Effect of ethylenediaminetetraacetic acid and hyaluronic acid on the viability and cytokine expression of periodontal ligament fibroblasts.

BACKGROUND/AIMS: Dental root cell proliferation following tooth avulsion has not been well researched. Understanding the effects of dry time and dentin treatment influences on cell proliferation is essential to provide evidence-based guidelines for tooth replantation. The study evaluated the viability of periodontal ligament fibroblasts (PLF) in contact with roots, submitted to surface treatments with ethylenediaminetetraacetic acid (EDTA) and hyaluronic acid (HA) at different times, including to quantify inflammatory cytokines interleukin (IL)-6, IL-8, IL-1ß and TNF-α expressed by PLF. The adhesion of fibroblasts to treated root surfaces was also evaluated. MATERIAL AND METHODS: One hundred and eight cementum discs from bovine teeth were randomly divided into groups according to time periods of being dry (n = 12) as follows: (i) fresh discs that were not kept dry, (WD); (ii) dry for 1 hour (1 hd); and (iii) dry for 24 hours (24 hd). The discs were subdivided into 3 subgroups (n = 12) according to surface treatments: (iv) no treatment, (v) treatment with EDTA, (vi) treatment with HA. The discs were placed in 96-well plates, and PLF were seeded and kept in contact with the discs for 48 hours. Cell viability on the surface of the discs was assessed by XTT, and PLF adhesion was evaluated using scanning electron microscopy (SEM). Quantification of cytokines was performed using enzyme-linked immunosorbent assay (ELISA). Data were submitted to ANOVA and Tukey's test (α = .05). RESULTS: Surface treatment had a statistically significant effect on the cell viability in groups WD (P = .03), 1 hd (P = .01) and 24 hd (P = .048). PLF in contact with dried root surfaces expressed more cytokines without treatment with IL-6 decreasing the expression when treated with HA for 24 hours. SEM also showed adhesion of PLF to the surface of all discs at all time periods. CONCLUSION: EDTA + HA is an alternative treatment for cases of avulsed teeth as it promoted adhesion and increased viability of PLF.

Adaptation of gutta percha to oval canals: a cone beam computed tomographic analysis.

This study evaluated the influence of different instrumentation techniques on the adaptation of gutta percha cones in oval-shaped root canals. A total of 90 human single-rooted teeth were divided into groups according to the instrumentation technique that was used: group M, manual; group OM, hybrid: oscillatory plus manual; group PT, rotary (ProTaper Universal); group OPT, hybrid: oscillatory plus rotary (ProTaper Universal); group MT, rotary (Mtwo); and group OMT, hybrid: oscillatory plus rotary (Mtwo). After a single gutta percha cone was adapted to the canal, cone beam computed tomography was utilized to perform assessments at areas of greater contact of gutta percha to root canal walls. The adaptation was rated as ideal, satisfactory, minimally acceptable, or unsatisfactory. The frequency of ideal adaptation of the cone to the 4 canal walls varied from 26.67% (groups OMT and PT) to 73.33% (group MT); the differences between group MT and groups PT and OMT were statistically significant (P < 0.05). The mean area of the canal preparation without adaptation (contact) of the cone varied from 0.06 mm² (group MT) to 0.41 mm² (group OMT). The results showed that an instrumentation technique using the Mtwo system provided the best adaptation of the .04 gutta percha cones to anatomically oval root canals.

Persistent infection by Staphylococcus epidermidis in endodontic flare-up: a case report.

Endodontic flare-ups are challenging situations and may result from selective growth of specific bacterial species; microbial cultures and antibiograms should be used to allow faster, successful management of refractory lesions. A 47-year-old man reported pain on percussion after uncomplicated retreatment of the maxillary left canine for prosthetic purposes. In the following days, pain dramatically increased, leading to removal of the filling and use of intracanal medication. After many unsuccessful attempts to resolve the problem, a microbial culture of the root canal detected the presence of Staphylococcus epidermidis. An antibiogram determined the best drug combination to control this infection: tetracycline (oxytetracycline hydrochloride, 500 mg orally) plus third-generation cephalosporin (ceftriaxone, 1 g intramuscularly). Once the infection was controlled, the root canal was obturated. There was a reduction in the area of radiolucency, and the patient reported no pain at a 2-year follow-up.

Efficacy of different techniques for removal of calcium hydroxide-chlorhexidine paste from root canals.

The purpose of this study was to evaluate the efficacy of different techniques for removal of combined calcium hydroxide [Ca(OH)2] and chlorhexidine paste from root canals. Fifty single-rooted human teeth were prepared by oscillatory and rotary systems and filled with a paste of Ca(OH)2 and 2% chlorhexidine gel. After incubation for 14 days, the specimens were divided into 5 groups (n = 10), and the medication was removed by 1 of 5 different procedures. In group 1 (control), removal procedures involved a master apical file, foraminal debridement, and 5 mL of saline solution applied with the NaviTip irrigation needle. Group 2 was treated the same as group 1, but in addition 0.5 mL of 17% ethylenediaminetetraacetic acid was used for 3 minutes. In group 3, ultrasonic agitation was performed for 1 minute. Group 4 was treated as group 2, but the NaviTip FX needle was used for irrigation. In group 5, a master apical file, foraminal debridement, and 3-minute application of 5 mL of citric acid were used. After the root-cleaning procedures, the crowns were removed at the cementoenamel junction, and the roots were split longitudinally into halves. The success of intracanal medicament removal was observed under stereoscopic microscope and scanning electron microscope. Remnants of Ca(OH)2 were found in all experimental groups, regardless of the removal technique used. There was no statistically significant difference in cleanliness in the apical third of the root canal among groups 1, 2, and 3. Group 4 showed the best and group 5 the worst results with statistically significant differences. Overall, the NaviTip FX irrigation needle technique was more efficient in removing a Ca(OH)2-chlorhexidine paste from the root canal.

In vitro antimicrobial and anti-endotoxin action of Zingiber Officinale as auxiliary chemical and medicament combined to calcium hydroxide and chlorhexidine.

OBJECTIVE: This study was conducted in vitro to compare the effectiveness of Zingiber Officinale as an auxiliary chemical substance followed by placement of different intra-canal medication in removing endotoxins and cultivable micro-organisms from infected root canals. MATERIALS AND METHODS: Seventy-two root canals were contaminated with Enterococcus faecalis, Candida albicans and Escherichia coli for 28 days. After, the teeth were instrumented using Zingiber Officinale and divided into six groups according to the intra-canal medication: chlorhexidine gel; calcium hydroxide + chlorhexidine gel; glycolic ginger extract; calcium hydroxide + glycolic ginger extract; calcium hydroxide + saline solution and saline solution (control). Sample collections were performed after root canal contamination (Baseline; S1), after instrumentation (S2), 7 days after instrumentation (S3), after 14 days with intra-canal medication (S4) and 7 days after removal of intra-canal medication (S5). The results were analyzed by the Kruskal-Wallis and Dunn tests. RESULTS: It was observed that in S2 and S3 there was significant reduction of the micro-organisms and the quantity of endotoxins after instrumentation. In samples S4 and S5 there was complete elimination of micro-organisms and significant reduction of endotoxins. CONCLUSION: It was concluded that Zingiber Officinale as an auxiliary chemical substance was effective on the micro-organisms tested, yet was unable to eliminate the endotoxins. Similarly, the intra-canal medication were effective on micro-organisms, yet did not completely eliminate the endotoxins.

Comparison of the effectiveness of 3 irrigation devices for the cleaning of root canal walls instrumented with oscillatory and rotary techniques.

In this study, scanning electron microscopy (SEM) was used to evaluate the smear layer removal of 3 irrigation devices. The study hypothesis was that more contemporary side-vented needles and brush-covered needles are more efficient for smear layer removal of root canals than traditional needles. The aim of this study was to compare the effectiveness of these irrigation devices in the cleansing of root canals instrumented with oscillatory and rotary systems. Sixty single-rooted teeth were divided according to instrumentation and irrigation techniques into 6 groups. The teeth were prepared for SEM analysis to evaluate the cleansing of cervical, middle, and apical thirds. For all groups, the cleansing of the cervical and middle thirds was better than that of the apical third (P < 0.05). Regardless of the instrumentation technique, no statistically significant differences were found between the groups regarding the cleansing of root canal walls (P > 0.05).

Evaluation of the bond strengths of 3 endodontic cements via push-out test.

In this study, the push-out method was used to evaluate the bond strengths of 3 types of endodontic cements according to their composite base: methacrylate, epoxy resin, and an experimental copaiba oil resin. The study hypothesis was that the methacrylate-based and experimental cements would have bond strengths equal to or greater than that of the epoxy resin-based cement. Thirty bovine tooth roots, 18 mm long, were divided into 3 groups (n = 10) based on the chosen cement treatment. After treatment, the specimens were sectioned and submitted to a push-out test. Results showed that there was no statistically significant difference (P < 0.05) between the cements used or between the middle and apical thirds of the roots. It could be concluded that the tested cements had satisfactory and similar bond strengths to dentin.

Peroxide penetration from the pulp chamber to the external root surface after internal bleaching.

PURPOSE: To quantify the amount of peroxide penetration from the pulp chamber to the external surface of teeth during the walking bleaching technique. METHODS: Seventy-two bovine lateral incisors were randomly divided over five experimental groups and one control (n = 12 per group): (1) 35% hydrogen peroxide (HP); (2) 35% carbamide peroxide (CP); (3) sodium perborate (SP); (4) (HP+SP); (5) (CP+SP) and (6) Control (CG), deionized water. All groups were treated according to the walking bleach technique. After 7 days at 37 degrees C in an acetate buffer solution, 100 microl violet leukocrystal coloring and 50 microl peroxidase was added, producing a blue stain that could be measured in a spectrophotometer and then converted into peroxide microg/ml. RESULTS: G5 exhibited the greatest penetration, while G2 and G3 produced the lowest values. All bleaching agents penetrated from the pulp chamber to the external root surface. There was a direct correlation between the presence of oxidative agents and penetration potential. Sodium perborate in distilled water was less oxidative and appeared to be the least aggressive bleaching agent.

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.

Penetration of 38% hydrogen peroxide into the pulp chamber in bovine and human teeth submitted to office bleach technique.

This study evaluated the pulp chamber penetration of peroxide bleaching agent in human and bovine teeth after office bleach technique. All the teeth were sectioned 3 mm apical of the cement-enamel junction and were divided into 2 groups, A (70 third human molars) and B (70 bovine lateral incisors), that were subdivided into A1 and B1 restored by using composite resin, A2 and B2 by using glass ionomer cement, and A3 and B3 by using resin-modified glass ionomer cement; A4, A5, B4, and B5 were not restored. Acetate buffer was placed in the pulp chamber, and the bleaching agent was applied for 40 minutes as follows: A1-A4 and B1-B4, 38% hydrogen peroxide exposure and A5 and B5, immersion into distilled water. The buffer solution was transferred to a glass tube in which leuco crystal violet and horseradish peroxidase were added, producing a blue solution. The optical density of the blue solution was determined by spectrophotometer and converted into microgram equivalents of hydrogen peroxide. Data were submitted to analysis of variance and Dunnett, Kruskal-Wallis, and Tukey tests (5%). A higher level of hydrogen peroxide penetrated into the pulp chamber in resin-modified glass ionomer cements in bovine (0.79 +/- 0.61 microg) and human (2.27 +/- 0.41 microg) groups. The bleaching agent penetration into the pulp chamber was higher in human teeth for any experimental situation. The penetration of the hydrogen peroxide depends on restorative materials, and under the conditions of this study human teeth are more susceptible to penetration of bleaching agent into the pulp chamber than bovine teeth.

Are Desensitizing Toothpastes Equally Biocompatible and Effective Against Microorganisms?

The aims of this study were evaluate cytotoxicity, genotoxicity, antimicrobial activity of desensitizing toothpastes compared to a common one and the surface roughness of tooth enamel submitted to brushing with these toothpastes. Samples of three desensitizing toothpastes (Colgate Sensitive, Sensodyne and Oral B Sensitive) and common toothpaste (Colgate) were placed in contact with gingival human fibroblasts. Cytotoxicity and genotoxocity were measured by MTT assay and micronucleus test. Antimicrobial activity of the toothpastes extracts against C. albicans, S. mutans and S. aureus were assessed. For surface roughness evaluation, bovine teeth were submitted to 10.000 brushing cycles. The results were analyzed statically using Mann-Whitney U, ANOVA and Z tests (p<0.05). All toothpastes caused cytotoxic effect to the cells (p<0.05), except Colgate Sensitive. The toothpastes did not increase the number of micronuclei compared to the untreated control group. Colgate eliminated all the evaluated microorganisms at lower concentrations compared to Colgate Sensitive and Oral B Sensitive, which were not able to eliminate S. aureus. Sensodyne did not reach the minimum microbicidal concentration. The surface roughness of tooth enamel increased after brushing with Colgate Sensitive and Oral B Sensitive, however the comparison between groups showed no difference on the enamel surface roughness presented by desensitizing toothpastes when compared with the common one (p>0.05). Based on these results, we can conclude that although none toothpaste has induced genotoxicity, Colgate Sensitive was also not cytotoxic. Colgate was the most effective against the microorganisms, and there were no differences on the enamel surface roughness between the groups.

The Antimicrobial Peptide LL-37 as a Possible Adjunct for the Proliferation and Differentiation of Dental Pulp Stem Cells.

INTRODUCTION: This study evaluated the biocompatibility of 5 and 10 µg/mL LL-37 in vitro and its effect on the differentiation of human dental pulp stem cells (DPSCs) into odontoblast-like cells. METHODS: Cell viability, genotoxicity, nitric oxide production, cell cycle, dentine sialophosphoprotein (DSPP) production, and DSPP gene expression. RESULTS: Concentrations of 5 and 10 µg/mL of LL-37 were not cytotoxic and generally increased cell viability, especially on the third day (P < .05). The tested concentrations did not induce genotoxicity (P < .05). LL-37 did not significantly alter nitrite production at either concentration. Cell cycle analysis revealed that 10 µg/mL of LL-37 arrested cells in G0/G1 (P < .05). The control group exhibited higher numbers of cells in other phases of the cell cycle (P < .05). The expression of the DSPP protein and gene was also higher in the 10 µg/mL of LL-37 group (P < .05). CONCLUSIONS: These results demonstrated that LL-37 was biocompatible at these concentrations and increased the number of viable cells, especially during the initial period. The 10 µg/mL concentration arrested the cell cycle and increased expression of the DSPP protein and gene, which indicates that this peptide contributes to odontoblastic differentiation.

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

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-1ß 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-1ß 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-1ß and TNF-α secreted by macrophages and fibroblast cells.

Effects of Nd:YAG laser irradiation on root canal dentin wall: a scanning electron microscopic study.

OBJECTIVE: The purpose of this study was to evaluate, by scanning electron microscopy (SEM), the effects of Nd:YAG laser irradiation applied perpendicular or parallel to the root canal dentin wall. METHODS: Thirty human teeth were divided into two groups: Group A (20 roots), laser application with circular movements, parallel to the dentin root surface; and Group B (10 roots), roots cut longitudinally and laser applied perpendicular to the root surface. Group A was subdivided into A1 (10 roots), laser application with 100 mJ, 15 Hz and 1.5 W; and A2 (10 roots) with 160 mJ, 15 Hz, and 2.4 W. Group B was subdivided into B1 (10 hemisections) and B2 (10 hemi-sections) with parameters similar to A1 and A2. Four applications of 7-sec duration were performed, with a total exposure of 28 sec. SEM evaluations were made in the cervical, middle, and apical thirds, with 500x and 2000x magnifications. Morphological changes scores were attributed, and the results were submitted to Kruskal Wallis statistical test (5%). RESULTS: Significant statistical differences were found between groups Aand B (p = 0.001). In groups A1 and A2, few areas of dentin melting were observed. In groups B1 and B2, areas of melting dentin covering dentin surface were observed. CONCLUSIONS: It was concluded that intracanal laser application with circular movements (parallel to the surface) produces limited morphological changes in root canal dentin wall.

Acting and knowledge of emergency rescue teams in dental trauma / Atuação e conhecimento de equipes de resgate de emergência em casos de trauma dental

Objetivo: O traumatismo dentário é regularmente freqüente em pacientes atendidos por equipes de resgate em locais de acidentes e por equipes de emergência em hospitais. No entanto, esses profissionais recebem pouca ou nenhuma informação sobre como cuidar e atuar em casos de lesões traumáticas em tecidos duros e moles. O objetivo deste estudo foi avaliar o nível de conhecimento dos profissionais das equipes de resgate e emergência (médicos, enfermeiros e paramédicos) sobre o atendimento do traumatismo dentário. Material e métodos: 196 profissionais das unidades de resgate e emergência receberam um questionário padronizado de 5 questões referentes ao atendimento do traumatismo dentário e preservação de tecido dentário avulsionado (um dente ou seu fragmento). Resultados: os resultados mostraram um alto nível de atendimento de pacientes de traumatismo dentário por paramédicos, enfermeiros e médicos, com conhecimento muito baixo sobre como lidar com esses casos, e com resultados moderados sobre o melhor meio para preservar o dente avulsionado, e as ações tomadas durante o processo de resgate. Conclusão: há uma alta ocorrência de casos de traumatismos dentários atendidos por equipes de resgate e emergência, com falta de conhecimento, o que sugere a necessidade de mais campanhas educativas para esses profissionais com acompanhamento a longo prazo. (AU)

Micro-CT assessment of two different endodontic preparation systems.

The aim of this study was to compare two endodontic preparation systems using micro-CT analysis. Twenty-four one-rooted mandibular premolars were selected and randomly assigned to two groups. The samples (n = 12) of Group 1 were prepared using the ProTaper Universal rotary system, while Group 2 (n = 12) was prepared using the EndoEZE AET system complemented by manual apical preparation with K-type hand files up to #30. A 2.5% sodium hypochlorite solution was used in both groups for irrigating. Both groups were scanned by high-resolution microcomputed tomography before and after preparation (SkyScan 1172, SkyScan, Kontich, Belgium). The root canal volume and surface area was measured before and after preparation, and the differences were calculated and analyzed for statistically significant differences using ANOVA complemented by the Tukey test (p < 0.05). The results showed no statistically significant differences between the mean volumes of dentin removal by the two systems. However, the EndoEZE AET system presented a significantly greater mean surface area compared to the ProTaper system (p < 0.05). The EndoEZE AET system enabled preparation of a greater root canal surface area when compared to the ProTaper Universal system. There seemed to be no difference in dentin volume loss between the two systems used.

Effect of Zingiber officinale and propolis on microorganisms and endotoxins in root canals.

UNLABELLED: The purpose of this study was to evaluate the effectiveness of glycolic propolis (PRO) and ginger (GIN) extracts, calcium hydroxide (CH), chlorhexidine (CLX) gel and their combinations as ICMs (ICMs) against Candida albicans, Enterococcus faecalis, Escherichia coli and endotoxins in root canals. MATERIAL AND METHODS: After 28 days of contamination with microorganisms, the canals were instrumented and then divided according to the ICM: CH+saline; CLX, CH+CLX, PRO, PRO+CH; GIN; GIN+CH; saline. The antimicrobial activity and quantification of endotoxins by the chromogenic test of Limulus amebocyte lysate were evaluated after contamination and instrumentation at 14 days of ICM application and 7 days after ICM removal. RESULTS AND CONCLUSION: After analysis of results and application of the Kruskal-Wallis and Dunn statistical tests at 5% significance level, it was concluded that all ICMs were able to eliminate the microorganisms in the root canals and reduce their amount of endotoxins; however, CH was more effective in neutralizing endotoxins and less effective against C. albicans and E. faecalis, requiring the use of medication combinations to obtain higher success.

Calcium Hydroxide improves epoxy sealer adhesion on root dentin / O hidróxido de cálcio melhora a adesão de selante resinoso à dentina radicular

Objetivo: O intuito deste estudo é avaliar a adesão de um cimento endodôntico (AH Plus) em canais radiculares após o uso de diferentes protocolos de EDTA 17% e o uso de medicação intracanal (ICM) à base de hidróxido de cálcio em veículo aquoso. Material e Métodos: Para isso, 72 dentes humanos unirradiculares foram instrumentadas até # 50 e divididos em seis grupos (n = 12). Grupo 1: EDTA por 3 min; Grupo 2: 3 mL de EDTA + 3 min de EDTA; Grupo 3: 3 ml de EDTA + 3 min de EDTA + 30 segundos de agitação ultra-sônica; Grupo 4: EDTA durante 3 min + ICM; Grupo 5: 3 ml de EDTA + 3 min de EDTA + ICM; Grupo 6: 3 ml de EDTA + 3 min de EDTA + 30 segundos de agitaçãosônica + ICM. Os canais radiculares foram preenchidos com cimento endodôntico após cada protocolo e, após 7 dias foram preparados para o teste de push-out. Os dados foram analisados utilizando ANOVA dois fatores (p < 0,05). Resultados: Não foi observado diferença estatística na resistência de união nos 3 diferentes protocolos de EDTA a 17%. No entanto, o uso de ICM aumentou significativamente a resistência de adesão. Conclusão: a medicação intracanal à base de hidróxido de cálcio melhorou a resistência de união do AH Plus às paredes dentinárias, independentemente do protocolo de EDTA. (AU)

Objective: The aim of this study is to evaluate the adhesion of an endodontic sealer (AHPlus) in root canals after the use of different protocols of 17% EDTA and the use of intracanal medication (ICM) based on calcium hydroxide in aqueous vehicle. Material and Methods: For this, 72 single-rooted human teeth were instrumented up to #50 and divided into six groups (n = 12). Group 1: EDTA for 3 min; Group 2: 3 mL of EDTA + 3 min of EDTA; Group 3: 3 mL of EDTA + 3 min of EDTA + 30 seconds of ultrasonic agitation; Group 4: EDTA for 3 min + ICM; Group 5: 3 mL of EDTA + 3 min of EDTA + ICM; Group 6: 3 mL of EDTA + 3 min of EDTA + 30 seconds of ultrasonic agitation + ICM. The root canals were filled with endodontic sealer after each protocol and after 7 days they were prepared to the push-out test. The data were analyzed using twoway ANOVA (p< 0.05). Results: It was observed no statistically difference in bond strength in the 3 different 17% EDTA protocols. However, the use of ICM increased significantly the resistance adhesion. Conclusion: Intracanal medication based on calcium hydroxide improved the bond strength of AHPlus to dentin walls, regardless of the EDTA protocol. (AU)

Are Desensitizing Toothpastes Equally Biocompatible and Effective Against Microorganisms?

Abstract The aims of this study were evaluate cytotoxicity, genotoxicity, antimicrobial activity of desensitizing toothpastes compared to a common one and the surface roughness of tooth enamel submitted to brushing with these toothpastes. Samples of three desensitizing toothpastes (Colgate Sensitive, Sensodyne and Oral B Sensitive) and common toothpaste (Colgate) were placed in contact with gingival human fibroblasts. Cytotoxicity and genotoxocity were measured by MTT assay and micronucleus test. Antimicrobial activity of the toothpastes extracts against C. albicans, S. mutans and S. aureus were assessed. For surface roughness evaluation, bovine teeth were submitted to 10.000 brushing cycles. The results were analyzed statically using Mann-Whitney U, ANOVA and Z tests (p<0.05). All toothpastes caused cytotoxic effect to the cells (p<0.05), except Colgate Sensitive. The toothpastes did not increase the number of micronuclei compared to the untreated control group. Colgate eliminated all the evaluated microorganisms at lower concentrations compared to Colgate Sensitive and Oral B Sensitive, which were not able to eliminate S. aureus. Sensodyne did not reach the minimum microbicidal concentration. The surface roughness of tooth enamel increased after brushing with Colgate Sensitive and Oral B Sensitive, however the comparison between groups showed no difference on the enamel surface roughness presented by desensitizing toothpastes when compared with the common one (p>0.05). Based on these results, we can conclude that although none toothpaste has induced genotoxicity, Colgate Sensitive was also not cytotoxic. Colgate was the most effective against the microorganisms, and there were no differences on the enamel surface roughness between the groups.

Resumo Os objetivos desse estudo foram avaliar a citotoxicidade, genotoxicidade, atividade antimicrobiana de dentifrícios dessensibilizantes em comparação com um comum e também a rugosidade superficial do esmalte dentário submetido à escovação com esses dentifrícios. Amostras de três dentifrícios dessensibilizantes (Colgate Sensitive, Sensodyne e Oral B Sensitive) e um dentifrício comum (Colgate) foram colocadas em contato com fibroblastos gengivais humanos e a citotoxicidade e genotoxidade foram mensuradas pelo ensaio MTT e teste do micronúcleo. A atividade antimicrobiana dos extratos dos dentifrícios contra C. albicans, S. mutans e S. aureus foi determinada. Para a avaliação da rugosidade superficial, espécimes de dentes bovinos foram submetidas à 10.000 ciclos de escovação. Os resultados foram analisados estatisticamente usando os testes Mann-Whitney U, ANOVA e Teste Z (P<0,05). Todos os dentifrícios causaram efeito citotóxico às células (P<0,05), exceto o Colgate Sensitive. Os dentifrícios não aumentaram o número de micronúcleos em comparação com o grupo não tratado. O Colgate foi capaz de eliminar todos os microorganismos avaliados em concentrações mais baixas em comparação com Colgate Sensitive e Oral B Sensitive, que não foram capazes de eliminar os S. aureus. O Sensodyne não atingiu a concentração microbicida mínima para qualquer microorganismo. A rugosidade superficial do esmalte dentário aumentou após a escovação com Colgate Sensitive e Oral B Sensitive, porém a comparação entre os grupos não mostrou diferença na rugosidade superficial do esmalte apresentada por dentifrícios dessensibilizantes quando comparados ao comum (p>0,05). Com base nesses resultados, podemos concluir que, embora nenhum dentifrício tenha induzido genotoxicidade, o Colgate Sensitive também não foi citotóxico. O Colgate foi o mais eficaz contra os microorganismos, e não houve diferença na rugosidade superficial do esmalte entre os grupos.

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.