoblasts, fibroblasts, lacunae of cementocytes, and acute and chronic inflammatory cells (macrophages, plasma cells and neutrophils). Its expression was correlated with the size of the periapical lesions., (© 2020 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Objectives: To investigate the regulation of inflammatory and osteoclastogenic signaling by 5-lipoxygenase (5-LO) in apical periodontitis induced by oral contamination of dental root canals in mice., Design: Apical periodontitis was induced in 5-lipoxygenase enzyme knockout (129-Alox5 tm1Fun ) and 129 wild-type mice (n = 96) by exposure of the dental root canal to the oral cavity. After 7, 14, 21, and 28 days, the animals were euthanized and the tissues removed (n = 12 teeth per period) for histopathological and histometric analyses (hematoxylin and eosin [HE]), evaluation of osteoclastogenic activity (tartrate-resistant acid phosphatase enzyme [TRAP]), and determination of inflammatory and osteoclastogenic signaling (qRT-PCR)., Results: Oral contamination of dental root canals induced recruitment of neutrophils and osteoclasts to the periodontal ligament, resulting in bone resorption. Absence of 5-LO did not impair neutrophil recruitment while osteoclastic formation was increased. Nonetheless, early bone resorption progressed similarly to lesions in wild-type animals. Interestingly, in the absence of 5-LO, the synthesis of mRNAs for cytokines, chemokines, and their receptors was significantly reduced while that of regulators of osteoclastogenesis (RANK, RANKL, and OPG) was increased in comparison with the corresponding levels in wild-type animals., Conclusions: The 5-LO pathway plays a role in the stimulation of inflammatory mediator synthesis and inhibition of osteoclastogenesis in apical periodontitis in mice. However, the paradoxical inflammatory-osteoclastogenic signaling did not impair inflammatory cell recruitment and bone resorption during early development of the disease., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Background: The objective was to analyze clinical, microbiological, and immunological periodontal parameters in patients in corrective orthodontic treatment., Materials and Methods: Twenty-eight patients were selected. Plaque index (PI), bleeding on probing (BOP), width of keratinized gingiva, levels of 40 bacterial species, and of 3 cytokines (IL-1β, MMP-8, and TNF-α) in gingival crevicular fluid (GCF) were evaluated at T0, before orthodontic treatment; T1, 6 months; and T2, 12 months post-treatment. Non-parametric, Friedman, Wilcoxon, ANOVA, and Spearman correlation coefficient tests were used for statistical analyses, with the significance level of 5%., Results: No significant difference was found for the width of keratinized gingiva, but PI presented a significant increase at T1 and T2 (p < 0.05) when compared with T0. The percentage of sites with BOP increased significantly from T0 to T1 (p < 0.05); however, at T2, the values decreased and did not differ anymore from T0 (p > 0.05). In the microbiological analysis, red complex pathogens were in significantly greater proportions in T2 compared with T0 (p < 0.05). There was no statistically significant difference in the cytokine levels between the periods but there was a positive correlation between BOP and IL-1β (r = 0.49 p = .01) and TNF-α (r = 0.39 and p = .05)., Conclusion: In conclusion, corrective orthodontic treatment caused clinical periodontal alterations regarding biofilm accumulation and gingival bleeding, with alteration of periodontopathogens.

Objectives: The objective of this study was to evaluate the efficiency of SB-20 M culture medium to perform differential morphological identification of S. mutans and S. sobrinus compared to biochemical identification and to proteomic identification by the MALDI-TOF mass spectrometry technique., Material and Methods: Unstimulated saliva samples from 266 dental students were seeded on SB-20 M culture medium by the wooden spatula technique. After incubation, S. mutans and S. sobrinus colonies were identified by stereomicroscopy based on their differential morphological characteristics. Following these procedures, 135 colonies with characteristic morphology of S. mutans (89 colonies) and S. sobrinus (46 colonies) were randomly selected, submitted to biochemical identification (biotyping) and proteomic identification by the MALDI-TOF mass spectrometry technique. The results were compared using the Kappa test, with a 5% significance level., Results: All (100%) S. mutans colonies were correctly identified after culture in SB-20 M medium compared to biotyping and proteomic identification. For S. sobrinus, morphological identification in SB-20 M medium was correct for 43 colonies (93.5%) compared to biotyping and proteomic identification. However, there was no statistically significant difference when comparing the capacity to identify S. mutans and S. sobrinus of the three techniques (p < 0.001; K = 0.951)., Conclusions: It was concluded that the SB-20 M culture medium for morphological identification of S. mutans and S. sobrinus was highly reliable, being comparable to the MALDI-TOF mass spectrometry technique., Clinical Relevance: The efficiency evaluation of identification methods of S. mutans and S. sobrinus is clinically relevant in order to determine caries risk and activity of patients., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Aim: To assess the efficacy of treatment using a minimally invasive approach (selective removal of carious tissue, restoration and preventive strategies) in immature permanent molars with MIH., Design: A total of 281 patients, aged 6-8 years, with carious lesions (ICDAS 5-6), severe MIH, and incomplete root formation (one tooth/patient) were included. After clinical and radiographic examinations, selective carious tissue removal was performed, and the teeth received interim restoration for 6 months and were then restored with composite resin. Clinical and radiographic follow-up was undertaken, 6, 12, 18, and 24 months. A protocol of preventive oral care measures was established and repeated at each follow-up, including diet counselling, oral hygiene instruction, dental plaque control, and topical application of fluoride varnish containing CPP-ACP. All clinical procedures and evaluations were done by a single operator., Results: Clinical and radiographic success was observed 24 months after treatment in 96.8% of the cases. Failures were due to enamel fracture at restoration margins, resulting in pulpitis and absence of apex closure., Conclusion: Selective removal of carious tissue, interim, and subsequently definitive restoration, combined with home and professional preventive measures, maintained marginal integrity of restorations in immature permanent molars with severe MIH, confirmed by pulp vitality and occurrence of apexogenesis., (© 2019 BSPD, IAPD and John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Objectives: To evaluate the in vitro effects of radiotherapy (RT) on the morphological surface of the enamel and dentin and to determine the best adhesive system and most appropriate time to restore teeth in head and neck cancer patients., Methods: Sixty third molars were cut into 120 enamel fragments and 120 dentin fragments and divided into four groups (n = 30): G1 (control): nonirradiated, only restorative procedure; G2: restorative procedure immediately before RT; G3: restorative procedure immediately after RT; and G4: restorative procedure 6 months after RT. Each group was divided into two subgroups: Adper™ Single Bond 2 (SB) and Clearfill SE Bond (CL) based on the material used. After RT and restorative procedures, the specimens were subjected to confocal microscopy and shear bond strength test. Data were analyzed using a two-way ANOVA followed by Tukey's test at a significance level of 5%., Results: Morphological changes were observed in both substrates after a cumulative dose of 40 Gy, and after 60 Gy, the changes were more evident in both substrates. CL had the highest strength values in both substrates (p < 0.05), and G2 had the lowest strength values for the enamel and dentin (p < 0.05)., Conclusions: Based on the in vitro study results, we can conclude that RT substantially changes the morphological surface of enamel and dentin and impairs the bond strength. The Clearfill system yielded better results than Adper Single Bond 2, and restoring teeth before RT resulted in the worst results in both substrates.

Background: Tooth eruption is a process that is not fully understood., Aim: To evaluate whether genetic polymorphisms for RANK/RANKL/OPG are associated with delayed tooth emergence. To evaluate whether the relative expression of this genes is associated with persistent primary teeth., Design: To evaluate whether genetic polymorphisms for RANK/RANKL/OPG could be involved in delayed tooth emergence, saliva samples from 160 children, aged 6-13 years old, were analysed. To test if there is correlation between gene expression of RANK/RANKL/OPG in children with delayed tooth emergence and persistent primary teeth, periapical tissue from 15 children with persistent primary teeth and from 15 control subjects were collected for qPCR analysis., Results: Fifty-six children with delayed tooth emergence (35%) had at least one permanent tooth with delayed emergence. The T allele in RANKL (rs9594738) increased the risk of delayed tooth emergence (P = 0.02; OR = 1.71, 95%CI 1.09-2.75). The relative gene expression for RANKL and the ratio RANKL/OPG in children with delayed tooth emergence and persistent primary teeth were lower compared to controls (P = 0.02 and P = 0.005, respectively)., Conclusions: Data suggest that the polymorphism rs9594738 in RANKL is associated with delayed permanent tooth emergence. Moreover, reduced relative gene expression of RANKL in periapical tissue is associated with persistent primary teeth., (© 2019 BSPD, IAPD and John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Objectives: To assess periodontal parameters and microbial species levels after orthodontic appliance placement in patients who received oral hygiene instructions and who were monitored and motivated throughout the study., Materials and Methods: The Periodontal Index was recorded and saliva collection was performed before (T0) and 30 (T1), 60 (T2), and 90 (T3) days after orthodontic appliance placement in 15 patients (mean age 17.53 ± 8.0 years). Analysis was carried out using checkerboard DNA-DNA hybridization. Nonparametric statistical analysis was performed., Results: The Periodontal Index did not change. The total amount of the purple and red complexes and Candida species showed a significant decrease from T2. The green, yellow, and orange complex showed a significant decrease at T3. The specific species analysis showed that Prevotella nigrescens, Pseudomonas putida, Fusobacterium periodonticum, Pseudomonas aeruginosa, Peptostreptococcus anaerobius, and Tanerella forsythia showed high incidence before bonding, and their levels decreased at T2 and T3. Only Porphyromonas gingivalis showed increased levels at T2 and displayed the highest level at T3. The Streptococcus group decreased their levels from T2 onward., Conclusions: A dynamic change in microbial levels was identified. The decrease in the levels of complexes present was only possible due to the mechanical method of oral hygiene implemented in this sample.

This work presents potential applications of low-cost fused deposition modeling 3D-printers to fabricate multiuse 3D-printed electrochemical cells for flow or batch measurements as well as the 3D-printing of electrochemical sensing platforms. Electrochemical cells and sensors were printed with acrylonitrile butadiene styrene (ABS) and conductive graphene-doped polylactic acid (G-PLA) filaments, respectively. The overall printing operation time and estimated cost per cell were 6 h and $ 6.00, respectively, while the sensors were printed within minutes (16 sensor strips of 1 × 2 cm in 10 min at a cost of $ 1.00 each sensor). The cell performance is demonstrated for the amperometric detection of tert-butylhydroquinone, dipyrone, dopamine and diclofenac by flow-injection analysis (FIA) and batch-injection analysis (BIA) using different working electrodes, including the proposed 3D-printed sensor, which presented comparable electroanalytical performance with other carbon-based electrodes (LOD of 0.1 μmol L -1 for dopamine). Raman spectroscopy and scanning electron microscopy of the 3D-printed sensor indicated the presence of graphene nanoribbons within the polymeric matrix. Electrochemical impedance spectroscopy and heterogeneous electron transfer constants (k 0 ) for the redox probe Ru(NH 3 ) 6 +3 revealed that a glassy-carbon electrode presented faster electron transfer rates than the 3D-printed sensor; however, the latter presented lower LOD values for dopamine and catechol probably due to oxygenated functional groups at the G-PLA surface., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Objectives: The aim of this study was to evaluate the subcutaneous response induced by Roeko Guttaflow2 (RG), Sealapex Xpress (SX), AH Plus (AHP) sealers., Methods: 100 BALB/c mice received implants in the subcutaneous tissue with the tested materials (10 animals per period for each evaluated sealer) and were evaluated after different experimental periods (7, 21 and 63 days), in each animal was placed a tube, the control group was an empty tube. Histological analysis evaluated semi-quantitatively the inflammatory infiltration, collagen fiber formation and tissue thickness. In addition, immunohistochemistry was performed for interleukin-6 (IL-6). Data were statistically analyzed (α = 0.05)., Results: RG promoted a greater collagen fiber formation at 7 days and 63 days compared to the CG (p = 0.004) and AHP (p = 0.005) respectively, while at 21 days, the SX promoted a greater reaction (p = 0.021). For the tissue thickness, there was a greater reaction at 7 days with CG (p = 0.0156) and with RG at 63 days (p = 0.03). Regarding the inflammatory infiltrate, there was no difference at 7 days and 63 days (p = 0.5; p = 0.27), while at 21 days, a statistically difference was found between SX, CG (p = 0.04) and RG (p = 0.027). In addition, the presence of IL-6 was observed in almost all groups, with a more intense marking at 7days., Significance: All cements evaluated presented a satisfactory tissue response, however, RG was the one that presented a more satisfactory tissue response., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

Objective: The aim of this study was to evaluate the association between genetic polymorphisms in RANK, RANKL and OPG with maxillary and mandibular dimensions in humans., Design: DNA extracted from saliva and the rs3826620, rs9594738 and rs2073618 polymorphisms in RANK, RANKL and OPG, respectively, were analysed by real-time PCR. Four linear measurements (Co-Gn, GoPg, Co-Go and PTM-A) from lateral cephalograms were examined for the evaluation of craniofacial measurements. ANOVA testing and a multivariate linear regression analysis, adjusted for age and gender, were used for statistical analysis, with an alpha of 5%. Hardy-Weinberg equilibrium was also evaluated using the chi-square test within each polymorphism., Setting: School of Dentistry of Ribeirão Preto, University of São Paulo., Participants: A total of 100 unrelated non-syndromic Brazilian Caucasian subjects were included in this study., Results: The polymorphism in RANK was associated with a higher Go-Pg measurement (p = .039). In the multivariate analysis, adjusted for age and gender, the polymorphism in RANK was associated with Go-Pg (p = .017) and Co-Gn (p = .043)., Conclusion: The polymorphism rs3826620 in RANK is associated with the mandibular size.

Objective: The aim of this study was to evaluate bone mineral density (BMD) and microarchitecture in femurs and maxillary bones of ovariectomized (OVX) rats treated or not treated with alendronate (ALD) or odanacatib (ODN)., Study Design: Twenty rats were divided into groups: SHAM, OVX, OVX/ALD, and OVX/ODN. After 12 weeks, the femurs and maxillae were removed and subjected to 3-dimensional analysis by micro-computed tomography. Results were analyzed with 1-way analysis of variance and Tukey's post hoc test (α = 0.05)., Results: OVX decreased maxillary and femoral BMD and altered femoral microarchitecture (P < .05). The drugs increased BMD of both types of bones, but only ALD maintained the phenotype similar to the SHAM group. The action of ALD was limited to the femoral trabecular separation (Tb.Sp). OVX and the drugs had no effect on the microarchitecture of the maxilla (P > .05)., Conclusions: ALD and ODN therapy increased BMD in both bones after ovariectomy. ALD was more successful than ODN in preserving the morphology of bone similar to the SHAM group. ALD maintained the phenotype for Tb.Sp in the femur, but ODN did not. In the maxillae, neither ovariectomy nor the 2 antiresorptive drugs had significant effects on microarchitecture., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Background: Dental treatment can cause symptoms such as fear and anxiety. Audiovisual distraction (AD) is a simple and low-cost technique that does not interfere with the dental treatment., Aim: The aim of this study was to perform a systematic review and meta-analysis to evaluate the effectiveness of AD methods for children who experience anxiety during dental treatment., Materials and Methods: Two reviewers performed a database search of the studies published between January 1950 and November 2015. The inclusion criteria were papers published in the English language, child samples aged 4-10 years, and use of AD. All potentially relevant studies were identified by the title and the abstract. After the full-text analysis of the potentially relevant studies, the selected studies were included in the systematic review. A meta-analysis calculation was performed for the overall data and the subgroup data., Results: Thirty-seven nonduplicated studies were found. However, after reviewing the articles, only five were included. A high variability was observed among the papers. Tools and questionnaires used to measure the anxiety during dental treatment presented the most common variability. Meta-analysis demonstrated a lower anxiety level in AD method groups when Modified Child Dental Anxiety Scale was used (P = 0.02) with a mean difference (confidence interval) of -8.72 (-16.7, -1.38)., Conclusion: The AD method is effective for controlling dental anxiety in children., Competing Interests: There are no conflicts of interest

Introduction: The purpose of this study was to evaluate a protocol for systemic administration of rosiglitazone in mice in order to stimulate apoptosis of osteocytes in the jaws and to evaluate the effect of osteocyte apoptosis induced by rosiglitazone in the progression of periapical lesions in mice at 7, 21, and 42 days., Methods: C57BL/6 mice at 4-5 weeks of age were used. In phase 1, mice (n = 24) were treated with rosiglitazone (gavage, 10 mg/kg dose) or without (phosphate-buffered saline + 10% dimethyl sulfoxide) for 1, 2, or 3 weeks. We used the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling and 4'-6-diamidino-2-phenylindole methods for quantification of apoptotic cells. In phase 2, mice (n = 30) received rosiglitazone for 2 weeks or just vehicle for 1 week (n = 30), and periapical lesions were induced for 7, 21, or 42 days. We performed the measurement of periapical lesions, tartrate-resistant acid phosphatase staining, dual-energy x-ray absorptiometry for the evaluation of bone mineral density (BMD) in long bone, and gene evaluation using real time quantitative polymerase chain reaction of osteocyte markers (Sost, Hyou1, and Dmp1) and receptor activator of nuclear factor kappa-B ligand (RANKL) (Tnfsf11)., Results: It was observed that systemic administration of rosiglitazone for 2 weeks showed apoptosis of osteocytes in a more expressive manner. In phase 2, in the groups that received rosiglitazone, a trend toward larger periapical lesions was observed (P > .05). Rosiglitazone also induced a greater number of osteoclasts and a greater expression of Sost and Hyou1 at 21 days of lesions. Moreover, there were no statistically significant differences in RANKL and Dmp1 expression or in the BMD of femurs., Conclusions: Rosiglitazone stimulated apoptosis of osteocytes, interfering in the progression of periapical lesions in mice., (Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

A strong association between celiac disease (CD) and dental enamel defects (DEDs) have been extensively reported, however, the nature of this relationship is still unclear. The aim of this study was to evaluate DEDs phenotype in CD individuals according to the time they were introduced to a gluten-free diet (GFD). Forty-five CD individuals were examined by a pediatric dentist. DEDs were classified according to the type of affected teeth. CD individuals were classified into two groups (with or without DEDs) and the differences between these groups were tested using chi-square or Fisher´s exact tests and t-test to compare differences between means. The Pearson coefficient test was used to evaluate the degree of the correlation between the age of GFD introduction and number of affected teeth. Individuals with MIH were introduced earlier to the GFD (p = 0.038). An association was also observed for molar DED (p = 0.013). In conclusion, our study suggested an association between a specific type of DED and the time that CD individuals were introduced to a GFD., (© 2017 Special Care Dentistry Association and Wiley Periodicals, Inc.)

Objectives: The aim of this study was to evaluate the association between tooth agenesis and skeletal malocclusions in Brazilian non-syndromic orthodontic patients., Material and Methods: Pretreatment orthodontic records of 348 patients of both genders and with various skeletal malocclusions were examined. Tooth agenesis was evaluated in panoramic radiographs. Angular measurements were taken from lateral cephalometric radiographs to classify the patient's malocclusion as skeletal Class I, Class II and Class III. Subjects were divided into 2 groups, "with tooth agenesis" and "without tooth agenesis". Chi-square or Fisher exact test was used to compare categorical data. ANOVA with Tukey's post-test was used for means comparisons. An alpha of 5% was established., Results: From 348 analysed patients, 28 presented tooth agenesis. There was no difference between genders (P = 0.27) nor mean age (P = 0.16). The most prevalent skeletal malocclusion was Class I (63.11%), followed by Class II (25.94%), and Class III (10.95%). The mean of congenitally missing teeth was 1.3 (SD 0.13). Thirteen subjects had premolar agenesis, 13 upper lateral incisor agenesis, 4 lower incisor agenesis and 2 molars agenesis. The group with tooth agenesis presented A point-nasion-B point (ANB) angle smaller (1.66 [SD 2.52]) than the group without tooth agenesis (2.86 [SD 2.49]) (P = 0.01). ANB angle had a negative correlation with the number of congenitally missing teeth (P = 0.039; r = -0.39)., Conclusions: Tooth agenesis is associated with a smaller A point-nasion-B point angle and is negatively correlated with the number of congenitally missing teeth.

Orthodontic appliances causes specific alterations in oral environment, including reduction of pH, increase of dental biofilm and elevation of salivary microbial levels, causing an increased risk for dental caries. This study evaluated, using microbial culture and scanning electron microscopy (SEM), the in situ contamination by mutans streptococci (MS) of different surfaces of Haas palatal expanders with and without use of chlorhexidine gluconate mouthrinses (CHX). Thirty-four patients were randomly assigned to two groups (n = 17/group), using placebo (Group I) and 0.12% CHX (Group II-Periogard ® ) mouthrinses twice a week. After 4 months, appliances were submitted to microbiological processing and after fragments were analyzed by SEM. Mann-Whitney U test (α = 5%) was used to assess differences between groups on the appliances' different surfaces and to compare the contamination on the free and nonfree surfaces of these components. There was no difference (p = 0.999) between groups regarding the number of MS colonies/biofilms on the nonfree surfaces, which showed intense contamination. However, free surfaces of Group II presented less contamination (p < 0.001) than those of Group I in all appliances' components. Results of the microbial culture were confirmed by SEM. Use of 0.12% CHX was effective in reducing the formation of MS colonies/biofilms on free surfaces of Haas expanders, in situ., (© 2016 Wiley Periodicals, Inc.)

Purpose: To evaluate in a systematic review whether the use of dental floss in primary teeth is associated with a reduction in incidence of proximal caries., Materials and Methods: Searches were performed using the following databases: PubMed MEDLINE, Web of Science, Bireme, Scopus and The Cochrane Library. Only 5 studies were eligible for inclusion. The quality assessment and bias control of the studies were carried out based on the Fowkes and Fulton Guideline. The study concept was first registered in the international prospective register of systematic reviews (PROSPERO)., Results: Flossing was professionally performed in one study and self-reported (by parents or by the children) through questionnaires in all other 4 studies. In the first study, the authors concluded that daily interdental flossing resulted in a significant reduction in the incidence of proximal caries in deciduous teeth during a 20-month period. Two cross-sectional studies found that the use of interdental floss did not show any relevant association with the prevalence of caries; one study found that a higher the frequency of flossing was associated with higher caries experience, and the other found an association between severe caries and the use of dental floss, independently of the flossing frequency., Conclusion: There is only one study in the current literature showing evidence of an association between the use of dental floss and proximal caries reduction on primary dentition. However, the use of dental floss should never be discouraged. Healthy habits acquired in childhood continue throughout adult life, with numerous oral- and general-health benefits.

Reactive oxygen species (ROS) play a central role in estrogen deficiency-induced bone loss. We previously identified and characterized a novel member of the Peroxiredoxin (PRX) like 2 family that we called PAMM: Peroxiredoxin Activated in M-CSF stimulated Monocytes, a redox regulatory protein that modulates osteoclast differentiation in vitro. In this study, we report increased PAMM expression in H2O2-treated cells and in bones from ovariectomized (OVX) mice 4 weeks after surgery, models for oxidative stress in vitro and in vivo, respectively. We also detected increased PAMM abundance and phosphorylated Akt in OVX mice treated with estrogen. In addition, Wortmannin, a specific PI3Kinase inhibitor and Rapamycin, an inhibitor of the PI3Kinase/Akt pathway, blocked Akt phosphorylation and stimulation of PAMM expression by M-CSF. These results indicate that M-CSF-induced PAMM expression is mediated by Akt phosphorylation. Our data also suggest that estrogen-induced PAMM expression is mediated by phosphorylation of Akt. These findings point to PAMM as a potential candidate for Akt-mediated protection against oxidative stress., (Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.)