Abstract Root canal infections are typically polymicrobial and involve strong bacterial interactions. The goal of endodontic treatment is to remove infected content from the root canal system to allow the healing of a pre-existing periapical lesion or to prevent infection of the periradicular tissues. Instrumentation alone is not capable of touching all of the root canal walls. Therefore, the irrigation process is an essential step in the endodontic treatment. However, due to the complex anatomy of the root canal system, this cleaning is very challenging. Although syringe and needle irrigation associated with the use of chemical substances is still the most used method, it does not guarantee optimal cleaning of the root canals. As a result, not only alternative irrigating substances but also numerous activation systems - which are technologies that aim to optimize the action of irrigating substances, both chemically and physically - have been developed. This work aimed to review the characteristics of both classic and current alternatives of irrigating substances and irrigation activation systems.

Resumo As infecções dos canais radiculares são tipicamente polimicrobianas e envolvem fortes interações bacterianas. O objetivo do tratamento endodôntico é remover o conteúdo infeccioso do sistema de canais radiculares, a fim de permitir a cicatrização de uma lesão periapical pré-existente ou prevenir a infecção dos tecidos perirradiculares. A instrumentação por si só não é capaz de tocar todas as paredes dos canais radiculares, desta forma a irrigação é uma etapa essencial no tratamento endodôntico. No entanto, devido à complexa anatomia do sistema de canais radiculares, essa limpeza é muito desafiadora. Embora a irrigação convencional com seringa e agulha, fazendo o uso de substâncias auxiliares seja o método mais utilizado, ela não garante a limpeza ideal dos canais radiculares. Como resultado, foram desenvolvidos inúmeros sistemas de ativação que visam otimizar a ação das substâncias irrigadoras, tanto química quanto fisicamente. O objetivo deste artigo foi revisar as características das substâncias irrigadoras (clássicas e alternativas) e dos sistemas de ativação destas substâncias.

Resumo Os cimentos endodônticos obturadores à base de silicato de cálcio foram desenvolvidos em formas pó/líquido ou pré-misturadas prontas para o uso. Foi realizada a avaliação das propriedades físico-químicas de um protótipo pó/líquido MTApex Sealer (Ultradent) em comparação com um material pronto para uso EndoSequence BC Sealer (Brasseler). O cimento obturador pasta/pasta à base de resina epóxi AH Plus (Dentsply) serviu de controle para as comparações. Os cimentos foram avaliados (n = 6) quanto a: tempo de presa (em ambientes secos e úmidos), escoamento e radiopacidade, seguindo a norma ISO-6876/2012; como também o pH. A superfície do material e a caracterização química foram realizadas utilizando a microscopia eletrônica de varredura (MEV) e espectrometria dispersiva de energia (EDS). ANOVA, Shapiro-Wilk, Levene e análise post-hoc com correção de Bonferroni foram realizadas com nível de significância de 5%. O MTApex Sealer exibiu um escoamento mais alto e o EndoSequence BC Sealer teve um tempo de presa significativamente mais longo em ambiente seco em comparação com o ambiente úmido; entretanto, para MTApex Sealer e AH Plus, nenhuma mudança significativa ocorreu quando umidade adicional foi fornecida. Todos os materiais excederam 7 mm de Al de radiopacidade e mostraram uma alcalinidade decrescente ao longo dos 21 dias de análise. A avaliação MEV/EDS resultou em picos de cálcio, silício e respectivo radiopacificador. O protótipo de pó/líquido MTApex Sealer teve o maior escoamento e tempo de presa semelhante em ambientes secos e úmidos; ao contrário, o EndoSequence BC Sealer foi influenciado de maneira crucial pela umidade externa. Isso sugere que a configuração dos materiais em pó/líquido parece ser mais previsível.

Abstract Calcium silicate-based root canal sealers have been developed in powder/liquid or premixed ready-to-use forms. The evaluation of the physico-chemical properties of a prototype powder/liquid MTApex Sealer (Ultradent) in comparison to a ready-to-use material EndoSequence BC Sealer (Brasseler) was performed. The paste/paste epoxy resin-based AH Plus (Dentsply) served as control for comparisons. The sealers were evaluated (n = 6) regarding setting time (in dry and moist environments), flow and radiopacity, following the ISO-6876/2012 standard. Also, the pH was assessed. Material’s surface and chemical characterization was evaluated using scanning-electron-microscopy (SEM) and energy-dispersive-spectrometry (EDS). Mixed ANOVA, Shapiro-Wilk, Levene, and post-hoc analysis with Bonferroni correction were performed at a significance level of 5%. MTApex Sealer exhibited the highest flow and EndoSequence BC Sealer had a significantly longer setting time in dry compared to the moist environment; however, for MTApex Sealer and AH Plus no significant changes occurred when additional moisture was provided. All materials exceeded 7 mm Al of radiopacity and showed a decreasing alkalinity over the 21 day-analysis. SEM/EDS evaluation resulted in peaks of calcium, silicon, and the respective radiopacifier. The prototype powder/liquid MTApex Sealer had the highest flow and similar setting time in both dry and moist environments; opposingly, EndoSequence BC Sealer was crucially influenced by external moisture. This suggests that the powder/liquid materials’ setting seems to be more predictable.

Resumo A descontaminação do sistema do canal radicular (CR) é essencial para o sucesso da terapia endodôntica. Neste estudo in vitro, foi avaliado o potencial de desinfecção de um dispositivo de eletrofulguração, o Endox® Endodontic System (EES), em CR infectado com Enterococcus faecalis. Sessenta e cinco pré-molares inferiores humanos foram instrumentados com o sistema MTwo®. As amostras foram distribuídas em seis grupos experimentais (n = 10), de acordo com o protocolo de irrigação: clorexidina gel a 2% (CHX); CHX + EES; Hipoclorito de sódio 5,25% (NaOCl); NaOCl + EES; solução salina (SS); e SS + EES. Cinco amostras não foram tratadas (controle). As amostras de CR foram coletadas antes (C1) e após o tratamento com EES (C2), após preparo químico-mecânico (PQM) (C3) e após o tratamento final com EES (C4). Todas as amostras foram plaqueadas para unidades formadoras de colônias (CFU / mL) em meio sólido. Os dados foram analisados pelos testes de Wilcoxon e Friedman para comparações intragrupos e pelo teste de Kruskal Wallis, seguidos pelo teste de Dunn para comparações intergrupos (α = 0,05). O tratamento com o EES não reduziu significativamente o número de CFU / mL em comparação com os níveis basais (C1 vs. C2, p> 0,05). Após PQM (C3), todos os grupos apresentaram uma quantidade significativamente reduzida de CFU / mL (p <0,05), sem diferença entre as amostras tratadas com CHX e NaOCl (p> 0,05). Por fim, o tratamento com o EES após PQM (C4) não reduziu significativamente a quantidade de CFU / mL (C3 vs. C4, p> 0,05). Concluindo, o uso do Endox® Endodontic System não resultou em redução bacteriana considerável em todos os momentos operatórios, enquanto os tratamentos com NaOCl e CHX foram igualmente eficientes para esse fim.

Abstract Decontamination of the root canal (RC) system is essential for successful endodontic therapy. In this in vitro study, it was evaluated the disinfection potential of an electrofulguration device, the Endox® Endodontic System (EES), in RC infected with Enterococcus faecalis. Sixty-five human lower premolars were instrumented with MTwo® system. The specimens were distributed into six experimental groups (n = 10), according to the irrigation protocol: 2% chlorhexidine gel (CHX); CHX + EES; 5.25% sodium hypochlorite (NaOCl); NaOCl + EES; saline solution (SS); and SS + EES. Five specimens were untreated (control). RC samples were collected before (C1) and after EES treatment (C2), after chemo-mechanical preparation (CMP) (C3), and after final EES treatment (C4). All samples were plated for colony forming units (CFU/mL) onto solid media. The data were analyzed by Wilcoxon and Friedman tests for intragroup comparisons and by Kruskal Wallis test followed by Dunn’s test for intergroup comparisons (α = 0.05). Treatment with the EES did not significantly reduce the number of CFU/mL as compared to baseline levels (C1 vs. C2, p> 0.05). After CMP (C3), all groups showed a significantly reduced amount of CFU/mL (p <0.05), with no difference between CHX- and NaOCl-treated samples (p >0.05). Lastly, treatment with the EES following CMP (C4) did not significantly reduce the amount of CFU/mL (C3 vs. C4, p> 0.05). To conclude, the use of the Endox® Endodontic System did not result in considerable bacterial reduction at all operative times, while treatment with NaOCl and CHX was equally efficient for this purpose.

Abstract Objective Pulp revascularization is an effective treatment for immature necrotic teeth. Calcium hydroxide has been used in pulp revascularization as an intracanal medication due to its antimicrobial action and the non-exhibition of crown discoloration and cytotoxicity for stem cells from the apical papilla. Our study aimed to investigate the clinical success and quantitative radiographic changes of root development in immature traumatized teeth using calcium hydroxide plus 2% chlorhexidine gel as intracanal medication. Methodology In this retrospective study, 16 patients were treated with a standardized pulp revascularization protocol. Calcium hydroxide and 2% chlorhexidine gel were manipulated in a 1:1 (v/v) ratio and inserted into root canals with Lentulo spirals (Dentsply Maillefer, Baillaigues, Switzerland). Patients were followed up for a period from 9 to 36 months for the evaluation of clinical and radiological data. Radiographic measurements of root length, root width, apical diameter, and MTA placement from the apex were quantified using software ImageJ. Wilcoxon test and t-test were used, according to nonparametric or parametric data, respectively, for changes over time in root length, root width, and apical diameter. Results Fifteen teeth survived during the follow-up period (93.75%) and met the criteria for clinical success. Although the changes seem to be very small in many cases, significant increases in the average root length (14.28%, p<0.0001), root width (8.12%, p=0.0196), and decrease in apical diameter (48.37%, p=0.0007) were observed. MTA placement from the apex and age at the time of treatment was not significantly associated with the quantitative radiographic outcomes. Conclusions Pulp revascularization in traumatized immature teeth treated with calcium hydroxide plus 2% chlorhexidine gel as intracanal medication had high success and survival rates, showing periodontal healing and resolution of signs and symptoms. However, concerning the continued root development, the outcomes can still be considered unpredictable.

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.

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 O objetivo deste estudo foi investigar a composição bacteriana de canais radiculares associados com abscesso apical agudo através de identificação molecular (16S rRNA) de bactérias cultiváveis. Duzentas e vinte cepas, de 20 casos, isoladas por cultura foram submetidas a extração de DNA e amplificação do gene 16S rRNA (PCR), seguido de sequenciamento. As sequências de nucleotídeos obtidas foram comparadas com o banco de dados (GenBank) do National Center of Biotechnology Information através do BLAST. Cepas não identificadas pelo sequenciamento foram submetidas à clonagem. Associação de achados microbiológicos e características clínicas e associação entre espécies bacterianas também foram investigadas. Cinquenta e nove bactérias cultiváveis diferentes foram identificadas pelo sequenciamento do gene 16S rRNA, pertencentes a 6 filos, numa média de 6 espécies por canal. Método molecular permitiu identificação de 99% das cepas isoladas. As bactérias mais frequentes foram Prevotella spp., Pseudoramibacter alactolyticus, Parvimonas micra, Dialister invisus, Filifactor alocis, Peptostreptococcus stomatis. Associação positiva foi encontrada entre Prevotella buccae e Pseudoramibacter alactolyticus, e entre Parvimonas micra e Prevotella nigrescens (p<0,05). Foi concluído que a microbiota de canais radiculares infectados associados com abscesso apical agudo é diversa e heterogênea, composta principalmente por anaeróbios Gram-negativos, pertencentes aos filos Firmicutes e Bacteroidetes.

Abstract The objective of this study was to investigate the bacterial composition present in root canals of teeth associated with acute apical abscess by molecular identification (16S rRNA) of cultivable bacteria. Two hundred and twenty strains isolated by culture from 20 root canals were subjected to DNA extraction and amplification of the 16S rRNA gene (PCR), followed by sequencing. The resulting nucleotide sequences were compared to the GenBank database from the National Center of Biotechnology Information through BLAST. Strains not identified by sequencing were submitted to clonal analysis. The association of microbiological findings with clinical features and the association between microbial species were also investigated. Fifty-nine different cultivable bacteria were identified by 16S rRNA gene sequencing, belonging to 6 phyla, with an average number of 6 species per root canal. Molecular approaches allowed identification of 99% of isolates. The most frequently identified bacteria were Prevotella spp., Pseudoramibacter alactolyticus, Parvimonas micra, Dialister invisus, Filifactor alocis, and Peptostreptococcus stomatis. Positive association was found between Prevotella buccae and Pseudoramibacter alactolyticus and between Parvimonas micra and Prevotella nigrescens (both p<0.05). It was concluded that the microbiota of infected root canals associated with acute apical abscess is diverse and heterogeneous, composed mainly of anaerobic Gram-negative bacteria, with the great majority belonging to the phyla Firmicutes and Bacteroidetes.

Resumo O objetivo deste estudo foi comparar a atividade biológica de lipopolissacarídeos (LPS) purificados a partir de linhagens de Fusobacterium nucleatum e Porphyromonas gingivalis, ambas isoladas de infecções endodônticas primárias (IEP) nos níveis de IL-1β e TNF-α produzidos por macrófagos. Adicionalmente, LPS foi purificado de F. nucleatum e P. gingivalis "American Type Collection" (ATCC) e sua atividade comparada às respectivas linhagens clinicamente isoladas. Linhagens de F. nucleatum e P. gingivalis isoladas clinicamente de IEP tiveram sua identificação confirmada por sequenciamento do gene 16S rRNA. LPS de F. nucleatum e P. gingivalis e das respectivas linhagens foram extraídos com o uso do método "Tri-reagent". Macrófagos (Raw 264.7) foram estimulados com LPS a 100 ng/mL por 4, 8 e 12 h. A secreção de IL-1β e de TNF-α foi determinada. Foram usados os testes t-pareado, ANOVA de medidas repetidas e ANOVA de um fator. Todos os LPS induziram a produção significante de IL-1β e TNF-α, sendo o primeiro secretado em mais altas concentrações que o último em todos os tempos avaliados. F. nucleatum induziu uma maior expressão de ambas as citocinas comparativamente ao P. gingivalis (p<0,05). Não foram observadas diferenças entre as linhagens clínica e ATCC, uma vez que ambas apresentaram o mesmo potencial de indução da resposta pró-inflamatória. Conclui-se que F. nucleatum e P. gingivalis possuem diferentes padrões de ativação dos macrófagos, como visto pela produção de IL-1β e TNF-α, o que pode contribuir para a imunopatogênese da periodontite apical. Ainda, linhagens clínica e ATCC mantidas no mesmo ambiente in vitro apresentaram ativação biológica semelhante.

Abstract The aim of this study was to compare the biological activity of lipopolysaccharides (LPS) purified from Fusobacterium nucleatum and Porphyromonas gingivalis strains, both isolated from primary endodontic infection (PEI) in the levels of IL-1β and TNF-α released by macrophage cells. Moreover, LPS was purified from F. nucleatum and P. gingivalis American Type Collection (ATCC) and its biological activity was compared to respectively clinical isolates strains. F. nucleatum and P. gingivalis strains clinically isolated from PEI had their identification confirmed by sequencing the 16S rRNA gene. LPS from F. nucleatum and P. gingivalis and their respective ATCC strains were extracted by using Tri-reagent method. Macrophages (Raw 264.7) were stimulated with LPS at 100 ng/mL for 4, 8 and 12 h. Secretion of IL-1 β and TNF-α was also determined. Paired t-test, repeated measures ANOVA and one-way ANOVA were employed. All LPS induced significant production of IL-1β and TNF-α, with the former being secreted at higher levels than the latter in all time-points. F. nucleatum induced a higher expression of both cytokines compared to P. gingivalis (p<0.05). No differences were observed between clinical and ATCC strains, as both presented the same potential to induce pro-inflammatory response. It was concluded that F. nucleatum and P. gingivalis LPS presented different patterns of activation against macrophages as seen by the IL-1β and TNF-α production, which may contribute to the immunopathogenesis of apical periodontitis. Moreover, clinical and ATCC strains grown under the same in vitro environment conditions presented similar biological activity.

The purpose of this study was to evaluate in vivo the biocompatibility of Endométhasone, Pulp Canal Sealer EWT and AH-Plus root canal sealers after implantation in rat connective tissue. Twenty-four Wistar-Furth rats were used. Polyethylene tubes were filled with the sealers and implanted into specific dorsal subdermal tissue sites of the rats. Implants were removed after 3, 7 and 30 days, fixed and processed for glycol methacrylate-embedding technique to be examined microscopically. On the 3rd day, there was a mild inflammatory reaction to Pulp Canal Sealer EWT implants, but a severe response to the other sealers with presence of acute inflammatory cells. On the 7th day, tissue organization was more evident with attenuation of the inflammatory reaction, especially for the AH-Plus implants. On the 30th day, connective tissue with few inflammatory cells was observed in contact with all sealer implants. In this time interval, the tissue in contact with Pulp Canal Sealer EWT implants was more organized, while the tissue close to Endométhasone and AH-Plus implants showed a mild persistent inflammatory reaction and had similar results to each other. In conclusion, the sealers had a similar pattern of irritation, which was more severe in the beginning and milder with time, in such a way that all sealers showed a persistent mild reaction. Pulp Canal Sealer EWT yielded better tissue organization than Endométhasone and AH-Plus, which, in turn, showed similar results to each other.

O objetivo deste estudo foi avaliar in vitro a atividade antimicrobiana do gluconato de clorexidina gel, como irrigante endodôntico, comparando-o ao hipoclorito de sódio (NaOCl) e ao gluconato de clorexidina líquido. A atividade antimicrobiana das substâncias testadas foi avaliada pelo teste de difusão em ágar. As zonas de inibição de crescimento bacteriano produzidas pela clorexidina gel a 0,2%; 1% e 2% foram observados frente a 5 espécies de bactérias anaeróbias facultativas e 4 espécies de anaeróbios estritos, Gram-negativos e produtores de pigmento negro; e comparados com os resultados obtidos pelo NaOCl e pela clorexidina líquida. As maiores zonas de inibição foram produzidas quando as bactérias testadas ficaram em contato com a clorexidina a 2% em gel (11,79 mm), apresentando diferença estatisticamente significante (p<0,05) quando comparados às zonas de inibição de crescimento bacteriano produzidas por todas as concentrações avaliadas de NaOCl, incluindo 5,25% (9,54 mm). No entanto, não houve diferença estatisticamente significante (p>0,05) comparando as zonas produzidas por concentrações equivalentes de clorexidina líquida ou gel. Os resultados indicaram que a clorexidina em gel tem grande potencial para ser usada como substância química auxiliar quanto às suas propriedades antimicrobianas.

The purpose of this study was to assess the in vitro the antimicrobial efficacy of chlorhexidine gluconate gel as an endodontic auxiliary chemical substance compared to sodium hypochlorite (NaOCl) and chlorhexidine gluconate solution. The antimicrobial efficacy of the tested substances was evaluated using the agar diffusion test. The growth inhibition zones produced by 0.2%, 1% and 2% chlorhexidine gel were evaluated against 5 facultative anaerobic bacteria and 4 pigmented Gram-negative anaerobes, and compared to the results obtained by NaOCl and chlorhexidine solution. The largest growth inhibition zones were produced when the test bacteria were in contact with 2% chlorhexidine gluconate gel (11.79 mm), being significantly different (p<0.05) from the growth inhibition zones produced by all NaOClconcentrations, including 5.25% (9.54 mm). However, there was no statistically significant difference (p>0.05) between the growth inhibition zones obtained with equal concentrations of chlorhexidine solution and gel. The results of this study indicate that, as far as its antimicrobial properties are concerned, chlorhexidine gel has a great potential to be used as an endodontic auxiliary chemical substance.