Recuento de Streptococcus mutans sobre restauraciones realizadas con adhesivo con y sin carga de nanopartículas de óxido de zinc y cobre / Streptococcus mutans count on restorations made with adhesive with and without loading zinc and copper oxide nanoparticles

Objetivo: Determinar las propiedades antimicrobianas de la incorporación de nanopartículas de óxido de zinc y cobre en un adhesivo de grabado y lavado total sobre Streptococcus mutans en pacientes con restauraciones de resina compuesta confeccionadas con adhesivo cargado. Métodos: Estudio experimental, randomizado, la muestra estuvo conformada por 25 pacientes, de ambos sexos, pertenecientes al posgrado de Ortodoncia de la Facultad de Odontología de la Universidad de Chile, en los cuales se confirmó presencia de Streptococcus mutans en saliva. Se confeccionaron restauraciones de resina compuesta oclusales, en premolares superiores con indicación de exodoncia por el tratamiento de ortodoncia, con adhesivo cargado (cuya composición fue 5/0,2 por ciento ZnO y Cu, respectivamente) y control (sin presencia de nanopartículas en su composición), según el listado de aleatorización. Se tomaron muestras microbiológicas en tres tiempos con la técnica de la cubeta (antes, 1 semana y 4 semanas posterior a la confección de las restauraciones). Se obtuvieron, aislaron e identificaron colonias de Streptococcus mutans a partir de las muestras obtenidas. Se usó el test de Mann-Whitney mediante el paquete estadístico SPSS v.21 Resultados: El promedio del recuento de UFC de Streptococcus mutans en el grupo experimental fue mayor posterior a la confección de las restauraciones de resina compuesta. Los resultados de la identificación molecular por PCR demuestran la presencia de Streptococcus mutans en 20 de 25 muestras. Conclusiones: No existen diferencias en el recuento de Streptococcus mutans antes y después de la aplicación del adhesivo sobre las restauraciones de resina compuesta(AU)

Objective: To determine the antimicrobial properties of the incorporation of zinc and copper oxide nanoparticles in an etching and total wash adhesive on Streptococcus mutans in patients with composite resin restorations made with loaded adhesive. Methods: Experimental and randomized trial, the sample were 25 patients, of both sexes, belonging to the FOUCH Orthodontic postgraduate program, in whom the presence of Streptococcus mutans in saliva was confirmed. Occlusal composite resin restorations were made in upper premolars with indication of extraction by orthodontic treatment, with loaded adhesive (whose composition is 5 / 0.2% ZnO and Cu respectively) and control (without the presence of nanoparticles in their composition), according to the scrambling listing. Microbiological samples were taken in three stages with the cuvette technique (before, 1 week and 4 weeks after the restoration was made). Colonies of Streptococcus mutans were obtained, isolated and identified from the samples obtained. The statistical analysis used the SPSS v.21 software, the data was analyzed by Mann Whitney test Results: The average CFU count of Streptococcus mutans in the experimental group (adhesive modified with zinc oxide and copper nanoparticles) was higher after the fabrication of composite resin restorations. The results of molecular identification by PCR demonstrate the presence of Streptococcus mutans in 20 of 25 samples. Conclusions: There are no differences in the count of Streptococcus mutans before and after the application of the adhesive on the composite resin restorations(AU)

Efecto antimicrobiano sobre Streptococcus mutans de sistema adhesivo con nanopartículas de óxido de zinc y cobre / Antimicrobial effect against Streptococcus mutans of an adhesive system with copper and zinc oxide nanoparticles

Objetivo: Determinar las propiedades antimicrobianas de la incorporación de nanopartículas de óxido de zinc y cobre en un adhesivo de grabado y lavado total sobre Streptococcus mutans en pacientes con restauraciones de resina compuesta confeccionadas con adhesivo cargado. Métodos: Este estudio clínico randomizado, participaron 25 pacientes, de ambos sexos, pertenecientes al posgrado de Ortodoncia de la Facultad de Odontología de la Universidad de Chile, en los cuales se confirmó presencia de Streptococcus mutans en saliva. Se confeccionaron restauraciones de resina compuesta oclusales, en premolares superiores con indicación de exodoncia por el tratamiento de ortodoncia, con adhesivo cargado (cuya composición es 5 / 0.2 por ciento ZnO y Cu respectivamente) y control (sin presencia de nanopartículas en su composición), según el listado de aleatorización. Se tomaron muestras microbiológicas en tres tiempos con la Técnica de la cubeta (antes, 1 semana y 4 semanas posterior a la confección de las restauraciones). Se obtuvieron, aislaron e identificaron colonias de Streptococcus mutans a partir de las muestras obtenidas. Los datos fueron analizados por el test de Mann Whittney. Resultados: El promedio del recuento de UFC (unidades formadoras de colonias) de Streptococcus mutans en el grupo experimental fue mayor posterior a la confección de las restauraciones de resina compuesta.Los resultados de la identificación molecular por PCR demuestran la presencia de Streptococcus mutans en 20 de 25 muestras. Conclusiones: No existen diferencias en el recuento de Streptococcus mutans antes y después de la aplicación del adhesivo sobre las restauraciones de resina compuesta(AU)

Objective: Determine the antimicrobial properties of the incorporation of copper and zinc oxide nanoparticles into a total rinse and etch adhesive against Streptococcus mutans in patients with composite resin restorations made with loaded adhesive. Methods: A randomized clinical study was conducted of 25 patients of both sexes from the orthodontics graduate course taught at the Dental School of the University of Chile, in whom the presence of Streptococcus mutans was confirmed in saliva. Occlusal composite resin restorations were performed in upper premolars with exodontia indicated as part of the orthodontic treatment, using loaded adhesive (composition 5 / 0.2 percent ZnO and Cu, respectively) and control (without nanoparticles in its composition), according to the randomization list. Microbiological samples were taken at three moments applying the tray technique (before, 1 week after and 4 weeks after the restorations). Streptococcus mutans were obtained, isolated and identified from the samples taken. Data analysis was based on the Mann-Whitney test. Results: Mean Streptococcus mutans CFU count in the experimental group was higher after the composite resin restorations were made. Results of PCR molecular identification show the presence of Streptococcus mutans in 20 of 25 samples. Conclusions: No differences were found in the Streptococcus mutans count before and after application of the adhesive over the composite resin restorations(AU)

Efecto Antibacteriano de la Infusión de Orégano ( Origanum vulgare ) sobre el Crecimiento in Vitro de Streptococcus mutans, 2015 / Antibacterial Effect of Oregano Infusion ( Origanum vulgare ) on in Vitro Growth of Streptococcus mutans, 2015

RESUMEN: El Streptococcus mutans es una de las principales bacterias que participa en el desarrollo de la caries dental, una enfermedad de alta prevalencia en la población mundial, y por ende un problema de salud pública. Hoy se intentan buscar alternativas para su prevención, una de ellas es la fitoterapia o uso de plantas medicinales con fines terapéutico beneficiosos para la salud. Evaluar efecto antibacteriano del Origanum vulgare a diferentes concentraciones sobre el crecimiento in vitro de Streptococcus mutans. Se utilizaron cepas bacterianas de Streptococcus mutans previamente aisladas, se realizó una siembra bacteriana en 24 placas Petri con agar mitis salivarius. Se prepararon infusiones de orégano a 8 concentraciones diferentes (1 %, 5 % y 10 %, 20 %, 40 %, 60 %, 80 % y 100 %) y se aplicaron en perforaciones realizadas previamente en las placas de agar (4 perforaciones por placa para las infusiones de orégano y 2 para las placas de controles). Se llevó a incubadora por 48 horas y posteriormente se realizó la medición de los halos de inhibición. Los resultados fueron negativos para las infusiones de orégano al 1 %, 5 % y 10 %, debido a que no presentaron halos de inhibición bacteriana; mientras que para las infusiones al 20 %, 40 %, 60 %, 80 % y 100 % los resultados fueron positivos. El orégano posee efecto antibacteriano sobre el crecimiento de Streptococcus mutans en infusiones sobre el 20 % de concentración, siendo la solución madre preparada a partir de 20 gramos de hojas secas de orégano (Origanum vulgare) y 200 ml de agua destilada hervida. Este efecto es antibacteriano es directamente proporcional a la concentración de la infusión. El orégano podría ser utilizado como una alternativa de colutorio, pasta dental u otros coadyuvantes de higiene bucal para prevenir la aparición de caries.

ABSTRACT: Streptococcus mutans is one of the main bacteria in the development of dental caries, a disease with high prevalence in the world population, and therefore a public health problem. There is current research to find prevention alternatives one of these is the use of medicinal plants for therapeutic purposes beneficial to health. To evaluate the antibacterial effect of Origanum vulgare at different concentrations on in vitro growth of Streptococcus mutans, previously isolated bacterial strains of Streptococcus mutans were used. Bacterial seeding was carried out in 24 petri dishes with agar Mitis salivarius. Oregano infusions were prepared at 8 different concentrations (1 %, 5 % and 10 %, 20 %, 40 %, 60 %, 80 % and 100 %) and applied in predrilled holes in the agar plates (4 perforations per plate for the oregano infusions and 2 for control plates). They were maintained in an incubator for 48 hours and measurement of the inhibition zones was subsequently carried out. The results were negative for infusions of oregano at 1 %, 5 % and 10 %, as they did not present halos of bacterial inhibition; while results were positive for infusions at 20 %, 40 %, 60 %, 80 % and 100 %. Results show that oregano has an antibacterial effect on the growth of Streptococcus mutans in infusion concentrations above 20 %, with the basic solution prepared from 20 g of dried oregano leaves (Origanum vulgare) and 200 ml of boiled distilled water. This antibacterial effect is directly proportional to the concentration of the infusion. Oregano could be used as an alternative mouthwash, toothpaste or other oral hygiene adjuvants to prevent the incidence of caries.

Presence and count of S. mutans in children with dental caries: before, during and after a process of oral health education

Dental caries is an infectious, multifactorial, localized, transmissible process that leads to the destruction of hard dental tissue. Streptococcus mutans is considered to be the main microorganism associated with its development. The aim of this study was to determine presence and count of S. mutans in saliva samples from children with dental caries before and after an educational process including interviews, lessons, lectures, educational workshops and recreational activities on the importance of oral care and hygiene. Twentythree 3 to 6 year old schoolchildren provided 3 unstimulated saliva samples: one before the educational process, one at 3 months and one at 6 months into the educational process. The samples were serially diluted and plated on Mitis Salivarius agar supplemented with bacitracin and 20% sucrose, and incubated anaerobically for 2 days at 37oC. Presumptive S. mutans isolates were identified with biochemical tests. Before the beginning of the educational process, and at 3 and 6 months into the educational process, S. mutans was found, respectively, in 22 (95.6%), 15 (65.2%) and 10 (43.5%) of the 23 children. The S. mutans count was reduced by 64.8% and 86.6% at 3 and 6 months, respectively, compared to the levels found before the educational process. These results indicate that educational intervention produced a significant reduction in S. mutans levels in the saliva of children with dental caries at 3 and 6 months into the educational process (AU)

La caries dental es un proceso infeccioso multifactorial, localizado y transmisible que se caracteriza por la destrucción del tejido dental duro. Streptococcus mutans es considerado el principal microorganismo asociado al desarrollo de esta enfermedad. El objetivo de este estudio fue determinar la presencia y recuento de S. mutans en saliva de niños con caries dental antes y después de un proceso educativo. Con este fin se tomó saliva no estimulada de 23 niños con caries dental pertenecientes a un centro educativo con edades de 3 a 6 años. En todos los niños se tomaron 3 muestras de saliva: antes del proceso educativo y a los 3 y 6 meses de iniciado el proceso educativo. El proceso educativo consistió en entrevistas, enseñanzas, conferencias, talleres educativos y actividades lúdicas sobre la importancia del cuidado e higiene oral. Después de su recolección, las muestras de saliva fueron serialmente diluidas y sembradas en Agar Mitis Salivarius con bacitracina y sacarosa al 20%. Los medios de cultivo sembrados se incubaron en anaerobiosis durante 2 días a 37ºC y los aislamientos presuntivos de S. mutans se identificaron con pruebas bioquímicas. Antes del inicio del proceso educativo, a los 3 y 6 meses de iniciado el proceso educativo se encontró S. mutans, respectivamente, en 22 de los 23 niños (95.6%), en 15 de los 23 niños (65.2%) y en 10 de los 23 niños (43.5%). En cuanto al recuento de S. mutans, se encontró una reducción de 64.8 % y 86.6% a los 3 y 6 meses, respectivamente, en comparación a los niveles encontrados antes del inicio del proceso educativo. En conclusión, los resultados indican que la intervención educativa realizada produjo una reducción significativa en los niveles de S. mutans en saliva de niños con caries dental después de 3 y 6 meses de iniciado el proceso educativo (AU)

Model resin composites incorporating ZnO-NP: activity against S. mutans and physicochemical properties characterization

Abstract Although resin composites are widely used in the clinical practice, the development of recurrent caries at composite-tooth interface still remains as one of the principal shortcomings to be overcome in this field. Objectives To evaluate the activity against S. mutans biofilm of model resin composites incorporating different concentrations of ZnO-nanoparticles (ZnO-NP) and characterize their physicochemical properties. Materials and Methods Different concentrations of ZnO-NP (wt.%): E1=0, E2=0.5, E3=1, E4=2, E5=5 and E6=10 were incorporated into a model resin composite consisting of Bis-GMA-TEGDMA and barium borosilicate particles. The activity against S. mutans biofilm was evaluated by metabolic activity and lactic acid production. The following physicochemical properties were characterized: degree of conversion (DC%), flexural strength (FS), elastic modulus (EM), hardness (KHN), water sorption (Wsp), water solubility (Wsl) and translucency (TP). Results E3, E4, E5 and E6 decreased the biofilm metabolic activity and E5 and E6 decreased the lactic acid production (p<0.05). E6 presented the lowest DC% (p<0.05). No significant difference in FS and EM was found for all resin composites (p>0.05). E5 and E6 presented the lowest values of KHN (p<0.05). E6 presented a higher Wsp than E1 (p<0.05) and the highest Wsl (p<0.05). The translucency significantly decreased as the ZnO- NP concentration increased (p<0.05). Conclusions The incorporation of 2 - 5 wt.% of ZnO-NP could endow antibacterial activity to resin composites, without jeopardizing their physicochemical properties.

Effect of γ-lactones and γ-lactams compounds on Streptococcus mutans biofilms

Abstract Considering oral diseases, antibiofilm compounds can decrease the accumulation of pathogenic species such as Streptococcus mutans at micro-areas of teeth, dental restorations or implant-supported prostheses. Objective To assess the effect of thirteen different novel lactam-based compounds on the inhibition of S. mutans biofilm formation. Material and methods We synthesized compounds based on γ-lactones analogues from rubrolides by a mucochloric acid process and converted them into their corresponding γ-hydroxy-γ-lactams by a reaction with isobutylamine and propylamine. Compounds concentrations ranging from 0.17 up to 87.5 μg mL-1 were tested against S. mutans. We diluted the exponential cultures in TSB and incubated them (37°C) in the presence of different γ-lactones or γ-lactams dilutions. Afterwards, we measured the planktonic growth by optical density at 630 nm and therefore assessed the biofilm density by the crystal violet staining method. Results Twelve compounds were active against biofilm formation, showing no effect on bacterial viability. Only one compound was inactive against both planktonic and biofilm growth. The highest biofilm inhibition (inhibition rate above 60%) was obtained for two compounds while three other compounds revealed an inhibition rate above 40%. Conclusions Twelve of the thirteen compounds revealed effective inhibition of S. mutans biofilm formation, with eight of them showing a specific antibiofilm effect.

Effect of Myracrodruon urundeuva and Qualea grandiflora extracts on viability and activity of microcosm biofilm and prevention of enamel demineralization in vitro / Efeito de extratos de Myracrodruon urundeuva All. e Qualea grandiflora Mart. sobre a viabilidade e atividade de biofilme microcosmo e na prevenção da desmineralização do esmalte in vitro

The objective of this study was to evaluate the antimicrobial and anti-caries effects of two plant extracts. The first chapter dealt with a review of the literature whose objective was to discuss the antimicrobial potential of Brazilian natural agents on the biofilm related to dental caries and gingivitis/periodontal disease. The research of the articles was carried out using PubMed. We found a total of 23 papers. Most of the studies were performed using planktonic microorganisms or under clinical trials. Nineteen articles were focused on cariogenic bacteria. From these nineteen articles, eleven were also about periodontopathogenic bacteria. Four studies addressed only periodontopathogenic bacteria. The most tested Brazilian natural agents were green propolis, essential oils of Lippia sidoides and Copaifera sp. Most of the tested agents showed similar results when compared to positive control (essential oils and extracts) or better effect than negative control (green propolis). More studies involving protocols closer to the clinical condition and the use of response variables that allows understanding the mechanism of action of natural agents are necessary before the incorporation of these natural agents into dental products. The second chapter aimed to test the effect of the hydroalcoholic extracts of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves on the viability of the microcosm biofilm and on the prevention of enamel demineralization. The microcosm biofilm was produced on bovine enamel, using human saliva pool mixed with McBain saliva (0.2% sucrose) for 14 days. The biofilm was treated daily with the extracts for 1 min. M. urundeuva at 100, 10 and 0.1 µg/ml and Q. grandiflora at 100 and 0.1 µg/ml reduced cell viability similarly to the positive control and significantly more than negative control. M. urundeuva at 1000, 100 and 0.1 µg/ml were able to reduce the counting formation unit-CFU counting of lactobacilli sp. and Streptococcus mutans, while Q. grandiflora at 1000 and 1.0 µg/ml significantly reduced the S. mutans CFU counting. On the other hand, the natural extracts did not reduce the production of extracellular polyssacharides, lactic acid and the development of enamel caries lesions. The third chapter aimed to evaluate the effect of hydroalcoholic extracts of M. urundeuva and Q. grandiflora (alone or combined) on the viability of S. mutans biofilm and the prevention of enamel demineralization. S. mutans strain (ATCC 21175) was reactivated in BHI broth. Minimum inhibitory concentration, minimum bactericidal concentration, minimum biofilm inhibitory concentration and minimum biofilm eradication concentration were determined to choose the concentrations to be tested under the biofilm model. S. mutans biofilm (5x105 CFU/ml) was produced on bovine enamel using McBain saliva with 0.2% sucrose for 3 days. The biofilm was treated daily with the extracts for 1 min. M. urundeuva (isolated or combined) at concentrations equal or higher than 0.625 mg/ml was able to reduce the bacteria viability, whereas Q. grandiflora extract alone showed antimicrobial effect at 5 mg/ml only (p<0.05). On the other hand, none of the extracts was able to reduce the development of enamel caries lesions. Despite the tested natural extracts have antimicrobial effect; they are unable to prevent caries in enamel.(AU)

O objetivo foi avaliar os efeitos antimicrobiano e anti-cárie de dois extratos de plantas. O primeiro capítulo se referiu a uma revisão da literatura cujo objetivo foi discutir o potencial antimicrobiano dos agentes naturais brasileiros sobre o biofilme relacionado à cárie dentária e à gengivite/doença periodontal. A pesquisa dos artigos foi realizada usando o PubMed. Foram encontrados 23 trabalhos. A maioria dos estudos foi realizada utilizando microorganismos na fase planctônica ou ensaios clínicos. Dezenove artigos foram focados em bactérias cariogênicas. Dos dezenove artigos, onze também eram sobre bactérias periodontopatogênicas. Quatro estudos abordaram apenas bactérias periodontopatogênicas. Os agentes naturais brasileiros mais testados foram própolis verde, óleos essenciais de Lippia sidoides e Copaifera sp. Os agentes testados apresentaram resultados similares quando comparados ao controle positivo (óleos essenciais e extratos) ou melhor efeito que o controle negativo (própolis verde). Mais estudos próximos da condição clínica e o uso de variáveis de resposta que permitam entender o mecanismo de ação são necessários, para permitir a incorporação desses agentes naturais em produtos odontológicos. O segundo capítulo teve como objetivo testar o efeito dos extratos hidroalcoólicos de Myracrodruon urundeuva All. e Qualea grandiflora Mart. sobre a viabilidade do biofilme microcosmo e na prevenção da desmineralização do esmalte. O biofilme microcosmo foi produzido em esmalte bovino, utilizando pool de saliva humana misturada à saliva de McBain (0,2% de sacarose) durante 14 dias. O biofilme foi tratado diariamente com os extratos durante 1 min. M. urundeuva a 100, 10 e 0,1 µg/ml e Q. grandiflora a 100 e 0,1 µg/ml reduziram a viabilidade dos microrganismos de forma semelhante ao controle positivo e significativamente maior do que o controle negativo. M. urundeuva a 1000, 100 e 0,1 µg/ml foi capaz de reduzir a contagem de Unidade formadora de colônia-UFC para Lactobacilos totais e Streptococcus mutans, enquanto a Q. grandiflora a 1000 e 1,0 µg/ml reduziu significativamente a contagem de UFC para S. mutans. Os extratos naturais não conseguiram reduzir a produção de polissacarídeos extracelulares-PEC, ácido lático e o desenvolvimento da lesão cariosa em esmalte. O terceiro capítulo teve como objetivo avaliar o efeito dos extratos hidroalcoólicos de M. urundeuva. e Q. grandiflora (sozinhos ou combinados) sobre a viabilidade do biofilme de S. mutans e na prevenção da desmineralização do esmalte. Cepa de S. mutans (ATCC 21175) foi reativada em caldo BHI. Concentração inibitória mínima, concentração bactericida mínima, concentração inibitória mínima de biofilme e concentração de erradicação mínima de biofilme foram determinadas para escolher as concentrações a serem testadas sob o modelo de biofilme. O biofilme de S. mutans (5x105 CFU/ml) foi produzido em esmalte bovino, utilizando saliva de McBain com 0,2% de sacarose durante 3 dias. O biofilme foi tratado diariamente com os extratos durante 1 min. M. urundeuva (isolada ou combinada) nas concentrações iguais ou superiores a 0,625 mg/ml foi capaz de reduzir a viabilidade das bactérias, enquanto que o extrato da Q. grandflora apresentou efeito antimicrobiano somente a 5 mg/ml (p<0,05). Nenhum dos extratos reduziu o desenvolvimento da lesão da cárie. Apesar dos extratos naturais terem efeito antimicrobiano, são incapazes de prevenir o desenvolvimento da lesão cariosa em esmalte.(AU)

In vitro and in vivo evaluations of glass-ionomer cement containing chlorhexidine for Atraumatic Restorative Treatment

Abstract Objectives: Addition of chlorhexidine has enhanced the antimicrobial effect of glass ionomer cement (GIC) indicated to Atraumatic Restorative Treatment (ART); however, the impact of this mixture on the properties of these materials and on the longevity of restorations must be investigated. The aim of this study was to evaluate the effects of incorporating chlorhexidine (CHX) in the in vitro biological and chemical-mechanical properties of GIC and in vivo clinical/ microbiological follow-up of the ART with GIC containing or not CHX. Material and Methods: For in vitro studies, groups were divided into GIC, GIC with 1.25% CHX, and GIC with 2.5% CHX. Antimicrobial activity of GIC was analyzed using agar diffusion and anti-biofilm assays. Cytotoxic effects, compressive tensile strength, microhardness and fluoride (F) release were also evaluated. A randomized controlled trial was conducted on 36 children that received ART either with GIC or GIC with CHX. Saliva and biofilm were collected for mutans streptococci (MS) counts and the survival rate of restorations was checked after 7 days, 3 months and one year after ART. ANOVA/Tukey or Kruskal-Wallis/ Mann-Whitney tests were performed for in vitro tests and in vivo microbiological analysis. The Kaplan-Meier method and Log rank tests were applied to estimate survival percentages of restorations (p<0.05). Results: Incorporation of 1.25% and 2.5% CHX improved the antimicrobial/anti-biofilm activity of GIC, without affecting F release and mechanical characteristics, but 2.5% CHX was cytotoxic. Survival rate of restorations using GIC with 1.25% CHX was similar to GIC. A significant reduction of MS levels was observed for KM+CHX group in children saliva and biofilm 7 days after treatment. Conclusions: The incorporation of 1.25% CHX increased the in vitro antimicrobial activity, without changing chemical-mechanical properties of GIC and odontoblast-like cell viability. This combination improved the in vivo short-term microbiological effect without affecting clinical performance of ART restorations.

Antimicrobial activity of ozone and NaF-chlorhexidine on early childhood caries

Abstract An early childhood carie (ECC) is an extremely destructive form of tooth decay. The aim of this study was to investigate the action of ozone (O3), and the association of sodium fluoride (NaF) with chlorhexidine (CHX) on bacteria related to ECC. Overnight culture of the bacteria was performed. On exponential phase the suspension was adjusted (101-108 CFU/mL). A drop (10μL) of each concentration of bacteria was applied on sheep blood agar plates and treated with O3 (2, 20, 200, and 2,000 ppm); after 18 hours, recovery analysis of CFU verified the reduction of bacterial activity. For NaF-CHX, sterile 96-well plates were prepared and divided into groups: G1 (150 µL TSB); G2 (20 µL of bacteria + 25 µL CHX + 25 µL NaF); and G3 (150 µL TSB + 20 µL of bacteria + 50 µL water). The plates were verified by analysis of the optical density (0, 12, 14, 16, and 18 hours). The data from O3 test were submitted to ANOVA and Tukey’s test (p < 0.05). For the data from NaF-CHX, the ANOVA 2-way and Bonferroni’s test (p < 0.05) were used. The number of CFU/mL showed death > 3log10 (99.9%) for all bacteria (ozone ≥ 20ppm), while the combination of NaF-CHX was more effective (p < 0.001) compared to each substance tested alone and the control group. The antimicrobial agents tested were able to inhibit all bacteria tested; O3 seemed to be a good alternative for controlling progression of carious lesions, while the association of NaF-CHX showed to be a good antimicrobial with easy and inexpensive application.

Effect of silver nanoparticles on the physicochemical and antimicrobial properties of an orthodontic adhesive

ABSTRACT Orthodontic treatment with fixed brackets plays a major role on the formation of white spot lesions. Objective This study aimed to incorporate silver nanoparticle solutions (AgNP) in an orthodontic adhesive and evaluate its physicochemical and antimicrobial properties. Material and Methods Silver nanoparticle solutions were added to a commercial adhesive in different concentrations (w/w): 0%, 0.11%, 0.18%, and 0.33%. Shear bond strength (SBS) test was performed after bonding metal brackets to enamel. Raman spectroscopy was used to analyze in situ the degree of conversion (DC) of the adhesive layer. The surface free energy (SFE) was evaluated after the measurement of contact angles. Growth inhibition of Streptococcus mutans in liquid and solid media was determined by colony-forming unit count and inhibition halo, respectively. One-way ANOVA was performed for SBS, DC, SFE, and growth inhibition. Results The incorporation of AgNP solution decreased the SBS (p<0.001) and DC in situ (p<0.001) values. SFE decreased after addition of 0.18% and 0.33% AgNP. Growth inhibition of S. mutans in liquid media was obtained after silver addition (p<0.05). Conclusions The addition of AgNP solutions to Transbond™ XT adhesive primer inhibited S. mutans growth. SBS, DC, and SFE values decreased after incorporation up to 0.33% AgNP solution without compromising the chemical and physical properties of the adhesive.

Influência da contaminação cavitária na adaptação marginal das restaurações de resina composta / Influence of cavity contamination on marginal adaptation of composite resin restorations

Objetivo: avaliar a influencia da contaminação cavitária na adaptação marginal das restauraçõesde resina composta e a associação da lisozima ao sistema adesivo. Materiais e Métodos:Foram selecionados 40 terceiros molares permanentes e confeccionados preparos cavitários classeV com pontas diamantadas cilíndricas 1090 (4 mm de largura, 6 mm de comprimento e 1,5 mmde profundidade). Todos os dentes foram esterilizados e divididos aleatoriamente em 4 grupos(n=10): G1: sistema adesivo (SA) e restauração com resina composta (RC); G2: cavidadescontaminadas com 10 μl de Streptococcus mutans (0,5 de Mc Farland), SA e RC; G3:sistema adesivo associado com lisozima (ADL) e RC e G4: cavidades contaminadas com 10 μlde Streptococcus Mutans (0,5 de Mc Farland), ADL e RC. A lisozima foi adicionada ao SA com ummicrobrush embebido no SA e na lisozima em pó. Os dentes foram impermeabilizados e imersosem azul de metileno por 4 horas a 37ºC. Os dentes foram seccionados e fotografados. Resultados:A avaliação da microinfiltração foi feita por dois avaliadores utilizando o Programa Tpsdig. Paraavaliar a calibração entre os examinadores, foi utilizado o teste de Correlação de Pearson. As médiasdas porcentagens de fendas e bolhas foram submetidas ao teste de Kruskal Wallis com 5% designificância. Conclusão: a contaminação cavitária com Streptococcus mutans acarretou na formaçãode fendas entre a resina composta e as margens cavitárias. A lisozima associada ao sistemaadesivo não influenciou na formação de fendas e bolhas nas restaurações de resina composta.

Objective: evaluate the influence of cavity contamination on marginal adaptation of compositerestorations and the association of lysozyme to the adhesive system. Materials and Methods:Were selected 40 permanent third molars and made cavity preparations class V with cylindricaldiamond burs 1090 (4 mm wide, 6 mm long and 2 mm deep). All teeth were sterilized and randomlydivided into 4 groups (n = 10): G1: adhesive system (SA) and restoration with compositeresin (CR); G2: cavity contaminated with 10 μl of Streptococcus mutans (0.5 McFarland), SA andRC; G3: adhesive system associated with lysozyme (ADL) and RC and G4: cavity contaminated with10 μl of Streptococcus mutans (0.5 Mc Farland), ADL and RC. Lysozyme was added to the SA witha microbrush embedded in the SA and powdered lysozyme. The teeth were sealed and immersedin a methylene blue for 4 hours at 37 ° C. The teeth were sectioned and photographed. Results:The evaluation of microleakage was made by two reviewers using the Tpsdig Program. To assessthe calibration of examiners, we used the Pearson correlation test. The mean percentages of cracksand bubbles were submitted to Kruskal Wallis test at 5% significance level. Conclusion: the cavityStreptococcus mutans contamination resulted in the formation of cracks between the compositeresin and the cavity margins. Lysozyme associated with the adhesive system did not influence theformation of cracks and bubbles in the composite restorations.

Avaliação dos métodos de higienização sobre S. mutans dos protetores bucais / Evaluation of cleaning methods on custom-made mouthguards of S. mutans

Objetivo: avaliar a eficácia de três diferentes métodos de higienização sobre cepas de S.mutans dos protetores bucais feitos sob medida: dentifrício fluoretado, detergente neutro eclorexidina. Materiais e Métodos: foram confeccionados 30 protetores bucais feitos sob medida com placas de silicone de 3 mm que foram contaminados em meio de sobrevivência BHI (BrainHeart Infusion) suplementado com 1% de glicose e sacarose e 0,5% de extrato de levedura e incubados em jarra de anaerobiose a 37°C em estufa bacteriológica por 15 dias. O meio foi renovado a cada 24 horas. Após a contaminação, foi feita a coleta dos protetores bucais, esfregaço e plaqueamento na superfície do meio de cultura mitis salivarius com bacitracina. Os protetores foram higienizados com escova dental, com três diferentes meios de higienização:dentifrício fluoretado, detergente neutro e clorexidina e, após a higienização foram submetidos a uma nova coleta, esfregaço e plaqueamento. Após incubação, foi feita a contagem do total de bactérias viáveis. Os resultados obtidos foram submetidos à análise estatística e ao teste não paramétrico de Wilcoxon. Resultados: houve redução microbiana significativa da contagem deS. mutans utilizando clorexidina e detergente (p<0.05). Não houve diferença significante na contagem antes e depois utilizando o dentifrício (p>0.05). Conclusão: a clorexidina e o detergente podem ser utilizados para desinfecção dos protetores bucais.

Objective: The effect of three different cleaning methods on custom-made mouth guards’strains of S. mutans: fluoridated tooth paste, neutral detergent and chlorhexidine. Materials and Methods: it were made 30 mouthguards custom-made with 3mm silicone slabs that have been subjected to a microbiological processing in supplemented BHI (Brain Heart Infusion). They were in cubated in anaerobic jar at 37 ° bacteriological green house for 15 days, changing the medium every 24 hours. After the contamination, samples were drawn and it was made the collection, smear and plating on surface of the culture medium mitis salivarius with bacitracin and then they were cleaned with tooth brush with three different modes of cleaning: chlorhexidine, neutral detergent and fluoridated tooth paste. After incubation, the count was made of total viable bacteria. The results were statistically analyzed by the Wilcoxon non-parametric test. Results: there wasa significant reduction in microbial counts before and after using chlorhexidine and detergent (p<0.05). There wasn’t significant difference in the count before and after using the tooth paste(p>0.05). Conclusion: chlorhexidine and detergent can be used for disinfection of mouth guards

Enamel and dentine demineralization by a combination of starch and sucrose in a biofilm - caries model

Abstract Sucrose is the most cariogenic dietary carbohydrate and starch is considered non-cariogenic for enamel and moderately cariogenic for dentine. However, the cariogenicity of the combination of starch and sucrose remains unclear. The aim of this study was to evaluate the effect of this combination on Streptococcus mutans biofilm composition and enamel and dentine demineralization. Biofilms of S. mutans UA159 were grown on saliva-coated enamel and dentine slabs in culture medium containing 10% saliva. They were exposed (8 times/day) to one of the following treatments: 0.9% NaCl (negative control), 1% starch, 10% sucrose, or 1% starch and 10% sucrose (starch + sucrose). To simulate the effect of human salivary amylase on the starch metabolization, the biofilms were pretreated with saliva before each treatment and saliva was also added to the culture medium. Acidogenicity of the biofilm was estimated by evaluating (2 times/day) the culture medium pH. After 4 (dentine) or 5 (enamel) days of growth, biofilms (n = 9) were individually collected, and the biomass, viable microorganism count, and polysaccharide content were quantified. Dentine and enamel demineralization was assessed by determining the percentage of surface hardness loss. Biofilms exposed to starch + sucrose were more acidogenic and caused higher demineralization (p < 0.0001) on either enamel or dentine than those exposed to each carbohydrate alone. The findings suggest that starch increases the cariogenic potential of sucrose.

Microbial contamination and disinfection methods of pacifiers

Objectives To evaluate the microbial contamination of pacifiers by Mutans Streptococci(MS) and the efficacy of different methods for their disinfection.Methods Twenty-eight children were assigned to a 4-stage changeover system with a 1-week interval. In each stage, children received a new pacifier and the parents were instructed to maintain their normal habits for 1 week. After this time, the pacifiers were subjected to the following 4 disinfection methods: spraying with 0.12% chlorhexidine solution, Brushtox® or sterile tap water, and immersion in boiling tap water for 15 minutes. Microbiological culture for MS and Scanning Electron Microscopy (SEM) were performed. The results were analyzed statistically by Friedman’s non-parametric test (a=0.05).Results The 0.12% chlorhexidine spray was statistically similar to the boiling water (p>0.05) and more effective than the Brushtox®spray and control (p<0.05). The analysis of SEM showed the formation of a cariogenic biofilm in all groups with positive culture.Conclusions Pacifiers become contaminated by MS after their use by children and should be disinfected routinely. Spraying with a 0.12% chlorhexidine solution and immersion in boiling water promoted better disinfection of the pacifiers compared with a commercial antiseptic toothbrush cleanser (Brushtox®).

Effects of probiotic fermented milk on biofilms, oral microbiota, and enamel

The aim of this study was to evaluate in vitro and in vivo the effects of 2 brands of probiotic fermented milk on biofilms, oral microbiota, and enamel. For the in situ experiment, ten volunteers wore palatine devices containing four blocks of bovine dental enamel over 3 phases, during which 20% sucrose solution, Yakult® (Treatment A), and Batavito® (Treatment B) were dropped on the enamel blocks. Salivary microbial counts were obtained and biofilm samples were analyzed after each phase. For the in vivo experiment, the same ten volunteers drunk Yakult® (Treatment C) and Batavito® (Treatment D) in two phases. Saliva samples were collected for microbial analysis after each phase. The in situ study showed that in comparison with Treatment A, Treatment B resulted in fewer total cultivable anaerobes and facultative microorganisms in biofilms, higher final microhardness, lower percentage change in surface hardness, and smaller integrated subsurface enamel hardness. In the in vivo study, Treatment D resulted in a reduction in the counts of all microorganisms. The results suggested that the probiotic fermented milk Batavito®, but not Yakult®, reduced the amount of oral microorganisms and mineral loss in bovine enamel.

Development of a novel resin with antimicrobial properties for dental application

The adhesion of biofilm on dental prostheses is a prerequisite for the occurrence of oral diseases. Objective: To assess the antimicrobial activity and the mechanical properties of an acrylic resin embedded with nanostructured silver vanadate (β-AgVO3). Material and Methods: The minimum inhibitory concentration (MIC) of β-AgVO3 was studied in relation to the species Staphylococcus aureus ATCC 25923, Streptococcus mutans ATCC 25175, Pseudomonas aeruginosa ATCC 27853, and Candida albicans ATCC 10231. The halo zone of inhibition method was performed in triplicate to determine the inhibitory effect of the modified self-curing acrylic resin Dencor Lay - Clássico®. The surface hardness and compressive strength were examined. The specimens were prepared according to the percentage of β-AgVO3 (0%-control, 0.5%, 1%, 2.5%, 5%, and 10%), with a sample size of 9x2 mm for surface hardness and antimicrobial activity tests, and 8x4 mm for the compression test. The values of the microbiologic analysis were compared and evaluated using the Kruskal-Wallis test (α=0.05); the mechanical analysis used the Shapiro-Wilk's tests, Levene's test, ANOVA (one-way), and Tukey's test (α=0.05). Results: The addition of 10% β-AgVO3 promoted antimicrobial activity against all strains. The antimicrobial effect was observed at a minimum concentration of 1% for P. aeruginosa, 2.5% for S. aureus, 5% for C. albicans, and 10% for S. mutans. Surface hardness and compressive strength increased significantly with the addition of 0.5% β-AgVO3 (p<0.05). Higher rates of the nanomaterial did not alter the mechanical properties of the resin in comparison with the control group (p>0.05). Conclusions: The incorporation of β-AgVO3 has the potential to promote antimicrobial activity in the acrylic resin. At reduced rates, it improves the mechanical properties, and, at higher rates, it does not promote changes in the control. .

Crecimiento bacteriano de Streptococcus mutans frente a la cetirizina y prednisona con y sin sacarosa: estudio in vitro / Bacterial growth of Streptococcus mutans exposed to cetirizine and prednisone with and without sucrose: in vitro study

Objetivos: Comparar el crecimiento de Streptococus mutans antes y después de la aplicación de cetirizina y prednisona con y sin sacarosa a las 6, 12, 24 y 36 horas. Métodos: Diseño experimental, para ello se utilizó una muestra de 84 tubos de ensayo, 6 tubos por cada una de las medicinas o controles utilizados. Las variables que se consideraron fueron medicinas pediátricas con edulcorantes divididas en dos grupos: cetirizinas y prednisonas. En el grupo de las cetirizinas se consideraron las cetirizinas comerciales Hisaler®, Lergium® y Rigotax®, así como los preparados de cetirizina con xilitol, sacarosa y aspartame. En el grupo de las prednisonas se consideraron las prednisonas comerciales: Cortiprex® y Nisona® así como los preparados de prednisona con sacarosa y otros edulcorantes. La técnica que se empleó fue la de turbimetría la cual nos da el indicador absorvancia. Resultados: los resultados fueron leídos a las 6, 12, 24 y 36 horas. Conclusiones: Las medicinas comerciales que presentaron menor crecimiento de Streptococcus mutans fueron Rigotax® y Nisona®. Los preparados que presentaron menor crecimiento de Streptoccoccus mutans fueron cetirizina con aspartame y prednisona con edulcorante. Las medicinas comerciales presentaron diferencias estadísticamente significativas con sus respectivos preparados, produciéndose menor crecimiento de Streptococcus mutans en las medicinas comerciales.

Objetives: To compare the growth of Streptococcus mutans, before and after the application of cetirizine and prednisone with and without sucrose at 6, 12, 24 and 36 hours. Methods: The type of study was experimental. To carry it on, a sample of 84 test tubes was used, 6 tubes for each of the medications or controls. The variables considered were pediatric medication with sweeteners, divided in two groups: cetirizines and prednisones. In the cetirizines groups, those over the counter were: Hisaler®, Lergium® and Rigotax®; those that in their formulas included cetirizine with xylitol, sucrose, and aspartame. In the prednisone groups, which are over the counter, the following were found: Cortiprex® and Nisona®, as well as those that in their formulas included prednisone with saccarose and other sweeteners. The method used was turbimetry with the spectrophotometer, to meassure: absorbency. Results: The results were read at 6, 12, 24 and 36 hours. Conclusions: The commercial medications that showed less growth of the Streptococcus mutans, were Rigotax® and Nisona®. The preparation that showed less growth of the Streptococcus mutans were cetirizine with aspartame and prednisone with sorbitol. Over the counter medications showed significant statistical differences with their respective preparation, producing less growth of the Streptococcus mutans in over the counter medications.

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 ...

Anti-Streptococcal activity of Brazilian Amazon Rain Forest plant extracts presents potential for preventive strategies against dental caries

Caries is a global public health problem, whose control requires the introduction of low-cost treatments, such as strong prevention strategies, minimally invasive techniques and chemical prevention agents. Nature plays an important role as a source of new antibacterial substances that can be used in the prevention of caries, and Brazil is the richest country in terms of biodiversity. Objective: In this study, the disk diffusion method (DDM) was used to screen over 2,000 Brazilian Amazon plant extracts against Streptococcus mutans. Material and Methods: Seventeen active plant extracts were identified and fractionated. Extracts and their fractions, obtained by liquid-liquid partition, were tested in the DDM assay and in the microdilution broth assay (MBA) to determine their minimal inhibitory concentrations (MICs) and minimal bactericidal concentrations (MBCs). The extracts were also subjected to antioxidant analysis by thin layer chromatography. Results: EB271, obtained from Casearia spruceana, showed significant activity against the bacterium in the DDM assay (20.67±0.52 mm), as did EB1129, obtained from Psychotria sp. (Rubiaceae) (15.04±2.29 mm). EB1493, obtained from Ipomoea alba, was the only extract to show strong activity against Streptococcus mutans (0.08 mg/mL<MIC<0.16 mg/mL; MBC=0.16 mg/mL) in the MBA. Conclusions: The active extracts, discovered in the Amazon rain forest, show potential as sources of new antibacterial agents for use as chemical coadjuvants in prevention strategies to treat caries. .

Influencia de los tipos de anclaje de los filamentos en la retención de streptococcus mutans en cepillos dentales: estudio preliminar / Influence of the types of filaments anchoring in the retention of streptococcus mutans on toothbrushes: preliminary study

Este estudio evaluó la retención del S. mutans (ATCC 25175) en los cepillos dentales Oral B Complete® y Sensodyne Esmalte Care®, que poseen distintas características de fijación y anclaje de los filamentos en sus partes activas. Nueve cepillos dentales de cada marca comercial fueron esterilizados en autoclave y, a continuación, inoculados con S. mutans (ATCC 25175), previamente desarrollado en caldo Brain Heart Infusion (BHI). Los análisis de retención bacteriana se realizaron en los tempos de 04, 12 y 24 horas de incubación, a 37ºC y en microaerofilia, en triplicata. En esas etapas, los cepillos fueron lavados con agua esterilizada e introducidos en tubos Falcon con caldo BHI, del cual fueron preparadas diluciones decimales hasta 10-3 para las cuatro primeras horas, hasta 10-4 para 12 horas, y hasta 10-5 para 24 horas de incubación. Alícuotas de esas diluciones fueron sembradas en la superficie del Ágar BHI, siendo las placas incubadas en microaerofilia por 24h a 37ºC. Tras la incubación, fue realizado un recuento de las unidades formadoras de colonia por mL (UFC/mL) y el análisis estadístico (Tests de Tukey pareado, p?0,05). Fueron observadas diferencias estadísticamente significantes (p?0,05) entre lo tiempos de evaluación de 12 horas. Concluyese que el cepillo dental Oral B Complete® presentó, comparativamente a Sensodyne Esmalte Care®, mayor retención del S. mutans (ATCC 25175).

This work evaluated the retention of S. mutans ATCC 25175 in the Oral B Complete and Sensodyne Esmalte Care toothbrushes that have distinct characteristics of insertion and anchorage of the filaments in its active parts. Nine toothbrushes of each trademark were sterilized in autoclave and, following this procedure, inoculated with S. mutans ATCC 25175, previously developed in Brain Heart Infusion (BHI).The analyses of bacterial retention occurred in the times of 04, 12 and 24 hours after incubation, at 37ºC and in microaerophilic, in triplicate. In these stages, the toothbrushes were washed with sterilized water and enclosed in Falcon tubes with BHI broth, of which decimal dilutions up to 10-3 were made for the four first hours, up to 10-4 for 12 hours, and up to 10-5 for 24 hours of incubation. Aliquots of these dilutions were sown in the surface of agar BHI, and the Petri dishes were incubated in microaerophilic for 24 hours, in a temperature of 37ºC. After the incubation, the counting of the colony forming units and the statistical analysis (Tukey Test paired, p?0.05) were performed. Significant statistical differences were observed (p?0.05) between the times of evaluation of 12 hours. It was concluded that the Oral B Complete toothbrush presents, compared with Sensodyne Esmalte Care, greater retention of S. mutans ATCC 25175.