Antibacterial Activity of Homeopathic Medications Lycopodium clavatum and Arsenicum album Against Periodontal Bacteria / Evaluación antibacteriana de los medicamentos homeopáticos sobre bacterias periodontales

Abstract There are several controversies regarding the efficacy of homeopathic substances; however, these remedies are used in many countries for the treatment of various pathological conditions. The purpose of this study was to evaluate the in vitro antibacterial activity of two homeopathic tinctures Arsenicum album (mineral extract) and Lycopodium clavatum (plant extract) on the periodontal bacteria Actinomyces israelii, Streptococcus sanguinis, Prevotella intermedia, Aggregatibacter actinomycetemcomitans and Phorphyromonas gingivalis (P. gingivalis). Materials and methods: Equal numbers of bacteria were seeded on agar plates containing enriched media with the homeopathic solutions at 1dH and 1cH dilutions. After 7 days of incubation under anaerobic conditions, colony forming units (CFUs) were counted. The antibacterial effect was calculated based on the total number of CFUs observed on non-tincture containing agar, and on the tincture containing plates. Results: No visible growth of any of the strains was observed on the plates containing Arsenicum album at any of the dilutions tested. In contrast, when Lycopodium clavatum at 1cH dilution was tested, only P. gingivalis was susceptible to this compound. Conclusions: The results suggest that the mineral extract tincture had a greater antibacterial activity than the plant extract tincture, also Lycopodium clavatum preparation could be an effective inhibitor of periodontal pathogens bacteria such as P. gingivalis.

Resumen Se necesita un mayor número de estudios in vitro e in vivo para validar estos resultados.

Effect of TiO2 nanoparticles incorporation on antibacterial properties and shear bond strength of dental composite used in Orthodontics

ABSTRACT Introduction: Plaque accumulation and bond failure are drawbacks of orthodontic treatment, which requires composite for bonding of brackets. As the antimicrobial properties of TiO2 nanoparticles (NPs) have been proven, the aim of this study was to evaluate the antimicrobial and mechanical properties of composite resins modified by the addition of TiO2 NPs. Methods: Orthodontics composite containing 0%, 1%, 5% and 10% NPs were prepared. 180 composite disks were prepared for elution test, disk agar diffusion test and biofilm inhibition test to collect the counts of microorganisms on three days, measure the inhibition diameter and quantify the viable counts of colonies consequently. For shear bond strength (SBS) test, 48 intact bovine incisors were divided into four groups. Composites containing 0%, 1%, 5% and 10% NPs were used for bonding of bracket. The bracket/tooth SBS was measured by using an universal testing machine. Results: All concentration of TiO2 NPs had a significant effect on creation and extension of inhibition zone. For S. mutans and S. sanguinis, all concentration of TiO2 NPs caused reduction of the colony counts. Composite containing 10% TiO2 NPs had significant effect on reduction of colony counts for S. mutans and S. sanguinis in all three days. The highest mean shear bond strength belonged to the control group, while the lowest value was seen in 10% NPs composite. Conclusions: Incorporating TiO2 nanoparticles into composite resins confer antibacterial properties to adhesives, while the mean shear bond of composite containing 1% and 5% NPs still in an acceptable range.

RESUMO Introdução: o acúmulo de placa e as descolagens de braquetes são algumas desvantagens presentes no tratamento ortodôntico, no qual se requer o uso de materiais compósitos para a colagem dos braquetes. Objetivo: tendo em vista que as propriedades antimicrobianas das nanopartículas (NPs) de TiO2 já foram confirmadas, o objetivo do presente estudo foi avaliar as propriedades antimicrobianas e mecânicas de resinas compostas modificadas pela adição de NPs de TiO2. Métodos: compósitos ortodônticos contendo 0%, 1%, 5% e 10% de NPs foram preparados. Cento e oitenta discos de compósito foram preparados para o teste de eluição, o ensaio de difusão em ágar por disco, e o ensaio de inibição da formação de biofilme, para se calcular as contagens de microrganismos ao longo de três dias, medir o diâmetro da inibição e, consequentemente, quantificar as contagens de colônias viáveis. Para o teste de resistência da colagem ao cisalhamento (SBS), 48 incisivos bovinos intactos foram divididos em quatro grupos, nos quais os compósitos contendo 0%, 1%, 5% e 10% de NPs foram utilizados para colagem dos braquetes. A SBS da interface braquete/dente foi medida em uma máquina universal de ensaios. Resultados: todas as concentrações de NPs de TiO2 apresentaram efeito significativo na formação e na extensão da zona de inibição. Para o S. mutans e o S. sanguinis, todas as concentrações de NPs de TiO2 causaram redução na contagem das colônias. O compósito contendo 10% de NPs de TiO2 apresentou uma diminuição significativa na contagem de colônias de S. mutans e S. sanguinis durante os três dias. A média mais alta da SBS foi observada no grupo controle, enquanto o valor mais baixo foi observado para o compósito com 10% de NPs. Conclusões: a incorporação de nanopartículas de TiO2 nas resinas compostas lhes conferiu propriedades antibacterianas, e o valor médio da SBS das resinas contendo 1% e 5% de NPs apresentou-se dentro de uma faixa aceitável.

Influence of time, toothpaste and saliva in the retention of Streptococcus mutans and Streptococcus sanguinis on different toothbrushes

Objectives: The intraoral transmission of cariogenic and periodontopathogenic species seems to be facilitated by contaminated toothbrushes and other oral hygiene devices. The aim of this investigation was to analyze the in vitro retention and survival rate of Streptococcus mutans and Streptococcus sanguinis on different toothbrushes. The impacts of human saliva and antimicrobial toothpaste on these parameters were further evaluated. Material and Methods: Part I: Four toothbrushes (Colgate 360°, Curaprox CS5460 ultra soft, elmex InterX, Trisa Flexible Head3) were contaminated by S. mutans DSM 20523 or S. sanguinis DSM 20068 suspensions for three minutes. Bacteria were removed from the toothbrushes after either three minutes (T0) or 24 hours (T24) of dry storage and grown on Columbia blood agar plates for the quantification of colony-forming units (CFUs). Part II: The effects of saliva from a caries-active or a caries-inactive person and of toothpaste containing 0.12% chlorhexidine digluconate were also tested. Results: Part I: After three minutes of dry storage, approximately one percent of the bacteria were still detectable on the toothbrushes. After 24 hours, S. sanguinis exhibited a more pronounced decrease in viable cell numbers compared with S. mutans but the differences were not significant (Kruskal-Wallis test, p>0.05). Part II: The addition of human saliva from a caries-active or caries-inactive person slightly increased the retention of both streptococcal species at T0. The use of toothpaste had no influence on the amount of viable streptococci at T0, but it reduced the microbial load after 24 hours of storage. There were only slight nonsignificant differences (p>0.05) between the four toothbrushes. Conclusions: In vitro bacterial retention and survival of S. sanguinis and S. mutans on different toothbrushes occurred. Within the limitations of this study, the use of human saliva or an antimicrobial toothpaste ...

In Vitro Effect of photodynamic therapy on Aggregatibacter actinomycetemcomitans and Streptococcus sanguinis

New periodontal disease treatments are needed to prevent infection progression. Photodynamic therapy (PDT) is one of the greatest pledges for this purpose. It involves the use of light of specific wavelength to activate a nontoxic photosensitizing agent in the presence of oxygen for eradication of target cells, and can be used for photokilling of microorganisms. This study evaluated in vitro the photodynamic effect of 0.01 percent toluidine blue-O (TBO) in combination with an AlGaInP diode laser light source on Aggregatibacter actinomycetemcomitans (A.a.) and Streptococcus sanguinis (S.s.). Suspensions (2 mL) containing A.a. and S.s. at 1.5 x 108 CFU/mL concentration were prepared and divided into 3 groups: Control group (no treatment), Dye group (inoculum and TBO for 5 min) and Dye/Laser group (inoculum, TBO for 5 min and laser for 3 min). Next, a dilution for subsequent subculture in 20 mL of Trypic Soy Agar (A.a) and Brucella Agar (S.s.) in Petri dishes (Pourplate Method) was done. Incubation of A.a. in microaerophilia and S.s. in aerobiosis at 35oC for 48 h was performed for subsequent visual counting of CFU/mL. Data were analyzed by one-way ANOVA and Tukey’s HSD test at 5 percent significance level. For both strains, the control group showed a significantly higher (p<0.05) bacterial growth (1.5 x 108 CFU/mL), while the Dye group presented no significant reduction (p>0.05) in the CFU counts. The Dye/Laser group presented a significant decrease in the CFU counts (p<0.05) compared with the Control group (61.53 percent for A.a. and 84.32 percent for S.s.). It may be concluded that PDT was effective in reducing the numbers of A.a. and S.s. in vitro.

Novos tratamentos são propostos para evitar a progressão da periodontite, sendo a terapia fotodinâmica (PDT) uma notória promessa. Sua aplicação associa o Azul de orto-toluidina a 0,01 por cento (TBO) e uma fonte luminosa a laser de diodo (TwinFlex, Mmoptics), liberando assim, toxinas às bactérias. O objetivo do estudo é avaliar in vitro a eficiência da PDT sobre Aggregatibacter actinomycetemcomitans (A.a.) e Streptococcus sanguinis (S.s.). Preparou-se suspensões distintas de 2 mL contendo A.a. e S.s. na concentração de 1,5x108 UFC/mL, e divisão de cada suspensão em 3 grupos: Controle (sem tratamento); Corante (suspensão e TBO por 5min) e Corante/Laser (suspensão, TBO por 5 min e laser por 3 min). Promoveu-se a diluição, a semeadura em 20 mL de TSA (A.a.) e de Ágar Brucella (S.s.), em placas de Petri (Método Pourplate), e a incubação da A.a. em microaerofilia e da S.s. em aerobiose, por 48 h a 35oC, para posterior contagem visual das UFC. Os grupos Controle mostraram ótimo crescimento bacteriano (1,5 x 108 UFC/mL). Os grupos Corante não apresentaram redução significativa para ambas bactérias. Os grupos Laser apresentaram redução em relação ao controle, 61,53 por cento para A.a. e 84,32 por cento para S.s. A análise estatística (ANOVA, p<0,05) corrobou que a PDT é eficaz na redução destas bactérias in vitro.

Antimicrobial effect and pH of chlorhexidine gel and calcium hydroxide alone and associated with other materials

The purposes of this study were to evaluate the effectiveness of 2 percent chlorhexidine (CHX) gluconate gel, calcium hydroxide [Ca(OH)2] and their combination with iodoform and zinc oxide powder as intracanal medications against select microorganisms, and to measure the pH changes caused by these medications. Antimicrobial activity was determined by the agar diffusion method. The zones of growth inhibition were measured and the results were analyzed statistically by Kruskal-Wallis test (p<0.05). The pH of the pastes was measured right after preparation, after 24 h and 1 week later. The largest mean zones of microbial inhibition were produced by 2 percent CHX gel, followed by Ca(OH)2 + 2 percent CHX gel + iodoform, Ca(OH)2 + 2 percent CHX gel, Ca(OH)2 + 2 percent CHX gel + zinc oxide, and Ca(OH)2 + water. The mean pH of all medications stayed above 12.0 during the whole experiment, except for CHX gel (pH=7.0). The results of this study showed that all medications had antimicrobial activity, but the most effective against the tested microorganisms were 2 percent CHX gel, followed by its combination with Ca(OH)2 and iodoform.

O objetivo do estudo foi avaliar, in vitro, a efetividade antimicrobiana da clorexidina gel 2 por cento (CHX) e hidróxido de cálcio, isoladamente e associados com iodofórmio e pó de óxido de zinco como medicamentos intracanais frente a microrganismos e medidos pHs das diferentes medicações. A atividade antimicrobiana foi determinada pelo método de difusão em ágar. As áreas de inibição de crescimento foram medidas e os resultados estatisticamente analisados utilizando-se o teste de Kruskal-Wallis (p<0,05). O pH das pastas foi mensurado após a manipulação, após 24 h e após uma semana. Os resultados mostraram que a maior zona de inibição foi da CHX gel 2 por cento, seguida pelo Ca(OH)2 + 2 por cento CHX gel, Ca(OH)2 + 2 por cento CHX gel + iodofórmio, Ca(OH)2 + 2 por cento CHX gel +óxido de zinco, Ca(OH)2 + água. A média de pH de todos os medicamentos intracanais foi de 12 durante todo o experimento, exceto com CHX gel 2 por cento (pH=7,0). Estes resultados permitiram concluir que todos os medicamentos tiveram atividade antimicrobiana, no entanto, a maior foi da CHX gel 2 por cento, seguido da associação com o Ca(OH)2. e iodofórmio.

The effect of mango and neem extract on four organisms causing dental caries: Streptococcus mutans, Streptococcus salivavius, Streptococcus mitis, and Streptococcus sanguis: an in vitro study.

BACKGROUND AND OBJECTIVES: Chewing twigs of the mango or neem tree is a common way of cleaning the teeth in the rural and semi-urban population. These twigs are also believed to possess medicinal properties. The present study was conducted to evaluate the antimicrobial effects of these chewing sticks on the microorganisms Streptococcus mutans , Streptococcus salivarius , Streptococcus mitis , and Streptococcus sanguis which are involved in the development of dental caries. An additional objective was to identify an inexpensive, simple, and effective method of preventing and controlling dental caries. MATERIALS AND METHODS: The sticks were sun dried, ground into a coarse powder, and weighed into 5 gm, 10 gm, and 50 gm amounts. These were added to 100 ml of deionized distilled water. After soaking for 48 h at 4 degrees C, the water was filtered. The filtrate was inoculated onto blood agar plates containing individual species of microorganisms and incubated at 37 degrees C for 48 h. Results: Mango extract, at 50% concentration, showed maximum zone of inhibition on Streptococcus mitis . Neem extract produced the maximum zone of inhibition on Streptococcus mutans at 50% concentration. Even at 5% concentration neem extract showed some inhibition of growth for all the four species of organisms. INTERPRETATION AND CONCLUSION: A combination of neem and mango chewing sticks may provide the maximum benefit. We recommend the use of both the chewing sticks.

Comparative study of the antimicrobial efficacy of chlorhexidine gel, chlorhexidine solution and sodium hypochlorite as endodontic irrigants

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

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

Minimum inhibitory concentration of adherence of Punica granatum Linn (pomegranate) gel against S. mutans, S. mitis and C. albicans

The purpose of this study was to investigate the antimicrobial effect of a Punica granatum Linn (pomegranate) phytotherapeutic gel and miconazole (Daktarin® oral gel) against three standard streptococci strains (mutans ATCC 25175, sanguis ATCC 10577 and mitis ATCC 9811), S. mutans clinically isolated and Candida albicans either alone or in association. The effect of minimum inhibitory concentrations of the gels on the adherence of these microorganisms to glass was assessed in the presence of 5 percent sucrose, using increasing and doubled concentrations of the diluted solution of the gels ranging from 1:1 to 1:1024. The minimum inhibitory concentrations of adherence of Punica granatum L. gel against the test organisms were: 1:16 for S. mutans (ATCC), S. mutans (CI) and S. sanguis; 1:128 for S. mitis and 1:64 for C. albicans. The minimum inhibitory concentrations of adherence of miconazole against the same organisms were: 1:512, 1:64, 1:4, 1:128 and 1:16, respectively. In experiments with three and four associated microorganisms, the Punica granatum L. gel had greater efficiency in inhibiting microbial adherence than the miconazole. The results of this study suggest that this phytotherapeutic agent might be used in the control of adherence of different microorganisms in the oral cavity.

O propósito deste estudo foi investigar a concentração inibitória mínima de aderência (CIMA) de três linhagens de estreptococos (mutans ATCC 25175, sanguis ATCC 10557 e mitis ATCC 9811), S. mutans isolado clinicamente e de cepas de Candida albicans, separadamente ou associadas, frente a um gel fitoterápico obtido da Punica granatum Linn. (romã) e ao agente antifúngico miconazol (Daktarin® gel oral). A concentração inibitória mínima de aderência das bactérias ao vidro foi determinada na presença de sacarose a 5 por cento, usando-se concentrações crescentes e dobradas da solução diluída do gel variando de 1:1 a 1:1024. Os valores de inibição do gel fitoterápico foram de 1:16 para S mutans (ATCC), S. mutans (IC) e S. sanguis; 1:128 para S. mitis e 1:64 para C. albicans. Sobre as mesmas linhagens, as concentrações inibitórias mínimas de aderência do miconazol foram: 1:512, 1:64, 1:4, 1:128, 1:16 respectivamente. O gel da romã apresentou maior eficácia sobre associações de três e quatro microrganismos do que o gel do miconazol. Os achados deste estudo sugerem o emprego desse agente fitoterápico pode ser uma opção no controle da aderência dos microrganismos testados na cavidade bucal.

Influencia de la prótesis parcial removible sobre el nivel de Streptococcus mutans y Streptococcus sanguis en la saliva / The influence of removable partial dentures on the level of Streptococcus mutans and Streptococcus sanguis in saliva

El presente trabajo, tiene como objetivo evaluar la influencia de la prótesis parcial removible sobre elnivel de Streptococcus mutans y Streptococcus sanguis en la saliva de un grupo de pacientes parcialmente edéntulos. Participaron 18 pacientes (16 femeninos y 2 masculinos) de la Facultad de Odontología de la Universidad Central de Venezuela. Las prótesis parciales removibles fueron confeccionadas con aleación de cromo-cobalto, bases protéticas metálicas y combinadas (metal y acrílico) y dientes de acrílico. El aislamiento de las cepas, se efectuó sembrando diluciones de las muestras de saliva en agar mitis salivarius; la incubación se realizó en condiciones anaeróbicas (Sistema Gas-Pak) a 37 grados C por 48 horas. Las muestras de saliva fueron obtenidas durante: a) el examen clínico inicial (muestra control); b) un mes antes de la instalación de la prótesis parcial removible; c) un mes después de la instalación de la ppr y d) tres meses después de la instalación de la ppr. El análisis estadístico se realizó mediante la prueba de chi-cuadrado. Los resultados fueorn estadísticamente significativos para un valor 0.01, lo que indica que posiblemente al aumentar el tiempo de instalación de la ppr, la concentración salival de los Streptococcus mutans y sanguis tiende a incrementarse. Se utilizaron diferentes pruebas bioquímicas para la identificación de los Streptococcus, tales como: fermentación de manitol, sorbitol, inulina, rafinosa y trehalosa; hidrólisis de la arginina y esculina y la producción de leván o dextrán en la sacarosa

Adherencia de candida modificada por la acción de antisépticos / Adherence of candida albicans modifiqued for the action the antisepsic

La capacidad de los microorganismos para adherirse tiene importancia en el mecanismo de la formación de placa "in vitro". El objetivo de este trabajo fue investigar la posible acción antiadherente de antisépticos, de uso oral; la influencia ejercida por la presencia de sacarosa y las modificaciones que se producirían si se asociara un microorganismo de la flora indígena bucal dominante con otro de la flora accesoria. Se prepararon suspensiones homologadas con el n§3 de la escala de Mac Farland de 10 cepas de Candida albicans y una suspensión de Streptococcus sanguis. Cada suspensión de Candida albicans fue distribuida en 12 tubos a razón de 0,5 ml en 10 de ellos y 0,25 en los tubos restantes; a estos se le completó el volumen con 0,25 ml de la suspensión de Streptococcus sanguis. En cada tubo agregamos 0,5 ml de los distintos antisépticos en estudio (Fluoruro de sodio (NaF) en distintas concentraciones, clorhexidina al 0,2 % floruro estañoso, povidona y agua destilada estéril (testigo)). El conjunto fue incubado 2 hs. en agitación constante a 37§C. Se lavaron las células una vez y se restituyó el volumen. 0,5 ml de cada uno se sembró en tubos que contenían 5 ml de medio base para la formación de placa "in vitro" con y sin el agregado de sacarosa. De los tapones pendían trocitos de diente estéril. Se incubó 5 días a 37§C. Transcurrido este tiempo procedimos a interpretar los resultados por el aspecto macroscópico de los cultivos y se procedió a la fijación de los dientes para su posterior observación por Microscopía electrónica de barrido (M.E.B.). Algunos de los antisépticos ensayados, ejercen una débil acción candicida, no se demostró influencia en la adherencia. La presencia de sacarosa en el medio es un factor importante para la formación de placa "in vitro"; la asociación de Candida albicans y Esteptococcus sanguis no influye en la misma