Interaction between polymorphisms in TNF-⍺ and RANKL genes is associated with the development of persistent apical periodontitis, in Brazilian subjects.

OBJECTIVES: To investigate the association between genetic polymorphisms in suppressor of cytokine signaling-1 (SOCS-1), tumor necrosis factor-⍺ (TNF-α) and its receptors 1 and 2 (TNFRSF1A and TNFRSF1B), receptor activator of nuclear factor kappa-b (RANK), receptor activator of nuclear factor-kappa B ligand (RANKL) and osteoprotegerin (OPG), and persistent apical periodontitis (PAP). METHODS: Patients with pulp necrosis and apical periodontitis at the time of non-surgical root canal treatment were followed up for at least one year. A total of 423 subjects were included, 172 with signs/symptoms of PAP and 251 with apical periodontitis healed. DNA was extracted from saliva and used for genotyping polymorphisms in SOCS1 (rs243327 and rs33977706), TNF-α (rs1800629), TNFRSF1A (rs1800693), TNFRSF1B (rs1061622), RANK (rs1805034), RANKL (rs1054016) and OPG (rs1032128) by real-time PCR. The frequency of genotypes and alleles was assessed using chi-squared test or Fisher's exact test and odds ratio. Interactions were also tested using multifactor dimensionality reduction (α = 5 %). RESULTS: In the polymorphism rs1800629 in TNF-α, carrying at least one A risk allele significantly decreased the risk to develop PAP (OR=0.47; 95 %CI: 0.28-0.79; p=0.004). For the polymorphism rs1054016 in RANKL carrying both T risk alleles significantly decreased the risk to develop PAP (OR=0.47; 95 %CI: 0.24-0.92; p=0.027). None of the other polymorphisms evaluated were associated with PAP (p>0.05). A strong interaction was observed among rs1800629, rs1061622 and rs1054016. CONCLUSIONS: Genetic polymorphisms rs1800629 (TNF-α) and rs1054016 (RANKL) had a strong interaction and were associated with a lower risk to develop PAP.

Abrasive challenge effects on enamel and dentin from irradiated human teeth: an in vitro study.

This study aimed to evaluate the effects of radiotherapy (RT) and chemoradiotherapy (CRT) on the wear and surface roughness of in vitro irradiated human enamel and dentin subjected to abrasive challenge. Enamel and dentin specimens (n = 42) were prepared from teeth donated by healthy patients and those with head and neck cancer who had received radiotherapy (RT) or chemoradiotherapy (CRT). The specimens were categorized into three groups: control, RT, and CRT (n = 14 per group for both enamel and dentin). These samples were subjected to an in vitro abrasive experiment using a brushing machine, followed by wear and surface roughness assessments with a confocal laser scanning microscope conducted before and after the abrasive challenge, considering both exposed and non-exposed areas. Statistical analysis used Shapiro-Wilk tests for normality, Wilcoxon tests for comparing two means, and Kruskal-Wallis tests. A significance level of 5% was adopted. In enamel specimens, wear profile values ​​of CRT and RT groups were not different from the control (p > 0.05). The RT group presents lower step values than the CRT and control groups (p < 0.001). No significant difference in final surface roughness was observed in all groups (p > 0.05). In dentin specimens, no significant difference in wear profile and step was observed in all groups (p > 0.05). However, CRT and RT groups present higher values in final surface roughness (p < 0.001). The exposure to ionizing radiation (associated or not to chemotherapy) influenced the surface roughness of dentin and the wear (step) of enamel after the in vitro abrasive challenge.Trial registration: Ethical procedures were approved by the FORP/USP Research Ethics Committee (CAAE: 61308416.4.0000.5419), and Hospital do Câncer de Barretos/Fundação Pio XII (CAAE: 61308416.4.3001.5437).

Implications of Histological and Ultrastructural Characteristics on the Chemical and Mechanical Properties of Hypomineralised Enamel and Clinical Consequences.

Molar incisor hypomineralisation (MIH) is a qualitative type of enamel defect, which occurs due to a failure in the biomineralisation process of the enamel organic matrix during amelogenesis. The tooth enamel affected by MIH shows changes in its chemical, structural, and mechanical properties, leading to different clinical repercussions. The color of MIH opacities varies from opaque white to yellow/brown, and elemental analyses of these lesions show a lower calcium and phosphate content, minerals that are more abundant in sound enamel. Furthermore, the incorporation of other molecules occurs, such as carbonate, a component that provides a greater degree of solubility, thus making hypomineralised enamel more susceptible to posteruptive fractures. At a structural level, the layer of hydroxyapatite crystals appears to be disorganized, with morphological changes, implying a greater degree of porosity in the structure. The increase in porosity of the structure may be associated with dental hypersensitivity, a common clinical repercussion among patients with MIH. Among the mechanical properties, a decrease in hardness and modulus of elasticity occurs, and this also makes the enamel more fragile. Deficiency in biomineralisation can be caused by changes in the function of ameloblasts or by failures at the intercellular junction that result in lower activity of proteases such as MMP-20 and KLK4. The increase in proteins in the organic matrix of enamel impairs the growth and incorporation of minerals into the hydroxyapatite crystals, so that the enamel becomes hypomineralised and has larger organic content, thus having an impact on its properties. These changes present in the enamel with MIH help to explain the clinical repercussions caused by this condition.

Acid challenge exacerbates activation of matrix metalloproteinases in permanent teeth undergoing radiotherapy.

The aim of this study was to investigate the effect of acid challenge on the activation of matrix metalloproteinases (MMPs) in the Dentinoenamel junction of primary and permanent teeth submitted to radiotherapy. For this purpose, a total of 178 dental fragments obtained from molars were used, and randomly divided into 2 groups (primary and permanent teeth) / 4 experimental subgroups (irradiated and non-irradiated, demineralized and non-demineralized). The fragments were exposed to radiation, with a dose fraction of 2 Gy, for 5 consecutive days, until a total dose of 60 Gy was reached, with a total of 30 cycles, for 6 weeks. To determine the activity of MMPs on the dentinoenamel junction (DEJ), in situ zymography assays on 0.6mm dental fragments were performed. To assess whether MMP activity would be impacted by an acidic environment, the fragments were placed in a demineralizing solution (pH of 4.8). The finding was that irradiation activated MMPs in DEJ and these effects were more evident in permanent when compared with primary teeth. When the effect of an acid challenge on MMPs activity was investigated, demineralization was observed not to increase MMPs activity in non-irradiated teeth, but it did increase MMPs activity in irradiated teeth. In conclusion, an acid challenge was found to exacerbate activation of MMPs in DEJ of permanent teeth submitted to irradiation, but not in primary teeth.

The effects of re-irradiation on the chemical and morphological properties of permanent teeth.

This study aimed to assess the in vitro effects of re-irradiation on enamel and dentin properties, simulating head and neck cancer radiotherapy retreatment. Forty-five human permanent molars were classified into five groups: non-irradiated; irradiated 60 Gy, and re-irradiated with doses of 30, 40, and 50 Gy. Raman spectroscopy, scanning electron microscopy (SEM), and energy dispersive x-ray spectroscopy (EDS) were employed for analysis. Raman spectroscopy assessed intensity, spectral area, and specific peaks comparatively. Statistical analysis involved Kolmogorov-Smirnov and One-Way ANOVA tests, with Tukey's post-test (significance level set at 5%). Significant changes in irradiated, non-irradiated, and re-irradiated enamel peaks were observed, including phosphate (438 nm), hydroxyapatite (582 nm), phosphate (960 nm), and carbonate (1070 nm) (p < 0.05). Re-irradiation affected the entire tooth (p > 0.05), leading to interprismatic region degradation, enamel prism destruction, and hydroxyapatite crystal damage. Dentin exhibited tubule obliteration, crack formation, and progressive collagen fiber fragmentation. EDX revealed increased oxygen percentage and decreased phosphorus and calcium post-reirradiation. It is concluded that chemical and morphological changes in irradiated permanent teeth were dose-dependent, exacerbated by re-irradiation, causing substantial damage in enamel and dentin.

Optical coherence tomography and gray scale digital analysis as noninvasive techniques for evaluating molar-incisor hypomineralization severity: A comparative study with microcomputed tomography.

Molar-incisor hypomineralization (MIH) is a qualitative defect of dental enamel characterized by demarcated opacities present in permanent first molars and other teeth. It is considered a major clinical challenge in dentistry because it makes affected teeth more susceptible to fractures and dental caries. Its diagnosis is mainly clinical and there are few technological resources that allow for a more accurate diagnosis, especially with respect to the depth of the defect in the dental enamel. In this context, optical coherence tomography (OCT), which is routinely used in ophthalmology, can produce images of the depth of the dental enamel, making it a promising method. In this study, 33 teeth with different MIH severities were evaluated using OCT and microcomputed tomography (microCT). Semi-quantitative methods of grayscale pattern analysis were used to compare images obtained from different severities of MIH with the mineral density obtained through microCT. MicroCT evaluation revealed that hypomineralized enamel had a significantly lower mineral density than intact enamel. However, this difference was not observed between the mild and severe MIH lesions. In the OCT evaluation, significant differences were observed between the intact and hypomineralized enamel, and the gray value comparison provided a method for quantitative differentiation between the two. This study suggests that OCT could be a useful adjunct to traditional diagnostic methods for MIH, offering a noninvasive approach to evaluate enamel defects. RESEARCH HIGHLIGHTS: Combining optical coherence tomography with grayscale digital analysis shows potential as a promising method for diagnosing molar-incisor hypomineralization and assessing its level of severity.

Analysis of the cytotoxicity and bioactivity of CeraSeal, BioRoot™ and AH Plus® sealers in pre-osteoblast lineage cells.

BACKGROUND: The objective of the present study was to evaluate in vitro the cytotoxicity and bioactivity of various endodontic sealers (CeraSeal, BioRoot™ and AH Plus®) in pre-osteoblast mouse cells (MC3T3 cells). METHODS: MC3T3 cells (ATCC CRL-2594) were plated in 1 × 104 cells/well in 96-well plates in contact with endodontic sealers at concentrations of 1:10 and 1:100. Cell viability was evaluated by MTT assay after 24 and 48 h. In addition, sealer bioactivity was measured by RT-PCR for mediator of inflammation (Tnf, Ptgs2) and mineralization (Runx2, Msx1, Ssp1 and Dmp1) after 24 h and by Alizarin Red S Assay of mineralization after 28 days. Data were analyzed using one-way ANOVA followed by the Tukey's post-test at a significance level of 5%. RESULTS: BioRoot™ presented 24-hour cytotoxicity (p < 0.05) at 1:10 concentration. In the period of 48 h, no endodontic cement was cytotoxic to the cells compared to the control (p > 0.05). TNF-α gene expression was induced by AH Plus® (p < 0.05), while Ptgs2 was induced by the CeraSeal and BioRoot™ (p < 0.05). The expression of Runx2 was stimulated by BioRoot™ and AH Plus® (p < 0.05). In contrast, the expression of Dmp-1 Dmp1 was higher for the CeraSeal and BioRoot™ (p < 0.05). Nonetheless, the sealers did not impact the formation of mineralization nodules (p > 0.05). CONCLUSION: CeraSeal, BioRoot™ and AH Plus® sealers were not cytotoxic to MC3T3 cells within 48 h, but differentially induced the expression of genes related to inflammation and mineralization without impacting biomineralization by the cells.

Acid challenge exacerbates activation of matrix metalloproteinases in permanent teeth undergoing radiotherapy

Abstract The aim of this study was to investigate the effect of acid challenge on the activation of matrix metalloproteinases (MMPs) in the Dentinoenamel junction of primary and permanent teeth submitted to radiotherapy. For this purpose, a total of 178 dental fragments obtained from molars were used, and randomly divided into 2 groups (primary and permanent teeth) / 4 experimental subgroups (irradiated and non-irradiated, demineralized and non-demineralized). The fragments were exposed to radiation, with a dose fraction of 2 Gy, for 5 consecutive days, until a total dose of 60 Gy was reached, with a total of 30 cycles, for 6 weeks. To determine the activity of MMPs on the dentinoenamel junction (DEJ), in situ zymography assays on 0.6mm dental fragments were performed. To assess whether MMP activity would be impacted by an acidic environment, the fragments were placed in a demineralizing solution (pH of 4.8). The finding was that irradiation activated MMPs in DEJ and these effects were more evident in permanent when compared with primary teeth. When the effect of an acid challenge on MMPs activity was investigated, demineralization was observed not to increase MMPs activity in non-irradiated teeth, but it did increase MMPs activity in irradiated teeth. In conclusion, an acid challenge was found to exacerbate activation of MMPs in DEJ of permanent teeth submitted to irradiation, but not in primary teeth.

Use of Optical Coherence Tomography for Differential Diagnosis of Severity of Molar Incisor Hypomineralization (MIH).

Purpose: To evaluate descriptively and quantitatively teeth affected by enamel hypomineralization (EH) using optical coherence microtomography (OCT). Methods: Twenty teeth were classified according to the European Academy of Pediatric Dentistry's molar incisor hypomineralization (MIH) index and separated into groups according to the degree of EH severity. For each tooth, scans were performed on both the affected and the non-affected areas, and their corresponding optical images were captured. Results: In the qualitative analyses, in most of the images bright lines were observed in relation to the enamel surface and a high level of photon scattering immediately below the enamel surface. This showed that the shading distribution can be identified as hypomineralized areas in which the scattering signal can be used as a diagnostic criterion. In the quantitative analyses, Tukey's test was performed to evaluate the means of the optical attenuation coefficient, which did not present significant differences. However, considering the correlation, homogeneity and contrast analyses, a statistically significant difference was observed between the groups. The group with severe MIH showed greater homogeneity and correlation, but less contrast. Conclusion: Currently, MIH has its severity measured by essentially clinical means. OCT processing techniques reveal advances in the diagnostic imaging of MIH, showing that image texture analysis can be a promising and useful method to aid in its diagnosis.

Activation of gelatinases in permanent human teeth after different experimental radiotherapy protocols.

The objective of this study was to compare the activation of gelatinases in dentin-enamel junction (DEJ) and underlying dentin of permanent teeth after experimental radiotherapy in conventional and hypofractionated modalities. Newly extracted third molars (n = 15) were divided into three experimental radiotherapy groups: control, conventional (CR), and hypofractionated (HR) (n = 5 per group). After in vitro exposure to ionizing radiation, following standardized protocols for each modality, a gelatinous substrate was incubated on the tooth slices (n = 10 per group). Activation of gelatinases was measured by in situ zymography, expressed in arbitrary fluorescence units (mm2) from three tooth regions: cervical, cuspal, and pit. Fluorescence intensity was compared among radiotherapy protocols and tooth regions in each protocol, considering a significance level of 5%. Considering all tooth regions, the fluorescence intensity of the CR group was higher than the HR and control groups, both in DEJ and underlying dentin (p <0.001). In addition, the fluorescence intensity was higher in underlying dentin when compared to DEJ in all groups (p <0.001). Considering each tooth region, a statistically significant difference between CR and HR was only observed in the pit region of underlying dentin (p <0.001). Significant and positive correlations between fluorescence intensities in DEJ and underlying dentin were also observed (p <0.001). Experimental radiotherapy influenced the activation of gelatinases, as well as exposure to the conventional protocol can trigger a higher activation of gelatinases when compared to hypofractionated, both in DEJ and underlying dentin.

Comparison between intraoral scanning and direct visual analysis for the assessment of developmental defects of enamel.

OBJECTIVE: To compare direct visual analysis (DVA) and intraoral scanning (IOS) for the assessment of developmental defects of the enamel (DDE). METHODS: Thirty-nine extracted permanent human teeth with DDE were selected by an experienced examiner and digitised using IOS. The scanning was recorded using the OBS Studio software parallel to the IOS software to obtain a coloured high-definition MP4 file of the process. Two other experienced, blinded, and calibrated examiners randomly analysed the same teeth through DVA and IOS. A third examiner resolved any disagreements between the two examiners. Descriptive statistics were used to analyse the frequencies of the scores. Cohen's kappa test was used to determine whether the DVA scores were different from those assigned using IOS. Spearman's test was used to verify non-random examiner errors. The Chi-square test was used to compare score frequencies. Statistical significance was set at p <0.05. RESULTS: Scores indicating more severe and extended DDE (p <0.05) were more frequently assigned with IOS than with DVA (IOS: 25.64%, 25.64%, 38.46%, and 35.90% between one-third to two-third of the lingual, occlusal, mesial, and distal surfaces, respectively; vs. DVA: 10.26%, 7.69%, 15.38%, and 10.26% for the respective aforementioned tooth surfaces). Contrarily, 'no visible enamel defect' was significantly less assigned for IOS than for DVA (IOS: 15.38%, 43.59%, 35.90%, 15.38%, and 17.95% for buccal, lingual, occlusal, mesial, and distal surfaces, respectively; vs. DVA: 38.46%, 66.67%, 56.41%, 51.28%, and 43.59% for the respective aforementioned tooth surfaces). Kappa agreement ranged from fair to moderate when comparing DVA and IOS; the correlation between both methods was positive, indicating that the examiners assigned the scores properly and the differences arose from employing different methods. CONCLUSION: The assessment of DDE differed depending on the method used. IOS scores indicated more severe and extended DDE than DVA scores. Clinical investigation is the next step in validating the use of IOS for DDE diagnosis. CLINICAL SIGNIFICANCE: This study showed that DDE can be assessed differently using IOS. It is clinically relevant as it directly affects the determination of the severity of the defect and dental treatment planning.

TNF-α-TNFR1 Signaling Mediates Inflammation and Bone Resorption in Apical Periodontitis.

INTRODUCTION: The aim of this study was to investigate the role of the proinflammatory axis TNF-α-TNFR1 in experimentally induced periapical inflammation and bone resorption in mice. METHODS: After receiving Ethics Committee Approval (2019.1.139.58.0), experimental apical periodontitis was induced by means of inoculating oral microorganisms into the root canals of molars of mice. Genetically deficient tumor necrosis factor-α receptor-1 mice (TNFR1-/-; n = 50) response was compared with that of C57Bl6 wild-type mice (wild-type; n = 50) after 7, 14, 28, and 42 days. The analyses performed were micro-computed tomographic, histopathologic, histomicrobiological, and histometric evaluation, tartrate-resistant acid phosphatase staining, immunohistochemistry, and quantitative reverse transcriptase polymerase chain reaction. Data were analyzed by using one-way analysis of variance, followed by Tukey or Bonferroni tests (α = 5%). RESULTS: TNFR1-/- mice exhibited lower recruitment of neutrophils at 14, 28, and 42 days (P < .05), which resulted in reduced area and volume of apical periodontitis at 42 days (P < .05). The number of osteoclasts was also lower in TNFR1-/- animals at 14 and 42 days (P < .01), along with reduced synthesis of CTSK, MMP-9, and COX-2. Expression of RANKL, but not OPG, was reduced at 14 and 42 days (P < .001). The highest RANKL expression over OPG (ratio > 1) was found in wild-type animals at 7 (P < .0001) and 42 days (P < .001). CONCLUSIONS: Periapical inflammation and bone resorption were exacerbated in wild-type animals compared with TNFR1-/- mice, demonstrating that the TNF-α-TNFR1 signaling pathway mediated catabolic events in bone after root canal contamination.

Reparative Dentin Formation Following Dental Pulp Capping is Mediated by TNFR1 In Vivo.

INTRODUCTION: Tumor necrosis factor (TNF)-α is a pro-inflammatory cytokine that promotes biomineralization in vitro in dental pulp cells. However, the role of TNF-α-TNF receptor 1 (TNFR1) signaling in reparative dentin formation and related inflammatory pathways is not known. Therefore, the aim of this study was to evaluate the role of the TNF-α-TNFR1 axis in dental pulp repair following pulp capping in vivo. METHODS: Dental pulp repair response of genetically deficient TNF-α receptor-1 mice (TNFR1-/-; n = 20) was compared with that of C57Bl6 mice (wild type [WT]; n = 20). Pulp capping was performed with mineral trioxide aggregate on the mandibular first molars of mice. After 7 and 70 days, tissues were collected and stained with hematoxylin and eosin for histopathological and histometric evaluation, and assessed by the Brown and Brenn methods for histomicrobiological analysis and by immunohistochemistry to localize TNF-α, Runt-related transcription factor 2, Dentin Sialoprotein (DSP) and Osteopontin (OPN) expression. RESULTS: Compared with WT mice, TNFR1-/- mice showed significantly decreased reparative dentin formation with a lower mineralized tissue area (P < .0001). Unlike WT mice, TNFR1-/- mice also exhibited significant dental pulp necrosis, neutrophil recruitment, and apical periodontitis formation (P < .0001) without bacterial tissue invasion. TNFR1-/- animals further exhibited decreased TNF-α, DSP, and OPN expression (P < .0001), whereas Runt-related transcription factor 2 expression was unchanged (P > .05). CONCLUSION: The TNF-α-TNFR1 axis is involved in reparative dentin formation following dental pulp capping in vivo. Genetic ablation of TNFR1 modified the inflammatory process and inhibited the expression of the DSP and OPN mineralization proteins, which culminated in dental pulp necrosis and development of apical periodontitis.

Detection of Apical Dental Resorption Caused by Endodontic Infection in Mice Using Fluorescence and Bright-Field Microscopy.

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.

Electromyographic analysis of the stomatognathic system of children with Molar-incisor hypomineralization.

INTRODUCTION: Molar-incisor hypomineralization (MIH) is a qualitative defect of dental enamel that affects one or more permanent first molars, with or without involvement of the incisor teeth. This condition leads to challenges to dental care and treatment planning. AIM: Based on the hypothesis that children who have MIH possibly present alterations in postural and masticatory activities and considering the absence of studies investigating these parameters, the present study evaluated the functionality of the stomatognathic system considering the mentioned aspects. MATERIALS: The comparison of individuals with (MIHG; n = 32) and without MIH (CG; n = 32) was evaluated by electromyographic activity of the masseter and temporal muscles (right and left), as well as evaluation of the masticatory cycles during habitual mastication. RESULTS: MIHG showed muscle hyperactivity in postural and dynamic conditions compared to the CG; higher electromyographic values for MIHG when compared to CG in the following postural conditions: at rest for the right temporal (p = 0.00) and left temporal muscles (p = 0.03); in the protrusion to the right temporal muscle (p = 0.02); in the right laterality for the right masseter (p = 0.00) and left temporal muscles (p = 0.01); in the left laterality for the right masseter (p = 0.03) and left temporal (p = 0.04) muscles. In dynamic conditions with consistent food, significance was observed for the left temporal (p = 0.01); and with soft food for the right (p = 0.01) and left temporal muscles (p = 0.04). CONCLUSIONS: Children with MIH seem to have impaired functionality of the stomatognathic system. Children with MIH have alterations in the stomatognathic system.

Systemic inhibition of 5-lipoxygenase by MK-886 exacerbates apical periodontitis bone loss in a mouse model.

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.

Genetic polymorphism in the tumour necrosis factor alpha gene (G-308A) is associated with persistent apical periodontitis in Brazilians.

AIM: To investigate if there was an association between genetic polymorphisms in tumour necrosis factor (TNF)-⍺ and its receptors TNFRSF1A and TNFRSF1B with persistent apical periodontitis (PAP) in Brazilian subjects. METHODOLOGY: Patients who had pulpal necrosis and apical periodontitis at the time of treatment, with at least 1-year of follow-up after non-surgical root canal treatment were recalled. Three hundred and seventy eight subjects were included, 150 subjects with signs/symptoms of PAP and 228 subjects with root canal-treated teeth exhibiting healthy perirradicular tissues (healed). Genomic DNA was extracted from saliva and used for TNF-⍺ (rs1800629), TNFRSF1A (rs1800693) and TNFRSF1B (rs1061622) genotyping by real-time PCR. Genotypes and alleles frequencies were evaluated by c2 or Fisher's exact tests and odds ratios were implemented (α = 5%). RESULTS: The genetic polymorphism in TNF-α (rs1800629) was associated as a protective factor for the development of PAP (p < .05), once subjects who presented at least one allele A (AA+AG X GG), had a higher chance to lesion repair (p < .05). The polymorphisms rs1800693 and rs1061622 in TNF receptors (TNFRSF1A and TNFRSF1B, respectively) were not associated with the development of PAP (p > .05). CONCLUSIONS: The observed results demonstrate that polymorphism in TNF-α but not in its receptors is associated with PAP.

Randomized Clinical Study of the Use of MTA and Biodentine™ for Pulpotomy in Primary Teeth

ABSTRACT Objective: To verify, through clinical and radiographic evaluations, the in vivo response of the dentin-pulpal complex of human primary teeth after pulpotomy with MTA and Biodentine™ in a follow-up period of 3, 6, and 12 months. Material and Methods: Thirty teeth were divided into MTA pulpotomy (n = 15) and Biodentine™ pulpotomy (n = 15) from children between 5 and 9 years of age, a randomized clinical trial with simple random sampling. The materials were inserted into the cavity after opening and removing the coronary pulp tissue. The cavity base consisted of glass ionomer cement and light-cured composite resin restoration. Clinical and radiographic analyses were performed after 3, 6, and 12 months. Statistical analysis by Fisher's exact test for dichotomous data at a 5% significance level was utilized. Results: Both materials caused color change after 12 months. However, MTA showed a higher percentage than Biodentine™ (p<0.0001). Pain was detected only with Biodentine™ at six months and mobility at 12 months (p=0.0013). Radiographically, after 12 months, periapical lesions, interradicular lesions, and internal resorption were evidenced in 13% of the cases for Biodentine™-treated teeth (p<0.0013). MTA induced pulp calcification in 13% of cases, unlike Biodentine™ (p<0.0013). Conclusion: BiodentineTM and MTA are suitable for clinical use in pulpotomy treatment, yet both materials lead to tooth discoloration.

Use of Restorative Materials in Primary Teeth - A Retrospective University-Based Study

ABSTRACT Objective: To investigate the types of restorative materials used for restorative treatment in primary teeth through a retrospective university-based study. Material and Methods: The sample consisted of all clinical records of children attended at the Pediatric Dentistry Clinic at the School of Dentistry of Ribeirão Preto at the University of São Paulo in Brazil. Inclusion criteria were primary anterior and posterior teeth that received dental restorations for treatment of dental caries lesions, dental trauma or dental development defects from 2013 to 2018. Restoration repairs and interim restorations during this period were also recorded. Descriptive analyzes were performed to assess the distribution according to the type of restorative material used over the years. Results: A total of 5,236 restorative procedures were performed in primary teeth, including restoration repair and interim restorations. Of those, 69% were done in posterior teeth and 31% in anterior teeth. Sixty percent of the procedures performed during this period were made of composite resin and a lower percentage of glass ionomer cement (18%) followed by silver amalgam (1%). The number of interim restorations was smaller but proportional to those of composite resin over the years. Conclusion: A tendency to carry out restorative treatment of primary teeth with composite resin during the 6 years of follow-up was observed.

Activation of gelatinases in permanent human teeth after different experimental radiotherapy protocols

Abstract The objective of this study was to compare the activation of gelatinases in dentin-enamel junction (DEJ) and underlying dentin of permanent teeth after experimental radiotherapy in conventional and hypofractionated modalities. Newly extracted third molars (n = 15) were divided into three experimental radiotherapy groups: control, conventional (CR), and hypofractionated (HR) (n = 5 per group). After in vitro exposure to ionizing radiation, following standardized protocols for each modality, a gelatinous substrate was incubated on the tooth slices (n = 10 per group). Activation of gelatinases was measured by in situ zymography, expressed in arbitrary fluorescence units (mm2) from three tooth regions: cervical, cuspal, and pit. Fluorescence intensity was compared among radiotherapy protocols and tooth regions in each protocol, considering a significance level of 5%. Considering all tooth regions, the fluorescence intensity of the CR group was higher than the HR and control groups, both in DEJ and underlying dentin (p <0.001). In addition, the fluorescence intensity was higher in underlying dentin when compared to DEJ in all groups (p <0.001). Considering each tooth region, a statistically significant difference between CR and HR was only observed in the pit region of underlying dentin (p <0.001). Significant and positive correlations between fluorescence intensities in DEJ and underlying dentin were also observed (p <0.001). Experimental radiotherapy influenced the activation of gelatinases, as well as exposure to the conventional protocol can trigger a higher activation of gelatinases when compared to hypofractionated, both in DEJ and underlying dentin.

Resumo O objetivo deste estudo foi comparar a ativação de gelatinases na junção dentina-esmalte (DEJ) e na dentina subjacente de dentes permanentes após a radioterapia experimental nas modalidades convencional e hipofracionada. Os terceiros molares recém-extraídos (n = 15) foram divididos em três grupos de radioterapia experimental: controle, convencional (CR) e hipofracionada (HR) (n = 5 por grupo). Após a exposição in vitro à radiação ionizante, seguindo protocolos padronizados para cada modalidade, um substrato gelatinoso foi incubado nas fatias de dente (n = 10 por grupo). A ativação das gelatinases foi medida por zimografia in situ, expressa em unidades arbitrárias de fluorescência (mm2) de três regiões do dente: cervical, cúspide e fossa. A intensidade da fluorescência foi comparada entre os protocolos de radioterapia e as regiões do dente em cada protocolo, considerando um nível de significância de 5%. Considerando todas as regiões do dente, a intensidade de fluorescência do grupo CR foi maior do que a dos grupos HR e controle, tanto no DEJ quanto na dentina subjacente (p <0,001). Além disso, a intensidade da fluorescência foi maior na dentina subjacente quando comparada à DEJ em todos os grupos (p <0,001). Considerando cada região do dente, uma diferença estatisticamente significativa entre CR e HR foi observada apenas na região da fossa da dentina subjacente (p <0,001). Também foram observadas correlações significativas e positivas entre as intensidades de fluorescência no DEJ e na dentina subjacente (p <0,001). A radioterapia experimental influenciou a ativação das gelatinases, assim como a exposição ao protocolo convencional pode desencadear uma maior ativação das gelatinases quando comparada ao hipofracionamento, tanto no DEJ quanto na dentina subjacente.