Antibacterial effect of low-temperature plasma on Enterococcus faecalis in dentinal tubules in vitro / 北京大学学报(医学版)

OBJECTIVE@#To construct a model of Enterococcus faecalis (E. faecalis) infection in dentinal tubules by gradient centrifugation and to evaluate the antibacterial effect of low-temperature plasma on E. faecalis in dentinal tubules.@*METHODS@#Standard dentin blocks of 4 mm×4 mm×2 mm size were prepared from single root canal isolated teeth without caries, placed in the E. faecalis bacterial solution, centrifuged in gradient and incubated for 24 h to establish the model of dentinal tubule infection with E. faecalis. The twenty dentin blocks of were divided into five groups, low-temperature plasma jet treatment for 0, 5 and 10 min, calcium hydroxide paste sealing for 7 d and 2% chlorhexidine gel sealing for 7 d. Scanning electron microscopy and confocal laser scanning microscope were used to assess the infection in the dentinal tubules and the antibacterial effect of low-temperature plasma.@*RESULTS@#The results of scanning electron microscopy and confocal laser scanning microscopy showed that after 24 h of incubation by gradient centrifugation, E. faecalis could fully enter the dentinal tubules to a depth of more than 600μm indicating that this method was time-saving and efficient and could successfully construct a model of E. faecalis infection in dentinal tubules. Low-temperature plasma could enter the dentinal tubules and play a role, the structure of E. faecalis was still intact after 5 min of low-temperature plasma treatment, with no obvious damage, and after 10 min of low-temperature plasma treatment, the surface morphology of E. faecalis was crumpled and deformed, the cell wall was seriously collapsed, and the normal physiological morphology was damaged indicating that the majority of E. faecalis was killed in the dentinal tubules. The antibacterial effect of low-temperature plasma treatment for 10 min exceeded that of the calcium hydroxide paste sealing for 7 d and the 2% chlorhexidine gel sealing for 7 d. These two chemicals had difficulty entering deep into the dentinal tubules, and therefore only had a few of antibacterial effect on the bacterial biofilm on the root canal wall, and there was also no significant damage to the E. faecalis bacterial structure.@*CONCLUSION@#Gradient centrifugation could establish the model of E. faecalis dentin infection successfully. Low-temperature plasma treatment for 10 min could kill E. faecalis in dentinal tubules effectively, which is superior to the calcium hydroxide paste sealing for 7 d and the 2% chlorhexidine gel sealing for 7 d.

Comparação da propriedade antimicrobiana da clorexidina e do hipoclorito de sódio como irrigantes endodônticos: revisão integrativa / Comparison between the antimicrobial properties of chlorhexidine and sodium hypochlorite as endodontic irrigants: an integrative review / Comparación de la propiedad antimicrobiana de la clorhexidina y el hipoclorito de sodio como irrigantes endodónticos: revisión integradora

Introdução:Durante o tratamento endodôntico, devido às complexidades anatômicas dos canais radiculares, a ação mecânica dos instrumentos não é suficiente para a completa desinfecção dos condutos. Dessa forma, se faz necessário o uso de soluções irrigadoras que possampotencializar a desinfecção do sistema de canais radiculares. Objetivo:Realizar uma revisão integrativa da literatura para comparar as propriedades antimicrobianas da clorexidina com o hipoclorito de sódio.Metodologia:A busca na literatura foi realizada no período de setembro de 2019 a agosto de 2021, nas seguintes bases de dados: PUBMED/MEDLINE, LILACS e SCIELO. Utilizando os descritores: clorexidina, hipoclorito de sódio, irrigante do canal radicular e limpeza. Utilizou-se como critérios de busca, trabalhos experimentais laboratoriais in vitro, publicados entre os anos de 2017 e 2021.Resultados:Foram encontrados 165 artigos, dos quais 15 foram selecionados ao final do processo. 8 trabalhos não encontraram diferença estatisticamente significativa entre a clorexidina e o hipoclorito,5 artigos apresentaram resultados superiores dohipoclorito de sódioem 2 a clorexidina foi superior. Conclusões:Após análise da literatura, observamos semelhança entre a ação antimicrobiana do hipoclorito de sódioe da clorexidina, e podemos concluir que ambas apresentam boa ação antimicrobiana, justificando seu uso clinicamente (AU).

Introduction:During endodontic treatment, due to the anatomical complexities of the root canals, the mechanical action of the instruments is not sufficient for the complete disinfection of the canals. Thus, it is necessary to use irrigating solutions that can makethe disinfection of the root canal system.Objective:Conduct an integrative literature review to compare the antimicrobial properties of chlorhexidine with sodium hypochlorite.Methodology:The literature search was carried out from September 2019 to August 2021, in the following databases: PUBMED/MEDLINE, LILACS and SCIELO. Using the descriptors: chlorhexidine, sodium hypochlorite, root canal irrigant and cleaning. As search criteria, in vitro laboratory experimental works published between 2017 and 2021 were used.Results:A total of 165 articles were found, of which 15 were selected at the end of the process. 8 studies did not find a statistically significant difference between chlorhexidine and hypochlorite, 5 articles showed superior results for NaOCl and in 2 chlorhexidine was superior.Conclusions:After analyzing the literature, we observed a similarity between the antimicrobial action of sodium hypochlorite and chlorhexidine, and we can conclude that both have good antimicrobial action, justifying their clinical use (AU).

Introducción:Durante el tratamiento endodóntico, debido a las complexidades anatómicas de los conductos radiculares, la acción mecánica de los instrumentos no es suficiente para la desinfección completa de los conductos. Por lo tanto, es necesario utilizar soluciones de irrigación que puedan mejorar la desinfección del sistema de conductos radiculares.Objetivo: Realice una revisión integradora de la literatura para comparar las propiedades antimicrobianas de la clorhexidina con el hipoclorito de sodio.Metodología: La búsqueda bibliográfica se realizó desde septiembre de 2019 hasta agosto de 2021, en las siguientes bases de datos: PUBMED/MEDLINE, LILACS y SCIELO. Usando los descriptores: chlorhexidine, sodium hypochlorite, root canal irrigant and cleaning. Como criterio de búsqueda se utilizaron trabajos experimentales de laboratorio in vitro publicados entre 2017 y 2021.Resultados: Se encontraron un total de 165 artículos, de los cuales 15 fueron seleccionados al final del proceso. 8 estudios no encontraron diferencia estadísticamente significativa entre clorhexidinae hipoclorito, 5 artículos mostraron resultados superiores para NaOCl y en 2 la clorhexidina fue superior.Conclusiones: Después de analizar la literatura, observamos una similitud entre la acción antimicrobiana del hipoclorito de sodio y la clorhexidina,y podemos concluir que ambos tienen una buena acción antimicrobiana, lo que justifica su uso clínico (AU).

Post preparation: cleanness achieved by different irrigating protocols / Preparación para poste: limpieza lograda por diferentes protocolos de irrigación

ABSTRACT The aim of this study was to evaluate the ability of different irrigation protocols to remove from the walls of the root canal and entrance to dentinal tubules the smear layer produced during preparation for a fiber post. Fifty decoronated human lower premolars were treated endodontically and the apical third of the canal was filled with a single gutta-percha point using warm vertical compaction. Ten millimeters were left free for post preparation, which was done by sequential use of a Largo #1 drill and Exacto #3 pilot drill (Angelus, Brazil), with irrigation with distilled water upon each instrument change. Samples were distributed randomly into 5 groups (n=10). G 1: No irrigation after post preparation. G 2: Distilled water activated for 60 s + distilled water. G 3: 5.25% NaOCl 15 s+ distilled water. G 4: 17% EDTA (Farmadental, Argentina) 60 s + 5.25% NaOCl 15 s + distilled water. G 5: 10% polyacrylic acid (Densell) 15 s + distilled water. Irrigant was activated mechanically with a low-speed conical brush. Roots were then split longitudinally in vestibular-lingual direction. Each surface was observed under SEM in its different thirds. Microphotographs were taken at 150X and 600X magnification and results analyzed statistically using Kruskall Wallis and Friedman tests (p<0.05). The results [mean (standard deviation)] were: for 150X: G1 11.00 (1.33), G2 7.50 (3.13), G3 6.30 (2.58), G4 2.20 (2.80), G5 4.30 (1.50), and for 600X G1 11.40 (0.84), G2 10.00 (1.94), G3 7.70 (3.33), G4 5.80 (3.70), G5 7.20 (2.65). The statistical analysis showed significant differences between irrigants (p<0.05) but not between root thirds (p>>0.05). The EDTA+NaOCl combination and polyacrylic acid showed greater capacity to remove the smear layer created during post preparation.

RESUMEN El objetivo del presente trabajo fue evaluar la capacidad de diferentes protocolos de irrigación para remover el barro dentinario de las paredes del conducto y entrada de los túbulos dentinarios, producido durante la preparación para anclaje intarradicular. Se utilizaron 50 premolares inferiores humanos a los cuales se les cortó la corona clínica. Luego se les realizó tratamiento endodóntico y obturación del tercio apical con cono único de conicidad aumentada y compactación vertical caliente. Se dejaron libres 10 mm para la preparación para poste que se realizó utilizando secuencialmente fresa de Largo # 1y fresa piloto Exacto #1(Angelus, Brasil), irrigando a cada cambio de instrumento con agua destilada. Las muestras fueron distribuidas aleatoriamente en 5 grupos (n=10). G 1: No se efectuó ningún lavaje. Los irrigantes fueron activados con cepillo mecánico cónico a baja velocidad. G 2: Agua destilada activada 60 s+ agua destilada G 3: NaOCl al 5,25%, 15 s+ agua destilada. G 4: EDTA al 17% (Farmadental, Argentina) 60 s +NaOCl al 5,25% 15 s , + agua destilada. G 5 : Ácido poliacrílico al 10% ( Densell) 15 s + agua destilada. Posteriormente, las raíces fueron seccionadas longitudinalmente en sentido vestíbulo lingual. Cada superficie fue observada al MEB en sus diferentes tercios. Las micrografías se realizaron a 150 y 600 X y los resultados analizados estadísticamente mediante test de Kruskall Wallis y Friedman (p<0,05), Los resultados [Medias (desviaciones estándar)] fueron: para 150X G1 11,00(1,33), G2 7,50 (3,13), G3 6,30 (2,58), G4 2,20 (2,80), G5 4,30 (1,50) y para 600X G1 11,40 (0,84), G2 10,00 (1,94), G3 7,70 (3,33), G4 5,80 (3,70), G5 7,20 (2,65). El análisis estadístico mostró diferencia significativa entre irrigantes (p<0,05) pero no entre tercios radiculares (p>0,05). La combinación de EDTA+NaOCl y el ácido poliacrílico tendrían mayor capacidad de remoción del barro dentinario creado durante la preparación para poste.

Decontamination of Gutta-percha Cones employed in Endodontics / Descontaminação de cones de guta-percha empregados em endodontia

ABSTRACT The guttapercha cones used in endodontic treatment are produced in aseptic conditions and their composition includes zinc oxide, which is responsible for antibacterial activity. However, there is the possibility of microbial contamination by manipulation, aerosol or during storage. Although several chemical agents have been tested for their decontamination, there is no consensus on the best disinfection protocol to be used. The aim of this study was to evaluate the decontamination of guttapercha cones contaminated with the bacteria Enterococcus faecalis, by using chlorhexidine digluconate (CHX) and sodium hypochlorite (NaClO) at different concentrations for short exposure times. For this purpose, guttapercha cones (size 40) were selected at random from a sealed box and immersed for 1 min in a microbial suspension. Then they were immersed in specific Petri dishes for different groups containing: CHX 2%, NaClO 1% or NaClO 2.5% for 30 s or 1 min, and subsequently placed in tubes containing BHI broth. After incubating the tubes for 48 h, it was observed that 1% and 2.5% NaClO and 2% CHX were effective for decontaminating the cones at those exposure time intervals. Microbial growth was detected in one of the replicates of the group with CHX applied for 30 s. To prevent the possibility of failures at this stage, the exposure time of guttapercha cones to the decontaminating agent should not be reduced.

RESUMO Os cones de gutapercha utilizados no tratamento endodôntico são produzidos em condições assépticas e possuem óxido de zinco em sua composição, responsável pela atividade antibac te riana. No entanto, existe a possibilidade de contaminação microbiana por manipulação, aerossol ou seu armazenamento. Embora vários agentes químicos já tenham sido testados para sua descontaminação, não há consenso sobre o melhor proto colo de desinfecção a ser usado. Nosso objetivo foi avaliar a descontaminação de cones de gutapercha contaminados com a bactéria Enterococcus faecalis, utilizando digluconato de clorexidina (CHX) e hipoclorito de sódio (NaClO) em diferentes concentrações e tempos de exposição curtos. Para esse fim, 40 cones de gutapercha foram selecionados aleatoriamente, de uma caixa selada e imersos por 1 min em uma suspensão microbiana. Em seguida, foram imersos em placas de Petri específicas para diferentes grupos contendo: CHX 2%, NaClO 1% ou 2,5%, nos tempos de exposição de 30s e 1min e subseqüentemente imersos em tubos contendo caldo BHI. Após incubação dos tubos por 48 h, observouse que NaClO 1% e 2,5% e CHX 2% foram eficazes para a descontaminação dos cones nesses intervalos de tempo de exposição. Em uma das réplicas do grupo com CHX aplicado por 30s foi detectado crescimento microbiano. O tempo de exposição dos cones de gutapercha ao agente de desconta minação não deve ser reduzido para evitar a possibilidade de falhas nesse estágio.

Antimicrobial activity of intracanal dressings composed by natural products associated to chlorhexidine and its influence on dentinal colour change / Avaliação da ação antimicrobiana de medicações intracanais à base de produtos fitoterápicos associados à clorexidina e sua influência na alteração de cor da estrutura dentinária

Purpose: evaluate the antimicrobial activity of intracanal dressings and their influence on dentinal colour changes. Material and methods: eighty single-rooted human extracted teeth were decoronated and divided into eight groups (n=10) according to intracanal dressing protocols inserted into the root canals: G1­distilled water (DW); G2­2% chlorhexidine gel (CHX); G3­calcium hydroxide (Ca[OH]2)+DW; G4­grape seed extract (GSE)+DW; G5­ginger extract (GE)+DW; G6­Ca(OH)2+CHX; G7­GSE+CHX; and G8­GE+CHX. The antimicrobial activity was evaluated by colony-forming units (CFUs) counting and dentinal colour changes was evaluated by digital spectrophotometry. Data were statistically analysed by One-way ANOVA followed by Tukey´s post hoc test (antimicrobial evaluation) and non-parametric Wilcoxon followed by the Mann- Whitney-U test (colour change evaluation) (α=0.05). Results: the highest bacterial reduction was observed in groups 4, 6, 7 and 8, with no significant difference between them (p<0.05). Groups 4 and 7 showed the highest medians of dentinal colour change (p<0.05). Conclusion: the addition of CHX improved the antimicrobial activity of GE-based intracanal dressing, with no effect in GSE-based intracanal dressing; moreover, these protocols induced significant dentinal colour changes. (AU)

Objetivo: avaliar a atividade antimicrobiana de medicações intracanais e sua influência na alteração da cor dentinária. Materiais e métodos: oitenta dentes humanos extraídos unirradiculares foram seccionados e divididos em oito grupos (n = 10), de acordo com os protocolos de medicação intracanal inseridos nos canais radiculares: água destilada G1 (DW); G2-2% de gel de clorexidina (CHX); hidróxido de cálcio G3 ­ (Ca [OH] 2) + DW; extrato de semente de uva G4 (GSE) + DW; extrato de gengibre G5 (GE) + DW; G6- Ca (OH) 2 + CHX; G7 ­ GSE + CHX; e G8-GE + CHX. A atividade antimicrobiana foi avaliada por contagem de unidades formadoras de colônias (UFCs) e as alterações de cor dentinária foram avaliadas por espectrofotometria digital. Os dados foram analisados estatisticamente por ANOVA one-way, seguida pelo teste post hoc de Tukey (avaliação antimicrobiana) e Wilcoxon não paramétrico, seguido pelo teste de Mann- Whitney-U (avaliação da mudança de cor) (α = 0,05). Resultados: a maior redução bacteriana foi observada nos grupos 4, 6, 7 e 8, sem diferença significativa entre eles (p < 0,05). Os grupos 4 e 7 apresentaram as maiores medianas da alteração da cor dentinária (p < 0,05). Conclusão: a adição de CHX melhorou a atividade antimicrobiana da medicação intracanal baseado em GE, sem efeito na medicação intracanal baseado em GSE; além disso, esses protocolos induziram alterações significativas na cor dentinária.(AU)

Effect of a calcium hydroxide-based intracanal medicament containing N-2-methyl pyrrolidone as a vehicle against Enterococcus faecalis biofilm

Abstract This study investigated the effect of a calcium hydroxide (CH) paste (CleaniCal®) containing N-2-methyl pyrrolidone (NMP) as a vehicle on Enterococcus faecalis (E. faecalis) biofilms compared with other products containing saline (Calasept Plus™) or propylene glycol (PG) (Calcipex II®). Methodology Standardized bovine root canal specimens were used. The antibacterial effects were measured by colony-forming unit counting. The thickness of bacterial microcolonies and exopolysaccharides was assessed using confocal laser scanning microscopy. Morphological features of the biofilms were observed using field-emission scanning electron microscopy (FE-SEM). Bovine tooth blocks covered with nail polish were immersed into the vehicles and dispelling was observed. The data were analyzed using one-way analysis of variance and Tukey tests (p<0.05). Results CleaniCal® showed the highest antibacterial activity, followed by Calcipex II® (p<0.05). Moreover, NMP showed a higher antibacterial effect compared with PG (p<0.05). The thickness of bacteria and EPS in the CleaniCal® group was significantly lower than that of other materials tested (p<0.05). FE-SEM images showed the specimens treated with Calasept Plus™ were covered with biofilms, whereas the specimens treated with other medicaments were not. Notably, the specimen treated with CleaniCal® was cleaner than the one treated with Calcipex II®. Furthermore, the nail polish on the bovine tooth block immersed in NMP was completely dispelled. Conclusions CleaniCal® performed better than Calasept Plus™ and Calcipex II® in the removal efficacy of E. faecalis biofilms. The results suggest the effect might be due to the potent dissolving effect of NMP on organic substances.

Penetration degree of sealer in artificial lateral canal after passive ultrasonic irrigation with EDTA for different times / Grau de penetração do cimento obturador em canais laterais simulados após ativação ultrassonica passiva do EDTA em diferentes tempos

The aim of this study was to evaluate the degree of penetration of obturation cement in artificial lateral canals after Passive Ultrasonic Irrigation (PUI) with ethylenediaminetetraacetic acid (EDTA) for different times. Fifty upper molar palatine roots were used, in which two artificial lateral canals were made at distances of 7 and 3 millimeters from the root apex. After instrumentation and drying the canal, the final toilet stage was performed on five groups (n = 10), as follows: G1 - EDTA 17% + PUI for 10 seconds; G2 - EDTA 17% + PUI for 20 seconds; G3 - EDTA 17% + PUI for 30 seconds; G4 - EDTA 17% + PUI for 60 seconds; G5 - EDTA 17% + activation by instrument R50 for 5 minutes (Control). The canals were sealed by the single cone technique, and after 72 hours, sectioned in two planes transverse to the artificial canal, to see the degree of penetration of the sealing cement. In the radiographic analysis, there was no statistical difference (p> 0.05) between groups in the two artificial lateral canals. However, PUI of EDTA for 60 seconds produced a significant difference in the degree of penetration of the sealing cement (p <0.05) at 7 mm from the apex. Therefore, PUI with EDTA for 60 seconds promoted a higher degree of penetration of the obturator cement in the artificial lateral canal.

O objetivo deste trabalho foi avaliar o grau de penetração do cimento obturador em canais laterais artficiais, após Irrigação Ultrassonica Passiva (IUP) do ácido etilenodiaminotetracético (EDTA), em diferentes tempos. Foram utilizadas 50 raízes palatinas de molares superiores, e em seguida confeccionados dois canais laterais artificiais a 7 e 3 milímetros do ápice radicular. Após a instrumentação e secagem dos canais, foi iniciada a etapa de toillet final, de acordo com os seguintes grupos (n=10): G1- EDTA 17%+IUP durante 10 segundos; G2 - EDTA 17%+IUP durante 20 segundos; G3 - EDTA 17%+IUP durante 30 segundos; G4- EDTA 17%+IUP durante 60 segundos; G5- EDTA 17%+ativação pelo instrumento R50 durante 5 minutos (Controle). Os canais foram obturados pela técnica do cone único, e após 72 horas, seccionados em dois planos transversais dos canais artificiais, para se visualizar o grau de penetração do cimento obturador. Na análise radiográfica, não houve diferença estatística (p>0,05) entre os grupos, nos dois canais laterais artificiais. Entretanto, a IUP do EDTA por 60 segundos conseguiu um obter resultado significativo, sobre o grau de penetração do cimento obturador (p<0,05) a 7 milímetros do ápice. Portanto, a IUP do EDTA no tempo de 60 segundos promoveu maior grau de penetração do cimento obturador nos canais laterais artifciais.

Root canal dressings for revascularization influence in vitro mineralization of apical papilla cells

Abstract Endodontic revascularization is based on cell recruitment into the necrotic root canal of immature teeth after chemical disinfection. The clinical outcome depends on the ability of surviving cells from the apical tissue to differentiate and promote hard tissue deposition inside the dentinal walls. Objective To investigate the effect of calcium hydroxide (CH) and modified triple antibiotic paste (mTAP - ciprofloxacin, metronidazole and cefaclor) on the viability and mineralization potential of apical papilla cells (APC) in vitro . Material and Methods APC cultures were kept in contact with CH or mTAP (250-1000 µg/mL) for 5 days, after which cell viability was assessed using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Next, APCs were subjected to CH or mTAP at 250 µg/mL for 5 days before inducing the differentiation assay. After 14 and 21 days, calcium deposition was assessed by the Alizarin Red S staining method, followed by elution and quantification using spectrophotometry. Data were analyzed using ANOVA followed by Tukey post hoc test. Results CH induced cell proliferation, whereas mTAP showed significant cytotoxicity at all concentrations tested. APC treated with CH demonstrated improved mineralization capacity at 14 days, while, for mTAP, significant reduction on the mineralization rate was observed for both experimental periods (14 and 21 days). Conclusion Our findings showed that CH induces cell proliferation and improves early mineralization, whereas mTAP was found cytotoxic and reduced the mineralization potential in vitro of APCs.

Antibacterial efficacy of synthetic and natural-derived novel endodontic irrigant solutions

Abstract The aim of this study was to compare the efficacy of grape seed extract (GSE), calcium hypochlorite [Ca(ClO)2], and sodium hypochlorite (NaOCl) irrigant solutions with rotary or reciprocating instrumentation for disinfection of root canals inoculated with Enterococcus faecalis. The mesiobuccal root canals of mandibular molars were prepared and inoculated with Enterococcus faecalis for 21 days. The roots were then randomly divided into the following eight experimental groups (n=11) according to the instrumentation technique and disinfection protocol: ProTaper Next or Reciproc R25 with sodium chloride (control group), 6% NaOCl, 6% Ca(ClO)2, or 50% GSE used for irrigation during instrumentation. The antimicrobial activity was determined on the basis of a reduction in colony-forming units (CFUs) counted on bacterial samples collected before and after root canal instrumentation and expressed as a percentage of reduction. Data were evaluated by two-way ANOVA followed by Tukey HSD post-hoc tests (p<0.05). No significant differences were observed in bacterial reduction between the ProTaper Next and Reciproc R25 systems (p>0.05), regardless of the irrigant solution used. Furthermore, all active solutions (6% NaOCl, 50% GSE, and 6% Ca(ClO)2) showed similar potential to reduce bacterial counts (p>0.05) and were significantly more effective than sodium chloride (control) (p<0.05). The results suggest that the GSE and Ca(ClO)2 have potential clinical application as irrigant solutions in endodontic therapy since they present bactericidal efficacy against Enterococcus faecalis.

Resumo O objetivo deste estudo foi comparar a eficácia do extrato de semente de uva (ESU), hipoclorito de cálcio [Ca(ClO)2] e hipoclorito de sódio (NaOCl) como soluções irrigadores quando utilizadas com instrumentos reciprocantes e rotatórios para desinfecção de canais radiculares infectados com Enterococcus faecalis. Raízes mesio-vestibulares de molares inferiores foram preparados e inoculados com E. faecalis por 21 dias. As raízes foram aleatoriamente divididas em 8 grupos (n=11) de acordo com a técnica de instrumentação e protocolo de irrigação: ProTaper Next ou Reciproc R25 associados com soro fisiológico (grupo controle), Ca(ClO)2 6%, NaOCl 6% ou ESU 50%. A atividade antimicrobiana foi determinada pela redução do número de Unidades Formadoras de Colonias (UFCs) coletadas antes e após a instrumentação e expressas em porcentagens de redução. Os dados foram analisados estatisticamente pelos testes ANOVA seguido pelo teste complementar de Tukey HSD (p<0,05). Não foi encontrado diferença estatisticamente significante na redução bacteriana entre os sistemas ProTaper Next e Reciproc R25 (p>0.05), independente da solução irrigadora usada. Além disso, todas as soluções ativas (NaOCl, ESU e Ca(ClO)2) mostraram similar potencial em reduzir a quantidade de bactérias (p>0.05) e foram significativamente mais efetivas que o soro fisiológico (p<0.05). Pode-se concluir que o ESU e o Ca(ClO)2 apresentam potencial para aplicação clínica como irrigantes endodônticos uma vez que apresentaram efetividade antimicrobiana contra o E. faecalis.

Avaliação da expressão de citocinas pró e anti-inflamatórias após estímulo celular com diferentes agentes irrigantes / Evaluation of the expression of pro and anti-inflammatory cytokines after cellular stimulation with different irrigating agents

O objetivo do trabalho foi investigar o efeito de diferentes soluções químicas auxiliares, em concentrações subcitotóxicas, na expressão de citocinas pró e anti-inflamatórias, quando em contato com células da linhagem de linfoma humano, diferenciadas em macrófagos, human macrophage-like (U937), através da adição de 125ng/mL de PMA. As concentrações, citotóxica e subcitotóxica, de cada solução, foram determinadas de acordo com padrão ISO, utilizando-se fibroblastos de camundongos (L929). As células L929 foram cultivadas em meio MEM completo e mantidas em placas de 96 poços. As células foram então colocadas em contato com as diluições das soluções químicas auxiliares (a partir das concentrações recomendadas para uso, NaOCL 5,25%; CHX 2%; Quitosana 0,2%; HEBP 18%; GSE de Vitis vinífera, 6,5%), por 24 horas a 37ºC em estufa de CO2. Em seguida, o meio de cultivo contendo 1mg/mL de MTT foi adicionado aos poços em triplicata, para avaliação da viabilidade celular e determinação das concentrações citotóxicas (30% de morte das células L929) e subcitotóxicas (15% de morte) por regressão linear por meio do programa GraphPad Prism versão 6.0. As concentrações subcitotóxicas foram colocadas em contato com as células human macrophage-like (U937) por 60 min. Os sobrenadantes da pré-incubação (controles sem substâncias químicas auxiliares), e incubação com as soluções químicas auxiliares em meio DMEM completo foram congelados em freezer ultrafrio (­ 80ºC) e avaliados para as concentrações de dezessete citocinas através do kit Bio-Plex Pro Human Th17 Cytokine Panel® pelo método Luminex®. Foi possível obter valores de expressão de 7 citocinas pró-inflamatórias: IL-1ß, TNFα, IL-6, IL-17A, IL-17F, IL-22 e IL-23. As substâncias com maior atividade citotóxica (padrão ISO para morte celular) foram em ordem de grandeza, a quitosana, a CHX, o NaOCl, o HEBP e o GSE de Vitis vinifera. As concentrações subcitotóxicas das soluções químicas auxiliares, CHX e NaOCl, induziram ao aumento da maioria das citocinas pró-inflamatórias, exceto para a IL-6. Quanto a IL-1ß, a quitosana e a CHX, estas foram as que mais induziram a expressão pelas células U937 (P<0,05),enquanto o NaOCl induziu maior expressão de TNFα (P>0,05). Dentre as citocinas relacionadas ao fenótipo Th17, a CHX foi a que mais induziu a expressão de IL-17A (P<0,05), IL-17F e IL-23, seguido do NaOCl(P<0,05). O NaOCl induziu maior expressão de IL22 seguido da CHX (p<0,05). Quanto a IL-6, todas as substâncias modularam a sua expressão pelas células U937(p>0.05). O HEBP atuou como um excelente modulador da expressão de citocinas pró-inflamatórias, induzindo a redução da expressão de todas as citocinas testadas (P<0,05). O GSE de Vitis vinifera, apesar de ser o menos citotóxico não apresentou diferença estatística para o controle em nenhuma das situações avaliadas, exceto para a redução da IL-6 (assim como o HEBP e a quitosana) (p<0,05). A quitosana comportou-se de forma similar ao GSE de Vitis vinífera (p>0,05) exceto para a IL-1ß. Conforme observado, as substâncias químicas auxiliares são capazes de induzir a expressão de citocinas pró-inflamatórias diversas. O HEBP foi o agente químico que melhor modulou a expressão de citocinas pró-inflamatórias.

The aim of this investigation was to analyze the effect of different chemical endodontic solutions, in subcytotoxic concentrations, on the expression of pro and anti-inflammatory cytokines when in contact with human lymphoma lineage differentiated into human macrophages, human macrophage-like (U937), by the addition of 125ng/ml PMA. The cytotoxic and subcytotoxic concentrations of each solution were determined according to ISO standard using mouse fibroblasts (L929). L929 cells were cultured in complete MEM medium and maintained in 96-well plates. Dilutions of the auxiliary chemical solutions, from the recommended concentrations for use: NaOCL 5,25%; CHX 2%; Chitosan 0.2%; Etidronic acid 18% and Grape seed extract, Vitis vinifera, 6.5%, were carried out in complete MEM medium, and placed in contact with the cells for 24 hours at 37ºC in CO2 cell incubator, in triplicate. Thereafter, the culture medium containing 1 mg/mL MTT was added to the wells. Cytotoxic concentrations (30% death of L929 cells) and subcytotoxic (15% death) were determined by linear regression using GraphPad Prism version 6.0. Subcytotoxic concentrations were obtained in complete DMEM medium and maintained in contact for 60 min. with U937 cells. Controls without chemical substances were also performed. Supernatants from the pre-incubation, incubation with the chemical solutions and complete DMEM were removed, frozen in ultra-cold freezer (-80ºC) and evaluated for the concentrations of 17 cytokines through the Bio-Plex Pro Human Th17 Cytokine Panel® kit by Luminex®. The substances with higher cytotoxic activity were in order of magnitude: chitosan, CHX, NaOCl, etidronic acid and Vitis vinifera extract. The values of 7 pro-inflammatory cytokines: IL-1ß, TNFα, IL-6, IL-17A, IL-17F, IL-22 and IL-23 were obtained. Subcytotoxic concentrations of the irrigant agents (chlorhexidine and NaOCl) generally induced the increase of most pro-inflammatory cytokines, except for IL-6. Chitosan and CHX were the agents that most induced the expression of IL-1ß by U937 cells, whereas NaOCl induced higher TNFα expression (P> 0,05). Among the cytokines related to the Th17 phenotype, CHX was the agent that induced the highest expression of IL-17A (P <0,05), IL-17F and IL-23, followed by NaOCl (P<0,05). NaOCl induced the highest expression of IL22 followed by CHX. All of the substances modulated the expression of IL-6 by U937 cells (P >0,05). HEBP was an excellent modulator of the expression of pro-inflammatory cytokines, inducing the reduction of the expression of all cytokines tested. Vitis vinifera extract, despite being the least cytotoxic agent, did not present statistical differences compared to the control for any of the evaluated cytokines, except for the reduction of IL-6 (as well as etidronic acid and chitosan) (P<0,05). Chitosan, except for IL-1 ß, was very similar to the Vitis vinifera (P>0,05) extract. As observed, the auxiliary substances (especially the irrigants) are capable of inducing the expression of various pro-inflammatory cytokines. Etidronic acid was the chemical that modulated the expression of pro-inflammatory cytokines.

Irrigation endodontique à l'hypochlorite de sodium : Types de solutions et modalités de préparations dans 102 cabinets dentaires de la ville d'Abidjan (Côte d'Ivoire)

Introduction : L'hypochlorite de sodium (NaOCl) représente à ce jour le "gold standard" pour l'irrigation endodontique. L'objectif de cette étude était d'identifier les solutions de NaOCl utilisées comme irrigant canalaire et les modes de préparations dans les cabinets dentaires. Matériels et méthodes : Une étude prospective a été réalisée dans 102 cabinets. Par la suite, les teneurs en chlore actif (c.a.) des solutions de NaOCl ont été évaluées par titrage iodométrique. Les analyses statistiques ont été effectuées par le logiciel SPSS version 22 sous Windows (SPSS Inc., Chicago, IL, USA). Résultats : Les sources commerciales de solution de NaOCl étaient utilisées à 72,5% pour l'irrigation endodontique. Ces solutions étaient concentrées à 2,53% c.a. ou 3,80% c.a. selon l'emballage. Dans l'ensemble, les titrages iodométriques ont révélé des rétrogradations en c.a. des solutions mères. Les dilutions au 1/5ième étaient généralement réalisées, ce qui signifiait une concentration à 0,5% c.a. pour l'irrigation endodontique. Une solution de NaOCl stabilisée à 0,5% c.a. et disponible en officine était principalement utilisée dans les cabinets privés. Discussion : Les concentrations des solutions de NaOCl pour l'irrigation en endodontie se situe entre 0,5% et 6% d'après la littérature. Cependant un compromis semble être trouvé autour des valeurs situées entre 2,5% c.a. et 3% c.a. Dans ce cadre, les solutions de NaOCl issues du commerce dans cette étude peuvent être considérées comme prêtes à l'emploi. Il faut cependant tenir compte de leur instabilité qui suggère des préparations finales à teneur en c.a. inférieure à la minimale préconisée en endodontie (0,5% c.a.). Conclusion : cette étude montre l'intérêt de l'usage de solutions de NaOCl stabilisées et conçues spécifiquement pour l'irrigation endodontique. Ce sont elles qui sont considérées comme dispositif médical par les autorités sanitaires

Demetallization of Enterococcus faecalis biofilm: a preliminary study

Abstract Objectives To determine the concentration of calcium, iron, manganese and zinc ions after the application of chelator to Enterococcus faecalis biofilms. Material and Methods Fifty bovine maxillary central incisors were prepared and inoculated with E. faecalis for 60 days. The following were used as irrigation solutions: 17% EDTA (pH 3, 7 and 10), 2.5% sodium hypochlorite (NaOCl) combined with 17% EDTA (pH 3, 7 and 10), distilled water (pH 3, 7 and 10), and 2.5% NaOCl. Each solution was kept in the root canal for five minutes. Fifteen uncontaminated root canals were irrigated with 17% EDTA (pH 3, 7 and 10). Six teeth were used as bacterial control. The number of calcium, iron, manganese and zinc ions was determined using flame atomic absorption spectrometry. Mean ± standard deviation (SD) values were used for descriptive statistics. Results Calcium chelation using 17% EDTA at pH 7 was higher than at pH 3 and 10, regardless of whether bacterial biofilm was present. The highest concentration of iron occurred at pH 3 in the presence of bacterial biofilm. The highest concentration of manganese found was 2.5% NaOCl and 17% EDTA at pH 7 in the presence of bacterial biofilm. Zinc levels were not detectable. Conclusions The pH of chelating agents affected the removal of calcium, iron, and manganese ions. The concentration of iron ions in root canals with bacterial biofilm was higher after the use of 17% EDTA at pH 3 than after the use of the other solutions at all pH levels.

Apical Negative Pressure irrigation presents tissue compatibility in immature teeth

Abstract Aim: To compare the apical negative pressure irrigation (ANP) with conventional irrigation in the teeth of immature dogs with apical periodontitis. Methods: Fifty-two immature pre-molar root canals were randomly assigned into 4 groups: ANP (n=15); conventional irrigation (n=17); healthy teeth (control) (n = 10); and teeth with untreated apical periodontitis (control) (n=10). After induction of apical periodontitis, teeth were instrumented using EndoVac® (apical negative pressure irrigation) or conventional irrigation. The animals were euthanized after 90 days. The sections were stained by HE and analyzed under conventional and fluorescence microscopy. TRAP histoenzymology was also performed. Statistical analyses were performed with the significance level set at 5%. Results: There was difference in the histopathological parameters between ANP and conventional groups (p<0.05). The ANP group showed a predominance of low magnitude inflammatory infiltrate, a smaller periodontal ligament, and lower mineralized tissue resorption. There were no differences in the periapical lesion extensions between the ANP and conventional groups (p>0.05). However, a lower number of osteoclasts was observed in the ANP group (p<0.05). Conclusion: The EndoVac® irrigation system presented better biological results and more advanced repair process in immature teeth with apical periodontitis than the conventional irrigation system, confirming the hypothesis.

Efetividade de soluções endodônticas na redução da viabilidade de Enterococcus faecalis cultivados em forma séssil e planctônica / Effectiveness of endodontic solutions to reduce the viability of sessile and planktonic Enterococcus faecalis

Este estudo teve como objetivo avaliar a sensibilidade dos biofilmes de E. faecalis a NaOCl, CHX e PAA após uma exposição de ácido cítrico 2,5%. O biofilme de E. faecalis foi formado em lamínulas de vidro circular de 13 mm Ø em placas de cultura de 24 poços. Os biofilmes foram tratados ou não durante 5 minutos com ácido cítrico, posteriormente, exposto a diferentes concentrações de NaOCl, CHX e PAA. A atividade antimicrobiana foi avaliada por meio da contagem de unidade formadora de colônias (UFCs). Ao analisar o percentual de redução bacteriana na forma planctônica em função do tempo e da solução/concentração, o teste de Anova demonstrou não haver diferença estatística entre CHX e NaOCl (p>0,05). Contudo a CHX, quando utilizada sozinha, apresentou menor efetividade que NaOCl (p<0,05) sobre o biofilme. O pré-tratamento do biofilme com ácido cítrico tornou as bactérias na forma séssil mais sensível ao NaOCl e ao PAA. Baseado nos resultados obtidos foi possível concluir que o tratamento com ácido cítrico aumentou a sensibilidade do biofilme de E. faecalis a irrigante utilizado em procedimento endodôntico e orienta o início do tratamento com irrigação prévia com o ácido cítrico.

The present study aimed to evaluate the sensitivity of E. faecalis biofilms to NaOCl, CHX and PAA after exposure to 2.5% citric acid. The E. faecalis biofilm was formed in 13-mm diameter circular glass microslides in 24-well culture plates. The biofilms were treated or not for 5 minutes with citric acid, and subsequently exposed to different concentrations of NaOCl, CHX and PAA. The antimicrobial activity was assessed by colony-forming unit (CFU) count. When analyzing the bacterial reduction percentage in planktonic form according to the time and solution/concentration, the Anova test demonstrated no statistical difference between CHX and NaOCl (p>0.05). However, CHX, when utilized alone, presented less effectiveness than NaOCl (p<0.05) on the biofilm. The citric acid pretreatment of the biofilm made the bacteria in the sessile form more sensitive to NaOCl and PAA. Based on the obtained results, it was possible to conclude that citric acid treatment increased the sensitivity of the E. faecalis biofilm to irrigants employed in endodontic procedures, guiding the beginning of treatment with prior citric acid irrigation.

Evaluation of methods for removal of filling material in endodontic retreatment and assessment of antimicrobial action of solutions used in final irrigation / Avaliação de métodos de desobturação do canal radicular no retratamento endodôntico e avaliação da ação antimicrobiana de soluções utilizadas na irrigação final

When endodontic treatment fails, an alternative could be root canal retreatment. During this procedure, all filling material should be removed to allow a new root canal preparation and new obturation of the root canal system. Bacteria are the main cause of endodontic treatment failure, and persistent infection may be related to microorganism ability to penetrate into dentinal tubules. Therefore, this requires the use of irrigating solutions with antimicrobial action and low toxicity. The aim of this study was to evaluate the performance of instruments made of different alloys in root canal re-instrumentation during endodontic retreatment of lateral incisors with apical curvature, using computed microtomography and assessment of these samples by scanning electron microscopy after irrigant activation. Furthermore, the antimicrobial action of a root canal irrigant containing silver nanoparticles, 2% chlorhexidine and 2.5% sodium hypochlorite was evaluated against Enterococcus faecalis biofilm and dentin infected with this microorganism. Thirty extracted maxillary lateral incisors with apical curvature were selected. The teeth were instrumented, filled and divided into three different groups according to the protocol for removal of filling material: Group 1: re-instrumentation with Reciproc R25 instruments, Mtwo 40 and ProDesign Logic 50.01; Group 2: re-instrumentation with ProDesign R, ProDesign Logic 40 and ProDesign Logic 50.01; Group 3: re-instrumentation with Gates-Glidden drills and manual instruments K-file and Hedstroem files. For filling material removal analysis, the samples were scanned in a microtomograph device SkyScan 1174, for comparison of images taken before and after removing the root canal filling, and in each sample the volume was calculated at four levels (apical 1, apical 2, middle and cervical). These samples were split and analyzed by scanning electron microscopy to visualize filling material residues before and after irrigant activation with an ultrasonic device and with the EasyClean system used in continuous rotary motion. Statistical analyses were performed using Kruskal-Wallis, Friedman, Wilcoxon and Dunn tests. Furthermore, the minimum inhibitory concentration of an irrigating solution containing silver nanoparticles was determined against strains of Enterococcus faecalis, by using the microdilution method. Additionally, the antimicrobial activity of silver nanoparticle solution, 2% chlorhexidine and 2.5% sodium hypochlorite was tested against Enterococcus faecalis biofilm in vitro. For biofilm formation, bovine dentin blocks were placed in 24-well culture plates and Enterococcus faecalis biofilm was developed for 21 days. The dentin blocks were divided into 9 experimental groups of 5 blocks each, according to the irrigating solution used and the time in contact with the irrigant (5, 15 and 30 minutes). The samples were stained with Live/Dead reagent for analysis by confocal laser scanning microscopy (CLSM). Finally, the antimicrobial action of these solutions was tested after dentinal tubules were contaminated with Enterococcus faecalis. Dentin tubes were made from bovine incisors, taken to a centrifuge and infected with Enterococcus faecalis. The dentin tubes were treated with silver nanoparticle solution, 2% chlorhexidine and 2.5% sodium hypochlorite, and analyzed by CLSM to assess the antimicrobial activity of these solutions against bacteria in the dentinal tubules. The results showed that residues of filling material were found after root canal reinstrumentation in all groups. No significant difference was observed in removal of filling material between the reciprocating instruments Reciproc and ProDesign R and between rotary instruments Mtwo 40 and ProDesign Logic 40. The ProDesign Logic 50/.01 instrument significantly improved the removal of filling material compared with the use of Reciproc and ProDesign R instruments. The apical levels presented greater amount of remnant filling material compared with middle and cervical levels. After canal reinstrumentation, the passive ultrasonic irrigation and irrigant agitation with EasyClean significantly improved the removal of residual filling material in all root canal thirds. There was no significant difference between the performance of ultrasonics and EasyClean regarding the removal of residual filling material, as well no significant difference was observed in the removal of these residues when comparing apical, middle and cervical thirds. Regarding the action of the irrigating solutions against Enterococcus faecalis, the minimum inhibitory concentration of silver nanoparticle solution capable of eliminating this microorganism in broth and agar plates was 94 ppm. After the irrigation of Enterococcus faecalis biofilm, the silver nanoparticle solution was significantly less effective in killing bacteria compared with chlorhexidine when used for time of contact of 5 minutes. The sodium hypochlorite solution presented antimicrobial activity significantly higher compared with the silver nanoparticle solution and chlorhexidine. This solution also presented higher ability to dissolve biofilm in all times tested, whereas the silver nanoparticle solution presented higher ability to dissolve biofilm compared with chlorhexidine in times of 5 and 15 minutes. In infected intratubular dentin with Enterococcus faecalis, the sodium hypochlorite solution presented significant higher effectiveness than the silver nanoparticle solutions and chlorhexidine, especially in middle third and deep areas of the root canal. When comparing the antimicrobial activity of these solutions in biofilm and infected intratubular dentin, it was shown that when the silver nanoparticle solution was used for shorter periods of time, it was more effective in intratubular dentin compared with biofilm. On the other hand, with longer time of 30 minutes, the number of viable bacteria was higher in intratubular dentin than in biofilm, which was also observed when using the sodium hypochlorite solution in this time of action. It was concluded that to increase the rate of success in endodontic retreatment, the combination of the use of reciprocating and rotary instruments in the removal of filling material, the agitation of irrigants and the use of antimicrobial agents could be used in an attempt to eliminate bacteria that resisted to endodontic treatment. (AU)

Em casos de insucesso do tratamento endodôntico, uma alternativa seria o retratamento do canal radicular. Durante este procedimento deve haver remoção de todo o material obturador para que seja realizada novo preparo biomecânico e nova obturação do sistema de canais radiculares. Bactérias são o principal fator etiológico em casos de fracasso da terapia endodôntica, e esta infecção persistente pode estar relacionada à capacidade dos microganismos em penetrar nos túbulos dentinários. Por este motivo é necessário o uso de soluções na irrigação do canal radicular com ação antimicrobiana e com boa tolerância tecidual. O objetivo deste trabalho foi avaliar o desempenho dos instrumentos de diferentes ligas metálicas na desobturação do canal radicular durante o retratamento endodôntico de incisivos laterais superiores com curvatura apical, por meio da microtomografia computadorizada e análise destas amostras no microscópio eletrônico de varredura após a ativação de irrigantes. Posteriormente foi avaliada a capacidade antimicrobiana de um irrigante do canal radicular contendo nanopartículas de prata, clorexidina a 2% e hipoclorito de sódio a 2,5% frente ao biofilme de Enterococcus faecalis e à dentina contaminada com este mesmo microrganismo. Foram selecionados trinta incisivos laterais superiores humanos extraídos que apresentavam curvatura apical. Os dentes foram instrumentados, obturados e divididos em três diferentes grupos de acordo com o protocolo de remoção do material obturador do canal radicular: no Grupo 1: a desobturação foi realizada com os instrumentos Reciproc R25, Mtwo 40 e ProDesign Logic 50.01; no Grupo 2: foram utilizados os instrumentos ProDesign R, ProDesign Logic 40 e ProDesign Logic 50.01; e no Grupo 3: a desobturação foi realizada com brocas de Gates-Glidden e instrumentos manuais tipo K e Hedstroem. Para a análise da remoção do material obturador, as amostras foram escaneadas em micrótomogafo SkyScan 1174 para que fossem comparadas as imagens antes e após a desobturação do canal radicular, e em cada amostra este volume foi calculado nos quatro níveis (apical 1, 2, médio e cervical). Estas amostras posteriormente foram clivadas e analisadas no microscópio eletrônico de varredura, para a visualização de resíduos de material obturador antes e após a ativação de irrigantes com o ultrassom e com o sistema EasyClean utilizado em rotação contínua. Para a análise estatística dos resultados foram utilizados os testes de Kruskal-Wallis, Friedman, Wilcoxon e Dunn. Posteriormente, foi determinada a concentração inibitória mínima de uma solução irrigadora do canal contendo nanopartículas de prata frente a cepas de Enterococcus faecalis, através do método de diluição em caldo. Em seguida, foi testada a atividade antimicrobiana das soluções de nanopartículas de prata, da clorexidina a 2% e do hipoclorito de sódio a 2,5% sobre o biofilme de Enterococcus faecalis in vitro. Para isso, foram utilizados blocos de dentina bovina colocados em placas de 24 poços e biofilme de Enterococcus faecalis foi formado durante 21 dias. Os blocos de dentina foram divididos em 9 grupos experimentais com 5 blocos cada um, em função dos irrigantes avaliados e do tempo de exposição à solução irrigadora (5, 10 e 15 minutos). As amostras foram coradas com corante Live/Dead para posterior análise no microscópio confocal de varredura a laser (MCVL). Por fim, foi testada a atividade antimicrobiana destas soluções irrigadoras após a contaminação de túbulos dentinários com Enterococcus faecalis. Foram confeccionados tubos de dentina a partir de incisivos bovinos que foram levados à centrífuga e contaminados com Enterococcus faecalis. Os tubos de dentina receberam tratamento com a solução de nanopartículas de prata, com clorexidina a 2% e com hipoclorito de sódio a 2,5%, e foram analisados no MCVL para avaliar a atividade antimicrobiana das soluções sobre bactérias presentes nos túbulos dentinários. Os resultados demonstraram que resíduos de material obturador foram encontrados após a desobturação do canal em todos os grupos. Não houve diferença significante na remoção de material obturador entre os instrumentos reciprocantes Reciproc e ProDesign R e entre os instrumentos rotatórios Mtwo 40 e ProDesign Logic 40. O instrumento ProDesign Logic 50/.01 melhorou significantemente a remoção de material obturador comparado com o uso dos instrumentos Reciproc e ProDesign R. Os níveis apicais apresentaram uma maior quantidade de material obturador remanescente comparados com os níveis médio e cervical. Após a desobturação do canal radicular, a irrigação ultrassônica passiva e agitação dos irrigantes com o EasyClean melhoraram significantemente a remoção de resíduos de material obturador em todos os terços do canal radicular. Não houve diferença significante no desempenho do ultrassom e do EasyClean em relação à remoção de resíduos de material obturador, assim como não foi observada diferença significante na remoção destes resíduos quando comparados terços apical, médio e cervical. No que diz respeito à ação das soluções irrigadoras sobre o Enterococcus faecalis, a concentração inibitória mínima da solução de nanopartículas de prata capaz de eliminar este microrganimo em meio de cultura e ágar foi de 94 ppm. Após a irrigação no biofilme de Enterococcus faecalis, a solução de nanopartículas de prata foi significantemente menos efetiva em matar bactérias comparada com a clorexidina quando utilizadas pelo tempo de 5 minutos. A solução de hipoclorito de sódio apresentou atividade antimicrobiana significantemente maior comparada com as soluções de nanopartículas de prata e clorexidina. Essa solução ainda apresentou maior capacidade de dissolução do biofilme em todos os tempos testados, enquanto que a solução de nanopartículas de prata apresentou maior capacidade de dissolver o biofilme comparada à clorexidina nos tempos de 5 e 15 minutos. Na dentina intratubular infectada com Enterococcus faecalis, a solução de hipoclorito de sódio apresentou efetividade significantemente maior que as soluções de nanopartículas de prata e clorexidina, principalmente no terço médio e na região profunda do canal radicular. Quando comparada a atividade antimicrobiana destas soluções no biofilme e na dentina intratubular infectada, verificou-se que quando a solução de nanopartículas de prata foi utilizada por um tempo mais curto, foi mais efetiva na dentina intratubular comparada com o biofilme. De maneira contrária, com tempo maior de 30 minutos, o número e bactérias viáveis foi maior na dentina intratubular do que no biofilme, o que ocorreu também com a solução de hipoclorito de sódio neste tempo de ação. Conclui-se que para se aumentar a chance de sucesso em tratamentos retratamentos endodônticos, a combinação do uso de instrumentos reciprocantes e rotatórios na desobturação do canal, agitação de irrigantes e uso de agentes com capacidade antimicrobiana podem ser utilizados na tentativa de se eliminar bactérias resistente ao tratamento endodôntico. (AU)

Effect of ultrasonic streaming on intra-dentinal disinfection and penetration of calcium hydroxide paste in endodontic treatment

ABSTRACT Objective The antimicrobial effect of ultrasonic agitation of calcium hydroxide (CH) pastes in infected bovine dentin and their penetrability were evaluated using confocal laser scanning microscopy (CLSM) and microbiological culture. Material and Methods Fifty-two bovine teeth were infected with Enterococcus faecalis using a new contamination protocol; then they received CH paste and were divided into groups with or without ultrasound. Ultrasonic agitation was conducted for 1 min with a plain point insert. After 15 d, the CLSM analyzed the viable and dead bacteria with Live and Dead assay. The dentinal wall debris was collected by burs, and the colony forming units (CFU/mL) were counted. The penetrability of the paste inside dentinal tubules was tested using the B-rodamine dye. Results The calcium hydroxide paste showed better results with the use of ultrasonic agitation (p<0.05). Conclusion The ultrasonic agitation of CH paste increased its antimicrobial action and was responsible for intradentinal penetration with the fulfilment of the tubules.

Exopolysaccharide dispelled by calcium hydroxide with volatile vehicles related to bactericidal effect for root canal medication

ABSTRACT Objective: Enterococcus faecalis is the dominant microbial species responsible for persistent apical periodontitis with ability to deeply penetrate into the dentin. Exopolysaccharides (EPS) contribute to the pathogenicity and antibiotic resistance of E. faecalis. Our aim was to investigate the antimicrobial activity of calcium hydroxide (CH), camphorated parachlorophenol (CMCP), and chlorhexidine (CHX) against E. faecalis in dentinal tubules. Material and Methods: Decoronated single-canal human teeth and semicylindrical dentin blocks were incubated with E. faecalis for 3 weeks. Samples were randomly assigned to six medication groups for 1 week (n=10 per group): CH + 40% glycerin-water solution (1:1, wt/vol); CMCP; 2% CHX; CH + CMCP (1:1, wt/vol); CH + CMCP (2:3, wt/vol); and saline. Bacterial samples were collected and assayed for colony-forming units. After dentin blocks were split longitudinally, confocal laser scanning microscopy was used to assess the proportion of viable bacteria and EPS production in dentin. Results: CMCP exhibited the best antimicrobial activity, while CH was the least sensitive against E. faecalis (p<0.05). CHX showed similar antimicrobial properties to CH + CMCP (1:1, wt/vol) (p>0.05). CH combined with CMCP inhibited EPS synthesis by E. faecalis, which sensitized biofilms to antibacterial substances. Moreover, increasing concentrations of CMCP decreased EPS matrix formation, which effectively sensitized biofilms to disinfection agents. Conclusion: The EPS matrix dispelled by CH paste with CMCP may be related to its bactericidal effect; the visualization and analysis of EPS formation and microbial colonization in dentin may be a useful approach to verify medicaments for antimicrobial therapy.

Antimicrobial activity and substantivity of Uncaria tomentosa in infected root canal dentin

Abstract The aim of this study was to analyze the antimicrobial activity and substantivity of Uncaria tomentosa Willd DC (cat's claw, CC) in root dentin contaminated with Enterococcus faecalis. Forty-eight human premolars were contaminated with E. faecalis (ATCC 29212) and randomly divided into four groups according to the irrigant used during chemomechanical preparation (CMP): CC group: 2% CC gel; CHX group: 2% chlorhexidine digluconate gel (CHX); NaOCl group: 5.25% sodium hypochlorite (NaOCl); and SS group: sterile saline (SS). Microbiological samples were collected before (S1) and after (S2) CMP and after 7 days (S3). Colony-forming units (CFU/mL) at the different sampling times and comparisons among the groups were statistically analyzed by Wilcoxon and Kruskal-Wallis tests (p < 0.05). Significant bacterial reduction was achieved in all groups after CMP (p < 0.05). Results show no significant difference between S3 and S2 (p > 0.05) in the CC and CHX groups. Bacterial load was higher in S3 than in S2 samples (p < 0.05) in the NaOCl and SS groups. Our results suggest antibacterial effect of 2% CC gel against E. faecalis in infected dentin, in addition to antibacterial substantivity of 2% CC and 2% CHX up to 7 days.

Tissue characterization following revascularization of immature dog teeth using different disinfection pastes

Abstract Revascularization of immature teeth with necrotic pulps traditionally involves the use of triple antibiotic paste, which may sometimes lead to undesirable complications. The objective of this study was to assess tissue repair in immature dog teeth with apical periodontitis subjected to revascularization, comparing two different pastes used for root canal disinfection. Apical periodontitis was induced in 30 dog premolars. Teeth were randomly divided into three experimental groups: root canals filled with triple antibiotic paste (n = 10); root canals filled with 1% propolis paste (n = 10); and no medication (n = 10). An additional group (n = 10, no intervention) was used as control. After 7 months, the jaws were histologically evaluated for the following variables: newly formed mineralized tissue (present/absent); vital tissue in the canal space (absent/periodontal ligament-like/pulp-like); apical extension of root (present/absent); and severity of inflammatory process (absent/mild/moderate/severe). There were no statistically significant differences among the experimental groups in new mineralized tissue formation and apical root development. The formation of vital tissue in the canal space, in turn, was statistically different between the triple paste and propolis groups: vital tissues were present in all revascularized teeth disinfected with propolis paste (100%), compared to 71% of those disinfected with the triple paste. Severity of inflammatory process was different between the triple paste and no medication groups. The new tissues formed onto canal walls and in the root canal space showed characteristics of cementum and periodontal ligament, respectively. Propolis may have some advantages over the triple paste for the revascularization of immature teeth.

Effect of EDTA on TGF-ß1 released from the dentin matrix and its influence on dental pulp stem cell migration

Abstract: Bioactive molecules stored in dentin, such as transforming growth factor beta1 (TGF-b1), may be involved in the signaling events related to dental tissue repair. The authors conducted an in vitro evaluation of the amount of TGF-b1 released from dentin slices after treatment with 10% ethylenediaminetetraacetic acid (EDTA), 2.5% sodium hypochlorite (NaOCl) or phosphate-buffered saline (PBS), and the effect of this growth factor on stem cell migration from human exfoliated deciduous teeth (SHED). Sixty 1-mm-thick tooth slices were prepared with or without the predentin layer, and treated with either 10% EDTA for 1 minute, 2.5% NaOCl for 5 days or kept in PBS. Tooth slice conditioned media were prepared and used for TGF-b1 ELISA and migration assays. Culture medium with different concentrations of recombinant human TGF-b1 (0.5, 1.0, 5.0 or 10.0 ng/mL) was also tested by migration assay. The data were evaluated by ANOVA and Tukey's test. Optical density values corresponding to media conditioned by tooth slices either containing or not containing the predentin layer and treated with 10% EDTA were statistically greater than the other groups and close to 1 ng/mL. Increased rates of migration toward media conditioned by tooth slices containing the predentin layer and treated with PBS, 10% EDTA or 2.5% NaOCl were observed. Recombinant human TGF-b1 also stimulated migration of SHED, irrespective of the concentration used. EDTA may be considered an effective extractant of TGF-b1 from the dentin matrix. However, it does not impact SHED migration, suggesting that other components may account for the cell migration.