Effect of EDTA Activation on Blood Clot Structure in Regenerative Endodontics: A Scanning Electron Microscopy Study.

INTRODUCTION: EDTA plays a crucial role in regenerative endodontic therapy (RET) because of its significant biological effects. However, EDTA is also recognized as the preferred anticoagulant for hematologic tests. Thus, this study aimed to assess the influence of different EDTA activation techniques on the morphology of blood clots after conditioning the root canal dentin. METHODS: Forty extracted human teeth were prepared to simulate immature teeth and divided into the following 5 groups: (1) saline solution (negative control), (2) EDTA 17% + saline solution (CNI), (3) CNI + ultrasonic activation, (4) CNI + Easy clean activation, and (5) CNI + XP-endo Finisher activation. After irrigation, the roots were cleaved, and the root canals were filled with human blood to clot formation. The morphology and density of erythrocytes, platelets, and the fibrin network were observed using a scanning electron microscope. The fibrin network density was classified using a 4-point scale. Data were analyzed using the Friedman test and the Kruskal-Wallis test with Bonferroni adjustment (α = 5%). RESULTS: All groups exhibited consistent blood clot morphology characterized by a high density of erythrocytes, platelets, and white blood cells throughout the entire length of the root canal. The negative control group showed statistically significant high scores of fibrin density compared with the CNI group in all root thirds (P < .05). However, there was no statistical difference in the scores for the fibrin network density between the groups irrigated with EDTA with and without activation (P > .05). CONCLUSIONS: EDTA may impair the fibrin network formation compared with the saline group. However, EDTA activation did not significantly change the effects on the blood clot in contact with the conditioned intraradicular dentin.

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.

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.

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.

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.

Calcium Silicate-Based Sealers Do Not Reduce the Risk and Intensity of Postoperative Pain after Root Canal Treatment when Compared with Epoxy Resin-Based Sealers: A Systematic Review and Meta-Analysis.

This review aims to evaluate whether root canal obturation with calcium silicate-based (CSB) sealers reduces the risk and intensity of endodontic postoperative pain when compared to epoxy resin-based (ERB) sealers. The review was registered at PROSPERO (CRD42020169255). Two independent reviewers conducted an electronic search in PubMed, Scopus, EMBASE, Web of Science, Cochrane Library and LILACS until November 2020 and included only randomized clinical trials with adult health participants undergoing root canal treatment. After selection, the JBI Critical Appraisal tool was used to assess the risk of bias. A fixed-effect meta-analysis was performed to summarize the results of pain risk and pain intensity at time intervals of 24 and 48 hours. Finally, the certainty of evidence was assessed using the GRADE approach. The search resulted in 1,206 records, of which five studies (n = 421 patients) met the eligibility criteria and presented moderate to low risk of bias. There was no significant difference between groups in the risk of pain in the first 24 hours (relative risk or RR = 0.83, 95% confidence interval or CI: 0.60, 1.16, I 2 =) or 48 hours (RR = 0.56, 95% CI: 0.26, 1.21, I 2 =). Silicate-based sealers led to lower pain intensity only at 48 hours (mean and standard deviation = 0.37, 95% CI: 0.69, 0.05). All analyses revealed low heterogeneity (I 2 < 25%). The evidence presented moderate level of certainty. Currently available evidence has shown that there is no difference between CSB and ERB sealers in the risk or intensity of postoperative pain.

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.

Stem cells of the apical papilla regulate trigeminal neurite outgrowth and targeting through a BDNF-dependent mechanism.

Regenerative endodontic procedures have become a valuable alternative for the treatment of immature teeth with pulp necrosis. In addition to resolution of periradicular pathosis and promotion of continued root development, positive vitality testing has been observed in some regenerative clinical cases. Importantly, the positive response to electric stimulation of the regenerated tissue requires targeting of periradicular axons into the previously empty root canal space. However, the mechanism by which this process occurs is largely unknown. Since stem cells of the apical papilla (SCAP) have been proposed to populate the root canal following regenerative endodontic procedures, we hypothesized that SCAP regulate neurite outgrowth and axonal targeting. To test this hypothesis, we established primary co-cultures of human SCAP and rat trigeminal neurons, and performed neurite outgrowth assays using ELISA and confocal microscopy to determine the effect of increasing concentration of SCAP on the total neurite outgrowth and axonal targeting. In addition, we evaluated whether SCAP evoked axonal targeting in vivo using a matrigel subcutaneous implant assay. Data were analyzed by ANOVA with Bonferroni's post hoc test, and significance was set at p<0.05. The results demonstrated that SCAP release a soluble factor that regulates neurite outgrowth from cultured trigeminal neurons. Next, we demonstrated that this effect is completely abolished by pretreatment with a neutralizing antibody to brain-derived neurotrophic factor (BDNF), but not by antibodies to other neurotrophins. Further, SCAP release BDNF in a concentration-dependent manner as detected by ELISA, and trigger directed axonal targeting both in vitro and in vivo as demonstrated by microfluidic and matrigel implant experiments, respectively. Collectively, these results suggest that SCAP may be responsible for the chemical signal driving axons to target regenerated tissue via a BDNF-dependent mechanism.

Concentration-dependent effect of sodium hypochlorite on stem cells of apical papilla survival and differentiation.

INTRODUCTION: Intracanal disinfection is a crucial step in regenerative endodontic procedures. Most published cases suggest the use of sodium hypochlorite (NaOCl) as the primary irrigant. However, the effect of clinically used concentrations of NaOCl on the survival and differentiation of stem cells is largely unknown. In this study, we tested the effect of various concentrations of NaOCl on the stem cells of the apical papilla (SCAPs) survival and dentin sialophosphoprotein (DSPP) expression. METHODS: Standardized root canals were created in extracted human teeth and irrigated with NaOCl (0.5%, 1.5%, 3%, or 6%) followed by 17% EDTA or sterile saline. SCAPs in a hyaluronic acid-based scaffold were seeded into the canals and cultured for 7 days. Next, viable cells were quantified using a luminescence assay, and DSPP expression was evaluated using quantitative real-time polymerase chain reaction. RESULTS: There was a significant reduction in survival and DSPP expression in the group treated with 6% NaOCl compared with the untreated control group. Comparable survival was observed in the groups treated with the lower concentrations of NaOCl, but greater DSPP expression was observed in the 1.5% NaOCl group. In addition, 17% EDTA resulted in increased survival and DSPP expression partially reversing the deleterious effects of NaOCl. CONCLUSIONS: Collectively, the results suggest that dentin conditioning with high concentrations of NaOCl has a profound negative effect on the survival and differentiation of SCAPs. However, this effect can be prevented with the use of 1.5% NaOCl followed by 17% EDTA. The inclusion of this irrigation regimen might be beneficial in regenerative endodontic procedures.

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.

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.

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.

Resumo Substâncias químicas auxiliares (SQA) são essenciais para o processo de limpeza e desinfecção dos canais radiculares, sendo utilizadas durante a instrumentação dos canais radiculares e, se necessário, como medicamentos intracanais. Diferentes SQA têm sido propostas e utilizadas, entre elas: hipoclorito de sódio (NaOCl), clorexidina (CHX), EDTA 17%, ácido cítrico, MTAD e solução de ácido fosfórico a 37%. CHX tem sido usada na endodontia como SQA ou medicação intracanal. CHX possui uma ampla gama de atividade antimicrobiana; substantividade (atividade antimicrobiana residual); menor citotoxicidade que NaOCl, demonstrando desempenho clínico eficiente; propriedades de lubrificação; ação reológica (presente na apresentação gel, mantendo os detritos em suspensão); inibe metaloproteinases; é quimicamente estável; não mancha tecidos; é inodora; solúvel em água; entre outras propriedades. CHX tem sido recomendada como uma alternativa ao NaOCl, especialmente em casos de ápice aberto, reabsorção radicular, perfuração radicular e durante a ampliação foraminal, devido à sua biocompatibilidade, ou em casos de alergia ao NaOCl. O objetivo deste trabalho é fazer uma revisão do uso da clorexidina na medicina e na odontologia; sua estrutura química; forma de apresentação e armazenamento; mecanismo de ação, atividade antimicrobiana, incluindo, substantividade, efeitos sobre biofilmes e endotoxinas; efeito sobre infiltração microbiana coronal e apical; capacidade de dissolução do tecido; interação com os irrigantes; efeitos sobre a união à dentina, metaloproteinases e fibrilas de colágeno; a sua utilização como medicamento intracanal e difusão ...