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).

Effects of different concentrations of sodium hypochlorite on dentine adhesion and the recovery application of sodium erythorbate / 中南大学学报(医学版)

OBJECTIVES@#Root canal therapy is the most effective and common method for pulpitis and periapical periodontitis. During the root canal preparation, chemical irrigation plays a key role. However, sodium hypochlorite (NaOCl), the widely used irrigation fluid, may impact the bonding strength between dentin and restorative material meanwhile sterilization and dissolving. Therefore, it's important to explore the influence of NaOCl on the adhesion between dentin and restoration materials to ensure clinical efficacy. This study aims to explore the effect of NaOCl on dentine adhesion and evaluate the effect of dentine adhesion induced by sodium erythorbate (ERY), and to provide clinical guidance on dentin bonding after root canal therapy.@*METHODS@#Seventy freshly complete extracted human third molars aged 18-33 years old, without caries and restorations were selected. A diamond saw was used under running water to achieve dentine fragments which were divided into 10 groups with 14 fragments in each group: 2 control [deionized water (DW)±10% ERY] and 8 experimental groups (0.5%, 1%, 2.5%, and 5.25% NaOCl±10% ERY). The dentine specimens in the control group (treated with DW) and the experimental groups (treated with 0.5% NaOCl, 1% NaOCl, 2.5% NaOCl, and 5.25% NaOCl) were immersed for 20 min using corresponding solutions which were renewed every 5 min. The other 5 groups were immersed in 10% ERY for 5 min after an initial washing with DW for 1 min. Then, we selected 4 dentine fragments from all 14 fragments in each group and the numbers and diameters of opening dentinal tubules were observed under scanning electron microscope (SEM). The other 10 dentine fragments from each group were used to make adhesive samples by using self-etch adhesive wand composite resin. All the above adhesive samples were sectioned perpendicular to the bonded interface into 20 slabs with a cross-sectional area of 1 mm×1 mm using a diamond saw under the cooling water, and then the morphology of 10 slabs in each group's bonding interface was observed from aspects of formation of resin tags, depth of tags in dentin, and formation of hybrid layer under SEM. The other 10 slabs of each group's microtensile bond strength and failure modes were also analyzed.@*RESULTS@#Among the 0.5% NaOCl, 1% NaOCl, 2.5% NaOCl, and 5.25% NaOCl groups, the number and diameter of patent dentinal tubules gradually increased with the rise of concentration of NaOCl solution (all P<0.05). Among the DW, 0.5% NaOCl, 1% NaOCl, 2.5% NaOCl, and 5.25% NaOCl groups, the number and diameter of patent dentinal tubules increased after using ERY, but without significant difference (all P>0.05). Among the DW, 0.5% NaOCl, 1% NaOCl, and 2.5% NaOCl groups, the scores of formation of resin tags under SEM gradually increased with the increase of concentration of NaOCl solution, while the score in the 5.25% NaOCl group decreased significantly compared with the score of the 2.5% NaOCl group (P<0.05). There was no significant difference between using 10% ERY groups and without using 10% ERY groups (all P>0.05). The scores of length of the tags under SEM in the 5.25% NaOCl group was significantly higher than the scores of DW, 0.5% NaOCl, and 1% NaOCl groups (all P<0.05), and it was also higher than the score of the 2.5% NaOCl group, but without significant difference (P>0.05). There was no significant difference between using 10% ERY groups and without using 10% ERY groups (P>0.05). The scores of formation of hybrid layer under SEM in the 2.5% NaOCl and 5.25% NaOCl groups significantly decreased compared with the score of the DW group (all P<0.05). There were significant differences between the 2.5% NaOCl±10% ERY groups and between the 5.25% NaOCl±10% ERY groups (all P<0.05). Microtensile bond strength was greater in the 0.5% NaOCl, 1% NaOCl, and 2.5% NaOCl groups, but lower in the 5.25% NaOCl group than that in the DW group (all P<0.05). There were significant differences between the 2.5% NaOCl±10% ERY groups and between the 5.25% NaOCl±10% ERY groups (all P<0.05). The incidence of type "Adhesive" of failure modes in the 5.25% NaOCl group was significantly higher than that in other groups (all P<0.05), while the incidence of type "Adhesive" in the 5.25% NaOCl+10% ERY group was lower than that in the 5.25% NaOCl group (P<0.05).@*CONCLUSIONS@#The bonding strength to dentine increases with the increase of NaOCl concentration when the concentration lower than 2.5%; whereas it is decreased at a higher concentration (such as 5.25%). 10% ERY has a definite recovery effect on attenuated bonding strength to 5.25% NaOCl-treated dentine.

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.

Influencia de la dentina en el efecto antibacteriano de 2 concentraciones de hipoclorito de sodio en el crecimiento de Enterococcus faecalis ATCC 29212 / Influence of the dentin on the antibacterial effect of 2 concentrations of sodium hypochlorite in the growth of Enterococcus faecalis ATCC 29212

Este estudio in vitro evaluó la influencia de la dentina sobre el efecto antibacteriano contra Enterococcus faecalis ATCC 29212 de dos concentraciones de Hipoclorito de Sodio (NaOCl) 2,5 % y 5 %. Se empleó polvo de dentina a partir de dientes humanos (84 µg/ml) y la supervivencia de la bacteria se evaluó realizando recuento de unidades formadoras de colonias (UFC) a los 10, 30 y 60 segundos. Los datos se analizaron con la prueba estadística ANOVA factorial no encontrándose diferencias estadísticamente significativas entre los grupos con dentina y sin dentina. En conclusión, la dentina en este estudio no influyó en el efecto antibacteriano del Hipoclorito de Sodio en ninguna concentración, ni en los tiempos.

This in vitro study evaluated the influence of dentin on the antibacterial effect against Enterococcus faecalis ATCC 29212 of two concentrations of Sodium Hypochlorite NaOCl 2.5 % and 5 %. Dentin powder was used from human teeth (84 mg/ml) and the survival of the bacteria was evaluated by counting colony forming units (CFU) at 10, 30 and 60 seconds. The data were analyzed with the statistical ANOVA factorial test, finding no statistically significant differences between the groups with and without dentin. In conclusion, the dentin in this study had no inhibitory effect on antibacterial activity of Sodium Hypochlorite and any concentration, nor over time.

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.

Role of extracellular DNA in Enterococcus faecalis biofilm formation and its susceptibility to sodium hypochlorite

Abstract Objective This study investigated the role of extracellular deoxyribonucleic acid (eDNA) on Enterococcus faecalis ( E. faecalis ) biofilm and the susceptibility of E. faecalis to sodium hypochlorite (NaOCl). Methodology E. faecalis biofilm was formed in bovine tooth specimens and the biofilm was cultured with or without deoxyribonuclease (DNase), an inhibitor of eDNA. Then, the role of eDNA in E. faecalis growth and biofilm formation was investigated using colony forming unit (CFUs) counting, eDNA level assay, crystal violet staining, confocal laser scanning microscopy, and scanning electron microscopy. The susceptibility of E. faecalis biofilm to low (0.5%) or high (5%) NaOCl concentrations was also analyzed by CFU counting. Results CFUs and biofilm formation decreased significantly with DNase treatment (p<0.05). The microstructure of DNase-treated biofilms exhibited less structured features when compared to the control. The volume of exopolysaccharides in the DNase-treated biofilm was significantly lower than that of control (p<0.05). Moreover, the CFUs, eDNA level, biofilm formation, and exopolysaccharides volume were lower when the biofilm was treated with DNase de novo when compared to when DNase was applied to matured biofilm (p<0.05). E. faecalis in the biofilm was more susceptible to NaOCl when it was cultured with DNase (p<0.05). Furthermore, 0.5% NaOCl combined with DNase treatment was as efficient as 5% NaOCl alone regarding susceptibility (p>0.05). Conclusions Inhibition of eDNA leads to decrease of E. faecalis biofilm formation and increase of susceptibility of E. faecalis to NaOCl even at low concentrations. Therefore, our results suggest that inhibition of eDNA would be beneficial in facilitating the efficacy of NaOCl and reducing its concentration.

Influence of surfactants addition on the properties of calcium hypochlorite solutions

Abstract Objectives The aim of this study was to evaluate the influence of surfactants 0.2% or 0.1% cetrimide (Cet) or 0.008% benzalkonium chloride (BAK) on 2.5% calcium hypochlorite (Ca(OCl)2), and compare to sodium hypochlorite (NaOCl), regarding the properties of pH, free chlorine content, surface tension, contact angle, pulp dissolution and antimicrobial activity. Material and Methods The pH and free chlorine content were evaluated by digital pHmeter and by titration, respectively. Surface tension was measured by the platinum ring technique with a Du Noüy tensiometer. The solution's contact angle in human dentin surfaces was checked by Drop Shape Analyzer software. Bovine pulps were used for pulp dissolution analysis and the dissolving capacity was expressed by percent weight loss. Antimicrobial activity over Enterococcus faecalis was evaluated by the agar diffusion method. Results Surfactants addition to Ca(OCl)2 and NaOCl did not alter the pH, free chlorine content and pulp dissolution properties. Ca(OCl)2 had the highest surface tension among all tested solutions. When surfactants were added to Ca(OCl)2 and NaOCl, there was a significant reduction of surface tension and contact angle values. The addition of 0.2% or 0.1% Cet enhanced antimicrobial activity of both Ca(OCl)2 and NaOCl. Conclusion Surfactant addition to 2.5% Ca(OCl)2 has shown acceptable outcomes for pH, free chlorine content, surface tension, contact angle, pulp dissolution and antimicrobial activity. Furthermore, the addition of 0.2% Cet showed better results for all tested properties.

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.

Biofilm formation by Staphylococcus aureus and Salmonella spp. under mono and dual-species conditions and their sensitivity to cetrimonium bromide, peracetic acid and sodium hypochlorite

Abstract The aim of this study was evaluated the biofilm formation by Staphylococcus aureus 4E and Salmonella spp. under mono and dual-species biofilms, onto stainless steel 316 (SS) and polypropylene B (PP), and their sensitivity to cetrimonium bromide, peracetic acid and sodium hypochlorite. The biofilms were developed by immersion of the surfaces in TSB by 10 d at 37 °C. The results showed that in monospecies biofilms the type of surface not affected the cellular density (p > 0.05). However, in dual-species biofilms on PP the adhesion of Salmonella spp. was favored, 7.61 ± 0.13 Log10 CFU/cm2, compared with monospecies biofilms onto the same surface, 5.91 ± 0.44 Log10 CFU/cm2 (p < 0.05). The mono and dual-species biofilms were subjected to disinfection treatments; and the most effective disinfectant was peracetic acid (3500 ppm), reducing by more than 5 Log10 CFU/cm2, while the least effective was cetrimonium bromide. In addition, S. aureus 4E and Salmonella spp. were more resistant to the disinfectants in mono than in dual-species biofilms (p < 0.05). Therefore, the interspecies interactions between S. aureus 4E and Salmonella spp. had a negative effect on the antimicrobial resistance of each microorganism, compared with the monospecies biofilms.

Efficacy of relacin combined with sodium hypochlorite against Enterococcus faecalis biofilms

Abstract Objective Relacin is a synthetic molecule that targets RelA, an essential protein in a conserved bacterial stress response system. It was shown to inhibit bacterial growth. The aims of this study were to evaluate the antimicrobial effect of relacin combined with sodium hypochlorite (NaOCl) on Enterococcus faecalis biofilms and to evaluate the cytotoxicity of relacin. Material and Methods 48-h E. faecalis OG1RF biofilms were treated by various concentrations of relacin in order to determine its inhibitory concentration. Then, the 48-h biofilms were treated either with 1-min NaOCl (0.01%, 0.05%) alone, or in combination of relacin. As a means of comparison, the biofilms of ΔrelA were also treated by 1-min NaOCl (0.01%, 0.05%, 0.25%). The treatment efficacy was determined by agar plate count assays. The cytotoxicity of relacin was examined on human gingival epithelial cells Ca9-22 and murine fibroblasts NIH-3T3 by a methyl thiazolyltetrazolium (MTT) assay and a lactate dehydrogenase assay. Statistical analysis was performed by one-way or two-way analysis of variance (ANOVA) with Bonferroni's post-hoc test and an independent Student's t-test. A significance level of p<0.05 was used. Results Relacin inhibited the growth of OG1RF biofilms partially at 8 mM and fully at 14 mM. The relacin (14 mM) and NaOCl combined treatment resulted in significantly higher treatment efficacy than NaOCl treatment alone. At 0.05% NaOCl, the combined treatment resulted in 5.65 (±0.19) log reduction in biofilm viability. The ΔrelA biofilms were more susceptible to NaOCl treatment than the wild type biofilms at 0.25% NaOCl. Relacin at 14 mM was not toxic to host epithelial cells and fibroblasts. Conclusions The combination of relacin with a low concentration of NaOCl was effective and not cytotoxic.

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.

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.

Effect of final irrigation protocols on microhardness reduction and erosion of root canal dentin

Abstract This study aimed to evaluate the effect of final irrigation protocols on microhardness reduction and erosion of root canal dentin. Sixty root canals from mandibular incisors were instrumented and randomly divided into six groups (n = 10) according to the irrigant used: QMiX, 17% EDTA, 10% citric acid (CA), 1% peracetic acid (PA), 2.5% NaOCl (solution control), and distilled water (negative control). The chelating solutions were used to irrigate the canal followed by 2.5% NaOCl as a final flush. After the irrigation protocols, all specimens were rinsed with 10 mL of distilled water to remove any residue of the chemical solutions. Before and after the final irrigation protocols, dentin microhardness was measured with a Knoop indenter. Three indentations were made at 100 µm and 500 µm from the root canal lumen. Afterwards, the specimens were prepared for scanning electron microscopic analysis and the amount of dentin erosion was examined. Wilcoxon and Kruskal-Wallis tests were used to analyze the results with a significance level set at 5%. At 100 µm, all protocols significantly reduced dentin microhardness (p < .05), while at 500 µm, this effect was detected only in the EDTA and QMiX groups (p < .05). CA was the irrigant that caused more extensive erosion in dentinal tubules, followed by PA and EDTA. QMiX opened dentinal tubules, but did not cause dentin erosion. Results suggest that QMiX and 17% EDTA reduced dentin microhardness at a greater depth. Additionally, QMiX did not cause dentin erosion.

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)

Ozone therapy as an adjuvant for endondontic protocols: microbiological – ex vivo study and citotoxicity analyses

ABSTRACT Objectives This study evaluated the antimicrobial efficacy of ozone therapy in teeth contaminated with Pseudomonas aeruginosa, Enterococcus faecalis, and Staphylococcus aureus using a mono-species biofilm model. Parallel to this, the study aimed to evaluate the cytotoxicity of ozone for human gingival fibroblasts. Material and Methods: One hundred and eighty single-root teeth were contaminated with a mono-species biofilm of Enterococcus faecalis, Pseudomonas aeruginosa, and Staphylococcus aureus. Groups were formed: Group I – control; Group II – standard protocol; Group III – standard protocol + ozone gas at 40 µg/mL; and Group IV – standard protocol + aqueous ozone at 8 µg/mL. In parallel, human gingival fibroblasts were submitted to the MTT test. Cells were plated, then ozone was applied as follows: Group I (control) – broth medium; Group II – aqueous ozone at 2 µg/mL; Group III – aqueous ozone at 5 µg/mL; and Group IV – aqueous ozone at 8 µg/mL. Data were submitted to the Kruskal Wallis test and Bonferroni post hoc analyses to assess microbiology and cytotoxicity, respectively (p<0.05%). Results The results revealed antimicrobial efficacy by Group IV with no CFU count. The cytotoxicity assay showed Groups III and IV to be the most aggressive, providing a decrease in cell viability at hour 0 from 100% to 77.3% and 68.6%, respectively. Such a decrease in cell viability was reverted, and after 72 hours Groups III and IV provided the greatest increase in cell viability, being statistically different from Groups I and II. Conclusion According to the applied methodology and the limitations of this study, it was possible to conclude that ozone therapy improved the decontamination of the root canal ex vivo. Ozone was toxic to the cells on first contact, but cell viability was recovered. Thus, these findings suggest that ozone might be useful to improve root canal results.

Influence of Temperature on the Antibacterial Activity of Sodium Hypochlorite

Abstract The aim of this study was to compare the antimicrobial activity of 5.25% NaOCl, Hypoclean and Chlor-Xtra at 20 °C and 45 °C in bovine root dentin. One-hundred-and-seventy dentin tubes prepared from bovine maxillary incisors were infected for 21 days with Enterococcus faecalis. The specimens were divided into the following groups: 1. 5.25% NaOCl 20 °C; 2. Hypoclean 20 °C; 3. Chlor-Xtra 20 °C; 4. 5.25% % NaOCl 45 °C; 5. Hypoclean 45 °C; 6. Chlor-Xtra 45 °C; 7. positive control; 8. negative control. Dentin chips were collected with round burs into Brain Heart Infusion (BHI) broth. After culturing, the number of colony-forming units (CFU) was counted. Statistical analyses were performed using descriptive statistics (mean, standard deviation, median), Shapiro-Wilk test, ANOVA and Tukey test. Significance level was set at p<0.05. In all experimental groups, CFU was minimum after treatment (day 0) and the obtained results were significantly different from each other at any period (p<0.05). After treatment, the Hypoclean and Chlor-Xtra showed the lowest numbers of CFU at 20 °C and 45 °C, whereas 5.25% NaOCl showed the highest number of CFU at both temperatures. In each group, the number of CFUs increased significantly with time (p<0.05). The antibacterial activity of Hypoclean and Chlor-Xtra at 45 °C were significantly greater than other tested solutions.

Resumo O objetivo deste estudo foi comparar a atividade antimicrobiana do hipoclorito de sódio a 5,25%, Hypoclean e Cloro-Xtra a 20 °C e 45 °C em dentina radicular bovina. Um total de 170 tubos de dentina foram preparados a partir de incisivos superiores bovinos infectados por 21 dias com Enterococcus faecalis. Os espécimes foram divididos nos seguintes grupos: 1. NaOCl - 5,25% a 20 °C; 2. Hypoclean 20 °C; 3. Cloro-Xtra 20 °C; 4. NaOCl - 5,25% a 45 °C; 5. Hypoclean 45 °C; 6. Cloro-Xtra 45 °C; 7. Controle positivo; 8. Controle negativo. Raspas de dentina foram coletadas com brocas esféricas e cultivadas em infusão cérebro coração (brain heart infusion - BHI). Após a cultura, o número de unidades formadoras de colônias (UFC) foi contado. As análises estatísticas foram realizadas utilizando estatística descritiva (média, desvio padrão, mediana), teste de Shapiro-Wilk, ANOVA e teste de Tukey. O nível de significância foi estabelecido p<0,05. Em todos os grupos experimentais, o número de UFC foi mínimo após o tratamento inicial. Os resultados obtidos foram significativamente diferentes nos períodos de tempos experimentais (p<0,05). O Hypoclean e o Cloro-Xtra mostraram o menor número de UFC a 20 °C e a 45 °C, enquanto que o NaOCl - 5,25% apresentou maior número de UFC em ambas as temperaturas. Em cada grupo, o número de CFUs foi significativamente aumentado por período de tempo (p<0,05). As atividades antibacterianas do Hypoclean e Chlor-Xtra a 45 °C foram significativamente maiores do que nas outras soluções testadas.

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.

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.

Antimicrobial activity of complete denture cleanser solutions based on sodium hypochlorite and Ricinus communis: a randomized clinical study

ABSTRACT To preserve oral health and to maintain the prosthetic devices, it is important not only to improve the properties of commonly known hygiene products, but also to investigate new materials with antimicrobial action. Objectives This study evaluated the antimicrobial activity of sodium hypochlorite (0.25% and 0.50%) and 10% Ricinus communis’ solutions against specific microorganisms. Material and Methods Sixty four maxillary complete denture wearers were instructed to brush their dentures three times a day and to soak them (20 min/day) in the solutions: SH1: 0.25% sodium hypochlorite; SH2: 0.5% sodium hypochlorite; RC: 10% R. communis oil; and C: 0.85% saline (control). The solutions were used for 7 days in a randomized sequence. Following each period of use, there was a 1-week washout period. Antimicrobial activity was determined by Colony Forming Units (CFU) counts of Streptococcus mutans, Candida spp., and gram-negative microorganisms. For collecting biofilm, the internal surface of maxillary dentures was brushed with saline solution, and biofilm suspension obtained. After dilutions (100 - 10-3), aliquots were seeded in Mitis salivarius, CHROMagar Candida®, and MacConkey agar for detecting S. mutans, Candida spp., or gram-negative microorganisms, respectively. After incubation, colonies were counted, and CFU/mL values were calculated. Then, transformation - log10 (CFU+1) - data were analyzed using the Friedman test (α=0.05). Results showed significant differences between the solutions (p<0.001). Results All three solutions showed antimicrobial activity against S. mutans. Against Candida spp., RC and SH1 solutions showed similar effect while SH2 showed superior activity. SH1 and SH2 solutions showed antimicrobial action against gram-negative microorganisms. The Candida species most frequently isolated was C. albicans, followed by C. tropicalis and C. glabrata. Conclusions The 0.5% sodium hypochlorite solution was the most effective and might be used to control denture biofilm. C. albicans was the most frequently isolated Candida sp.