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Rev. habanera cienc. méd ; 20(4): e3971, 2021. tab
Article in Spanish | LILACS, CUMED | ID: biblio-1289617


Introducción: El aceite esencial de Minthostachys mollis ha demostrado poseer importantes propiedades antimicrobianas. Objetivo: Caracterizar químicamente las fracciones obtenidas del aceite esencial de Minthostachys mollis y evaluar la actividad antimicrobiana sobre Streptococcus mutans, Lactobacillus acidophilus, Enterococcus faecalis, Porphyromonas gingivalis y Candida albicans. Material y Métodos: Las fracciones de éter de petróleo, diclorometano y metanol del AE de M. mollis fueron caracterizadas químicamente por cromatografía de gases acoplada a espectrometría de masas. Las repeticiones del ensayo antimicrobiano se calcularon con el programa EPIDAT v.4.2. La actividad antimicrobiana se realizó por el método de difusión de disco y se calculó la concentración mínima inhibitoria por el método de microdilución. Los datos fueron analizados empleando la prueba ANOVA (p=0,05). Resultados: Los principales constituyentes de las fracciones de éter de petróleo, diclorometano y metanol fueron cis-Menthone (39,8 por ciento, thymol (31,2 por ciento) y α-Terpineol (43,6 por ciento), respectivamente. Todas las cepas fueron sensibles a las tres fracciones, aunque C. albicans fue la cepa más sensible, registrando halos de inhibición de 14,73±0,57 mm para la fracción de metanol, 20,91±0,55 mm para éter de petróleo y 20,38±0,58 mm para diclorometano, se encontraron diferencias significativas cuando se compararon frente a Clorhexidina al 0,12 por ciento y Nistatina (p<0,05). Las concentraciones mínimas inhibitorias de las fracciones variaron de 0,2 a 3,2 µg/mL. Conclusiones: Los principales constituyentes de las fracciones de éter de petróleo, diclorometano y metanol fueron cis-Menthone, thymol y α-Terpineol. Las fracciones de éter de petróleo y diclorometano fueron altamente efectivas para inhibir el crecimiento de S. mutans, L. acidophilus, E. faecalis, P. gingivalis y C. albicans(AU)

Introduction: The essential oil of Minthostachys mollis has demonstrated to have important antimicrobial properties. Objective: To chemically characterize the fractions obtained from the essential oil of Minthostachys mollis and to evaluate the antimicrobial activity against Streptococcus mutans, Lactobacillus acidophilus, Enterococcus faecalis, Porphyromonas gingivalis and Candida albicans. Material and Methods: The petroleum ether, dichloromethane and methanol fractions of the AE of M. mollis were chemically characterized by gas chromatography coupled to mass spectrometry. The repetitions of the antimicrobial test were calculated using the EPIDAT v.4.2 program. The antimicrobial activity was performed by the disk diffusion method and the minimum inhibitory concentration was calculated by the microdilution method. The data were analyzed using the ANOVA test (p=0.05). Results: The main constituents of the petroleum ether, dichloromethane and methanol fractions were cis-Menthone (39,8 percent), thymol (31,2 percent)) and α-Terpineol (43,6 percent)), respectively. All strains were sensitive to the three fractions, although C. albicans was the most sensitive strain, registering inhibition halos of 14,73±0.57 mm for the methanol fraction, 20,91±0.55 mm for petroleum ether and 20.38±0.58 mm for dichloromethane, finding significant differences when compared to 0,12 percent) Chlorhexidine and Nystatin (p<0,05). The minimum inhibitory concentrations of the fractions ranged from 0,2 to 3,2 µg/mL. Conclusions: The main constituents of the petroleum ether, dichloromethane and methanol fractions were cis-Menthone, thymol and α-Terpineol. The petroleum ether and dichloromethane fractions were highly effective in inhibiting the growth of S. mutans, L. acidophilus, E. faecalis, P. gingivalis, and Calbicans(AU)

Humans , Oils, Volatile/therapeutic use , Microbial Sensitivity Tests , Enterococcus faecalis , Porphyromonas gingivalis , Lactobacillus acidophilus , Analysis of Variance , Chromatography, Gas
Braz. dent. j ; 32(3): 1-9, May-June 2021. tab, graf
Article in English | LILACS, BBO | ID: biblio-1345506


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

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

Humans , Root Canal Irrigants , Dental Pulp Cavity , Sodium Hypochlorite , Chlorhexidine , Enterococcus faecalis , Root Canal Preparation
Braz. dent. j ; 32(3): 10-20, May-June 2021. tab, graf
Article in English | LILACS, BBO | ID: biblio-1345504


Abstract The host defense response to microbial challenge emerging from the root canal system leads to apical periodontitis. The aim of this study was to evaluate the expression of inflammatory cytokines and Nitric Oxide (NO) by macrophages after interaction with Enterococcus faecalis in the: plankton and dislodged biofilm mode; intact biofilm mode stimulated by calcium hydroxide (CH), CH and chlorhexidine (CHX) or Triple Antibiotic Paste (TAP). For this purpose, culture of macrophages from monocytes in human peripheral blood (N=8) were exposed to the different modes of bacteria for 24 hours. Subsequently, the cytokines, such as, Tumor Necrotic Factor- alfa (TNF-α), interleukin (IL)-1β, IL-6, IL-10; and NO were quantified by Luminex xMAP and Greiss reaction, respectively. In addition to the potential therapeutic effects of the intracanal medication, their antimicrobial activity against Enterococcus faecalis biofilm were also tested in vitro by confocal microscopy. The experiments` data were analyzed by the Kruskal-Wallis test with the Dunn post hoc test (α < 0.05). Bacteria in dislodged biofilm mode were shown to be more aggressive to the immune system than bacteria in plankton mode and negative control, inducing greater expression of NO and TNF-α. Relative to bacteria in intact biofilm mode, the weakest antimicrobial activity occurred in Group CH. In Groups CH/CHX and TAP the percentage of dead bacteria was significantly increased to the same extent. Interestingly, the biofilm itself did not induce the release of pro-inflammatory cytokines - except for NO - while the biofilm treated with TAP and CH based pastes enhanced the levels of IL-6 and TNF-α; and IL-1 β, respectively. In contrast, the levels of a potent anti-inflammatory (IL-10) were increased in Group TAP.

Resumo A resposta de defesa do hospedeiro ao desafio microbiano que emerge do sistema de canais radiculares leva à periodontite apical. Os objetivos deste estudo foram avaliar a expressão de citocinas pró e anti-inflamatórias e Óxido Nítrico (NO) por macrófagos após interação com Enterococcus faecalis no modo: planctônio e de biofilme desalojado; biofilme intacto estimulado por hidróxido de cálcio (CH), CH e clorexidina ou Pasta Tri Antibiótica (TAP). Para isto, a cultura de macrófagos originados de monócitos do sangue periférico de humanos (N=8) foi exposta aos diferentes tipos de bactéria por 24 horas. Então, a quantificação da produção de of Fator de Necrose Tumoral- alfa (TNF-α), interleucina (IL)-1β, IL-6, IL-10 e NO por macrófagos se deu por meio do Luminex xMAP e reação de Greiss, respectivamente. Além dos potenciais efeitos terapêuticos desses compostos, sua atividade antimicrobiana contra E. faecalis também foi testada através microscopia confocal. Os dados dos experimentos foram analisados através do teste de Kruskal-Wallis com Dunn`s post hoc (α < 0.05). Bactéria em modo de biofilme desalojado se mostrou mais agressivo ao sistema imune que as bactérias planctônicas e controle negativo induzindo a maior excreção de NO e TNF-α. Em relação ao biofilme intacto, a atividade antimicrobiana mais fraca ocorreu no grupo de CH. Os grupos CHX e TAP aumentaram significativamente a porcentagem de bactérias mortas na mesma extensão. Interessantemente, o biofilme por ele mesmo não induziu a liberação de citocinas pro-inflamatórias - exceto por NO - enquanto que o biofilme tratado com TAP ou pastas a base de CH aumentaram os níveis de IL-6; e TNF-α e IL-1 β respectivamente. Em contraste, os níveis da potente citocina anti-inflamatória (IL-10) foram aumentados pelo grupo TAP.

Humans , Plankton , Biofilms , Root Canal Irrigants , Bacteria , Calcium Hydroxide , Chlorhexidine , Enterococcus faecalis , Anti-Bacterial Agents
Rev. cuba. estomatol ; 58(2): e3026, 2021. graf
Article in Portuguese | LILACS, CUMED | ID: biblio-1289394


Introdução: Os fitoconstituintes são moléculas naturais que apresentam atividade antimicrobiana satisfatória e devem ser estudados quanto ao seu uso como novas substâncias para irrigação dos canais radiculares. Objetivo: Avaliar o efeito inibitório dos fitoconstituintes cinamaldeído e α-terpineol frente a biofilmes monoespécie e duoespécie de microrganismos envolvidos na infecção endodôntica. Métodos: Trata-se de um estudo experimental na área de microbiologia aplicada, in vitro, cego quanto às análises e randomizado. Foram selecionados os fitoconstituintes cinamaldeído e α-terpineol. A atividade antimicrobiana frente Candida albicans e Enterococcus faecalis foi avaliada por meio da análise da capacidade metabólica com o uso da resazurina e análise da viabilidade celular pelo plaqueamento. O meio de cultura e a clorexidina 1 porcento serviram de controle negativo e positivo, respectivamente. Resultados: Observou-se ausência de crescimento para exposição dos biofilmes nas concentrações de 10 e 5 mg/mL de ambos os fitoconstituintes. Na concentração de 2,5 mg/mL de terpineol, constatou-se crescimento somente nos biofilmes monoespécie de C. albicans e duoespécie. Já na concentração de 1mg/mL de terpineol e cinamaldeído, verificou-se crescimento para todos os biofilmes. Conclusão: O cinamaldeído e α-terpineol apresentaram atividade inibitória frente biofilmes monoespécie e duoespécie de Candida albicans e Enterococcus faecalis, nas concentrações de 10 e 5 mg/mL(AU)

Introducción: Los fitoconstituyentes son moléculas naturales que presentan actividad antimicrobiana satisfactoria y deben ser estudiados en cuanto a su uso como nuevas sustancias para irrigación de los canales radiculares. Objetivo: Evaluar el efecto inhibitorio de fitoconstituyentes cinamaldehído y α-terpineol frente a biopelículas monoespecies y duoespecies de microorganismos involucrados en la infección endodóntica. Métodos: Estudio experimental en el campo de la microbiología aplicada, in vitro, ciego al análisis y aleatorizado. Se seleccionaron los fitoconstituyentes cinamaldehído y α-terpineol. La actividad antimicrobiana frente Candida albicans y Enterococcus faecalis fue evaluada por medio del análisis de la capacidad metabólica con el uso de la resazurina y análisis de la viabilidad celular por el plaqueamiento. El medio de cultivo y la clorexidina 1 por ciento sirvieron de control negativo y positivo, respectivamente. Resultados: Se observó ausencia de crecimiento para exposición de las biopelículas en las concentraciones de 10 y 5 mg/mL de ambos fitoconstituyentes. En la concentración de 2,5 mg/mL de terpineol se constató crecimiento solo en los biofilmios monoespecies de C. albicans y duoespecies. En la concentración de 1 mg/mL de terpineol y cinamaldehído se verificó crecimiento para todas las biopelículas. Conclusiones: Cinamaldehído y α-terpineol presentaron actividad inhibitoria frente a biofilmes monoespecies y duoespecies de Candida albicans y Enterococcus faecalis, en las concentraciones de 10 y 5 mg/mL(AU)

Introduction: Phytoconstituents are natural molecules displaying satisfactory antimicrobial activity. Studies should be conducted about their use as new root canal irrigants. Objective: Evaluate the inhibitory effect of the phytoconstituents cinnamaldehyde and α-terpineol against mono- and duo-species biofilms of microorganisms involved in endodontic infection. Methods: An experimental applied microbiology blind randomized in vitro study was conducted. The phytoconstituents selected were cinnamaldehyde and α-terpineol. Antimicrobial activity against Candida albicans and Enterococcus faecalis was evaluated by metabolic capacity analysis with resazurin and cell viability analysis by the plaque. The culture medium and 1 percent chlorhexidine served as negative and positive controls, respectively. Results: An absence of growth was observed for exposure of the biofilms at concentrations of 10 and 5 mg/ml of both phytoconstituents. At a concentration of 2.5 mg/ml terpineol displayed growth only in the mono-species biofilms of C. albicans and duo-species biofilms. At a concentration of 1 mg/ml terpineol and cinnamaldehyde displayed growth in all biofilms. Conclusions: Cinnamaldehyde and α-terpineol displayed inhibitory activity against mono- and duo-species biofilms of Candida albicans and Enterococcus faecalis at concentrations of 10 and 5 mg/ml(AU)

Humans , Biological Products/adverse effects , Candida albicans , Cell Survival , Biofilms , Enterococcus faecalis , Anti-Infective Agents/adverse effects
Rev. Eugenio Espejo ; 15(1): 12-21, 20210102.
Article in Spanish | LILACS | ID: biblio-1145479


La elevada incidencia del Enterococcus faecalis en procedimientos odontológicos es un tópico de interés para el área de la salud, esta bacteria resiste a varios antimicrobianos y su proliferación aumenta debido a su baja susceptibilidad a sustancias de uso convencional como el hidróxido de calcio, convirtiéndose en una de las principales causas del fracaso de los tratamientos de conduc- to. La presente investigación evaluó la susceptibilidad in vitro del Enterococcus faecalis cepa ATCC-29212 frente a la combinación de sustancias y antibióticos con el hidróxido de calcio. Se utilizó la técnica de difusión en agar, inoculando el microorganismo en cajas de Petri con agar Cerebro Corazón (BHI), tratado con discos de papel filtro impregnados con 1µl de cada trata- miento por triplicado; T1 (Hidróxido de Calcio + propilenglicol), T2 (Hidróxido de Calcio + paramonoclorofenol), T3 (Hidróxido de Calcio + ampicilina gentamicina + propilenglicol), T4 (Hidróxido de Calcio + Solución Salina Fisiológica), Control positivo (medicamento comercial a base de Hidróxido de Calcio), se incubó a 35°C durante 24h, los halos formados alrededor de cada disco fueron medidos y comparados con la escala de Duraffourd y procesados mediante ANOVA de un factor. Se obtuvo una medida del halo de inhibición de 22,50±3,3 mm, con el trata- miento T3, siendo sumamente sensible. Concluyendo que este resultó el más efectivo en compa- ración el resto de la pruebas in vitro en comparación con el resto de los fármaco investigados.

The high incidence of Enterococcus faecalis in dental procedures is a topic of interest for the health area. This bacterium resists various antimicrobials and its proliferation increases due to its low susceptibility to substances of conventional use such as calcium hydroxide, becoming a of the main causes of root canal failure. The present investigation evaluated the in vitro suscepti- bility of Enterococcus faecalis strain ATCC-29212 to the combination of substances and antibio - tics with calcium hydroxide. The agar diffusion technique was used, inoculating the microorga- nism in Petri dishes with Brain Heart agar (BHI), treated with filter paper disks impregnated with 1µl of each treatment in triplicate; T1 (Calcium Hydroxide + propylene glycol), T2 (Cal- cium Hydroxide + paramonochlorophenol), T3 (Calcium Hydroxide + ampicillin gentamicin + propylene glycol), T4 (Calcium Hydroxide + Physiological Saline Solution), Positive control (Hydroxide-based commercial medicine Calcium), incubated at 35 ° C for 24h, the halos formed around each disc were measured and compared with the Duraffourd scale and processed by one-factor ANOVA. A measure of the inhibition halo of 22.50 ± 3.3 mm was obtained with the T3 treatment, being extremely sensitive. It was concluded that this was the most effective in comparison with the rest of the in vitro tests of the rest of the investigated drugs.

Calcium Hydroxide , Enterococcus faecalis , Ampicillin , Pharmaceutical Preparations , Gentamicins , Health
Article in English | LILACS, BBO | ID: biblio-1154996


ABSTRACT Objective: To determine the level of clinical contamination in the clinic and laboratory of the prosthodontics department of Kerman Dental School. Material and Methods: Clinical surfaces of the dental units, the laboratory, and the professors' lounge of the prosthodontics department were randomly sampled. The sampled surfaces included the dental units' console, light switch, light handle, headrest, and air-water spray syringe in the clinic, plastering tables, buttons of the vibrator, polishing, and trimmer machines, acryl tables, handles of pressure pot and press machine, handpiece holders, work desks, and drawer handles in the laboratory, and desks, computer mouse and keyboard, telephone sets, and doorknob in the professor's lounge. The samples were examined for the type and growth of microorganisms. The data were entered into SPSS, where they were analyzed using the chi-square test at the 0.05 significance level. Results: Of all the samples taken, 89.9% showed microbial contamination. The most common type of contamination was fungus (34.8%) and the least common types were Enterococcus faecalis and Staphylococcus epidermidis (1.1%). The second and third most common types of bacteria in the samples were Staphylococcus aureus (18%) and Pseudomonas aeruginosa (12.4%), respectively. There was no significant difference between the frequencies of microbial contamination in the clinic, the laboratory, and the professors' lounge. Conclusion: Given the strong chance of cross-infection in the examined department and laboratory, it is necessary to enforce protocols for proper disinfection of surfaces before, between and after treatments.

Schools, Dental , Disinfection/instrumentation , Enterococcus faecalis , Environmental Pollution , Microbiology , Chi-Square Distribution , Epidemiology, Experimental , Analysis of Variance , Protocols/methods , Iran/epidemiology
J. appl. oral sci ; 29: e20200733, 2021. graf
Article in English | LILACS | ID: biblio-1154616


Abstract Enterococcus faecalis (E. faecalis), one of the main pathogens responsible for refractory periapical periodontitis and nosocomial infections, exhibits markedly higher pathogenicity in biofilms. Objectives Studies have shown that caseinolytic protease P (ClpP) is involved in biofilm formation. However, to date, few studies have investigated the role of ClpP in the survival of E. faecalis, and in enhancing biofilm formation. Therefore, we investigated the role of ClpP in the formation of E. faecalis biofilms. Methodology In our study, we used homologous recombination to construct clpP deleted and clpP complement strains of E. faecalis ATCC 29212. A viable colony counting method was used to analyze the growth patterns of E. faecalis. Crystal violet staining (CV) and confocal scanning laser microscopy (CLSM) were used to characterize biofilm mass formation and scanning electron microscopy (SEM) was used to observe the biofilm microstructure. Data was statistically analyzed via Student's t-test or one-way analysis of variance (ANOVA). Results The results exhibited altered growth patterns for the clpP deletion strains and depleted polysaccharide matrix, resulting in reduced biofilm formation capacity compared to the standard strains. Moreover, ClpP was observed to increase biofilm formation in E. faecalis. Conclusion Our study shows that ClpP can increase biofilm formation in E. faecalis and emphasizes the importance of ClpP as a potential target against E. faecalis.

Humans , Enterococcus faecalis , Biofilms , Peptide Hydrolases , Virulence , Microscopy, Electron, Scanning , Microscopy, Confocal , Endopeptidase Clp
Rev. Fac. Odontol. (B.Aires) ; 36(84): 21-26, 2021. tab
Article in Spanish | LILACS | ID: biblio-1363852


La terapia endodóntica tiene como uno de sus objetivos lograr la completa desinfección del sistema de conductos radiculares. Por esto, se deben seleccionar sustancias irrigantes que tengan la capacidad de eliminar todo el contenido de dicho sistema. La acción antimicrobiana es una de las características más importantes a tener en cuenta en la elección. El hipoclorito de sodio (NaOCl) tiene capacidad bactericida sobre muchos de los microorganismos de la flora endodóntica. El Enterococcus faecalis es una bacteria altamente resistente a antibacterianos que sobrevive en condiciones extremas. El ácido hipocloroso (HOCl) es una molécula derivada del NaOCl que ha demostrado tener alto poder bactericida sobre cepas patogénicas bucales. El objetivo de este trabajo fue evaluar y comparar la efectividad antimicrobiana in vitro del NaOCl 2.5% y el HOCl al 5% frente a Enterococcus faecalis. Una suspensión de Enterococcus faecalis (ATCC29212), de turbidez 0.5 en escala de McFarland, fue inoculada en varios tubos de ensayo, los cuales contenían cada antimicrobiano. Se dejaron actuar durante 1, 5 y 10 minutos para luego neutralizarlos e inclubarlos a 37º C en condiciones de capnofilia durante 48 hs. Todo el procedimiento se realizó por quintuplicado. Los resultados se midieron mediante recuento de UFC/ml. No se evidenció presencia de Enterococcus faecalis en las placas que contenían la solución de NaOCl al 2.5% como tampoco en aquellas que contenían HOCl al 5%. In vitro, el HOCl y el NaOCl en las concentraciones probadas, eliminaron completamente las cepas de Enterococcus faecalis (AU)

Sodium Hypochlorite/therapeutic use , Enterococcus faecalis/drug effects , Hypochlorous Acid/therapeutic use , Anti-Bacterial Agents/therapeutic use , Root Canal Irrigants/therapeutic use , In Vitro Techniques , Colony Count, Microbial , Culture Media , Dental Pulp Cavity/microbiology
Article in English | LILACS, BBO | ID: biblio-1143399


ABSTRACT Objective: To evaluate the antibacterial effect and the solubility of experimental root canal filling pastes containing the phytoconstituents terpineol and cinnamaldehyde. Material and Methods: Minimum Inhibitory Concentration (MIC) of each phytoconstituent was determined against Enterococcus faecalis. Five groups of antibiotic pastes based on zinc oxide were obtained by mixing: only terpineol, only cinnamaldehyde, terpineol and cinnamaldehyde combined, chlorhexidine (antibiotic control), and CTZ paste (control paste). Antibacterial activity was analyzed through direct contact test within 24 and 72 hours. Solubility was evaluated by spectrophotometry within 48 and 144 hours. Antibacterial activity data were analyzed descriptively, and solubility data was analyzed using ANOVA and Tukey tests (p<0.05). Results: The MIC obtained for terpineol and cinnamaldehyde were, respectively, 2000 µg/mL and 500 µg/mL. After 24h, only the terpineol paste did not inhibit E. faecalis growth. After 72h, all groups inhibited E. faecalis growth. After 48h, the highest solubility was verified in the terpineol paste (p<0.05), and no differences were detected among other groups (p>0.05). After and 144h, highest solubility was observed in the terpineol paste (p<0.05), followed by the CTZ paste (p<0.05). No differences were detected for cinnamaldehyde, terpineol+cinnamaldehyde and chlorhexidine pastes (p>0.05). Conclusion: Pastes containing cinnamaldehyde or terpineol+cinnamaldehyde showed antibacterial activity against E. faecalis similar to CTZ paste, with lower solubility.

Root Canal Obturation/instrumentation , Tooth, Deciduous , Plant Extracts , Enterococcus faecalis , Anti-Bacterial Agents , Solubility , Spectrophotometry , Brazil/epidemiology , Analysis of Variance , Statistics, Nonparametric
Braz. dent. sci ; 24(1): 1-7, 2021. tab, ilus
Article in English | LILACS, BBO | ID: biblio-1145548


Though aloe vera extract, green tea extract and coriander oil are proven antimicrobial agents, very little information is available regarding its effects on oral bacteria, Streptococcus mutans, which is responsible for initiating caries and Enterococcus faecalis, responsible for failure of root canal treatment. Objective: To find the antimicrobial activity of aloe vera extract, black tea extract and coriander oil against S. mutans and E. faecalis. Materials and Methods: The agar well diffusion method was used to determine the antibacterial activity of Aloe vera extract, black tea extract and coriander oil. Different concentration of prepared plant extracts and coriander seed oil (50 & 100 µl) was incorporated into the wells and the plates containing S. mutans and E. faecalis were incubated at 37 °C for 24 h. The antibiotic (amoxicillin 30 µl) was used as positive control. Zone Of Inhibition (ZOI) was recorded in each plate. Results: For S. mutans, the maximum ZOI was created by coriander oil with a diameter of 25.00±0.58 mm at 50 µl and for E. faecalis, maximum ZOI was created by aloe vera extract 16.00±0.58 mm at 100 µl concentration which were far better than the control: amoxicillin 30 µl concentration. Conclusion: The extracts of Aloe vera, black tea and coriander oil, showed significant activity against the investigated microbial strains, Streptococcus mutans and Enterococcus faecalis which further helps in the development of new topical agents that help in reducing the numbers of these organisms present in the oral cavity. (AU)

Embora o extrato de aloe vera, extrato de chá verde e óleo de coentro sejam agentes antimicrobianos comprovados, há pouca informação disponível sobre seus efeitos nas bactérias orais, Streptococcus mutans, que é responsável por iniciar cáries e Enterococcus faecalis, responsável pela falha do tratamento de canal radicular. Objetivo: Avaliar a atividade antimicrobiana do extrato de aloe vera, extrato de chá preto e óleo de coentro contra S. mutans e E. faecalis. Materiais e Métodos: O método de difusão em agar foi usado para determinar a atividade antibacteriana do extrato de Aloe vera, extrato de chá preto e óleo de coentro. Diferentes concentrações dos extratos de plantas e óleo de semente de coentro (50 e 100 µl) foram preparados e colocados nos poços e nas placas contendo S. mutans e E. faecalis e foram incubadas a 37°C por 24 h. O antibiótico (amoxicilina 30 µl) foi utilizado como controle positivo. A zona de inibição (ZOI) foi registrada em cada placa. Resultados: Para S. mutans, a ZOI máxima foi obtida com o óleo de coentro com um diâmetro de 25,00 ± 0,58 mm a 50 µl e para E. faecalis, a ZOI máxima foi obtiada pelo extrato de aloe vera 16,00 ± 0,58 mm na concentração de 100 µl, as quais foram melhores do que o controle: concentração de 30 µl de amoxicilina. Conclusão: Os extratos de Aloe vera, chá preto e óleo de coentro apresentaram atividade significativa contra as cepas microbianas investigadas, Streptococcus mutans e Enterococcus faecalis auxiliando no desenvolvimento de novos agentes tópicos visando a redução do número desses organismos presentes no cavidade oral. (AU)

Streptococcus mutans , Tea , Enterococcus faecalis , Aloe , Microbiota
Pesqui. vet. bras ; 41: e06706, 2021. tab
Article in English | LILACS, VETINDEX | ID: biblio-1346697


Contamination of the veterinary hospital environment with multiresistant pathogens endangers not only hospitalized animals, but also the workplace safety of veterinarians and nurses, animal guardians and, when in case of a teaching hospital, veterinary students. The objective of this study was to map the main points of bacterial contamination of a veterinary teaching hospital in Brazil to identify multiresistant microorganisms and their antimicrobial resistance genes. Samples were collected from 39 different locations of a veterinary school hospital which comprised a pool according to each hospital environment. In certain environments, more than one pool has been collected. All samples were collected in quadruplicates for the selective isolation of the main multiresistant microorganisms: methicillin-resistant Staphylococcus (MRS), vancomycin resistant Enterococcus (VRE), cephalosporinases and/or extended-spectrum beta-lactamase-producing Gram-negative bacteria (ESBL) and Carbapenemase-producing (CP). After isolation and identification of isolates, multiplex-PCR reactions were performed to detect the main genes for each microorganism and antimicrobial susceptibility tests with the main antibiotics used for each bacterial group according to CLSI. Of the 39 veterinary teaching hospital sites collected, all (100%) had at least one of the microorganisms surveyed, and 17.95% (n=7) of the sites were able to isolate the four pathogens. From the 94 pools collected, it was possible to isolate MRS in 81.91% (n=77), VRE in 12.77% (n=12), cephalosporinases and/or ESBL in 62.77% (n=59) and CP in 24.47%. (n=23). Regarding MRS, the mecA gene was detected in all isolates. All isolated VREs were identified as Enterococcus faecalis and presented the vanA gene. Regarding cephalosporinases and/or ESBL, 89.83% (n=53) of the isolates presented the blaTEM gene, 57.63% (n=34) the blaOXA-1 gene, 37.29% (n=22) blaCTX-M gene from some group (1, 2, 9 ou 8/25) and 20.34% (n=12) the blaSHV gene. It was possible to identify the main microorganisms responsible for causing nosocomial infections in humans (VRE, MRS, ESBL and CP) in the veterinary hospital environment, suggesting a source of infection for professionals and students of veterinary medicine, placing a high risk for public health.(AU)

A contaminação do ambiente hospitalar veterinário com patógenos multirresistentes coloca em perigo não apenas os animais hospitalizados, mas também a segurança no local de trabalho de veterinários e enfermeiros, responsáveis por animais e, quando se tratar de um hospital de ensino, estudantes de veterinária. O objetivo deste estudo foi mapear os principais pontos de contaminação bacteriana de um hospital veterinário de ensino no Brasil, identificando microorganismos multirresistentes e seus genes de resistência antimicrobiana. As amostras foram coletadas em 39 locais diferentes de um hospital de escola veterinária, que compreendia um pool de acordo com o ambiente de cada hospital. Em certos ambientes, mais de um pool foi coletado. Todas as amostras foram coletadas em quadruplicados para o isolamento seletivo dos principais microorganismos multirresistentes: Staphylococcus resistente à meticilina (MRS), Enterococcus resistente à vancomicina (VRE), bactérias Gram-negativas produtoras de cefalosporinases e/ou beta-lactamase de espectro estendido (ESBL) e produtoras de carbapenemase (PC). Após o isolamento e identificação dos isolados, foram realizadas reações de PCR multiplex para detectar os principais genes de cada microorganismo e testes de susceptibilidade a antimicrobianos com os principais antibióticos utilizados para cada grupo bacteriano de acordo com o CLSI. Dos 39 locais do VCH coletados, todos (100%) possuíam pelo menos um dos microrganismos pesquisados e 17,95% (n=7) dos locais foram capazes de isolar os quatro patógenos. Dos 94 pools coletados, foi possível isolar MRS em 81,91% (n=77), VRE em 12,77% (n=12), ESBL em 62,77% (n=59) e carbapenemases em 24,47% (n=23). Em relação ao MRS, o gene mecA foi detectado em todos os isolados. Todos os VREs isolados foram identificados como Enterococcus faecalis e apresentaram o gene vanA. Em relação às cefalosporinases e/ou ESBL, 89,83% (n=53) dos isolados apresentaram o gene blaTEM, 57,63% (n=34) o gene blaOXA-1, 37,29% (n=22) o gene blaCTX-M de algum grupo e 20,34% (n=12) o gene blaSHV. Foi possível identificar os principais microrganismos responsáveis por causar infecções nosocomiais em humanos (VRE, MRS, ESBL e CP) no ambiente hospitalar veterinário, sugerindo uma fonte de infecção para profissionais e estudantes de medicina veterinária, colocando alto risco para a saúde pública.(AU)

Staphylococcus , Cross Infection , Methicillin Resistance , Enterococcus faecalis , Multiplex Polymerase Chain Reaction , Anti-Infective Agents , Anti-Bacterial Agents , beta-Lactamases , Hospitals, Animal
Dent. press endod ; 10(3): 36-42, Sept-Dec.2020. Ilus
Article in English | LILACS | ID: biblio-1344767


Introdução: Pinos intrarradiculares são recomendados para melhorar a retenção de coroas artificiais e distribuir forças intrabucais ao longo da raiz. Se o espaço criado pela remoção parcial da obturação não for preenchido adequadamente, pode ocorrer uma infiltração maciça de microrganismos da cavidade bucal. Objetivo: O objetivo do presente estudo foi avaliar a eficácia contra a infiltração bacteriana de uma barreira intracanal colocada diretamente sobre o remanescente da obturação do canal radicular, após o preparo de espaço para o pino. Métodos: Setenta e dois dentes humanos unirradiculares foram instrumentados, obturados e divididos aleatoriamente em três grupos experimentais e dois grupos controle. O grupo 1 não recebeu tratamento adicional após a obturação e o preparo do espaço, enquanto os Grupos 2 e 3 receberam uma barreira composta por material de selamento temporário com 1,0 e 2,0 mm de espessura, respectiva- mente. Uma cultura de Enterococcus faecalis foi inoculada nos espaços preparados para receber o pino intrarradicular, a cada três dias, por um período de 60 dias. A infiltração foi avaliada diariamente pela turbidade do meio de cultura. Resultados e Conclusão: Houve infiltração bacteriana nos três grupos experimentais, sendo significativamente maior e ocorrendo mais rapidamente no G1 (p<0,05), em comparação aos G2 e G3. Houve infiltração em todos os grupos controles positivos; porém, nenhuma infiltração foi observada nos grupos controles negativos. Não foi encontrada diferença significativa (p>0,05) entre o G2 e o G3, em relação à taxa e ao período de infiltração. Pode-se concluir que a barreira reduziu a incidência de infiltração e atrasou o tempo de ocorrência (AU).

Introduction: Intraradicular posts are recommended to improve the retention of artificial crowns and to distribute intraoral forces along the root. If the space created by partially removing the obturation is not adequately filled, it can lead to a massive infiltration of microorganisms from the oral cavity. Aim: The aim of this study was to assess the influence on bacterial infiltration of an intracanal barrier placed directly over the remaining root canal filling following post space preparation. Material and Methods: Seventy-two human single-rooted teeth were instrumented, filled, and then randomly divided into three experimental groups and two control groups. Group 1 received no additional treatment after filling and post space preparation, whereas Groups 2 and 3 received a barrier composed of temporary sealing material 1.0- and 2.0-mm thick, respectively. A culture of Enterococcus faecalis was inoculated into the spaces prepared to receive the intraradicular post, every three days over a 60-day period. Infiltration was evaluated daily by observing the turbidity of the culture medium. Results and Conclusion: Bacterial infiltration was found in all three experimental groups, but was significantly greater and occurred more quickly in G1 (p < 0.05), compared with G2 and G3. Infiltration was observed in all positive controls, whereas no infiltration was found in the negative controls. No significant difference (p > 0.05) was found between G2 and G3, regarding the rate and time frame of infiltration. It can be concluded that the barrier reduced the incidence of infiltration, and delayed the time of occurrence (AU).

Humans , Root Canal Obturation , Enterococcus faecalis , Dental Pins , Efficacy , Crowns
Dent. press endod ; 10(3): 43-48, Sept-Dec.2020. Tab
Article in English | LILACS | ID: biblio-1344778


Introdução: A eliminação do Enterococcus faecalis dos canais radiculares é fundamental para o sucesso do tratamento endodôntico, uma vez que esses microrganismos são de difícil eliminação, principalmente quando organizados em forma de biofilmes. A busca por drogas ou suas combinações que possam eliminar esses microrganismos é um dos principais objetivos terapêuticos. Objetivo: O presente estudo avaliou a ação antimicrobiana de medicações intracanal experimentais sobre biofilmes de Enterococcus faecalis. Métodos: Quarenta dentes bovinos unirradiculares foram utilizados; suas coroas foram seccionadas e as raízes foram instrumentadas e esterilizadas. As raízes foram contaminadas com suspensão contendo Enterococcus faecalis, mantidas em estufa a 37°C por 30 dias e divididas em quatro grupos, de acordo com a medicação intracanal: GI) medicação experimental 1 (clorexidina [CHX] 0,2%/metronidazol, doxiciclina); GII) medicação experimental 2 (CHX 0,2%/metronidazol, minociclina); GIII) clorexidina a 2% (CHX 2%); e GIV) solução salina. As raízes foram seladas e mantidas em estufa por 7 dias, em tubos contendo TSB. Dentina foi coletada e semeada por 24 h, para formação de UFCs. Os valores obtidos foram comparados pelos testes ANOVA e Tukey (p<0,05). Quando comparados os resultados, não houve diferenças entre os Grupos I, II e III; no entanto, eles foram significativamente diferentes do Grupo IV. Conclusão: As medicações intracanal experimentais exerceram ação antimicrobiana sobre biofilmes de Enterococcus faecalis (AU).

Background: The elimination of Enterococcus faecalis of the root canals is fundamental for endodontic success, since these microorganisms are difficult to killed, especially when organized in biofilms. The search for drugs or their combinations that can eliminate these microorganisms is one of the main therapeutic aim. This study evaluated the antimicrobial action of experimental intracanal medications on Enterococcus faecalis biofilms. Methods: Forty uniradicular bovine teeth were used; their crowns were removed, and the roots were instrumented and sterilized. The roots were contaminated with suspension containing Enterococcus faecalis and kept in an oven at 37°C for 30 days. The roots were divided into 4 groups according to the intracanal medication: I- experimental medication 1 (0.2% CHX/metronidazole/ doxycycline); II- experimental medication 2 (0.2% CHX/ metronidazole/minocycline), III- 2% chlorhexidine (2% CHX), and IV- saline solution. The roots were sealed and kept in tubes containing TSB in an oven for 7 days. Dentin was collected and seeded for 24 h for perform of CFUs. The values obtained were compared using ANOVA and Tukeys tests (p<0.05). When comparing the results, there were no differences among groups I, II and III; however, they were significantly different from group IV. Conclusion: The experimental intracanal medications exerted an antimicrobial action on Enterococcus faecalis biofilms (AU).

Animals , Cattle , In Vitro Techniques , Enterococcus faecalis , Biofilms , Anti-Infective Agents , Methods
Int. j. odontostomatol. (Print) ; 14(4): 632-638, dic. 2020. tab, graf
Article in English | LILACS | ID: biblio-1134550


ABSTRACT: The aim of this in vitro study was to investigate the influence of ethylenediaminetetraacetic acid (EDTA) associated with the benzalkonium chloride (BAK) on the adhesion and formation of Enterococcus faecalis biofilms attached to coated dentin. Discs standard bovine dentin blocks were treated with the coating materials evaluated: Saline solution (control), 17 % EDTA, 17 % EDTA associated with 1 % BAK for 5 minutes and subsequently washed with saline solution. Afterwards, biofilms of E. faecalis (ATCC 29212) were grown on the surface of coated dentin blocks for time intervals of 1 hour and 7 days (n = 20) and were subsequently washed with phosphate-buffered saline (PBS). Bacterial viability and total biovolume were analyzed by confocal laser scanning microscopy (CLSM) using the Live/Dead technique. Nonparametric Kruskal-Wallis followed by Dunn tests were used to determine statistical differences (a = 5 %). The 17 % EDTA + 1 % BAK group showed significantly lower biovolume and bacterial viability values at the end of 1 hour (p < 0.05). After 7 days of contamination, the 17 % EDTA and 17 % EDTA + 1 % BAK groups showed similar results that differed statistically from those of the control group (p < 0.05). The saline solution group showed higher values. The use of BAK associated with EDTA on dentin blocks surfaces before exposure to contamination was able to interfere in the adhesion of E. faecalis to dentin. Also, dentin treatment by BAK associated with a chelating agent influences the secondary biofilm formation, which could have important effects on the long-term success of root canal treatment.

RESUMEN: El objetivo del estudio consistió en investigar in vitro, la influencia del ácido etilendiamino-tetraacético (EDTA) con cloruro de benzalconio (BAK) en la adhesión y formación de biopelículas de Enterococcus faecalis a la dentina. Discos de dentina bovina fueron tratadas con solución salina (control), 17 % de EDTA, 17% de EDTA asociado con 1 % de BAK durante 5 minutos y lavadas con solución salina. Las biopelículas de E. faecalis (ATCC 29212) se cultivaron sobre los discos de dentina durante intervalos de tiempo de 1 hora y 7 días (n = 20), lavados con solución salina tamponada con fosfato (PBS). La viabilidad bacteriana y el biovolumen total se analizaron mediante microscopía de barrido por láser (CLSM) utilizando la técnica Live / Dead. Se realizó prueba no paramétrica de Kruskal-Wallis, seguida por Dunn con una diferencia estadística (a = 5 %). El grupo de 17 % EDTA + 1 % BAK mostró valores significativamente menores de biovolumen y viabilidad bacteriana al final de 1 hora (p < 0,05). Después de 7 días de contaminación, los grupos de 17 % EDTA y 17 % EDTA + 1 % BAK mostraron resultados similares que diferían estadísticamente del grupo control (p < 0,05). La solución salina mostró valores más altos. La asociación de BAK con EDTA antes de la contaminación interfirió en la adhesión de E. faecalis. Además, el tratamiento de la dentina por BAK asociado con EDTA influye en la formación de biopelículas secundarias, lo que podría tener efectos importantes sobre el éxito a largo plazo del tratamiento del conducto radicular.

Animals , Cattle , Bacterial Adhesion/drug effects , Edetic Acid/pharmacology , Enterococcus faecalis/growth & development , Enterococcus faecalis/drug effects , Biofilms/drug effects , Dentin/microbiology , Benzalkonium Compounds/pharmacology , Statistical Analysis , Microscopy, Confocal , Saline Solution
Braz. dent. j ; 31(6): 611-616, Nov.-Dec. 2020. tab, graf
Article in English | LILACS, BBO | ID: biblio-1132345


Abstract The aim of this study was to evaluate the antibacterial potential of a calcium silicate-based sealer (Bio-C Sealer, Angelus) against common bacteria in primary and secondary endodontic infections. Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Streptococcus mutans were exposed to fresh Bio-C Sealer for 24 h by the agar diffusion method (n=5). Additionally, the antibacterial activity was investigated against E. faecalis and S. mutans biofilms (48 h old) grown in discs with 4 mm in diameter and 2 mm in height. (n=3) of set discs of Bio-C Sealer (Angelus), EndoFill (Dentsply-Mallefer), Sealer 26 (Dentsply), AH Plus (Dentsply), Sealapex (Sybron-Endo) and EndoSequence BC Sealer (Brasseler). The antibacterial activity was evaluated by colony forming unity (CFU) counting using ImageJ software. Data were compared by one-way ANOVA followed by Holm-Sidak test (a=5%). Fresh Bio-C Sealer exhibited antimicrobial activity against all bacteria evaluated by agar diffusion method, except for S. mutans. Set discs of all endodontic sealers tested showed similar CFU values for E. faecalis (p>0.05). S. mutans in biofilms showed higher susceptibility to EndoFill compared with the other sealers (p<0.05). In conclusion, the results indicate that fresh Bio-C Sealer does not inhibit S. mutans growth, but exhibits antibacterial activity against E. faecalis, S. aureus, P. aeruginosa and E. coli. After setting, the Bio-C Sealer exhibits an antimicrobial potential comparable to that of the other sealers evaluated in E. faecalis biofilm, but lower than that of EndoFill for S. mutans biofilm.

Resumo O objetivo deste estudo foi avaliar o potencial antibacteriano do novo cimento biocerâmico (Bio-C Sealer, Angelus) contra bactérias comuns em infecções endodônticas primárias e secundárias. Culturas de Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus e Streptococcus mutans foram expostos a amostras frescas do Bio-C sealer durante 24 h pelo método de difusão em agar (n=5). A atividade antibacteriana de amostras dos cimentos Bio-C Sealer (Angelus), EndoFill (Dentsply-Mallefer), Sealer 26 (Dentsply), AH Plus (Dentsply), Sealapex (Sybron-Endo) e EndoSequence BC Sealer (Brasseler) após a presa também foi investigada em biofilmes de 48 h das bactérias E. faecalis e S. mutans, crescidos em discos com 4 mm de diâmetro e 2 mm de altura. A atividade antibacteriana foi avaliada por contagem das unidades formadoras de colônias (UFC) utilizando o software ImageJ. Os dados foram comparados por ANOVA a um critério seguido pelo pós-teste Holm-Sidak (a=5%). Amostras frescas do Bio-C Sealer exibiram atividade antimicrobiana contra todas as bactérias avaliadas pelo método de difusão em ágar, exceto para S. mutans. A análise da formação de biofilme mostrou que todos os cimentos endodônticos testados apresentaram valores similares de UFC para E. faecalis (p> 0,05), enquanto biofilmes de S. mutans foram mais suscetíveis ao EndoFill em comparação com os demais cimentos (p<0,05). Conclui-se que o cimento Bio-C Sealer fresco exibe atividade antibacteriana para E. faecalis, S. aureus, P. aeruginosa e E. coli, mas não inibe o crescimento de S. mutans. Após a presa, o cimento Bio-C Sealer exibe potencial antimicrobiano similar ao dos demais cimentos avaliados em biofilme de E. faecalis, mas inferior ao do EndoFill para S. mutans.

Root Canal Filling Materials/pharmacology , Epoxy Resins , Staphylococcus aureus , Materials Testing , Enterococcus faecalis , Silicates/pharmacology , Calcium Compounds , Escherichia coli , Anti-Bacterial Agents/pharmacology
Rev. Asoc. Odontol. Argent ; 108(2): 46-51, mayo-ago. 2020. tab
Article in Spanish | LILACS | ID: biblio-1121108


Objetivos: Comparar ex vivo la eficacia del instrumento XP-endo Finisher y del sistema EndoActivator en la reducción/eliminación del biofilm microbiano en conductos radiculares infectados. Materiales y métodos: Se utilizaron 23 premolares inferiores humanos extraídos cuya longitud fue estandarizada en 17 mm. Todos los conductos se prepararon con el sistema WaveOne Gold Medium (#35.06). Los dientes se esterilizaron, se inocularon con Enterococcus faecalis y se separaron en dos grupos experimentales de 10 piezas cada uno. De los 3 dientes remanentes, 1 fue utilizado como control positivo y 2, como controles negativos. En el grupo 1, las soluciones irrigantes se agitaron con XP-endo Finisher. En el grupo 2, se utilizó EndoActivator. Se tomaron muestras antes de la contaminación, luego de esta y después de la agitación de los irrigantes mediante conos de papel estériles. La carga microbiana fue sembrada en agar sangre y los conos se cultivaron en caldo tripteína de soja. La remoción de la carga microbiana se determinó por la presencia o ausencia de turbiedad del medio. Las unidades formadoras de colonias (UFC) remanentes se cuantificaron y los resultados se categorizaron como R1 (≤10 UFC) o R2 (>10 UFC). Los datos fueron analizados mediante la prueba de Fisher. Resultados: No hubo diferencias significativas entre XP-endo Finisher y EndoActivator (P>0,05). El número de usos no influyó sobre la capacidad operativa de ambos instrumentos (AU)

Aim: To compare ex vivo the effectiveness of the XP-endo Finisher and the EndoActivator in biofilm reduction/ removal from infected root canals. Materials and methods: Twenty three extracted human single-rooted lower premolars were selected and standardised to 17 mm in length. All the canals were prepared with WaveOne Gold Medium reciprocating files (#35.06). The teeth were autoclaved and inoculated with Enterococcus faecalis. The infected teeth were then assigned to 2 experimental groups of 10 teeth each according to the final irrigation/agitation protocol. Of the three remaining teeth, one was used as a positive control, and the other two were used as negative controls. In Group 1 the irrigating solutions were agitated with XP-endo Finisher while in Group 2 the EndoActivator was used. All root canals were sampled before and after contamination, and again after irrigant agitation with sterile paper points. The microbial load was spread on blood agar plates and the paper points were cultured in sterile trypticase soy broth. The removal of the microbial load was determined by visual observation of the turbidity of the media and by quantification of the number of colony-forming units (UFC). The results were categorized as R1 (≤10 UFC) or R2 (>10 UFC). Data were analysed by the Fisher's exact test at P<0.05. Results: No significant differences was found between XP-endo Finisher and EndoActivator (P>0.05) regarding their effectiveness in the reduction/removal of the microbial biofilm. The number of uses of both instruments did not affect their operative performance (AU) Conclusion: XPF and EA were both equally effective for microbial biofilm reduction/removal from ex vivo infected root canals (AU)

Root Canal Irrigants/chemistry , Dental High-Speed Equipment , Biofilms , Dental Instruments , Dental Pulp Cavity/microbiology , In Vitro Techniques , Colony Count, Microbial/methods , Efficacy , Statistical Analysis , Enterococcus faecalis/isolation & purification , Culture Media
Braz. j. biol ; 80(2): 393-404, Apr.-June 2020. tab
Article in English | LILACS | ID: biblio-1132371


Abstract This paper reports the in vitro antiproliferative effects, antiprotozoal, anti-herpes and antimicrobial activities of 32 organic extracts of 14 marine sponges and 14 corals collected in northeast Brazilian coast. The ethanolic extracts of the sponges Amphimedon compressa and Tedania ignis, and the acetone extract of Dysidea sp. showed relevant results concerning the antiproliferative effects against A549, HCT-8, and PC-3 cell lines by sulforhodamine B assay, but also low specificity. Concerning the antiprotozoal screening, the ethanolic extract of Amphimedon compressa and the acetone and ethanolic extracts of Dysidea sp. were the most active against Leishmania amazonensis and Trypanosoma cruzi expressing β-galactosidase in THP-1 cells. In the preliminary anti-HSV-1 (KOS strain) screening, the ethanolic extracts of the sponges Amphimedon compressa, Haliclona sp. and Chondrosia collectrix inhibited viral replication by more than 50%. The most promising anti-herpes results were observed for the ethanolic extract of Haliclona sp. showing high selective indices against HSV-1, KOS and 29R strains (SI> 50 and >79, respectively), and HSV-2, 333 strain (IS>108). The results of the antibacterial screening indicated that only the ethanolic extract of Amphimedon compressa exhibited a weak activity against Enterococcus faecalis, Pseudomonas aeruginosa and Escherichia coli by the disk diffusion method. In view of these results, the extracts of Amphimedon compressa, Tedania ignis and Dysidea sp. were selected for further studies aiming the isolation and identification of the bioactive compounds with antiproliferative and/or antiprotozoal activities. The relevant anti-herpes activity of the ethanolic extract of Haliclona sp. also deserves special attention, and will be further investigated.

Resumo Este artigo reporta as atividades in vitro antiproliferativa, atiprotozoárica, anti-herpética e antimicrobiana de 32 extratos orgânicos provenientes de 14 esponjas marinhas e 14 corais coletados no litoral nordestino brasileiro. Os extratos etanólicos das esponjas Amphimedon compressa e Tedania ignis, e o extrato acetônico de Dysidea sp. demonstraram resultados promissores em relação aos efeitos antiproliferativos frente as linhagens celulares A549, HCT-8, PC-3 pelo método da sulforrodamina B, mas sem especificidade. Em relação à atividade antiprotozárica, os extratos etanólico de Amphimedon compressa e acetônico e etanólico de Dysidea sp. apresentaram atividade contra Leishmania amazonensis e Trypanosoma cruzi através do método de expressão de β-galactosidase em células THP-1. Na investigação preliminar de atividade antiviral frente ao vírus Herpes simplex tipo 1 (cepa KOS), os extratos etanólicos das esponjas Amphimedon compressa, Haliclona sp. e Chondrosia collectrix inibiram mais de 50% da replicação viral. O extrato etanólico da esponja Haliclona sp. demonstrou resultados promissores para atividade anti-herpética com altos índices de seletividade para as cepas KOS (IS >50) e 29R (IS>79) frente ao VHS-1 e cepa 333 (IS>108) frente ao VHS-2. O extrato etanólico da esponja Amphimedon compressa exibiu uma pequena atividade contra Enterococcus faecalis, Pseudomonas aeruginosa and Escherichia coli pelo método de difusão em disco. De acordo com os resultados apresentados, os extratos das esponjas Amphimedon compressa, Tedania ignis e Dysidea sp. serão selecionados para futuros estudos de isolamento e identificação dos compostos bioativos para as atividades antiproliferativa e antiprozoárica. O extrato etanólico de Haliclona sp. será investigado por possuir atividade relevante anti-herpética.

Animals , Porifera , Brazil , Plant Extracts , Enterococcus faecalis , Anti-Bacterial Agents
Biosci. j. (Online) ; 36(2): 568-577, 01-03-2020. tab
Article in English | LILACS | ID: biblio-1146421


In the Myrtaceae family, the species Eugenia involucrata DC., popularly known as "cerejeira-do-mato", is traditionally used for the antidiarrheal and digestive action of its leaves. However, no studies were found in the literature regarding its antimicrobial and antioxidant potential. In this context, the objective of the present study was to determine the chemical composition by gas chromatography coupled to mass spectrometry (GC-MS) to evaluate the antimicrobial activity by the broth microdilution technique and the antioxidant activity by the 2,2-diphenyl-1-picryl-hydrazila (DPPH) method of the essential oil of E. involucrataleaves. GC-MS identified 28 compounds, all sesquiterpenes, corresponding to 89.41% of the essential oil. The antimicrobial activity of the essential oil was observed for all Gram-positive bacteria tested (Staplylococcus epidermidis, Enterococcus faecalis,Bacillus subtilis and Staplylococcus aureus) and for yeast Candida albicans. The essential oil presented a reduction capacity of DPPH up to 66.81%, evidencing its antioxidant potential. It is suggested that the antimicrobial and antioxidant action of E. involucrata essential oil is related to the presence of the major compounds, elixene (26.53%), ß-caryophyllene (13.16%), α-copaene (8.41%) and germacrene D (7.17%).

Na família Myrtaceae, a espécieEugenia involucrata DC. popularmente denominada "cerejeira-do-mato" é conhecida tradicionalmente pela ação antidiarreica e digestiva de suas folhas. Contudo, na literatura não foram encontrados trabalhos referentes ao seu potencial antimicrobiano e antioxidante. Neste contexto, o objetivo do presente estudo foi determinar a composição química por cromatografia gasosa acoplada a espectrometria de massas (CG-EM) e avaliar a atividade antimicrobiana pela técnica de microdiluição em caldo e a atividade antioxidante pelo método do 2,2-difenil-1-picril-hidrazila (DPPH) do óleo essencial das folhas de E. involucrata. A CG-EM identificou 28 compostos, todos sesquiterpenos, correspondendo a 89,41% do óleo essencial. A atividade antimicrobiana do óleo essencial foi observada para todas as bactérias Gram-positivas testadas (Staplylococcus epidermidis, Enterococcus faecalis,Bacillus subtilise Staplylococcus aureus) e para a levedura Candida albicans. O óleo essencial apresentou capacidade redutora de radicais DPPH de até 66,81%, evidenciando sua potencialidade antioxidante. Sugere-se que a ação antimicrobiana e antioxidante do óleo essencial de E. involucrata esteja relacionada à presença dos compostos majoritários, elixeno (26,53%), ß-cariofileno (13,16%), -copaeno (8,41%) e germacreno D (7,17%).

Myrtaceae , Eugenia , Anti-Infective Agents , Antioxidants , Mass Spectrometry , Bacillus , Bacteria , Biological Products , Candida albicans , Oils , Distillation , Chromatography, Gas , Enterococcus faecalis , Volatile Organic Compounds
J. oral res. (Impresa) ; 9(1): 36-48, feb. 28, 2020. ilus, graf
Article in English | LILACS | ID: biblio-1151442


Endodontic treatment consists of the cleaning and disinfecting the root canal system, which is achieved using adequate mechanical instru-mentation and chemical irrigation. Endodontic microorganisms are present in root canals in the form of a biofilm, and their elimination ensures the success of endodontic treatment. Irrigation is a key factor contributing to the elimination of this intraconduct biofilm, and different irrigator agents and irrigation techniques, such as irrigation with negative apical pressure, a novel automated irrigation mechanism based on suction intraconduct, have been used. In this study, we evaluated the ability of a negative apical pressure system with different concentrations of sodium hypochlorite and durations to reduce the microbial load. Materials and Methods: An intraradicular biofilm composed of Enterococcus faecalis and Candida albicans was generated during twenty-one days of static culture on one hundred mesio-vestibular roots of upper molars with complex curvatures greater than 30°C, and the roots were classified in six groups with different concentrations and contact times of sodium hypochlorite. Subsequently, the reduction in the microbial load was measured with McFarland scale and the enumeration of colony forming units and was evaluated with scanning electronic microscopy. Results: We observed a significant difference in the reduction of the microbial load prior to instrumentation compared with postinstrumentation between the groups treated with 2.25% and 5.25% NaOCl for 30, 60 and 90 seconds of contact time (p<0.05), but we did not observe differences in the reduction of microbial load between different contact times and concentrations of sodium hypochlorite employed (p>0.05). Conclusion: Negative apical pressure is a good option for irrigation in endodontics, as it allows the passage of the irrigation fluid along the total length of the root canal and produces a better antimicrobial effect.

El tratamiento de endodoncia consiste en la limpieza y desinfección del sistema de conducto radicular, lo que se logra utilizando instrumentación mecánica adecuada y riego químico. Los microorganismos endodónticos están presentes en los conductos radiculares en forma de una biopelícula, y su eliminación asegura el éxito del tratamiento endodóntico. La irrigación es un factor clave que contribuye a la eliminación de esta biopelícula intraconductora, y se han utilizado diferentes agentes irrigadores y técnicas de irrigación, como la irrigación con presión apical negativa, un nuevo mecanismo automatizado de irrigación basado en la intraconducción de succión. En este estudio, evaluamos la capacidad de un sistema de presión apical negativa con diferentes concentraciones de hipoclorito de sodio y duraciones para reducir la carga microbiana. Material y Métodos: Se generó una biopelícula intraradicular compuesta por Enterococcus faecalisy Candida albicans durante veintiún días de cultivo estático en cien raíces mesio-vestibulares de molares superiores con curvaturas complejas superiores a 30°C, y las raíces se clasificaron en seis grupos con diferentes concentraciones y tiempos de contacto de hipoclorito de sodio. Posteriormente, la reducción en la carga microbiana se midió con la escala de McFarland y la enumeración de las unidades formadoras de colonias y se evaluó con microscopía electrónica de barrido. Resultado: Observamos una diferencia significativa en la reducción de la carga microbiana antes de la instrumentación en comparación con la postinstrumentación entre los grupos tratados con NaOCl 2.25% y 5.25% durante 30, 60 y 90 segundos de tiempo de contacto (p<0.05), pero lo hicimos No se observan diferencias en la reducción de la carga microbiana entre los diferentes tiempos de contacto y las concentraciones de hipoclorito de sodio empleado (p>0.05). Conclusión:La presión apical negativa es una buena opción para el riego en endodoncia, ya que permite el paso del líquido de riego a lo largo de todo el conducto radicular y produce un mejor efecto antimicrobiano.

Humans , Root Canal Therapy/methods , Candida albicans/physiology , Candidiasis , Enterococcus faecalis/physiology , Biofilms , Sodium Hypochlorite , In Vitro Techniques , Endodontics , Therapeutic Irrigation
Int. j. odontostomatol. (Print) ; 14(3): 448-456, 2020. tab, graf
Article in English | LILACS | ID: biblio-1114920


Enterococci are important nosocomial pathogens due to their intrinsic multiresistance and the acquisition of new antibiotic resistance genes (ARG). Enterococcus faecalis has been shown to be one of the main pathogens in persistent endodontic infections, therefore, the main objective of this study was to evaluate the phenotype and resistance genotype of strains of E. faecalis isolated from teeth with persistent endodontic lesions, to the most commonly prescribed antibiotics in dentistry. Thirteen strains of E. faecalis of different pulsotype were analyzed to evaluate the susceptibility to antibiotics, amoxicillin, amoxicillin/clavulanic acid, tetracycline, erythromycin and metronidazole, using the Epsilometer test (E- test) and the presence of beta-lactamases with nitrocefin test. Finally, the detection of ARG was performed with a molecular polymerase chain reaction (PCR) technique and confirmed by the sequencing of the amplification products. Fisher's exact test was used, using 95 % confidence. Regarding the phenotype of resistance, the evaluated strains, independent of the pulsotype, were totally resistant to the action of metronidazole. Antibiotics with higher minimum inhibitory concentration (MIC) after metronidazole include tetracycline and erythromycin. In contrast, lower MIC are applied to the combination of amoxicillin with clavulanic acid. The nitrocefin test was positive only in one strain. Genotypically, two genetically distant strains isolated from a single patient, presented a genotype of resistance to erythromycin, determined by the presence of the ermB gene. No statistically significant relationship was found between phenotypic resistance and the presence of ARG in relation to erythromycin (p> 0.05). It was concluded that isolates of E. faecalis from persistent endodontic infections showed phenotypes of resistance to several antimicrobial agents, all of which were susceptible to amoxicillin/clavulanic acid. Periodic evaluation of susceptibility to antibiotics is suggested as an important practice for the surveillance of antibiotic resistance in oral strains.

Los enterococos son importantes patógenos nosocomiales debido a su multi resistencia intrínseca y la adquisición de nuevos genes de resistencia a los antibióticos (ARG). Enterococcus faecalis es uno de los principales patógenos en infecciones endodónticas persistentes, por lo tanto, el objetivo principal de este estudio fue evaluar el fenotipo y el genotipo de resistencia de cepas de E. faecalis aisladas de dientes con lesiones endodóncicas persistentes, a los antibióticos comúnmente recetados en odontología. Se analizaron 13 cepas de E. faecalis de diferentes pulsotipos para evaluar la susceptibilidad a los antibióticos, amoxicilina, amoxicilina / ácido clavulánico, tetraciclina, eritromicina y metronidazol, utilizando la prueba de Epsilometría (E-test) y la presencia de beta-lactamasas con prueba de nitrocefina. Finalmente, la detección de ARG se realizó con una técnica molecular de reacción en cadena de la polimerasa (PCR) y se confirmó mediante la secuenciación de los productos de amplificación. Se utilizó la prueba exacta de Fisher, con un 95 % de confianza. En cuanto al fenotipo de resistencia, las cepas evaluadas, independientes del pulsotipo, fueron totalmente resistentes a la acción del metronidazol. Los antibióticos con los valores más altos de concentración mínima inibitoria (CMI) después del metronidazol incluyen tetraciclina y eritromicina. En contraste, las CMI mas bajas se aplican a la combinación de amoxicilina con ácido clavulánico. La prueba de nitrocefina fue positiva solo en una cepa. Genotípicamente, dos cepas distantes genéticamente, aisladas de un mismo paciente fueron positivas para el gen ermB. No se encontró una relación estadísticamente significativa entre la resistencia fenotípica y la presencia de ARG en relación con la eritromicina (p> 0,05). Se concluyó que los aislamientos de E. faecalis de infecciones endodónticas persistentes mostraron fenotipos de resistencia a varios agentes antimicrobianos, todos los cuales fueron susceptibles a amoxicilina / ácido clavulánico. Se sugiere una evaluación periódica de la susceptibilidad a los antibióticos como una práctica importante para la vigilancia de la resistencia a los antibióticos en las cepas orales.

Humans , Enterococcus faecalis/drug effects , Enterococcus faecalis/genetics , Dental Pulp Cavity/microbiology , Anti-Bacterial Agents/pharmacology , Tetracycline , Microbial Sensitivity Tests , Erythromycin , Polymerase Chain Reaction , Clavulanic Acid/pharmacology , Drug Resistance, Bacterial/genetics , Amoxicillin/pharmacology , Metronidazole