Side effects of different head and neck radiotherapy doses on wistar rat's behavior.

Radiotherapy (RT) is a common treatment for head and neck tumors. However, it causes several physical and behavioral side effects, and no study has assessed the emotional effects in rats. Therefore, the present study evaluated the influence of head and neck RT on the behavior and body weight gain in Wistar rats. Fifty-four male Wistar rats were allocated into six groups (n = 9) according to the irradiation dose, which was applied at the first day of the experiment: RT-7.5 (single dose of 7.5 Gy); RT-10 (single dose of 10 Gy); RT-15 (single dose of 15 Gy); RT-30 (single dose of 30 Gy); Control (without RT). The animals were irradiated in the region of the right face, and behavioral tests and weighing were performed on days one, seven, and 28. The open field and Y-maze tests were undertaken to analyze the animal's behavior. The dose of 30 Gy was lethal when applied to the head and neck region. The irradiated animals had less weight gain when compared to the control ones, but there was no statistical difference. In the open field and Y-maze tests, lower mobility of animals in the RT groups was observed both on day seven and at the end of the experiment (day 28) when compared to the control rats (p < 0.05). It was possible to conclude that the different doses of radiation induced depressive behavior in the animals, and that the weight gain tended to be lower in the irradiated groups, however, without statistical difference.

Clinical efficacy of endodontic protocols on reducing cultivable bacteria and endotoxin in infected root canal in patients submitted to head and neck radiotherapy: a randomised clinical trial.

OBJECTIVES: Assess the efficacy of biomechanical preparation using a reciprocating system followed by final irrigation protocols, then intracanal medication, on reducing endotoxins and cultivable bacteria of infected teeth in irradiated patients. MATERIALS AND METHODS: Twenty-two infected single-rooted canals in patients submitted to head and neck radiotherapy were prepared by reciprocating motion and 2.5% NaOCl. Patients were randomly divided into two groups of 11 patients before the final irrigation protocol: apical positive pressure (APP) or passive ultrasonic activation (PUA). Both groups were treated in two sessions, using Ca(OH)2 as intracanal medication for 14 days. Root canal content sampling was performed after canal access (S1), after biomechanical preparation plus the irrigation protocol (S2), and after intracanal medication (S3). Chromogenic limulus amoebocyte lysate assay measured endotoxin levels (EU/mL), and bacterial load was determined by culture techniques (CFU/mL). RESULTS: Treatment protocols reduced bacterial counts after S2 in both groups (p = 0.01). S3 differed from S1 (p = 0.01), but not from S2 (p = 0.4). Endotoxin levels were reduced in both groups after S2 (P = 0.03) and were lower in S3 than in S2, with significant differences in the APP group (p = 0.03). CONCLUSIONS: Biomechanical preparation using a reciprocating system and 2.5% NaOCl in irradiated teeth, followed by the irrigation protocol (APP or PUA), demonstrated efficacy in reducing endodontic contaminants. Ca(OH)2 as intracanal medication should be performed in irradiated patients with infected root canals. CLINICAL RELEVANCE: This clinical study demonstrated that endodontic treatment in irradiated patients is efficacious at reducing bacterial load and endotoxin levels.

Detoxification of LTA by intracanal medication: analysis by macrophages proinflammatory cytokines production.

The aim of this study was to evaluate in vitro the effect of calcium hydroxide [Ca(OH)2], 2% chlorhexidine gel (CHX) on macrophages (RAW 264.7) to produce pro-inflammatory cytokines and nitric oxide after pretreatment with lipoteichoic acid (LTA) of Enterococcus faecalis. Forty-eight human single-rooted teeth were instrumented with R25.08 (RECIPROC) and sterilized by gamma irradiation. LTA was inoculated in the root canal of each specimen for 96 hours. Specimens were instrumented with 40.06 and 50.05 (RECIPROC) and medicated with: I) Pyrogen-free saline solution (SS); II) 2% CHX gel; III) Ca(OH)2 + SS; or IV) Ca(OH)2 + CHX for 14 days. Three samples (S) were performed of the root canal of each specimen at: S1) immediately after instrumentation; S2) after Ethylenediaminetetraacetic acid (EDTA); S3) after intracanal medication removal. Subsequent quantification of cytokines (IL-1ß, TNF-α, MIP-1α, IP-10, G-CSF and IL-6) by immunosorbent assay (ELISA) and nitric oxide by the Griess method was carried-out. Data were submitted to a normality test and then analyzed with one-way ANOVA and Tukey test with a significance level of 5% using GraphPad Prism 6. Ca(OH)2 + SS and Ca(OH)2 + CHX presented lower levels of TNF-α, TNF-α, IL-6, G-CSF and nitric oxide. Ca(OH)2 + SS was the most effective in reducing MIP-1α. CHX was effective in reducing IL-6 and G-CSF. Therefore, the combined intracanal medication of calcium hydroxide and chlorhexidine is effective in reducing the cytokines TNF-α, IL-1ß, IL-6, G-CSF and nitric oxide.

Detoxification of LTA by intracanal medication: analysis by macrophages proinflammatory cytokines production

Abstract The aim of this study was to evaluate in vitro the effect of calcium hydroxide [Ca(OH)2], 2% chlorhexidine gel (CHX) on macrophages (RAW 264.7) to produce pro-inflammatory cytokines and nitric oxide after pretreatment with lipoteichoic acid (LTA) of Enterococcus faecalis. Forty-eight human single-rooted teeth were instrumented with R25.08 (RECIPROC) and sterilized by gamma irradiation. LTA was inoculated in the root canal of each specimen for 96 hours. Specimens were instrumented with 40.06 and 50.05 (RECIPROC) and medicated with: I) Pyrogen-free saline solution (SS); II) 2% CHX gel; III) Ca(OH)2 + SS; or IV) Ca(OH)2 + CHX for 14 days. Three samples (S) were performed of the root canal of each specimen at: S1) immediately after instrumentation; S2) after Ethylenediaminetetraacetic acid (EDTA); S3) after intracanal medication removal. Subsequent quantification of cytokines (IL-1β, TNF-α, MIP-1α, IP-10, G-CSF and IL-6) by immunosorbent assay (ELISA) and nitric oxide by the Griess method was carried-out. Data were submitted to a normality test and then analyzed with one-way ANOVA and Tukey test with a significance level of 5% using GraphPad Prism 6. Ca(OH)2 + SS and Ca(OH)2 + CHX presented lower levels of TNF-α, TNF-α, IL-6, G-CSF and nitric oxide. Ca(OH)2 + SS was the most effective in reducing MIP-1α. CHX was effective in reducing IL-6 and G-CSF. Therefore, the combined intracanal medication of calcium hydroxide and chlorhexidine is effective in reducing the cytokines TNF-α, IL-1β, IL-6, G-CSF and nitric oxide.

Resumo O objetivo deste estudo foi avaliar in vitro o efeito do hidróxido de cálcio [Ca(OH)2], clorexidina gel a 2% (CHX) em macrófagos (RAW 264.7) na produção de citocinas pró-inflamatórias e óxido nítrico após pré-tratamento com ácido lipoteicóico (LTA) de Enterococcus faecalis. Quarenta e oito dentes humanos uniradiculares foram instrumentados com R25.08 (RECIPROC) e esterilizados por irradiação gama. LTA foi inoculado no canal radicular de cada espécime por 96 horas. Os espécimes foram instrumentados com 40.06 e 50.05 (RECIPROC) e medicados com: I) solução salina apirogênica (SS); II) CHX 2%; III) Ca(OH)2 + SS; ou IV) Ca(OH)2 + CHX durante 14 dias. Três amostras (S) foram realizadas do canal radicular de cada espécime em: S1) imediatamente após a instrumentação; S2) após ácido etilenodiaminotetracético (EDTA); S3) após a retirada da medicação intracanal. Foi realizada a quantificação subsequente de citocinas (IL-1β, TNF-α, MIP-1α, IP-10, G-CSF e IL-6) por imunoensaio (ELISA) e de óxido nítrico pelo método de Griess. Os dados foram submetidos a um teste de normalidade e então analisados com ANOVA one-way e teste de Tukey com nível de significância de 5% usando GraphPad Prism 6. Ca(OH)2 + SS e Ca(OH)2 + CHX apresentaram níveis mais baixos de TNF -α, TNF-α, IL-6, G-CSF e óxido nítrico. Ca(OH)2 + SS foi o mais eficaz na redução de MIP-1α. CHX foi eficaz na redução de IL-6 e G-CSF. Sendo assim a associação de hidróxido de cálcio e clorexidina é eficaz na redução das citocinas TNF-α, IL-1β, IL-6, G-CSF e óxido nítrico.

Filling ability of three bioceramic root-end filling materials: A micro-computed tomography analysis.

This study aimed at evaluating the volume of gaps and voids, and the total porosity percentage of three calcium-silicate-based materials in mandibular molars apicoectomy by Micro-CT analysis. Thirty-three mesial roots of extracted human mandibular molars were instrumented and obturated. The apical 3mm of each root was resected and prepared. Root-end cavities were filled with EndoSequence BC Putty (BC); ProRoot MTA (MTA) and Biodentine (BIO). Samples were scanned using a Micro-CT scanner and the tridimensional images reconstructed. Percentage of gaps (VG%) and of voids (VV%) were obtained. Porosity percentage (Po%) was also assessed. Data were analysed using Student's t-test (P < 0.05). All materials presented gaps and voids. VG% was 2.006 (BC), 1.882 (MTA) and 1.450 (BIO), and VV% was 0.039 (BC), 0.021 (MTA) and 0.024 (BIO) with no statistical difference. Po% were 56.73 (BC), 51.94 (MTA) and 50.45 (BIO), with BC being statistically (P > 0.05) more porous.

Primary versus post-treatment apical periodontitis: microbial composition, lipopolysaccharides and lipoteichoic acid levels, signs and symptoms.

OBJECTIVES: To compare the microbial load and composition and to determine the lipopolysaccharides (LPS) and lipoteichoic acid (LTA) concentrations found in primary apical periodontitis (PAP) and post-treatment apical periodontitis (PTAP), correlating these findings with clinical/tomographic features. MATERIAL AND METHODS: Sixty patients with PAP (31) and PTAP (29) were submitted to clinical and tomographic assessment. Samples were collected from each root canal using paper points for microbiological assessment (culture technique and Checkerboard DNA-DNA hybridization) and determination of LPS and LTA levels (limulus amebocyte lysate and enzyme-linked immunosorbent assays, respectively). Data were correlated with clinical/tomographic findings and statistically analyzed using the Mann-Whitney and Pearson correlation tests (α = 5%). RESULTS: A higher number of cultivable bacteria and LPS were found in PAP (p < 0.05). The median number of species per root canal found in PAP and PTAP was 9 and 22, respectively (p < 0.05). LPS was positively correlated with a larger periapical lesion volume (p < .05). LTA levels were similar in both infections and had no correlation with signs and symptoms. In PAP, gram-positive bacteria were correlated with spontaneous pain (p < .05) and exudate (p < .05). Tenderness to percussion and pain on palpation were correlated to the presence of both gram-positive and negative bacteria. In PTAP, a positive correlation was observed between both gram-positive and gram-negative bacteria with exudate and periapical lesion volume (p < .05). CONCLUSIONS: PAP had higher contents of microbial load and LPS compared with PTAP. However, PTAP presented a more diverse microbiota compared with PAP. Higher content of LPS was positively correlated with larger periapical bone destruction, whereas signs and symptoms with specific microorganisms. CLINICAL RELEVANCE: It was verified that PAP and PTAP are polymicrobial infections with predominance of gram-negative bacteria and a more diverse bacterial population found in PTAP. A wide interaction of specific microbial species resulted in different clinical features in both infections.

Anti-inflammatory and anti-resorptive efficacy of adrenergic blockers on late replanted rat incisors.

BACKGROUND/AIM: In addition to their anti-inflammatory and anti-osteoclastogenesis properties, adrenergic blockers may also have promising anti-resorptive effects that can prevent root resorption when teeth are replanted because of avulsion. The aim of this study was to investigate the effects of phentolamine (Ph) and propranolol (Pr) in gels on the repair process of late replanted rat incisors. A further aim was to evaluate the biocompatibility of both drugs to human periodontal ligament fibroblasts (HPDLFs). MATERIALS AND METHODS: Forty-eight maxillary right incisors were extracted from male Wistar rats, stored in paper napkins for 60 minutes, and randomly allocated to one of eight groups (n = 6). The root canal, root surface, and alveolus were treated with 0.75 µg/mL Ph gel (Ph0.75), 10 µg/mL Ph gel (Ph10), 100 µg/mL Ph gel (Ph100), 2.5 µg/mL Pr gel (Pr2.5), 10 µg/mL Pr gel (Pr10), 100 µg/mL Pr gel (Pr100), or sodium carboxymethylcellulose gel (CMC) before replantation. In the control group (CH), only the root canal was treated with calcium hydroxide paste. Thirty days following surgery, the animals were euthanized, and the right hemimaxilla was removed to perform micro-CT and histomorphometric analysis to determine osteoclastic activity. Ethanolic solutions of Ph10 and Pr10 were selected based on the in vivo study, and the viability of HPDLFs stimulated with lipopolysaccharide was determined by MTT assays. RESULTS: The micro-CT and histomorphometric analysis revealed no significant differences among the treatments (P > .05). The presence of active osteoclasts was significantly decreased in the Ph10 and Pr10 groups (P < .05). Ph10 and Pr10 produced statistically similar cell survival rates compared to the control group (P > .05). CONCLUSIONS: Ph10 and Pr10 significantly decreased osteoclastogenesis in delayed replanted rat teeth and were not cytotoxic toward HPDLFs.

The influence of adrenergic blockade in rats with apical periodontitis under chronic stress conditions.

OBJECTIVE: To investigate the influence of chronic stress and adrenergic blockade in a rat model of apical periodontitis. METHODS: Thirty-two Wistar rats were submitted to an animal model of periapical lesion and randomly divided into 4 groups (n = 8): no stress (NS); stress + saline solution (SS); stress + ß-adrenergic blocker (Sß); stress + α-adrenergic blocker (Sα). The SS, Sß and Sα groups were submitted to an animal model of chronic stress for 28 days and received daily injections of saline solution, propranolol (ß adrenergic blocker) and phentolamine (α adrenergic blocker), respectively. After 28 days the animals were euthanized and the following analyses were carried out: a) serum corticosterone levels through Radioimmunoassay; b) measurement of serum levels of IL-1B, IL-6, IL-10 and IL-17 by enzyme-linked immunosorbent assay (ELISA); c) volume of periapical bone resorption by micro-computed tomography; d) histomorphometric analysis by staining with hematoxylin and eosin; e) expression of ß-AR, α-AR, receptor activator of nuclear factor kappa-B ligand (RANKL) and osteoprotegerin (OPG) by immunohistochemistry; f) tartrate-resistant acid phosphatase (TRAP) staining; g) ex-vivo cytokine release followed by the stimulation with LPS in superfusion system, by ELISA. RESULTS: SS group displayed significantly higher corticosterone levels than NS group (non-stressed). Higher IL-1ß serum level was observed in the NS group (p < .05); compared to all stressed groups. Other cytokines were present in similar amounts in the serum of all groups. All groups presented similar periapical lesions. All groups presented moderate inflammatory infiltrate, without statistically significant differences between them. No differences were observed regarding ß-AR, α-AR, Rank-L and OPG expression. The number of TRAP-positive cells was significantly decreased in the groups that received daily injections of adrenergic blockers. The IL-1ß release followed LPS stimulation was significantly suppressed when the superfusion media contained propranolol (p < .05). Perfusion containing phentolamine induced a greater release of IL-10. TGF-ß was significantly suppressed by phentolamine perfusion in the NS group (p < .05). CONCLUSIONS: Chronic stress can significantly change the inflammatory cytokines release. Rank-L/OPG system and periapical lesion volume were not affected following the current method applied. The administration of adrenergic blockers was not able to modulate the inflammatory response but presented effectivity in reducing the number of osteoclasts in the periapical region.

Effect of passive ultrasonic activation on microorganisms in primary root canal infection: a randomized clinical trial

Abstract Objective: This clinical study sought to evaluate the effectiveness of passive ultrasonic activation (PUA) in eliminating microorganisms in primary endodontic infection (PEI) after instrumentation of root canals using microbiological culture and checkerboard DNA-DNA hybridization. Methodology: Twenty root canals with PEI and apical periodontitis were selected. The root canals were instrumented and then randomly divided into 2 groups, according to the irrigation method: PUA and conventional needle irrigation (CNI). Microbiological samples were collected before instrumentation (S1), after instrumentation (S2) and after irrigation with 17% EDTA (S3). The samples were subjected to anaerobic culture technique and checkerboard DNA-DNA hybridization analysis. Results: A statistically significant difference was found between CNI (23.56%) and PUA (98.37%) regarding the median percentage values for culturable bacteria reduction (p<0.05). In the initial samples, the most frequently detected species was S. constellatus (50%), and after root canal treatment was E. faecalis (50%). Conclusion: Both treatments significantly decreased the number of bacterial species compared with the initial sample. However, no statistical difference in the total microbial load between PUA and CNI groups was detected. The number of cultivable anaerobic bacteria reduced significantly using PUA, and the bacterial composition and number of bacterial species after using either CNI or PUA was similar.

Incorporating N-acetylcysteine and tricalcium phosphate into epoxy resin-based sealer improved its biocompatibility and adhesiveness to radicular dentine.

OBJECTIVE: This in vitro study was designed to evaluate the biocompatibility, adhesiveness, and antimicrobial activity of epoxy resin-based sealer associated with N-Acetylcysteine (NAC) or beta-tricalcium phosphate nanoparticles (ß-TCP) as an experimental retro-filling material. METHODS: Cytotoxicity was assessed using 2,3-Bis-(Methoxy-4-Nitro-5-Sulphophenyl)-2H-Tetrazolium-5-Carboxanilide (XTT) and Sulforhodamine B (SRB) assays after exposing human periodontal ligament fibroblasts to extracts of the materials for 1, 3, or 7 days. For the adhesive resistance test, root canals (48 single-root teeth) were instrumented with Reciproc #40 files (VDW GmbH, Germany) and obturated. After 7 days, the apices were sectioned and a retrograde cavity prepared and filled with the experimental materials (Mineral trioxide aggregate, Epoxy sealer, Epoxy sealer+NAC, and Epoxy sealer+ß-TCP). For the push-out test, one 2-mm thick slice was obtained from the apical third of each specimen. Antimicrobial activity was performed using agar diffusion method. Biofilms were grown in microplates and exposed to the extracts of retro-filled materials, followed by analysis of growth inhibition on agar plates. RESULTS: Epoxy sealer in association with ß-TCP or NAC showed better bond strength while Mineral trioxide aggregate allowed for the lowest adhesion. Mineral trioxide aggregate, Epoxy sealer+ß-TCP, and Epoxy sealer+NAC showed low cytotoxicity. Epoxy sealer was the most cytotoxic. In antimicrobial activity assays, all materials had no effect on Candida albicans. Addition of NAC improved the antimicrobial property of Epoxy sealer against Enterococcus faecalis compared to unmodified Epoxy sealer (P<0.05). SIGNIFICANCE: Incorporating ß-TCP or NAC with Epoxy sealer could improve the adhesiveness and biocompatibility for better use in endodontic therapy.

The wear performance of glazed and polished full contour zirconia

Abstract This study evaluated the influence of zirconia surface finishes on the wear of an enamel analogue. 40 zirconia discs were divided into four groups: control (without finish); glazed; polished; polished and glazed. All samples were subjected to wear against steatite antagonists. The specimens underwent roughness, topographic, scanning electron microscopy (SEM) and wettability analyses. Quantitative wear measurements were performed on both steatites and discs. To measure wear of steatites the weight before and after the test and the diameter after the test were used. Profilometer measurements were performed to determine the wear on discs. Roughness, volumetric wear and mass loss were analyzed using the Kruskal-Wallis and Dunn tests (5%), while contact angle values were analyzed with One-way ANOVA and Tukey tests (5%). The polished group had the lowest roughness means, being statistically different from the other groups (p-value=0.0001). The glazed group presented the lowest steatite volumetric wear (p-value=0.0001), but not statistically different from the polished and glazed group, whereas these groups presented the highest zirconia volumetric wear, with statistically different (p-value=0.0002) compared to the others. SEM showed irregularities on the control groups surface, grooves on the polished group, and a homogeneous surface for the glazed group with a few pores. All groups presented contact angles lower than 90 degrees, characterizing hydrophilic surfaces. It can be concluded that just glazed zirconia caused less wear on the antagonist when compared to no finish and polished zirconia.

Resumo Este estudo avaliou a influência dos acabamentos superficiais da zircônia no desgaste de um análogo de esmalte. 40 discos de zircônia foram divididos em quatro grupos: controle (sem acabamento); glazeado; polido; polido e glazeado. Todas as amostras foram submetidas ao desgaste contra antagonistas de esteatita. Os espécimes foram submetidos à análise de rugosidade, topografia, microscopia eletrônica de varredura (MEV) e análise de molhabilidade. Medidas quantitativas de desgaste foram realizadas nos antagonistas e nos discos de zircônia. Para medir o desgaste das esteatitas, foi mensurado o peso antes e depois do desgaste e o diâmetro após o teste. A análise de perfilometria mensurou o desgaste dos discos. Rugosidade, desgaste volumétrico e perda de massa foram analisados pelos testes de Kruskal-Wallis e Dunn (5%), enquanto os valores de ângulo de contato foram analisados com os testes One-way ANOVA e Tukey (5%). O grupo polido apresentou as menores médias de rugosidade, sendo estatisticamente diferente dos demais grupos (p-valor=0,0001). O grupo glazeado apresentou o menor desgaste volumétrico do antagonista (p-valor=0,0001), mas não foi estatisticamente diferente do grupo polido e glazeado, enquanto que esses grupos apresentaram o maior desgaste volumétrico da zircônia, com diferença estatisticamente diferente (p-valor=0,0002) em comparação com os outros. MEV mostrou irregularidades na superfície do grupo controle, sulcos no grupo polido e uma superfície homogênea para o grupo glazeado, com poucos poros. Todos os grupos apresentaram ângulos de contato menores que 90 graus, caracterizando superfícies hidrofílicas. Dentro das limitações deste estudo in vitro, é possível concluir que zircônia glazeada causou menos desgaste ao antagonista quando comparada a zircônia sem tratamento ou polida. Além disso, não foi encontrada diferença no desgaste do antagonista para os grupos de zirconia polida e controle.

Microbial profile and endotoxin levels in primary periodontal lesions with secondary endodontic involvement

Abstract This study was carried out to investigate the microbial profile and endotoxin levels of endodontic-periodontal lesions of periodontal origin. Periodontal and endodontic samples were taken from periodontal pockets and necrotic root canals of 10 teeth with endodontic-periodontal lesions. Evidencing of 40 different bacterial species were determined in each endodontic and periodontal sample using the checkerboard DNA-DNA hybridization method and Kinetic chromogenic LAL assay was used for quantification of endotoxins. Fisher's exact test correlated the bacterial species with the endodontic or periodontal microbiota. The endotoxin levels (EU/mL) found in samples of the root canal and periodontal pocket were compared by the Wilcoxon test (p<0.05). Bacteria and LPS units were found in 100% of the endodontic and periodontal samples. The species E. faecium, P. acnes, G. morbillorum, C. sputigena and L. buccalis were strongly correlated with the endodontic microbiota and P. nigrescens with the periodontal microbiota. P. intermedia, P. endodontalis and V. parvula were more prevalent in both endodontic and periodontal microbiots. The endotoxin levels in the periodontal pocket (89600 EU/mL) were significantly higher than in the root canal (2310 EU/mL). It was concluded that the microbiota present in the periodontal and endodontic tissues is similar, with a higher prevalence of species of the orange complex and a higher level of endotoxin in the periodontal pockets.

Resumo Este estudo foi realizado para investigar o perfil microbiano e os níveis de endotoxina de lesões endoperiodontais de origem periodontal. Amostras periodontais e endodônticas foram obtidas de bolsas periodontais e canais radiculares necróticos de 10 dentes com lesões endoperiodontais. A investigação de 40 espécies bacterianas diferentes foram determinadas em cada amostra endodôntica e periodontal usando o método de hibridização de DNA-DNA (checkerboard) e o ensaio cinético cromogênico LAL foi usado para quantificação de endotoxinas. O teste exato de Fisher correlacionou as espécies bacterianas com a microbiota endodôntica ou periodontal. Os níveis de endotoxina (EU/mL) encontrados nas amostras do canal radicular e na bolsa periodontal foram comparados pelo teste de Wilcoxon (p<0,05). Bactérias e unidades de LPS foram encontradas em 100% das amostras endodônticas e periodontais. As espécies E. faecium, P. acnes, G. morbillorum, C. sputigena e L. buccalis foram fortemente correlacionadas com a microbiota endodôntica e P. nigrescens com a microbiota periodontal. P. intermedia, P. endodontalis e V. parvula foram mais prevalentes em ambas microbiotas endodôntica e periodontal. Os níveis de endotoxina na bolsa periodontal (89600 EU/mL) foram significativamente maiores do que no canal radicular (2310 EU/mL). Concluiu-se que a microbiota presente nos tecidos periodontal e endodôntico é semelhante, com maior prevalência de espécies do complexo laranja e maior nível de endotoxina nas bolsas periodontais.

Effect of age-related alterations on the biomechanics of teeth affected by non-carious cervical lesions / Efeito de alterações relacionadas à idade na biomecânica de dentes afetados por lesões cervicais não-cariosas

Objective: The prevalence of non-carious cervical lesions (NCCLs) has increased in the recent years, especially in the elderly population. The successful prevention and treatment of those lesions requires an understanding of the biomechanics of aged teeth. Considering the importance of such aspect, the impact of the age-related dentin deposition on the stress distribution of NCCLs was evaluated by means of finite element analysis. Material and Methods: A 2-dimensional model of a sound maxillary first premolar was created using CAD software. Two tooth geometries (sound, aged) and two lesion shapes (wedge, saucer) were simulated to the model. The mesh was built with 35,000 triangle and square elements of 0.1 mm in length. All tissues were considered isotropic, homogeneous and linear. Occlusal surfaces were loaded with 300 N for simulating normal chewing forces. The stress distribution was analyzed by a color scale and by the maximum principal stress at the cavosurface line angle. Results: The aged models presented lower stress concentration in the overall system in comparison to sound models. The sharp angle of wedge shaped lesions promoted higher stress concentration at the center of cavosurface angle, favoring the lesions progression. Conclusion: Considering the limitations of the current methodology, it is possible to conclude that aged tooth is a more compact structure that can better respond to stress loadings. This protective intrinsic mechanism should be considered when adopting preventive and restorative measures for NCCLs for the elderly. (AU)

Objetivo: A prevalência de lesões cervicais nãocariosas (NCCLs) tem aumentado nos últimos anos, especialmente em idosos. A prevenção e o tratamento bem-sucedidos dessas lesões requerem um entendimento da biomecânica dos dentes envelhecidos. Considerando a importância de tal aspecto, o impacto da deposição de dentina que ocorre com a idade na distribuição de estresse de NCCLs foi avaliado por meio da análise de elementos finitos. Material e Métodos: Um modelo bidimensional de um primeiro pré-molar superior foi criado usando o software CAD. Duas geometrias de dentes (padrão, envelhecido) e duas formas de lesão (cunha, pires) foram simuladas ao modelo. A malha foi construída com 35.000 elementos triangulares e quadrados de 0,1 mm de comprimento. Todos os tecidos foram considerados isotrópicos, homogêneos e lineares. Superfícies oclusais foram carregadas com 300 N para simular forças normais de mastigação. A distribuição das tensões foi analisada por uma escala de cores e pela tensão principal no ângulo da linha cavo-superficial. Resultados: Os modelos envelhecidos apresentaram menor concentração de tensão no sistema geral em comparação aos modelos padrão. O ângulo agudo das lesões em cunha promoveu maior concentração de tensão no centro do ângulo cavo-superficial, favorecendo a progressão das lesões. Conclusão: Considerando as limitações da metodologia atual, é possível concluir que o dente envelhecido é uma estrutura mais compacta que pode responder melhor às cargas de estresse. Este mecanismo intrínseco de proteção deve ser considerado ao adotar medidas preventivas e restaurativas para as NCCLs em idosos. (AU)

Influence of Zingiber officinale Extract on Push-Out Bond Strength of Glass-Fiber Post

Abstract The aim of this study was to evaluate in vitro the influence of 20% glycolic ginger extract on the bond strength of glass-fiber post cemented with etch-and-rinse or self-etching resin cement. Forty-eight bovine roots were standardized (17±0.5 mm) and randomly divided into two groups, according to irrigant used during biomechanical preparation: NaOCl: 1% sodium hypochlorite and GEN: 20% glycolic ginger extract. Root canal was prepared and randomly assigned to one of two subgroups (n=12), according to luting protocol: self-etching (RelyX U200) and etch-and-rinse (RelyX ARC). After 48 h, the roots were sectioned perpendicularly (4 in cervical third and 3 in middle third) and submitted to push-out bond strength test (50 kgf load cell, 1.0 mm/min). Failure mode was analyzed in SEM and stereomicroscopy. Data were analyzed by three-way ANOVA and Tukey tests (p<0.05). ANOVA revealed that there were no significant differences between U200 (2.01±0.17)B and ARC (1.93±0.12)B in GEN group, and at the cervical third for both irrigants, 2.69±0.16A and 2.54±0.17A, respectively. Middle third showed lower bond strength than cervical third, regardless the irrigant. Overall, the cervical third regardless the irrigant employed and the association between NaOCl and ARC, presented better adhesive performance.

Resumo O objetivo deste estudo foi avaliar in vitro a influência do extrato de gengibre 20% na resistência adesiva de pinos de fibra de vidro cimentados com cimentos resinosos convencional ou autocondicionantes. Quarenta e oito raízes bovinas foram padronizadas (17±0.5mm) e aleatoriamente divididas em dois grupos (n=24), de acordo com o irrigante utilizado durante o preparo biomecânico: NaOCl: hipoclorito de sódio 1%; e GEN: extrato glicólico de gengibre 20%. Os canais radiculares foram preparados e aleatoriamente distribuídos em dois subgrupos (n=12), conforme o protocolo de cimentação: autocondicionante (RelyX U200) e convencional (RelyX ARC). Após 48 h, as raízes foram seccionadas perpendicularmente (4 no terço cervical e 3 no terço médio) e submetidas ao teste de resistência adesiva push-out (50 kgf carga, 1.0 mm/min). O modo de falha foi analisado em MEV e estereomicroscopia. Os dados foram analisados pelos testes ANOVA 3-fatores e Tukey (p<0.05). ANOVA mostrou que não houve diferenças significativas entre U200 (2.01±0.17)B e ARC (1.93±0.12)B no grupo GEN, e no terço cervical para ambos os irrigantes (2.69±0.16A and 2.54±0.17A, respectivamente). O terço médio apresentou menor resistência adesiva que o terço cervical, independente do irrigante. Em geral, o terço cervical, independentemente do irrigante empregado, e a associação entre NaOCl e ARC apresentaram melhores desempenhos adesivos.

Effect of thermocycling aging on the flexural strength of feldspathic ceramic / Efeito da termociclagem na resistência flexural de uma cerâmica feldspática

Objective: The aim of the study was to evaluate the effect of aging through different thermocycling protocols on the flexural strength of a feldspathic ceramic. Material and Methods: Fifty ceramic bars, Vitablock Mark II (VITA), with dimensions of 18X4X2mm were prepared. The bars were randomly distributed to the groups (n = 10), which were defined according to the number of thermal cycles (TCy): G0 - no TCy; G500 - 500 cycles of TCy; G6000 - 6000 TCy; G10000 - 10000 TCy; G15000 - 15000 TCy. After aging, the specimens were subjected to the three point bending test in a universal testing machine (EMIC DL 1000), under 0.5mm/min speed and loading of 50kgf, until catastrophic failure. A sample of each group was evaluated for topographic morphology under Scanning Electron Microscopy. Results: For mechanical testing, the following mean values and standard deviation (MPa) were obtained: G0 (99.78 ± 5.07); G500 (101.64 ± 5.59); G6000 (98.13 ± 4.95); G10000 (91.77 ± 9.68); G15000 (101.51 ± 4.22). An analysis of variance by regression equation (p = 0.387) was performed, demonstrating a weak and non-significant correlation between flexural strength and number of thermal cycles. Conclusion: It can be concluded that aging in water solely by different numbers of temperature cycles did not influence on the flexural strength of a feldspathic ceramic. (AU)

Objetivo: O objetivo do estudo foi avaliar o efeito do envelhecimento por meio de diferentes protocolos de ciclos térmicos na resistência à flexão de uma cerâmica feldspática. Material e Métodos: Cinquenta barras de cerâmica, Vitablock Mark II (VITA), com dimensões de 18X4X2mm foram preparadas. As barras foram distribuídas aleatoriamente pelos grupos (n = 10), os quais foram definidos de acordo com o número de ciclos térmicos (CT): G0 ­ nenhum; G500 - 500 CT; G6000 - 6000 CT; G10000 ­ 10000 CT; G15000 - 15000 CT. Após envelhecimento, as amostras foram submetidas ao ensaio de flexão de três pontos em uma máquina de ensaios universal (EMIC DL 1000), sob a velocidade de 0,5 mm / min e célula de carga de 50kgf, até uma falha catastrófica. Uma amostra de cada grupo foi avaliada quanto à morfologia topográfica sob microscopia eletrônica de varredura. Resultados: Para os testes mecânicos, foram obtidos os seguintes valores médios e desvio padrão (MPa): G0 (99,78 ± 5,07); G500 (101,64 ± 5,59); G6000 (98,13 ± 4,95); G10000 (91,77 ± 9,68); G15000 (101,51 ± 4,22). Uma análise de variância pela equação de regressão (p = 0,387) foi realizada, o que demonstra uma correlação fraca e não significativa entre a resistência à flexão e do número de ciclos térmicos. Conclusão: Pode-se concluir que o envelhecimento somente em água por diferentes números de ciclos térmicos não influenciou na resistência à flexão de uma cerâmica feldspática (AU)

Effects of porcelain thickness on the flexural strength and crack propagation in a bilayered zirconia system

Abstract Objective: This study evaluated the influence of porcelain (VM9, VITA Zahnfabrik, Germany) thickness on the flexural strength and crack propagation in bilayered zirconia systems (YZ, VITA Zahnfabrik, Germany). Material and Methods: Thirty zirconia bars (20.0x4.0x1.0 mm) and six zirconia blocks (12.0x7.5x1.2 mm) were prepared and veneered with porcelain with different thickness: 1 mm, 2 mm, or 3 mm. The bars of each experimental group (n=10) were subjected to four-point flexural strength testing. In each ceramic block, a Vickers indentation was created under a load of 10 kgf for 10 seconds, for the propagation of cracks. Results: The results of flexural strength were evaluated by One-way ANOVA and Tukey's test, with a significance level of 5%. The factor "thickness of the porcelain" was statistically significant (p=0.001) and the l-mm group presented the highest values of flexural strength. The cracks were predominant among the bending specimens with 1 and 2 mm of porcelain, and catastrophic failures were found in 50% of 3-mm-thick porcelain. After the indentation of blocks, the most severe defects were observed in blocks with 3-mm-thick porcelain. Conclusion: The smallest (1 mm) thickness of porcelain on the zirconia infrastructure presented higher values of flexural strength. Better resistance to defect propagation was observed near the porcelain/ zirconia interface for all groups. Higher flexural strength was found for a thinner porcelain layer in a bilayered zirconia system. The damage caused by a Vickers indentation near and far the interface with the zirconia shows that the stress profiles are different.

Effectiveness in the Removal of Endotoxins and Microbiological Profile in Primary Endodontic Infections Using 3 Different Instrumentation Systems: A Randomized Clinical Study.

INTRODUCTION: This clinical study was conducted to correlate the microbiological profile and levels of endotoxins found in primary endodontic infection with the presence of clinical features and to evaluate the removal of microorganisms and endotoxins using rotary, reciprocating, and hybrid systems for biomechanical preparation. METHODS: Thirty single root canals with primary endodontic infection were evaluated with signs and symptoms and were randomly divided into 3 groups according to the instrumentation system used (n = 10) as follows: rotary Mtwo instruments (VDW, Munich, Germany) with 8 files, the reciprocating Reciproc system (VDW) with a single file, and Genius hybrid instruments with 3 files (1 rotary and 2 reciprocating files) with irrigation using 24 mL 2.5% sodium hypochlorite. Samples were collected before (S1) and after instrumentation (S2) before being submitted to microbiological culture (colony-forming units/mL) and the checkerboard DNA-DNA hybridization test. Endotoxins were quantified using the limulus amebocyte lysate assay. RESULTS: Microbiological culture showed statistical differences in the reduction of colony-forming units/mL with all systems tested (P < .05), but no statistical difference was found among the groups. The most frequently detected species were Capnocytophaga ochracea (53%) and Fusobacterium nucleatum (53%) at S1 and F. nucleatum (50%) and Leptotrichia buccalis (50%) at S2. As for the reduction of endotoxins at S2, Mtwo presented the best results (95.05%) followed by the Genius (91.85%) and Reciproc (64.68%) groups, but no statistical difference was found among the groups. Previous pain, tenderness to percussion, and presence of a sinus tract were associated with specific microorganisms (P < .05). CONCLUSIONS: Signs and symptoms were correlated with microorganisms. Endodontic treatment was effective in reducing bacteria and endotoxins but was not capable of completely removing them from the root canal.

Configuration of canal system in the mesiobuccal root of maxillary first molars / Configuración del sistema de canales en la raíz mesiovestibular de los primeros molares superiores

Knowledge on internal dental morphology is extremely important to the correct execution of the treatment and modeling of root canals. The aim of the present study was to analyze, "in vitro", the configuration of the canal system in the mesial root of 180 maxillary first molars obtained from a Brazilian sample. The teeth diaphanized in order to confirm the presence of the MB2 canal and to determine the internal anatomy. Only in 41 % of the teeth the orifice of MB2 canal emerged from the pulp chamber floor. Among teeth two apical foramina were found in 23.3 %, a single apical foramen was observed in 15,5 % and three apical foramina were present in 2,2 %. Considering the hole sample, the MB2 canal was observed in 65.4 %, merged with the MB1 or branching from the MB1 or even independent all the way. In 55.1 % of the teeth a single orifice stemmed from the pulp chamber floor. A single apical foramen was found in 43,5 % of these cases, two apical foramina were present in approximately 12 % and apical delta was found in 0,5 % of the specimens. Three orifices which emerged from the pulpal floor were observed in 1.1 % of the sample. Paths considered very atypical were observed at 2.8 % of the teeth. The MB root canal anatomy was complex. The frequency of occurrence of the bifurcated or double canal, isthmus, accessory canals and apical delta must be taken into consideration as a possible cause of otherwise unexplained failure during endodontic treatment of first maxillary molars.

El conocimiento de la morfología dental interna es muy importante para la correcta ejecución del tratamiento de conductos radiculares. El objetivo del presente estudio fue analizar "in vitro", la configuración del sistema de canales en la raíz mesiobucal del 180 primeros molares maxilares obtenidos de una muestra brasileña. Mediante el uso de la diafanización de los dientes se confirmó la presencia del canal MB2 y su anatomía interna. Solamente 41 % de los dientes el canales MB2 emergió desde el piso de la cámara pulpar. De estos dientes fueron encontrados dos forámenes apicales en 23,3 %, un solo foramen apical en 15,5 % y tres forámenes apicales estaban presentes en el 2,2 %. Teniendo en cuenta la totalidad de la muestra, 65,4 % presentaron el MB2; que se fusionó con el MB1, a veces se observó como una rama de este canal, o sigue siendo independiente hasta el final. En 55,1 % de los dientes un solo foramen provenía del piso de la cámara pulpar. Un solo foramen apical se encontró en 43,5 % de estos casos, dos forámenes apicales estaban presentes en aproximadamente el 12 % y el delta apical se encontró en 0,5 % de los especímenes. Tres canal a partir del piso pulpar se observaron en 1,1 % de la muestra. Caminos considerados muy atípicos se observaron a 2,8 % de los dientes. La morfología del sistema de canales radiculares en la raíz MB de los primeros molares maxilares es compleja y variable. La alta frecuencia de canal MB2, bien como la presencia del canal bifurcado o doble, istmo, conductos accesorios y delta apical deben tenerse en cuenta como posible causa de la falla de otro modo inexplicable durante el tratamiento endodóntico de los primeros molares maxilares.

Limewater and Polymyxin B Associated with NaOCl for Endotoxin Detoxification in Root Canal with Necrotic Pulp

Abstract This clinical study investigated the effects of endodontic treatment by using different irrigants (limewater + NaOCl and polymyxin B + NaOCl) and intracanal medication on endotoxins in teeth with primary endodontic infection and radiographically visible apical periodontitis. Thirty-three teeth with necrotic pulp and periapical lesions from different patients were selected for this study. Samples were collected after the coronal opening (S1) and after instrumentation (S2). Root canals were divided in 3 groups (n = 11) according to the irrigant combination used: NaOCl + LW: 2.5% NaOCl + calcium hydroxide solution (0.14%, limewater); NaOCl + PmB: 2.5% NaOCl + 10.000 UI/mL polymyxin B; 2.5% NaOCl (control). The third sampling (S3) was performed after ethylenediaminetetraacetic acid and the fourth (S4) after samples got 14 days with intracanal medication with 2% chlorhexidine gel + calcium hydroxide. Endotoxins (lipopolysaccharide) were quantified by chromogenic Limulus amebocyte lysate (LAL). Endotoxins were detected in all root canals after the coronal opening (S1). NaOCl + PmB group presented the greatest endotoxin reduction after instrumentation (76.17%), similar to NaOCl + LW group (67.64%, p<0.05) and different from NaOCl group (42.17%, p<0.05). After intracanal medication period (S4), there was significant increase of endotoxins neutralization. It was concluded that NaOCl + PmB promoted the greatest reduction of endotoxin levels, followed by NaOCl + LW. Intracanal medications had no significant complementary role in the reduction of endotoxins at the end of the treatment

Resumo Este estudo clínico investigou os efeitos do tratamento endodôntico com uso de diferentes irrigantes (NaOCl + água de cal e NaOCl + polimixina B) e medicação intracanal sobre endotoxinas em dentes com infecção endodôntica primária e presença de lesão periapical visível radiograficamente. Foram selecionados para o estudo trinta e três dentes de pacientes que apresentavam necrose pulpar e presença de lesão periapical. As amostras foram coletadas após a abertura coronária (S1) e após a instrumentação (S2). Os canais radiculares foram divididos em 3 grupos (n = 11) de acordo com a combinação de irrigantes utilizada: NaOCl + LW:- hipoclorito de sódio 2,5% + solução de hidróxido de cálcio (água de cal 0,14%); NaOCl + PmB: hipoclorito de sódio a 2,5% + polimixina B 10.000 UI/mL; NaOCl (controle): hipoclorito de sódio a 2,5%. A terceira coleta (S3) foi realizada após aplicação do ácido etilenodiamino tetra acético (EDTA) e a quarta coleta (S4) após 14 dias de medicação intracanal de hidróxido de cálcio + clorexidina gel 2%. Endotoxinas (lipopolissacarídeos) foram quantificadas pelo ensaio cromogênico do lisado de amebócitos de Limulus (LAL). Endotoxinas foram detectadas em todos os canais radiculares após abertura coronária (S1). Grupo NaOCl + PmB apresentou a maior redução de endotoxinas após a instrumentação (76,17%), sendo similar ao grupo NaOCl + LW (67,64%, P >.05) e diferente do grupo NaOCl (42,17%, P <.05). Após o período de medicação intracanal, houve aumento significativo da neutralização de endotoxinas. Concluiu-se que NaOCl + PmB promoveu a maior redução dos níveis de endotoxinas, seguido de NaOCl + LW. A medicação intracanal não teve um papel complementar significativo na redução de endotoxinas no final do tratamento.

Investigation of Bacterial Contents From Persistent Endodontic Infection and Evaluation of Their Inflammatory Potential

Abstract This clinical study investigated and quantified cultivable bacteria and their levels of endotoxins in persistent endodontic infection, determining their antigenicity against macrophages and fibroblast cells by IL-1β and TNF-α secretion and evaluating their relationship with clinical and radiographic features. Samples from the root canals were obtained after root filling removal. Culture techniques were used to determine the bacterial count and the endotoxins were determined by LAL-assay. PCR analysis (16S rDNA) was used for bacterial detection. Raw 264.5 macrophages and V79 fibroblast were stimulated with endodontic contents. ELISA assay measured the amounts of IL-1ß/TNF-?#61537; secretion. Bacteria and endotoxin medians were 1.24x105 CFU/mL and 9.62 EU/mL, respectively. Porphyromonas endodontalis was the most frequently detected species. Higher levels of endotoxins were found in teeth with pain on palpation (23.56 EU/mL) rather than in its absence (8.21 EU/mL). Larger areas of bone destruction were related to higher levels of endotoxins and IL-1β and TNF-α secretion. The study findings revealed the presence of Gram-negative bacteria species in persistent endodontic infection, with their endotoxins related to both severity of bone destruction and development of symptomatology. Moreover, larger areas of bone destruction were related to higher levels of IL-1β and TNF-α secreted by macrophages and fibroblast cells.

Resumo Este estudo clínico investigou e quantificou bactérias cultiváveis e seus níveis de endotoxinas na infecção endodôntica persistente, determinando a sua antigenicidade contra macrófagos e células de fibroblastos através de IL-1β e TNF-α; e avaliando sua relação com características clínicas e radiográficas. As amostras dos canais radiculares foram obtidas após a desobturação. Técnicas de cultura foram utilizadas para determinar a contagem de bactérias e a quantificação de endotoxinas foram determinadas por ensaio de LAL. Análise por PCR (16S rDNA) foi utilizada para a detecção bacteriana. Células 264,5 macrófagos e fibroblastos V79 foram estimuladas com conteúdo endodôntico. IL-1β e TNF-α produzidas pelas células avaliadas foram medidas por ensaio de ELISA. As medianas de bactérias e endotoxinas foram 1,24x105 UFC/mL e 9,62 EU/mL, respectivamente. Porphyromonas endodontalis foi a espécie mais frequentemente detectada. Níveis mais elevados de endotoxinas foram encontrados em dentes com dor à palpação (23,56 EU/mL) quando comparado a sua ausência (8,21 EU/mL). Maiores áreas de destruição óssea foram relacionados com níveis mais elevados de endotoxinas e IL-1β e TNF-α. O estudo revelou presença de espécies de bactérias Gram-negativas em infecção endodôntica persistente, com níveis elevados de endotoxinas relacionados a maior destruição óssea periapical e presença de sintomatologia. Além disso, grandes áreas de destruição óssea foram relacionados com níveis mais elevados de IL-1β e TNF-α secretadas por macrófagos e fibroblastos.