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INTRODUCTION: The objective of this study was to monitor the effects of chemomechanical preparation (CMP) performed with 6% sodium hypochlorite and calcium hydroxide-based intracanal medication (ICM) on the levels and diversity of bacteria, endotoxins (lipopolysaccharides [LPS]), and lipoteichoic acid (LTA) in root canals of teeth with symptomatic irreversible pulpitis. METHODS: Samples were collected from 10 teeth with symptomatic irreversible pulpitis before CMP (S1), after CMP (S2), and after ICM (S3). The levels of bacteria, LPS, and LTA were assessed by using checkerboard DNA-DNA hybridization, LAL Pyrogent 5000, and enzyme-linked immunosorbent assay, respectively. Wilcoxon test, repeated-measures analysis of variance, and Tukey post hoc test were used for statistical analysis at a significance level of 5%. RESULTS: Forty species were detected at S1. Two species were eliminated after CMP and 5 after ICM. Resistant and pain-related species were detected in the root canals. Higher levels of culturable bacteria were detected at S1. However, CMP and ICM effectively reduced the microbial load in the root canals. Higher levels of LPS and LTA were detected at S1. CMP was effective in reducing both LPS and LTA (P < .05). ICM produced additional reduction in the levels of LPS (P > .05) and LTA (P < .05). CONCLUSIONS: Chemomechanical preparation using 6% sodium hypochlorite and calcium hydroxide-based intracanal medication were effective in reducing the levels of bacteria, LPS, and LTA in teeth with vital pulp and irreversibly inflamed pulp.

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OBJECTIVE: To characterize the microbiota of teeth with endodontic treatment failure by 16S ribosomal RNA genetic sequencing (GS) and PCR at the different phases of the endodontic retreatment and to associate the presence of specific bacteria with clinical and radiographic features in teeth with apical periodontitis. MATERIALS AND METHODS: Twenty infected root canals of single-rooted teeth were selected. Samples were collected with sterile paper points before chemo-mechanical preparation (CMP) (S1), after CMP (S2) and after 30 days of intracanal medication (ICM) (S3). Microbial identification was performed using GS and PCR. Tukey-Kramer post hoc test and post hoc ANOVA were used for intergroup analysis. Paired t test and repeated-measures ANOVA were applied for intragroup analysis, at a significance level of 5%. RESULTS: A total of 89 strains were identified using GS. Sixty-five strains were recovered in S1 and 15 strains in S2, and 9 strains remained in S3. Enterococcus faecalis was the most predominant bacteria. Gram-positive cocci bacteria predominated. Gram-negative species were also detected. Using species-specific PCR primers to detect seven species, the most prevalent ones at all the phases of the endodontic retreatment were E. faecalis and Porphyromonas gingivalis. However, Parvimonas micra and P. gingivalis were associated with previous pain, P. gingivalis was associated with tenderness to percussion and E. faecalis, Fusobacterium nucleatum and P. gingivalis were associated with periapical lesion > 3 mm. CONCLUSIONS: In conclusion, the microbiota of persistent infection is polymicrobial with predominance of E. faecalis and P. gingivalis in all phases of the endodontic retreatment, regardless of the method used for microbial identification. Associations were found between specific bacteria and clinical/radiographic features. CLINICAL RELEVANCE: The characterization of the bacteria present at all phases of the endodontic retreatment is important for the monitoring of the effectiveness of the techniques used and to better understand the susceptibility of these species to the disinfection agent used during the procedures.

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OBJECTIVE: The present clinical study investigated the microbiota of teeth with persistent secondary endodontic infection in the different phases of root canal retreatment. METHODS: Twenty filled single-rooted teeth with apical periodontitis were included. Samples were collected with sterile paper points before chemo-mechanical preparation (CMP) (S1), after CMP (S2) and after 30 days of calcium hydroxide-based intracanal medication (ICM) (S3). Cultivable bacteria were assessed by colony forming units count (CFU/mL). DNA was extracted and assessed by using nested PCR. Paired t-test and repeated measures ANOVA were applied for intragroup analysis in the stages of endodontic therapy at a significance level of 5%. RESULTS: Cultivable bacteria were detected in all initial samples. CMP reduced bacteria by 99.4% and ICM by 99.5%. The most prevalent species found in the initial samples were E. faecalis (20/20), P. gingivalis (20/20), F. nucleatum (17/20) and A. actinomycetemcomitans (10/20), whereas D. pneumosintes, F. alocis, P. nigrescens and T. socranskii were not detected. After CMP, A. israelii, A. naeslundii, G. morbillorum, T. forsythia and T. denticola were not detected (P<0.05) either. E. faecalis and P. gingivalis had a low reduction (P>0.05) and F. nucleatum had its DNA significantly reduced after CMP (P<0.05). ICM had no additional effect on microbial reduction. CONCLUSION: The microbiota of teeth with persistent/secondary endodontic infection consists of a polymicrobial community with Gram-positive and Gram-negative species, bacillus and cocci, facultative and strict anaerobes. E. faecalis and P. gingivalis were frequently detected in all stages of root canal retreatment, evidencing their great resistance to endodontic procedures. The endodontic procedures were effective in reducing the levels of bacteria from teeth presenting with persistent/secondary endodontic infection.

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AIM: The objective of this study was to evaluate the presence of microorganisms in the root canals (RC) of teeth submitted to endodontic retreatment due to prosthetic reasons and without evidence of apical periodontitis. MATERIAL AND METHODS: Twenty teeth referred to endodontic retreatment due to prosthetic reasons and without evidence of apical periodontitis were included in this study. Gutta-percha (GP) was collected from each third of the RC. After GP removal, microbial samples were also collected from the full extension of RC using paper points and files. The samples were cultivated in Fastidious Anaerobe Agar and the colony-forming units (CFU/mL) were counted. They were also semi-quantitatively analyzed by checkerboard DNA-DNA hybridization using 40 DNA bacterial probes. RESULTS: Microorganisms were found in all samples. The coronal third of GP was more contaminated than the apical third (p ≤ 0.05). There was prevalence of Enterococcus hirae and Enterococcus faecalis in all RC thirds and also in the samples collected from the full extension of RCs. CONCLUSION: Microorganisms were found in all cases referred to endodontic retreatment due to prosthetic reasons and without evidence of apical periodontitis. Enterococcus was the genus most frequently detected. CLINICAL SIGNIFICANCE: An endodontic retreatment should be considered before replacing a prosthesis.

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The present study evaluated the amount of apically extruded debris after chemo-mechanical preparation (CMP) associated with passive ultrasonic irrigation (PUI) using four different root canal irrigants, namely, 6% sodium hypochlorite (NaOCl), 2% chlorhexidine gel + saline solution (2% CHXg+SS), 2% chlorhexidine solution (2% CHXs) and SS alone. Sixty mandibular premolars with single straight root canals were selected and randomly assigned into 4 groups (n=15) according to the root canal irrigant used as follows: G1 (PUI + NaOCl), G2 (PUI + CHXg+SS), G3 (PUI+CHXs) and G4 (PUI+SS). Reciproc® R25 files (25/.08) were used during CMP and the debris extruded from each tooth were collected in pre-weighted Eppendorf tubes and dried. The average weight of debris was assessed by using an analytical microbalance. Data were statistically analysed by using ANOVA and post-hoc Tukey's test (a=0.05). Debris extrusion was observed in all groups, irrespective of the root canal irrigating, with 2% CHXg + SS being associated with lower debris extrusion compared to other irrigants (p<0.05). No significant differences were observed between 6% NaOCl, 2% CHXs and SS. In conclusion, passive ultrasonic irrigation did not completely prevent apically extrusion of debris. PUI performed with 2% chlorhexidine gel + saline solution significantly minimized debris extrusion compared to 6% sodium hypochlorite, chlorhexidine solution and saline solution.

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Abstract The present study evaluated the amount of apically extruded debris after chemo-mechanical preparation (CMP) associated with passive ultrasonic irrigation (PUI) using four different root canal irrigants, namely, 6% sodium hypochlorite (NaOCl), 2% chlorhexidine gel + saline solution (2% CHXg+SS), 2% chlorhexidine solution (2% CHXs) and SS alone. Sixty mandibular premolars with single straight root canals were selected and randomly assigned into 4 groups (n=15) according to the root canal irrigant used as follows: G1 (PUI + NaOCl), G2 (PUI + CHXg+SS), G3 (PUI+CHXs) and G4 (PUI+SS). Reciproc® R25 files (25/.08) were used during CMP and the debris extruded from each tooth were collected in pre-weighted Eppendorf tubes and dried. The average weight of debris was assessed by using an analytical microbalance. Data were statistically analysed by using ANOVA and post-hoc Tukey's test (a=0.05). Debris extrusion was observed in all groups, irrespective of the root canal irrigating, with 2% CHXg + SS being associated with lower debris extrusion compared to other irrigants (p<0.05). No significant differences were observed between 6% NaOCl, 2% CHXs and SS. In conclusion, passive ultrasonic irrigation did not completely prevent apically extrusion of debris. PUI performed with 2% chlorhexidine gel + saline solution significantly minimized debris extrusion compared to 6% sodium hypochlorite, chlorhexidine solution and saline solution.

Resumo O presente estudo avaliou a quantidade de debris extruídos apicalmente após o preparo químico-mecânico (PQM) associado à irrigação ultrassônica passiva (IUP) em associação com quatro diferentes irrigantes - hipoclorito de sódio 6% (NaOCl), clorexidina gel 2% + solução salina (CLXg 2% + SS), solução de clorexidina 2% (CLXs 2%) e SS. Sessenta pré-molares inferiores com canais radiculares únicos e retos foram selecionados e aleatoriamente distribuídos em 4 grupos (n=15) de acordo com o irrigante utilizado: G1 (IUP + NaOCl), G2 (IUP + CLXg + SS), G3 (IUP + CLXs) e G4 (IUP + SS). Limas Reciproc® R25 (25/.08) foram utilizadas durante o PQM e os debris extruídos de cada dente foram coletados em tubos Eppendorf pré-pesados e secos. O peso médio de debris foi avaliado através de microbalança analítica, e os dados foram analisados estatisticamente utilizando ANOVA e teste de Tukey post hoc (a=0.05). Extrusão de debris foi observada em todos os grupos, independente do irrigante. CHXg 2% + SS foram associados a menor extrusão de debris comparado aos demais irrigantes (p<0.05). Não foram observadas diferenças estatisticamente significativas entre NaOCl 6%, CLXs 2% e SS. Concluindo, irrigação ultrassônica passiva não preveniu completamente a extrusão apical de debris, entretanto, IUP realizada com CLXg 2% + SS minimiza significativamente a extrusão de debris comparado ao NaOCl 6%, CLXs 2% e SS.

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OBJECTIVE: The objective of this work was to investigate in vivo the effects of calcium hydroxide-based intracanal medication (ICM) on the levels of bacteria, pro-inflammatory cytokines (PICs), and matrix metalloproteinases (MMPs) in root canals and periradicular tissues of teeth with failure of the root canal treatment and apical periodontitis. MATERIALS AND METHODS: Twenty infected root canals of single-rooted teeth were randomly assigned into two groups according to the irrigant used for chemomechanical preparation (CMP) (n = 10 per group): G1 - 2% chlorhexidine (CHX) gel and G2 - 6% sodium hypochlorite (NaOCl). Root canal contents were taken by using paper points before CMP (S1) and after 30 days of calcium hydroxide-based ICM (S2). Microbial reduction was calculated by means of colony-forming unit count (CFU/mL), with PICs and MMPs (pg/mL) being measured by using enzyme-linked immunosorbent assay (ELISA). RESULTS: Culturable bacteria (101.2 ± 79.2), PICs (IL-1ß 1.2 ± 0.4 and TNF-α 8.8 ± 4.7), MMP-2 (803.7 ± 96.4), MMP-3 (453.9 ± 229.3), MMP-8 (245.9 ± 122.4), MMP-9 (129.4 ± 29.6), and MMP-13 (70.8 ± 12.8) were present in all S1 samples. After 30 days of ICM (S2), a 99.5% microbial reduction was observed, together with a significant reduction of PICs in all groups. Overall, it was observed a decrease in the levels of MMPs (S2), except MMP-13, which was found in increased levels after ICM (P < .05), independently of the groups. CONCLUSIONS: Calcium hydroxide-based intracanal medications have had a positive effect on the microbial reduction by decreasing the levels of PICs and MMPs. Both auxiliary chemical substances (i.e., 2% CHX and 6% NaOCl) presented similar effects when calcium hydroxide was used as intracanal medication. CLINICAL RELEVANCE: Teeth with failure of the root canal treatment and apical periodontitis, and consequently with high levels of bacteria, PIC, and MMP, may present a better prognosis after a 30 days of a calcium hydroxide-based ICM.

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This study evaluated the amount of apically extruded debris after chemo-mechanical preparation (CMP) using positive and negative pressure irrigation systems [Conventional irrigation (CI) and EndoVac (EV)] in association with different irrigants [6% Sodium Hypochlorite (NaOCl), 2% Chlorhexidine gel + saline solution (CHXg + SS), 2% Chlorhexidine solution (CHXs) or Saline solution (SS)]. Eighty mandibular premolars with single root canals were selected and randomly assigned into 8 groups (n = 10) according to the irrigation system and the irrigant used during CMP: G1 (EV + NaOCl), G2 (EV + CHXg + SS), G3 (EV + CHXs), G4 (EV + SS), G5 (CI + NaOCl), G6 (CI + CHXg + SS), G7 (CI + CHXs) and G8 (CI + SS). Reciproc® R25 files (25/.08) were used during the CMP and the extruded debris from each tooth was collected in pre-weighted Eppendorf tubes and dried. The average weight of debris was assessed using a microbalance, and the data were statistically analyzed using ANOVA and the post hoc Tukey's test (a = 0.05). All groups were associated with debris extrusion. EV was the irrigation system with less extruded debris (p < 0.05). No differences were observed regarding the irrigant when EV was used. When CI was used, CHXg + SS were associated with lower debris extrusion (p < 0.05). It was concluded that no irrigation protocol succeeded in preventing debris extrusion. EV resulted in lower levels of debris extrusion than CI. The use of CHXg + SS resulted in lower debris extrusion.

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Abstract This study evaluated the amount of apically extruded debris after chemo-mechanical preparation (CMP) using positive and negative pressure irrigation systems [Conventional irrigation (CI) and EndoVac (EV)] in association with different irrigants [6% Sodium Hypochlorite (NaOCl), 2% Chlorhexidine gel + saline solution (CHXg + SS), 2% Chlorhexidine solution (CHXs) or Saline solution (SS)]. Eighty mandibular premolars with single root canals were selected and randomly assigned into 8 groups (n = 10) according to the irrigation system and the irrigant used during CMP: G1 (EV + NaOCl), G2 (EV + CHXg + SS), G3 (EV + CHXs), G4 (EV + SS), G5 (CI + NaOCl), G6 (CI + CHXg + SS), G7 (CI + CHXs) and G8 (CI + SS). Reciproc® R25 files (25/.08) were used during the CMP and the extruded debris from each tooth was collected in pre-weighted Eppendorf tubes and dried. The average weight of debris was assessed using a microbalance, and the data were statistically analyzed using ANOVA and the post hoc Tukey's test (a = 0.05). All groups were associated with debris extrusion. EV was the irrigation system with less extruded debris (p < 0.05). No differences were observed regarding the irrigant when EV was used. When CI was used, CHXg + SS were associated with lower debris extrusion (p < 0.05). It was concluded that no irrigation protocol succeeded in preventing debris extrusion. EV resulted in lower levels of debris extrusion than CI. The use of CHXg + SS resulted in lower debris extrusion.

Resumo Este estudo avaliou a quantidade de debris extruídos apicalmente após o preparo químico-mecânico (PQM) utilizando sistemas de irrigação com pressão positiva e negativa [irrigação convencional (IC) e EndoVac (EV)] em associação com diferentes irrigantes [hipoclorito de sódio 6% (NaOCl), clorexidina gel + solução salina (CLXg + SS), solução de clorexidina 2% (CLXs) ou solução salina (SS)]. Oitenta pré-molares inferiores com único canal radicular foram selecionados e aleatoriamente alocados em 8 grupos (n=10) de acordo com o sistema de irrigação e irrigante utilizado durante o PQM: G1 (EV + NaOCl), G2 (EV + CLXg + SS), G3 (EV + CLXs), G4 (EV + SS), G5 (IC + NaOCl), G6 (IC + CLXg + SS), G7 (IC + CLXs) e G8 (IC + SS). Limas Reciproc® R25 foram utilizadas durante o PQM e os debris extruídos de cada dente foi coletado em tubos pré-pesados e secos. O peso médio de debris foi avaliado por meio de microbalança, e os dados foram analisados estatisticamente utilizando ANOVA e teste de Tukey (a = 0.05). Todos os grupos foram associados com extrusão de debris. EV foi o sistema de irrigação com menos debris extruídos (p<0.05). Não foram observadas diferenças entre os irrigantes quando o EV foi utilizado. Quando foi utilizada IC, CLXg + SS foram associados a menor extrusão de debris (p<0.05). Concluiu-se que nenhum protocolo de irrigação conseguiu prevenir extrusão de debris. EV resultou em menores níveis de extrusão de debris que a IC. A utilização da CLXg + SS resultou em menor extrusão de debris.

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Introduction: Blood is a biological material with high potential of infectious transmission in dental environments, including herpes simplex, hepatitis and AIDS. Aim: To investigate the efficacy of luminol in detecting blood in endodontic files before and after the sterilization process. Material and method: Luminol was used to investigate the presence or absence of traces of blood tissue in 50 endodontic files, visible to naked eye or not, after performing endodontic treatment and after the cleaning/sterilization process. The results obtained were tabulated and statistically analyzed by using the Friedman's test at a significance level of 5% (p<0.05). Result: By naked eye, it was found that 31/50 files showed no trace of blood, 8/50 showed a slight presence of blood and 11/50 showed a considerable presence of blood after endodontic treatment. After the use of luminol, however, 16/50 endodontic files showed no trace of blood, 19/50 showed a slight presence of blood and 15/50 showed a considerable presence of blood. After the cleaning and sterilization process, no blood was detected in the files. Conclusion: It was concluded that the luminol solution is effective in detecting blood tissue in endodontic files as well as in validating the cleaning/sterilization process.

Introdução: Sangue é um material biológico com alto potencial de transmissão de infecção em ambientes odontológicos, incluindo herpes simples, hepatites e AIDS. Objetivo: Investigar a eficácia do luminol em detector sangue em limas endodônticas antes e após o processo de esterilização. Material e método: Luminol foi utilizado para investigar a presença ou ausência de vestígios tecido sanguíneo em 50 limas endodônticas, visíveis ou não à olho nu, após a realização do tratamento endodôntico e após o processo de limpeza/esterilização. Os resultados obtidos foram tabulados e analisados estatisticamente utilizando o teste de Friedman com nível de significância de 5% (p<0,05). Resultado: A olho nú, foi observado que 31/50 limas não apresentaram vestígios de sangue, 8/50 apresentaram uma leve presença de sangue e 11/50 apresentaram uma presença considerável de sangue após o tratamento endodôntico. Após a utilização do luminol, entretanto, 16/50 limas endodônticas não apresentaram vestígios de sangue, 19/50 apresentaram uma leve presença de sangue e 15/50 apresentaram uma presença considerável de sangue. Após o processo de limpeza e esterilização não foi detectado sangue nas limas endodônticas. Conclusão: A solução de luminol é efetiva na detecção de tecido sanguíneo em limas endodônticas, validando o processo de limpeza/esterilização.

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