Inflammatory response to bioceramic and epoxy resin-based endodontic sealers implanted in mice subcutaneous tissue: An in vivo study.

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.

Territorial Disparities in Dental Care for Disabled Persons and Oral Health-Related Indicators: A Population-Level Approach in Brazil's Public Healthcare System from 2014 to 2023.

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.

Root canal conicity determination of primary second molars using nanotomography: An in vitro study.

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.

The effects of NLRP3 inflammasome inhibition or knockout in experimental apical periodontitis induced in mice.

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-1ß, 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-1ß 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-1ß, 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.

TLR2 agonist prevents the progression of periapical lesions in mice by reducing osteoclast activity and regulating the frequency of Tregs.

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.

Bifidobacterium animalis subsp. lactis HN019 has antimicrobial activity against endodontic pathogens in vitro.

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.

Estimation of root canal conicity of deciduous canines evaluated by nano-CT.

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.

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.

In vitro study of structural and mechanical properties of latex and non-latex intermaxillary orthodontic elastics.

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.

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.

Use of the Probiotic Bifidobacterium animalis subsp. lactis HN019 in Oral Diseases.

The oral cavity is one of the environments on the human body with the highest concentrations of microorganisms that coexist harmoniously and maintain homeostasis related to oral health. Several local factors can shift the microbiome to a pathogenic state of dysbiosis. Existing treatments for infections caused by changes in the oral cavity aim to control biofilm dysbiosis and restore microbial balance. Studies have used probiotics as treatments for oral diseases, due to their ability to reduce the pathogenicity of the microbiota and immunoinflammatory changes. This review investigates the role of the probiotic Bifidobacterium animalis subsp. lactis (B. lactis) HN019 in oral health, and its mechanism of action in pre-clinical and clinical studies. This probiotic strain is a lactic acid bacterium that is safe for human consumption. It mediates bacterial co-aggregation with pathogens and modulates the immune response. Studies using B. lactis HN019 in periodontitis and peri-implant mucositis have shown it to be a potential adjuvant treatment with beneficial microbiological and immunological effects. Studies evaluating its oral effects and mechanism of action show that this probiotic strain has the potential to be used in several dental applications because of its benefit to the host.

Absence of Tumor Necrosis Factor Receptor 1 Inhibits Osteoclast Activity in Apical Dental Resorption Caused by Endodontic Infection in Mice.

INTRODUCTION: The aim of this study was to evaluate osteoclastogenesis and dental resorption resulting from endodontic infection in wild-type (WT) and tumor necrosis factor receptor 1 genetically deficient (TNFR1 KO) mice. METHODS: After approval by the ethics committee on the use of animals, 40 mice were distributed into 2 experimental groups based on time periods: 14 days (n = 10 WT mice and n = 10 TNFR1 KO mice) and 42 days (n = 10 WT mice and n = 10 TNFR1 KO mice). After these periods, morphometric analysis was performed using bright field and fluorescence microscopy and tartrate-resistant acid phosphatase histoenzymology to identify osteoclasts. One-way analysis of variance followed by the Tukey post hoc test was used for the statistical analysis (α = 0.05). RESULTS: WT mice in the 42-day period had a greater apical dental resorption in the distal root of the first molar than TNFR1 KO mice (P < .05). On the other hand, TNFR1 KO mice showed a smaller number of osteoclasts on the dental surface than WT mice (P < .05). CONCLUSIONS: WT mice with apical periodontitis had more extensive apical dental resorptions and a larger number of osteoclasts on the tooth surface than TNFR1 KO mice.

Leukotriene B4 loaded in microspheres regulate the expression of genes related to odontoblastic differentiation and biomineralization by dental pulp stem cells.

BACKGROUND: Leukotriene B4 (LTB4) is a potent lipid mediator that stimulate the immune response. Because dental pulp inflammation and dentin repair are intrinsically related responses, the aim of this research was to investigate the potential of LTB4 in inducing differentiation of dental pulp stem cells. METHODS: Microspheres (MS) loaded with LTB4 were prepared using an oil emulsion solvent extraction evaporation process and sterility, characterization, efficiency of LTB4 encapsulation and in vitro LTB4 release assay were investigated. Mouse dental pulp stem cells (OD-21) were stimulated with soluble LTB4 or MS loaded with LTB4 (0.01 and 0.1 µM). Cytotoxicity and cell viability was determined by lactate dehydrogenase and methylthiazol tetrazolium assays. Gene expression were measured by quantitative reverse transcription polymerase chain reaction after 3, 6, 24, 48 and 72 h. Mineralized nodule formation was assessed after 28 days of OD-21 cell stimulation with LTB4 in mineralized media or not. Groups were compared using one-way ANOVA test followed by Dunnett's post-test (α = 0.05). RESULTS: Treatment with LTB4 or MS loaded with LTB4 (0.01 and 0.1 µm-µM) were not cytotoxic to OD-21 cells. Treatment with LTB4 modulated the expression of the Ibsp (integrin binding sialoprotein) and Runx2 (runt-related transcription factor 2) genes differently depending on the experimental period analyzed. Interestingly LTB4 loaded in microspheres (0.1 µM) allowed long term dental pulp cell differentiation and biomineralization. CONCLUSION: LTB4, soluble or loaded in MS, were not cytotoxic and modulated the expression of the Ibsp and Runx2 genes in cultured OD-21 cells. When LTB4 was incorporated into MS, odontoblast differentiation and mineralization was induced in long term culture.

Self-ligating brackets exhibit accumulation of high levels of periodontopathogens in gingival crevicular fluid.

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.

Gene expression data of inflammatory mediators in apical periodontitis in 129 (wild type) and 5-lipoxygenase knockout mice.

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" .

Ex vivo evaluation of the effectiveness of XP-endo Finisher on the removal of smear layer from the root canal / Avaliação ex vivo da eficácia do XP-endo Finisher na remoção de smear layer do canal radicular

ABSTRACT Objective: The aim of this study was to evaluate the effectiveness of XP-endo Finisher (XP) on removal of the smear layer in root canals by comparing different irrigation protocols. Methods: Seventy-two human single-rooted teeth were similarly instrumented using R25 Reciproc files (VDW, Munich, Germany) applied in reciprocating mode with a VDW GOLD endo motor (VDW, Munich, Germany). The working length was determined at 1 mm short of the apical foramen. The canals were irrigated with 5 mL of 2.5% sodium hypochlorite during instrumentation. The teeth were divided at random into six groups (n=12). A control group, which was not submitted to the final irrigation protocol, and five experimental groups with different irrigants and agitation techniques: EDTA/File, EDTA/XP, EDTA/Passive Ultrasonic Irrigation (PUI), Distilled Water (DW)/XP, and DW/PUI). Smear layer removal quality scores were assessed in the apical, middle, and cervical thirds of the root canal based on images obtained by scanning electron microscopy. Data were analyzed using the Kruskal-Wallis test, followed by two-by-two comparisons with the Dunn test (α=5%). Results: EDTA/File, EDTA/PUI, and EDTA/XP groups demonstrated significantly lower scores than the other groups (P<0.05) in all thirds evaluated. No significant difference was observed between the groups in which distilled water was used and the control group in all thirds evaluated (P> 0.05). Conclusion: The XP-endo Finisher file did not increase the efficiency of EDTA in removal of the smear layer in root canals.

RESUMO Objective: Avaliar a eficácia do XP-endo Finisher (XP) na remoção da smear layer em canais radiculares, comparando diferentes protocolos de irrigação. Métodos: Setenta e dois dentes humanos unirradiculares foram similarmente instrumentados usando limas R25 (VDW, Munich, Germany) aplicadas no modo reciprocrantes em um motor endodôntico VDW GOLD (VDW, Munich, Germany). O comprimento de trabalho foi determinado a 1 mm aquém do forame apical. Os canais foram irrigados com 5 mL de hipoclorito de sódio 2,5% durante a instrumentação. Os dentes foram divididos aleatoriamente em seis grupos (n=12). Um grupo controle, que não foi submetido ao protocolo final de irrigação, e cinco grupos experimentais com diferentes irrigantes e técnicas de agitação: EDTA/Lima, EDTA/XP, EDTA/Irrigação Ultrassônica Passiva (IUP), Água destilada (AD)/XP, e AD/IUP). Os escores de qualidade de remoção da camada de smear layer foram avaliados nos terços apical, médio e cervical do canal radicular com base em imagens obtidas por microscopia eletrônica de varredura. Os dados foram analisados pelo teste de Kruskal-Wallis, seguido de comparações dois a dois pelo teste de Dunn (α = 5%). Resultados: Os grupos EDTA/lima, EDTA/PUI e EDTA/XP demonstraram escores significativamente menores que os outros grupos (P <0,05). Não foi observada diferença entre os grupos que utilizaram água destilada e o grupo controle em todos os terços avaliados (P> 0.05). Conclusão: A lima XP-endo Finisher não aumentou a eficiência do EDTA na remoção da smear layer em canais radiculares.

M1 and M2 macrophages phenotypes modulation after stimuli with materials used in endodontic treatment.

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-1ß 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.

An Epigallocatechin-3-gallate Formulation Developed for Endodontic Use: A Physicochemical and Biological Evaluation.

INTRODUCTION: Although epigallocatechin-3-gallate (EGCG) from green tea has been successfully used in the prevention and treatment of several infectious and immunoinflammatory diseases because of its proven anti-inflammatory, antioxidant, antimicrobial, and antiresorptive role, its use as an intracanal dressing has not been proposed. The aim of this study was to develop a formulation based on EGCG for endodontic use by assessing its physicochemical and biological properties. METHODS: Initially, physicochemical characterization of EGCG was performed by ultraviolet-visible and fluorescence spectroscopy to evaluate if the properties were maintained in acidic pH and time (1-6, 24, and 27 hours). After that, biological studies evaluated the developed formulation of EGCG at different concentrations (1.25, 5, 10, and 20 mg/mL). The tissue compatibility with subcutaneous tissue of mice was evaluated by plasma leakage after 24 hours and the examination of macroscopic and microscopic features at 7, 21, and 63 days after the insertion of polyethylene tubes containing the formulations. The repair of experimentally induced periapical lesions in dog's teeth by radiographic and histopathologic analysis was also evaluated. The scores were statistically analyzed by the chi-square and Fisher exact test. Analysis of variance followed by the Tukey posttest were used for the quantitative analysis. The significance level was 5%. RESULTS: The physicochemical characterization performed under ultraviolet-visible spectrophotometry showed that the EGCG properties remained unaltered in acid pH and function of time, keeping its wavelength to 274 nm. Macroscopic parameters evaluated at 7, 21, and 63 days showed that all concentrations presented no epithelial ulceration or presence of mild superficial tissue necrosis, edema, or vascularization with no significant difference in the control group. During all periods of microscopic examination, all groups presented the absence of abscess foci and edema and the presence of fibrous capsule and neovascularization. The presence of reparative tissue with a gentle presence of neutrophilic inflammatory cells was also observed for all groups, except for the calcium hydroxide paste group, which presented a more pronounced inflammation and tissue necrosis at days 7 and 21 (P < .001). At day 63, all groups presented an absence of inflammatory infiltrate and necrosis. The evaluation of dog teeth showed that treatment with the EGCG formulation provided a reduction of the periapical radiolucent area and allowed the repair of apical and periapical tissues (P > .05). CONCLUSIONS: The developed formulation based on EGCG from green tea presented physicochemical stability and tissue compatibility and provided the repair of periapical lesions when used as an intracanal dressing.

M1 and M2 macrophages phenotypes modulation after stimuli with materials used in endodontic treatment

Abstract 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.

Resumo O objetivo deste estudo foi avaliar a modulação dos macrófagos M1 e M2 após estímulos com diferentes materiais utilizados durante o tratamento endodôntico. Em cultura de células de macrófagos derivados da medula óssea de camundongos machos C57BL/6 wild-type (WT), após a exposição à cinco cimentos endodônticos: AH Plus, Sealapex Xpress, Endosequence BC Sealer, BioRoot RCS e pasta à base de hidróxido de cálcio foi realizada a análise da expressão gênica dos marcadores para macrófagos M1 e M2 por qRT-PCR (Cxcl10, CxCL9, iNOS, Arg1, Chil3, Retnla e MRC1) e quantificação de citocinas por Luminex® (GM -CSF, IL-10, IL-6, IL-1β e TNF-α). Para valores normais, foi utilizado o teste ANOVA, seguido do pós-teste de Tukey. Para valores não normais, foi utilizado o teste de Kruskall-Wallis. BioRootTM RCS e EndoSequence BC SealerTM estimularam maior expressão de marcadores para macrófagos M1, enquanto a pasta à base de hidróxido de cálcio estimulou expressão mais baixa desses marcadores gênicos. Para o marcador de proteínas para M2, BioRootTM RCS apresentou a maior estimulação, enquanto a pasta à base de hidróxido de cálcio também apresentou menor estimulação. Concluiu-se que os materiais obturadores avaliados aumentaram a expressão genética de marcadores pró- e anti-inflamatórios: TNF-α e IL-10 respectivamente. Os demais marcadores pró inflamatórios mostraram diferenças em relação aos materiais obturadores. No entanto, esse processo não induziu a polarização da resposta inflamatória, resultando em um macrófago híbrido.