Microbiological profile of root canals indicated for endodontic retreatment due to secondary endodontic infections or for prosthetic reasons.

OBJECTIVE: The objective of this study was to evaluate the microbiological profile of root canals indicated for endodontic retreatment due to secondary endodontic infections evidenced by the presence of chronic apical periodontitis (G1) or for prosthetic reasons, without clinical and radiographic signs of endodontic reinfection (G2). METHODS: Microbiological samples were collected from thirty teeth (N=30) out of which 15 were indicated for retreatment due to the presence of chronic apical periodontitis (G1) and 15 were indicated exclusively for prosthetic reasons (G2). Samples were collected from root canals before (S1), after chemomechanical preparation (S2), and after 30 days of intracanal medication composed of calcium hydroxide and 2% chlorhexidine gel (S3). The molecular analysis was performed using Nested-PCR for the detection of 17 bacterial species. The efficacy of each stage of the retreatment in reducing the microbial load was verified by counting colony-forming units (CFU). The statistical analysis considered a significance level of 5%. RESULTS: The results showed that bacteria were detected in 100% of the cases in S1, in both groups, by Nested PCR. The most frequently found species in S1 in both groups were Enterococcus faecalis, Fusobacterium nucleatum, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, and Parvimonas micra. The microbial load of G1 was higher than G2 in the initial samples (S1). Endodontic retreatment was effective in reducing the microbial load in G1 and G2. Statistically significant associations were found between specific bacteria and clinical signs and symptoms. CONCLUSION: It was concluded that the microbial community present in the root canal of teeth indicated for endodontic retreatment is mixed and heterogeneous. G1 and G2 differ in the number of species per case and microbial load. CLINICAL RELEVANCE: Endodontic retreatment was effective in reducing the microbial load. Statistically significant associations were found between specific bacteria and clinical signs and symptoms.

Comparative analysis of bacterial content, levels of lipopolysaccharides and lipoteichoic acid in symptomatic and asymptomatic endodontic infections at different stages of endodontic treatment.

OBJECTIVES: This study aimed to detect the presence of specific bacteria and to evaluate the levels of lipopolysaccharides (LPS) and lipoteichoic acid (LTA) in symptomatic necrotic root canals associated with acute apical abscess (symptomatic group - GI). It also aimed to compare the findings with those presented by asymptomatic necrotic root canals (asymptomatic group - GII) in the different stages of the endodontic treatment. MATERIALS AND METHODS: Microbiological samples were collected from 20 root canals, including purulent collection from acute apical abscesses, before and after chemo-mechanical preparation (CMP) preparation (CMP) with chlorhexidine gel 2% and after 30 days of intracanal medication (ICM) with (Ca[OH]2) + chlorhexidine. The culture method was used to evaluate the efficacy of CMP and ICM. Nested PCR was used to detect the presence of 17 specific bacteria. Levels of LPS were measured by using limulus amebocyte lysate, whereas enzyme-linked immunosorbent assay was used for the quantification of LTA. RESULTS: CMP was effective in reducing the microbial load in both groups (P < 0.05). LPS levels were higher in GI than in GII (P < 0.05). There was a significant reduction in the LPS levels after CMP and ICM (P < 0.05) in GI and GII. LTA levels were significantly reduced in GI after ICM and in GII after CMP and ICM (both P < 0.05). Fusobacterium nucleatum and Enterococcus faecalis were frequently identified in both groups, alone or in combination with each other. CONCLUSION: Different species were detected in all stages of the endodontic treatment. CMP was able to reduce bacterial content and the levels of LPS, but not of LTA in the symptomatic group. High levels of LPS were correlated with spontaneous pain and pain to percussion in the symptomatic group. CLINICAL RELEVANCE: This clinical study showed that chemo-mechanical preparation was able to reduce bacterial load and levels of LPS, but not of LTA in the symptomatic group. Elevated levels of LPS were correlated with the presence of symptomatology.

Evaluation of the presence of microorganisms from root canal of teeth submitted to retreatment due to prosthetic reasons and without evidence of apical periodontitis.

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.

Bond strength to radicular dentin and sealing ability of AH Plus in combination with a bonding agent.

OBJECTIVE: To evaluate the sealing ability and bond strength of AH Plus sealer associated with the hybridization protocol of radicular dentin with Scotchbond Multi Purpose (SB). MATERIALS AND METHODS: Ninety palatal roots of maxillary molars were selected and divided into three groups (n = 30) according to filling protocol (G1, AH Plus/Resilon; G2, SB/AH Plus/Resilon; and G3, AH Plus/Gutta-Percha). In groups in which AH Plus + resin cones were used, dentin was hybridized before applying sealing material. For the bond strength test, 60 roots of bovine teeth were selected. Six holes were made in each root, two in the cervical, middle and apical third of the root. The roles were filled with AH Plus sealer with or without an adhesive system and submitted to push out test and the fracture mode was examined using a stereomicroscope (×32). RESULTS: Statistical analysis showed that ScotchBond Multi Purpose (SB) + Resilon cone + AH Plus group promoted higher sealing ability than the gutta-percha + AH Plus group (p < 0.05). Bond strength was lower with SB application than without it (p < 0.05). A reverse correlation was found between bond strength and sealing ability. The fracture mode methodology revealed 22.77% of adhesive, 11.67% of cohesive and 65.55% of mixture fractures for SBMP/AH Plus protocol, whereas the AH Plus protocol indicated 86.11% of cohesive and 13.89% of mixed fractures. CONCLUSION: It was concluded that the use of the adhesive system Scotchbond Multi Purpose improved coronal sealing ability of AH Plus, but bond strength of sealer was reduced when adhesive was applied.

Epidemiology and severity of traumatic dental injuries in permanent teeth: A 20-year retrospective study.

This study aimed to assess the traumatic dental injuries (TDIs) in permanent dentition among patients who attended at the outpatient clinic of a Brazilian dental school, during the last 20 years, and to investigate factors associated with the severity of these injuries. Clinical records of patients who attended a specialized center for dental trauma care in Brazil presenting at least one TDI in a permanent tooth, between the years 2000 and 2019, were reviewed. The data recorded were sex, age, affected arch, etiology, number, and type of the teeth affected, and classification and severity of the TDIs. The diagnosis and classification of the TDIs were based on the guidelines of the International Association of Dental Traumatology (IADT). The severity of each patient's injuries was defined as mild, moderate, or severe. Descriptive statistics, chi-square and multinomial regression analyses were used to evaluate the results. The significance level was set at 5%. A total of 837 clinical records were included, totaling 2357 teeth. Males were more prevalent than females. The patients' age ranged from 5 to 71 years. The most common traumas were avulsion (n=512) and uncomplicated enamel-dentin fracture (n=488). Univariate analyses showed that there was a statistically significant association between age group (p=0.004), etiology (p=0.000) and number of teeth affected (p=0.000) with severity of dental trauma. In conclusion, TDIs that occurred in Piracicaba and region are epidemiologically similar to those found worldwide, and that more severe injuries are related to age range, etiology and number of teeth affected.

Clinical evaluation of culturable bacteria, endotoxins and lipoteichoic acid in teeth with vital normal pulp tissues.

The objective of this study was to investigate the presence of culturable bacteria, endotoxins (LPS) and lipoteichoic acid (LTA) levels in teeth with normal vital pulp (NVP) with intact crowns (IC) and those with coronal restoration (CR) limited to the enamel level. A total of 20 teeth indicated for endodontic treatment due to prosthetic reasons were selected. Samples were collected from the root canals. The levels of cultivable bacteria, LPS and LTA were assessed. Statistical analyses were performed at significance level set at 5%. None of the teeth presented microbial growth. In the IC group, the LPS levels were limited to the lowest concentration of LPS. On the contrary, higher LPS and LTA levels were detected in teeth with CR. It was concluded that teeth with NVP and IC were negative for bacteria, LPS and LTA; while teeth with CR were positive for bacterial virulence factors.

Efficacy of 6% Sodium Hypochlorite on Infectious Content of Teeth with Symptomatic Irreversible Pulpitis.

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.

Clinical and Radiographic Outcomes of Regenerative Endodontic Procedures in Traumatized Immature Permanent Teeth: Interappointment Dressing or Single-Visit?

INTRODUCTION: More data are needed before affirming that single-visit approaches are effective and safe for regenerative endodontic procedures (REPs). This study compared clinical and radiographic outcomes of REPs between interappointment dressing or single-visit protocols. METHODS: Twenty young patients presenting traumatized immature teeth with pulp necrosis were divided into 2 groups. Teeth were submitted to 6% sodium hypochlorite and 2% chlorhexidine irrigation and to 17% EDTA use before blood clot induction. In the interappointment dressing group (n = 11), calcium hydroxide was used with 2% chlorhexidine gel for 21 days before scaffold induction. In the single-visit group (n = 9), the scaffold was induced during the only appointment. Patients were followed up for 24 months. Primary, secondary, and tertiary outcomes were assessed by 3 independent evaluators. Preoperative and follow-up radiographs were assessed for quantitative measurements of the apical diameter, root width, root length, and cervical barrier placement and compared between groups using the t test or Mann-Whitney U test. Categoric variables were assessed with the G test and the Fisher exact test and continuous variables with the Mann-Whitney U test. Statistical significance was set at 5%. RESULTS: One tooth showed persistence of infection. No difference was observed in cervical barrier placement (P > .05); between groups regarding primary, secondary, and tertiary outcomes (P > .05); and in parameters of quantitative radiographic outcomes (P > .05). CONCLUSIONS: Completing REPs with an interappointment dressing or a single-visit protocol presented similar clinical and radiographic outcomes. A single-visit protocol of REPs using 6% sodium hypochlorite, 2% chlorhexidine, and 17% EDTA permitted satisfactory outcomes in necrotic immature permanent teeth.

Epidemiology and severity of traumatic dental injuries in permanent teeth: A 20-year retrospective study

Abstract This study aimed to assess the traumatic dental injuries (TDIs) in permanent dentition among patients who attended at the outpatient clinic of a Brazilian dental school, during the last 20 years, and to investigate factors associated with the severity of these injuries. Clinical records of patients who attended a specialized center for dental trauma care in Brazil presenting at least one TDI in a permanent tooth, between the years 2000 and 2019, were reviewed. The data recorded were sex, age, affected arch, etiology, number, and type of the teeth affected, and classification and severity of the TDIs. The diagnosis and classification of the TDIs were based on the guidelines of the International Association of Dental Traumatology (IADT). The severity of each patient's injuries was defined as mild, moderate, or severe. Descriptive statistics, chi-square and multinomial regression analyses were used to evaluate the results. The significance level was set at 5%. A total of 837 clinical records were included, totaling 2357 teeth. Males were more prevalent than females. The patients' age ranged from 5 to 71 years. The most common traumas were avulsion (n=512) and uncomplicated enamel-dentin fracture (n=488). Univariate analyses showed that there was a statistically significant association between age group (p=0.004), etiology (p=0.000) and number of teeth affected (p=0.000) with severity of dental trauma. In conclusion, TDIs that occurred in Piracicaba and region are epidemiologically similar to those found worldwide, and that more severe injuries are related to age range, etiology and number of teeth affected.

Resumo Esse estudo objetivou avaliar as injúrias dentárias traumáticas (IDTs) na dentição permanente entre os pacientes que compareceram ao ambulatório de uma faculdade de odontologia brasileira, durante os últimos 20 anos, e investigar fatores associados à severidade dessas lesões. Os registros clínicos dos pacientes que compareceram a um centro especializado de atendimento em traumatismo dentário no Brasil apresentando pelo menos uma IDT em dente permanente, entre os anos de 2000 e 2019, foram revisados. Os dados registrados foram sexo, idade, arco dental afetado, etiologia, número e tipo dos dentes afetados, e classificação e severidade das IDTs. O diagnóstico e a classificação das IDTs foram baseados nas diretrizes da Associação Internacional de Traumatologia Dentária (AITD). A gravidade das lesões de cada paciente foi definida como leve, moderada ou severa. Estatísticas descritivas, teste qui-quadrado e análises de regressão multinomial foram usadas para avaliar os resultados. O nível de significância foi fixado em 5%. Um total de 837 registros clínicos foi incluído, totalizando 2357 dentes. O sexo masculino foi mais prevalente que o feminino. A idade dos pacientes variou de 5 a 71 anos. Os traumas mais comuns foram avulsão (n=512) e fratura não-complicada do esmalte-dentina (n=488). As análises univariadas mostraram que houve associação estatisticamente significativa entre a faixa etária (p=0,004), etiologia (p=0,000) e número de dentes afetados (p=0,000) com a gravidade do traumatismo dentário. Em conclusão, as IDTs que ocorreram em Piracicaba e região são epidemiologicamente semelhantes aos encontrados em todo o mundo, e que lesões mais graves estão relacionadas à faixa etária, etiologia e número de dentes afetados.

Residual effects and surface alterations in disinfected gutta-percha and Resilon cones.

The aim of this study was to evaluate alterations on the surface of gutta-percha and Resilon cones when exposed to 2% chlorhexidine gel (CHX) and 5.25% sodium hypochlorite (NaOCl) and to evaluate their possible antibacterial residual effects against Enterococcus faecalis and Porphyromonas gingivalis. A total of 270 medium tapered cones (135 gutta-percha and 135 Resilon) were selected and divided into three groups, according to the substance (NaOCl, CHX, and sterile saline solution). The cones were immersed in each substance (for 1, 5, 10, 20, and 30 min), rinsed in saline solution, and dried. They were placed onto inoculated agar plates and incubated under specific conditions. Scanning electron microscopy analysis was performed on additional 270 disinfected cones. Another 18 cones were used as controls. Based on the results, it was concluded that Resilon cones exposed to CHX for 10, 20, and 30 min demonstrated residual antibacterial action and that substances did not cause alterations to the cones' surface.

The outcome of endodontic treatment: a retrospective study of 2000 cases performed by a specialist.

The purpose of this study was to evaluate the treatment outcome of initial endodontic treatment and nonsurgical retreatment performed by an endodontic specialist in his private office. A total of 2,000 teeth were examined clinically and radiographically and the results were analyzed statistically by Pearson or Fisher's Exact test and multivariate logistic regression. The multivariate analysis evaluated joint associations among various factors, using logistic regression models. The dependent variable for this analysis was the dichotomous outcome: healed versus disease. The overall endodontic success rate was 91.45%, and the healed rate was significantly higher for initial endodontic treatments than for nonsurgical retreatments; teeth without lesion than for those with lesions; teeth treated without complications than for those with complications; recall period of 18-24 months than for other periods, and teeth with final coronal restoration than for those without. Of the 1376 teeth treated in the initial endodontic treatment sample, the success rate was 94.0%. Multivariate analysis identified the presence of procedural complications (file breakage, perforation and flare-up), as well as the absence of the restorations at follow-ups as the significant predictors of outcome, showing lower rates of success. Of the 624 teeth in the nonsurgical retreatment sample, 85.9% were successful. Stepwise logistic regression analysis revealed that preoperative radiolucency was a strong statistically significant factor to determine lower rates of success than in its absence. Two additional variables (age and tooth type) were found to have a significant influence on the outcome of the retreatment sample. A higher healed rate was observed for the 50-59 years age groups than others, while multirooted (molars) teeth revealed a significantly lower percentage of success than pre-molars and anterior teeth.

Molecular analysis of Filifactor alocis, Tannerella forsythia, and treponema denticola associated with primary endodontic infections and failed endodontic treatment.

The aim of this study was to investigate the presence of strict anaerobes such as Filifactor alocis, Tannerella forsythia, and Treponema denticola in primary and secondary root-infected canals with periapical lesions by molecular analysis and the association of these species with specific endodontic signs and symptoms. Microbial samples were taken from 100 root canals, 50 with necrotic pulp tissues (NPT, primary infection), and 50 with failed endodontic treatment (FET, secondary infection). DNA was extracted from the samples, which were analyzed for the presence of three endodontic pathogens using species-specific primers and PCR. F. alocis were isolated from 23 canals with NPT and 12 canals with FET; T. forsythia from 12 canals with NPT and three canals with FET; T. denticola from 19 canals with NPT and 12 canals with TEP. Suggested associations were found between primary infection and the presence of F. alocis and T. forsythia (both p < 0.05). In particular, associations were found between: pain and F. alocis; swelling and F. alocis; tenderness to percussion and T. forsythia; mobility and T. forsythia and T. denticola; wet canals and F. alocis, T. forsythia, and T. denticola; purulent exsudate and F. alocis, T. forsythia and T. denticola; abscess and F. alocis, T. forsythia, and T. denticola (all p < 0.05). The findings of this study indicated that F. alocis, T. forsythia, and T. denticola seem to be associated with endodontic signs and symptoms. Additionally, F. alocis and T. forsythia were detected more frequently in teeth with necrotic pulp than in teeth with failing endodontic treatment.

Quantification of Lipoteichoic Acid Contents and Cultivable Bacteria at the Different Phases of the Endodontic Retreatment.

INTRODUCTION: The infectious content of root canals, including bacteria and lipoteichoic acid (LTA), cause injuries to the periapical tissues. The purpose of this clinical study was to quantify the levels of both LTA and cultivable bacteria at the different phases of endodontic retreatment (ER) of teeth with post-treatment apical periodontitis. It also aimed to investigate the presence of gram-positive microorganisms before and after chemomechanical preparation (CMP) and intracanal medication (ICM). METHODS: Twenty infected root canals of single-rooted teeth were randomly assigned into 2 groups according to the chemical substance used for CMP (n = 10 per group): chlorhexidine (CHX) group, 2% CHX gel, and the sodium hypochlorite (NaOCl) group, 6% NaOCl. Root canal samples were taken using paper points before (S1) and after CMP (S2) and after 30 days of ICM with calcium hydroxide + 2% CHX gel (S3). Microorganisms were identified by the culture technique using biochemical tests. Cultivable bacteria were determined by counting the colony-forming unit. LTA levels were measured using the enzyme-linked immunosorbent assay (pg/mL). RESULTS: A total of 70 gram-positive species, out of 102 species isolated, were found in the root canals (54 in S1, 4 in S2, and 12 in S3). Enterococcus faecalis was the most frequent isolated taxon in all phases of the ER. LTA (574.0 ± 94.7) and cultivable bacteria (101.2 ± 79.2) were present in all S1 samples. CMP decreased the overall levels of cultivable bacteria by 99.4% and LTA by 24.8% (P < .05), whereas the total overall reduction level of ICM on viable bacteria was 99.5% and on LTA it was 38.6% (P < .05). CMP with 2% CHX gel (CHX group, 99.3%) was more effective (P < .05) than 6% NaOCl (NaOCl group, 92.1%) on bacterial reduction. Likewise, ICM showed a 100% reduction in the CHX group and 98.5% in the NaOCl group. Regarding the reduction of LTA, CMP with 2% CHX gel (CHX group, 26.9%) was more effective (P < .05) than 6% NaOCl (NaOCl group, 22.6%). In addition, ICM showed a 43.2% reduction in the CHX group and 36.2% in the NaOCl group (P > .05). CONCLUSIONS: The reduction rates of bacteria were higher than the LTA. Moreover, gram-positive microorganisms were present in all phases of the endodontic retreatment.

Antimicrobial Susceptibility and Characterization of Virulence Genes of Enterococcus faecalis Isolates from Teeth with Failure of the Endodontic Treatment.

INTRODUCTION: The aim of this study was to investigate the prevalence of virulence factors and the antimicrobial resistance of Enterococcus faecalis isolates of teeth with failure of the endodontic treatment. METHODS: Twenty root canal samples were collected from teeth with apical periodontitis. E. faecalis was firstly identified based on phenotypic features and then by 16S ribosomal RNA gene sequencing. The antimicrobial susceptibility was determined by the minimum inhibitory concentration (MIC) of amoxicillin, amoxicillin + clavulanate, azithromycin, benzylpenicillin, ciprofloxacin, clindamycin, chloramphenicol, doxycycline, erythromycin, gentamicin, metronidazole, moxifloxacin, rifampicin, tetracycline, and vancomycin using the E test method. Virulence factors (ace, asa, asa373, cylA, efaA, esp, and gelE) were detected by polymerase chain reaction assay. RESULTS: Amoxicillin + clavulanate was effective against all strains. Intermediate and total resistance was found against the majority of the tested antimicrobials. The susceptibility of some microorganisms to some antimicrobial agents changed according to the evaluation time. MIC50 and MIC90 also varied according to the evaluation time. In relation to the virulence factors of the E faecalis isolates, ace was detected in 100% of the strains, asa (60%), asa373 (15%), efaA (95%), esp (70%), and gelE (75%), whereas cylA was not detected. CONCLUSIONS: It was concluded that E. faecalis isolates from persistent endodontic infections showed varied degrees of intermediate/total resistance to several antimicrobial agents, with amoxicillin + clavulanate being the most effective agent. Moreover, the strains showed different patterns for virulence gene detection.

Evaluation of endodontic treatments performed by students in a Brazilian Dental School.

The aims of this study were to evaluate the clinical outcomes of root canal treatments performed by final-year students in the Dental School of Piracicaba, State University of Campinas, SP, Brazil, during the year 2000 and to evaluate the success rate of these treatments at follow-ups conducted one and three years later (2001-03). All 579 endodontic treatments performed by final-year students were selected for this investigation. Detailed personal and dental history was obtained from the patient's records. Clinical and radiographic follow-up examinations were performed. For 78.8 percent of the cases, dental caries was recorded as the cause for performing endodontic treatment; for 8.8 percent, prosthetic reasons were given; and for 12.4 percent, failure of the endodontic treatment was the reason. The largest percentage (30.7 percent) of recalled patients was examined after one year; the success rate at this follow-up evaluation ranged from 83 percent to 96 percent depending on the pulp status prior to the root canal treatment. After three years, only 8 percent of treated patients returned; assessment for them revealed a 75.5 percent success rate. Dental caries is still the main reason for endodontic treatment. This level of success of the root canal treatment in a Brazilian dental school is similar to other reports in the literature.

Efficacy of ethylene-diamine-tetra-acetic acid associated with chlorhexidine on intracanal medication removal: a scanning electron microscopy study.

The aim of this study was to evaluate the effectiveness of 17% ethylene-diamine-tetra-acetic acid (EDTA) used alone or associated with 2% chlorhexidine gel (CHX) on intracanal medications (ICM) removal. Sixty single-rooted human teeth with fully formed apex were selected. The cervical and middle thirds of each canal were prepared with Gates Glidden drills and rotary files. The apical third was shaped with hand files. The specimens were randomly divided into two groups depending on the ICM used after instrumentation: calcium hydroxide Ca(OH)(2) +CHX or Ca(OH)(2) +sterile saline (SS). After seven days, each group was divided into subgroups according to the protocol used for ICM removal: instrumentation and irrigation either with EDTA, CHX+EDTA, or SS (control groups). All specimens were sectioned and processed for observation of the apical thirds by using scanning electron microscopy. Two calibrated evaluators attributed scores to each specimen. The differences between the protocols for ICM removal were analyzed with Kruskal-Wallis and Mann-Whitney U tests. Friedman and Wilcoxon signed rank tests were used for comparison between the score of debris obtained in each root canal third. Remains of Ca(OH)(2) were found in all specimens independently of the protocol and ICM used (P > 0.05). Seventeen percent EDTA showed the best results in removing ICM when used alone (P < 0.05), particularly in those associated with CHX. It was concluded that the chelating agent 17% EDTA significantly improved the removal of ICM when used alone. Furthermore, the type of the vehicle associated with Ca(OH)(2) also plays a role in the ICM removal.

Microbial evaluation of traumatized teeth treated with triple antibiotic paste or calcium hydroxide with 2% chlorhexidine gel in pulp revascularization.

INTRODUCTION: Revascularization outcome depends on microbial elimination because apical repair will not happen in the presence of infected tissues. This study evaluated the microbial composition of traumatized immature teeth and assessed their reduction during different stages of the revascularization procedures performed with 2 intracanal medicaments. METHODS: Fifteen patients (7-17 years old) with immature teeth were submitted to the revascularization procedures; they were divided into 2 groups according to the intracanal medicament used: TAP group (n = 7), medicated with a triple antibiotic paste, and CHP group (n = 8), dressed with calcium hydroxide + 2% chlorhexidine gel. Samples were taken before any treatment (S1), after irrigation with 6% NaOCl (S2), after irrigation with 2% chlorhexidine (S3), after intracanal dressing (S4), and after 17% EDTA irrigation (S5). Cultivable bacteria recovered from the 5 stages were counted and identified by means of polymerase chain reaction assay (16S rRNA). RESULTS: Both groups had colony-forming unit counts significantly reduced after S2 (P < .05); however, no significant difference was found between the irrigants (S2 and S3, P = .99). No difference in bacteria counts was found between the intracanal medicaments used (P = .95). The most prevalent bacteria detected were Actinomyces naeslundii (66.67%), followed by Porphyromonas endodontalis, Parvimonas micra, and Fusobacterium nucleatum, which were detected in 33.34% of the root canals. An average of 2.13 species per canal was found, and no statistical correlation was observed between bacterial species and clinical/radiographic features. CONCLUSIONS: The microbial profile of infected immature teeth is similar to that of primarily infected permanent teeth. The greatest bacterial reduction was promoted by the irrigation solutions. The revascularization protocols that used the tested intracanal medicaments were efficient in reducing viable bacteria in necrotic immature teeth.

Treponema diversity in root canals with endodontic failure.

OBJECTIVE: This study sought to investigate the prevalence of eight oral Treponemas (Treponema denticola, T. amylovorum, T. maltophilum, T. medium, T. pectinovorum, T. socranskii, T. vicentii and T. lecithinolyticum) in teeth with endodontic treatment failure and periapical lesion. METHODS: Samples were taken from 40 root canals presenting endodontic failure and periapical lesion. DNA extraction was performed and Nested-PCR technique was used for the detection of Treponema species using specific primers. RESULTS: Treponemas was detected in 56.5% of the samples analyzed (22/39). Individual root canals yielded a maximum of 6 target Treponema species. T. denticola (30.8%) and T. maltophilum (30.8%) were the most frequently detected species followed by T. medium (20.5%), T. socranskii (20.5%), T. pectinovorum (17.9%) and T. vicentii (17.9%). Positive association was verified between T. denticola and T. maltophilum such as T. medium (P<.05). T. lecithinolyticum was positively associated with intraradicular post (P<.05). CONCLUSION: The present study revealed that a wide variety of Treponema species plays a role in persistent/secondary infection turning the root canal microbiota even more complex than previously described by endodontic literature.

Chlorhexidine in endodontics.

Chemical auxiliary substances (CAS) are essential for a successful disinfection and cleanness of the root canals, being used during the instrumentation and if necessary, as antimicrobial intracanal medicaments. Different CAS have been proposed and used, among which sodium hypochlorite (NaOCl), chlorhexidine (CHX), 17% EDTA, citric acid, MTAD and 37% phosphoric acid solution. CHX has been used in Endodontics as an irrigating substance or intracanal medicament, as it possesses a wide range of antimicrobial activity, substantivity (residual antimicrobial activity), lower cytotoxicity than NaOCl whilst demonstrating efficient clinical performance, lubricating properties, rheological action (present in the gel presentation, keeping the debris in suspension); it inhibits metalloproteinase, is chemically stable, does not stain cloths, it is odorless, water soluble, among other properties. CHX has been recommended as an alternative to NaOCl, especially in cases of open apex, root resorption, foramen enlargement and root perforation, due to its biocompatibility, or in cases of allergy related to bleaching solutions. The aim of this paper is to review CHX's general use in the medical field and in dentistry; its chemical structure, presentation form and storage; mechanism of action; antimicrobial activity including substantivity, effects on biofilms and endotoxins, effects on coronal and apical microbial microleakage; tissue dissolution ability; interaction with endodontic irrigants; effects on dentin bonding, metalloproteinases and collagen fibrils; its use as intracanal medicament and diffusion into the dentinal tubules; its use as disinfectant agent of obturation cones; other uses in the endodontic therapy; and possible adverse effects, cytotoxicity and genotoxicity.

Quantitative and qualitative analysis of microorganisms in root-filled teeth with persistent infection: Monitoring of the endodontic retreatment.

OBJECTIVE: The aim of this study was to investigate in vivo microorganisms detected in root-filled teeth with post-treatment apical periodontitis and quantify colony-forming units (CFU) during endodontic retreatment. MATERIALS AND METHODS: Fifteen root-filled teeth had their previous gutta-percha removed and were randomly instrumented before being divided into three groups and medicated with either [Ca(OH)2 + 2% CHX gel], [Ca(OH)2 + 0.9% NaCl] or 2% CHX gel. Samples were taken after removal of gutta-percha (S1), after chemomechanical preparation using 2% CHX gel (S2), and after inter-appointment dressing (S3) for 7 or 14 days later. Cultivable bacteria recovered from infected root canals at the three stages were counted and identified by means of culture and PCR assay (16S rDNA). Quantitative data were statistically analyzed by using Mann-Whitney test in which pairs of groups were compared (P < 0.05). RESULTS: CFU counts decreased significantly from S1 to S2 (P < 0.05). No significant difference was found between S2 and S3 (P = 0.3093) for all three experimental groups. Chemomechanical preparation and intra-canal dressing promoted significant median reductions of 99.61% and 99.57%, respectively, in the number of bacteria compared to S1 samples. A total of 110 cultivable isolates were recovered by culture technique from 32 different species and 7 different genera. Out of the 13 target species-specific primer of bacteria analyzed, 11 were detected during endodontic retreatment. CONCLUSION: The great majority of taxa found in post-treatment samples were Gram-positive bacteria, although Gram-negative bacteria were found by molecular methods. Moreover, our results showed that gutta-percha removal and chemomechanical preparation are effective for root canal disinfection, whereas additional intra-canal dressing did not improve disinfection.