Effect of sodium hypochlorite solution and gel with/without passive ultrasonic irrigation on Enterococcus faecalis, Escherichia coli and their endotoxins.

Background: Sodium hypochlorite (NaOCl) is the most commonly used irrigant in endodontics . The purpose of this study was to evaluate the effect of NaOCl solution (2.5%) and gel (3%) with/without passive ultrasonic irrigation (PUI) on Enterococcus faecalis, Escherichia coli, and their endotoxins, lipopolysaccharide (LPS) and lipoteichoic acid (LTA). Methods: 40 human lower premolars were contaminated with E. coli (ATCC 25922) for 28 days and E. faecalis (ATCC 29212) for 21 days. Specimens were randomly divided into four groups: (1) 2.5% NaOCl irrigating the canals without PUI activation; (2) 2.5% NaOCl with PUI; (3) 3% NaOCl gel irrigating the canals without PUI; and (4) 3% NaOCl gel with PUI. 40 mL of irrigant was used for each group. PUI activation was carried out using E1-Irrisonic stainless-steel tip at 10% frequency. After treatment, all specimens were filled with 3mL of 17% ethylenediaminetetraacetic acid (EDTA) for 3min and then washed with nonpyrogenic saline solution. Three samples were collected from the canals: S1, at baseline to confirm biofilm formation; S2 after treatment; and S3 after EDTA. Samples were assessed for E. coli and E. faecalis colony forming units, and LPS and LTA were assessed using chromogenic kinetic LAL assay and ELISA, respectively. Data were analyzed by Kruskal-Wallis, Friedmann and Dunn tests with α≤0.05. Results: All groups were effective in reducing the microbial load of E. coli and E. faecalis after treatment without a significant difference among the groups. NaOCl and NaOCl gel groups had no significant difference in reducing LPS and LTA. Statistically increased reduction was seen for NaOCL + PUI and NaOCl gel + PUI compared for groups without PUI. Conclusions: NaOCl gel has the same antimicrobial action of NaOCl solution and can partially detoxify endotoxins. PUI improves NaOCl (gel or solution) action over E. faecalis and E. coli and their endotoxins.

Influência da ativação ultrassônica do gel de hipoclorito de sódio 3 por cento, e da medicação intracanal com hidróxido de cálcio sobre micro-organismos e ácido lipoteicóico / Influence of activation by ultrasound on sodium hypochlorite 3% gel and an intracanal medication of calcium hydroxide on microrganisms and lipotheicoic acid

O objetivo deste estudo foi avaliar in vitro a ação do gel de hipoclorito de sódio (NaOCl) 3 % e da medicação intracanal (MIC) de hidróxido de cálcio (CaOH2), agitados ou não por ultrassom sobre Enterococcus faecalis, Escherichia coli e ácido lipoteicóico (LTA). Para isso 80 dentes humanos unirradiculares tiveram suas coroas removidas padronizando seu comprimento em 16 mm +- 0,5 mm, sendo seus canais instrumentados inicialmente até instrumento R25 (Reciproc). Os canais foram contaminados com suspensões de E. faecalis e E. coli. Os canais foram instrumentados utilizando-se instrumento R40, sob irrigação com 2 ml de NaOCl 3% gel seguida de irrigação com 10 ml de solução salina estéril e apirogênica. Após os canais foram irrigados com EDTA 17%, seguido de irrigação com 5 ml de solução salina estéril e apirogênica, sendo por último preenchidos com medicação intracanal de hidróxido de cálcio mantida durante 7 ou 14 dias. Os espécimes foram divididos inicialmente em 2 grupos (n=40) de acordo com a agitação ultrassônica (US) ou não da substância química auxiliar. Sendo novamente divididos de acordo com a agitação ultrassônica ou não da medicação intracanal (MIC) e o tempo de ação desta (n=10): 1) NaOCl + Ca(OH)2 (7 dias). 2) NaOCl + Ca(OH)2 (14 dias). 3) NaOCl + Ca(OH)2 com US (7 dias) 4) NaOCl + Ca(OH)2 com US (14 dias). 5) NaOCl com US + Ca(OH)2 (7 dias). 6) NaOCl com US + Ca(OH)2 (14dias) 7) NaOCl com US + Ca(OH)2 com US (7dias). 8) NaOCl com US + Ca(OH)2 com US (14 dias). Foram realizadas coletas do canal radicular 28 dias após o início da contaminação dos espécimes (1ª coleta), logo após o preparo biomecânico (PBM) (2ª coleta), logo após o preenchimento com EDTA (3ª coleta) e após o tempo de ação da MIC (4ª coleta). Para todas as coletas foram avaliadas a atividade antimicrobiana (UFC/ml) e quantificação de LTA pelo teste de Elisa. Os resultados foram submetidos aos testes estatísticos Kruskal-Wallis e Dunn (5%). Verificou-se pela análise microbiana e quantificação de LTA, que o NaOCl 3% gel foi capaz de eliminar quase completamente os micro-organismos dos canais radiculares, mas não o ácido lipoteicóico, independente da ativação ultrassônica. A ativação da MIC de Ca(OH)2 não exerceu efeito sobre micro-organismos. Sobre LTA, a ativação da MIC de Ca(OH)2 não foi eficaz, sendo os grupos com maior percentual de redução os que não sofreram agitação da MIC, nem do NaOCl. Conclui-se que o NaOCl 3% gel boa tem capacidade antimicrobina, independente da ativação ultrassônica. O PBM não foi capaz de detoxificar o LTA dos canais radiculares. A MIC com ativação ultrassônica não foi eficiente na redução de LTA(AU)

The aim of this study was to evaluate in vitro the action of sodium hypochlorite (NaOCl) 3% gel and calcium hydroxide as an intracanal medication, both activated by ultrasound on the reduction of E. faecalis e E. coli and lipotheicoic acid (LTA). Eight human single-rooted teeth with standardized size of 16 mm were prepared initially to R25 instrument (Reciproc) and distributed in microplate (n = 10). After sterilization (Co60 gamma radiation), the infection was carried out 8 microlitres of E. coli suspension, and after 7 days 8 microlitres of E. faecalis suspension, maintained for 21 days. Following the collection confirmation was performed (1st collection), then the root were instrumented using R40 instrument, irrigation with 2 ml of NaOCl 3% gel followed by washing with 10 ml of saline. The specimens were divided into 8 groups (n = 10) according to different ultrasonic irrigation protocols and different periods exposed to intracanal medication: 1) NaOCl + Ca(OH)2(7 days). 2) NaOCl + Ca(OH)2 (14 days). 3) NaOCl + Ca(OH)2 with PUI (7 dias) 4) NaOCl + Ca(OH)2 with PUI (14 days). 5) NaOCl with PUI + Ca(OH)2 (7 days). 6) NaOCl with PUI + Ca(OH)2 (14 days) 7) NaOCl with PUI + Ca(OH)2 (7 days). 8) NaOCl with PUI + Ca(OH)2 with PUI (14 days). Were made to sample colections of content of root canals, after instrumentation (2nd sample), after the use of EDTA (3rd sample) and after the Ca(OH)2 (4th Collection) . Microbiological culture and LTA quantification revealed that 3 % NaOCl gel was capable to eliminate E. faecalis and E. coli almost completely from root canals, but not lipotheicoic acid, regardless the use of ultrasonic activation. The ultrasonic activation of intracanal medication (Ca(OH)2) was not effective in removing neither microorganisms nor LTA. In addition, the groups with greatest reduction were the ones with no ultrasonic activation either of intracanal medication or NaOCl. It was concluded that 3% NaOCl gel is effective on antimicrobial activity regardless the use of ultrasonic activation. Biomechanical preparation was not capable to detoxify LTA from root canals. Ultrasonic activation of intracanal medication was not effetive on LTA reduction(AU)