Validation of an adherence assay to detect group mutans streptococci in saliva samples / Validación del Test de adherencia para el recuento de estreptococcos del grupo mutans en muestras de saliva

The aim of the present study was to validate and establish a cut off point and the predictive value of an adhesion test (AA-MSMG), as a microbiological method for evaluating cariogenic risk. The study is based on a variant (20% sucrose) of a selective medium descripted by Gold et al. (MSMG). This method differentiates mutans group streptococci (MGS) by exacerbating the production of insoluble extracellular polysaccharide which gives adhesion to surfaces such as glass, plastic and dental enamel. Caries assessment according to ICDAS was conducted in 154 patients (aged >21 years) who were attended at Preventive and Community Dentistry Department, School of Dentistry, University of Buenos Aires, Argentina, between August 2017 to August 2018. The study population was assigned to groups according to the presence/ absence of caries lesions: Group A: ICDAS lesion code = 0 (L=0) on all dental surfaces (n=23); and Group B: L>1 (n=131). After mouth-rinsing with distilled water, saliva samples were collected with fasting and hygiene protocol, and sent immediately to the Microbiological Diagnosis Laboratory, Microbiology Department, School of Dentistry, University of Buenos Aires. Samples were homogenized and serially diluted to the tenth. 100 pl of the dilutions were cultured in 25 cm² sterile plastic flasks containing 9.9 ml of modified selective medium described by Gold (MSMG-selective and differential medium). Cultures were incubated in an anaerobic atmosphere at 36 ± 1°C for 48 hours. The supernatants were eluted and the samples washed with sterile distilled water. Colony forming unit counts were performed by calibrated researchers (Kappa >0.75) using a stereoscopic microscope at 50X. Mutans group streptococci (MGS) counts ranged from 1x10(4) to 1x10(5) CFU/ml in group A, and were higher than 1x10(6) CFU/ml in Group B. Statically analysis of results (ROC) showed that the AAMSMG has a satisfactory predictive value (91%) and established a cutoff point in 1.68x10(5) UFC / ml. This would indicate that individuals whose MGS saliva counts are higher than the cutoff value would be 5 times more likely to develop dental caries. Adherence assay could be a useful microbiological predictor of caries risk.

El objetivo del presente estudio fue validar, establecer el punto de corte y valor predictivo de una técnica microbiológica para evaluar el nivel de estreptococos del grupo mutans en saliva. La técnica consiste en un test de adherencia que emplea un medio selectivo modificado (20% sacarosa) descripto por Gold et al. (TA-MSMG). Este método permite diferenciar a los estreptococos del grupo mutans (SGM) exacerbando la producción del polisacárido extracelular insoluble que le confiere adhesión a superficies como vidrio, plástico y esmalte dental. De acuerdo con los criterios de ICDAS se sembraron 154 salivas de pacientes mayores de edad, que asistieron al Servicio de Odontología Preventiva y Comunitaria de la Facultad de Odontología de la Universidad de Buenos Aires entre los meses de agosto de los años 2017 y 2018. La población estudiada fue asignada a dos grupos según la presencia / ausencia de lesiones de caries: Grupo A: código de lesión ICDAS = 0 (L = 0) en todas las superficies dentales (n = 23); y Grupo B: L> 1 (n = 131). Después de realizar un enjuague bucal con agua destilada, las muestras de saliva se recogieron según protocolo (ayuno de 4 horas y suspensión de higiene dental de 12 hs). Las muestras se remitieron de inmediato al Laboratorio de Diagnóstico Microbiológico, Departamento de Microbiología de la Facultad de Odontología, Universidad de Buenos Aires. Para su procesamiento, las muestras fueron homogeneizadas y diluidas al décimo. Se cultivaron 100 pl de las diluciones en botellas de plástico estériles de 25 cm² que contenían 9,9 ml de medio de Gold modificado (MSMG-20% sacarosa). Los cultivos se incubaron en atmósfera anaeróbica a 36 ± 1°C durante 48 horas. El sobrenadante se eluyó y las muestras se lavaron con agua destilada estéril. Los recuentos de unidades formadoras de colonias SGMfueron realizados por investigadores calibrados (Kappa >0.75) utilizando un microscopio estereoscópico a 50X. Los recuentos de SGM presentaron una variación entre 1x10(4)y 1x10(5) UFC/ml en el grupo A, mientras que en el Grupo B fueron superiores a 1x10(6) UFC/ml. El análisis estadístico de los resultados determinó una curva ROC que establece para el TA-MSMG un valor predictivo del 91% y un punto de corte en 1.68x10(5) UFC SGM / ml. Esto indicaría que los individuos cuyos recuentos en saliva de SGM sean superiores al valor de corte, tendrían 5 veces más posibilidades de desarrollar caries (5:1). Este método podría ser un instrumento útil al momento de evaluar (indicador microbiológico) el riesgo cariogénico del paciente.

Does the bracket composition material influence initial biofilm formation.

Context: Orthodontic treatment has been reported to contribute to the development and accumulation of dental biofilm, which is commonly found on bracket and adjacent surfaces. Aims: The aim of this work is to test the hypothesis if there are differences in dental biofilm formation on the surface of orthodontic brackets according to the type of composition material. Subjects and Methods: Three bracket types (metallic, composite, and ceramic) had been evaluated. Subjects wore acrylic palatal orthodontic appliances, containing 6 brackets each, for two 3‑day cycles. On the end of first cycle, the amount of dental biofilm formed on the samples was extracted using 1.0‑M NaOH and analyzed by spectrophotometry for quantification. An additional cycle was carried out to verify the dental biofilm formation using scanning electronic microscope analysis. Statistical Analysis Used: Three‑way ANOVA was used to analyze the difference among the materials (metallic, ceramic and composite) concerning the dental biofilm absorbance spectrum. Multiple comparisons were made using the Tukey’s test (α =0.05). Results: Composite brackets showed greater values concerning biofilm formation, when compared with the metallic and ceramic ones, both of which presented similar scores. The hypothesis is accepted. There are differences on the biofilm formation according to the type of material. Conclusions: The in situ model tested was found to be effective in evaluating the accumulation and development of biofilm on orthodontic brackets. In the quantitative analysis, composite brackets showed greater biofilm adhesion values while metallic and ceramic presented similar biofilm absorbance spectrum.

In vitro assessment of antimicrobial activity of Pothomorphe umbellata extracts against Enterococcus faecalis.

Background : Due to the complex anatomy of the root canal system, biomechanical preparation is not able to completely eliminate all microorganisms present in the endodontic infections, making it necessary the use of an intracanal medication. Aim: The aim of this study was to assess the antimicrobial activity of an intracanal medication containing the ethyl-acetate fraction of Pothomorphe umbellata against Enterococcus faecalis. Materials and Methods: Fifty seven human maxillary canine teeth were used, of which 54 were infected with E. faecalis every 72 h, for 28 days, and cultured for 24 h. Contaminated teeth were randomly separated into three groups (n = 18) and treated as follows: Group I - calcium hydroxide-based medication; Group II - P. umbellata-based medication; Group III - contaminated teeth without medication. Three teeth were used as negative control. After 7, 14 and 28 days of treatment, six teeth from each group were assessed for the level of microbial growth after each period of treatment. Results: The intracanal medication containing P. umbellata was effective against E. faecalis after 7, 14, and 28 days of treatment without statistically significant difference in comparison to calcium hydroxide treatment (Kruskal-Wallis test, P > 0.05). Conclusion: Ethyl-acetate fraction of P. umbellate was efficient against E. faecalis, making this phytotherapy a viable option for endodontic treatment.

The cariogenic dental biofilm: good, bad or just something to control?

This paper discusses the role of dental biofilm and adjunctive therapies in the management of dental caries. Dental biofilm is a site of bacterial proliferation and growth, in addition to being a location of acid production. It also serves as a reservoir for calcium exchange between the tooth and saliva. The salivary pellicle, a protein-rich biofilm layer, regulates the reaction between tooth surface, saliva and erosive acids. The protective effects of this pellicle on enamel are well established. However, understanding the effects of the pellicle/biofilm interaction in protecting dentin from erosive conditions requires further research. Saliva interacts with the biofilm, and is important in reducing the cariogenic effects of dental plaque as acidogenic bacteria consume fermentable carbohydrates producing acids that may result in tooth demineralization. Adequate supplies of healthy saliva can provide ingredients for successful remineralization. Strategies for managing the cariogenic biofilm are discussed with emphasis on the effectiveness of over-the-counter (OTC) products. However, since many toothpaste components have been altered recently, new clinical trials may be required for true validation of product effectiveness. A new generation of calcium-based remineralizing technologies may offer the ability to reverse the effects of demineralization. Nevertheless, remineralization is a microscopic subsurface phenomenon, and it will not macroscopically replace tooth structure lost in a cavitated lesion. Optimal management of cavitations requires early detection. This, coupled with advances in adhesive restorative materials and microsurgical technique, will allow the tooth to be restored with minimal destruction to nearby healthy tissue.

Antimicrobial activities of the volatile oils of Ocimum Bacilicum L. and Ocimum Gratissimum l. [lamiaceae] against some aerobic dental isolates

Essential or volatile oils of plants have been variously reported to have many medicinal applications. Their probable uses against oral microbes have received little attention. Oral swabs obtained from eighteen dental patients at the University of Benin teaching Hospital, Benin City, Nigeria, led to the isolation of twenty nine bacteria. Using standard methods, the microorganisms were identified as Streptococcus viridians [16, representing 55.17%], Staphylococcus albus [9, 31.04%], Klebisiella pneumonia [2, 6.90%], Pseudomonas aeruginosa [1, 3.45%] and Proteus vulgaris [1, 3.45%]. The antimicrobial activities of the volatile oils of Ocimum basilicum L. and O. gratissimum L. were evaluated on the twenty nine organisms using agar diffusion and agar dilution methods. In the susceptibility tests, the volatile oils of O. basilicum and O. gratissimum independently inhibited the growth of Klebisiella pneumonia at a concentration of 0.51% in the agar, Streptococcus viridians and Staphylococcus albus at 1.10% and Pseudomonas aeruginosa at 10.0%. Proteus vulgaris was inhibited at 0.53% by the volatile oil of O. gratissimum and 0.67% by O. basilicum. Separate incorporation of the volatile oils into tooth pastes [2 and 5%], the volatiles oils showed antibacterial activities comparable to a commercial tooth paste [which contains O. basilicum 0.01% among others] against most resistant organisms. As components of mouth washes, the volatile oils completely inhibited the growth of organisms at a concentration of 0.5%

Histopathological and histomicrobiological study of root canal therapy medication, comparison of calcium hydroxide versus gutta-percha with zinc oxide/eugenol in the teeth of dogs

The presence of microorganisms in dental structures with experimentally induced necrosis was evaluated. The materials were tested to evaluate their antimicrobial activity and tissue repair efficacy. Four dogs were used in this experiment, with a total of 64 roots of premolar teeth, divided into three groups. The root canals of Group I were filled with gutta-percha and zinc oxide/eugenol cement; Group II were filled with calcium hydroxide, and Group III were not filled. All animals were clinically and radiographically examined 15 days after surgery andthen again every subsequent 15 days until 120 days, when the teeth were extracted en bloc.Histopathological analysis showed inflammatory infiltration, cement and bone resorption andnecrotic tissue in the apical delta in different proportions. Histomicrobiological analysis showedthe presence of microorganisms inside the teeth structures, with different concentrationsaccording to the treatment used. There was statistical significance between the groups(p>0.05). Gutta-percha with zinc oxide/eugenol demonstrated good antimicrobial activity;calcium hydroxide was not efficient. The conclusion of this study is that gutta-percha withzinc oxide/eugenol is the better protocol for filling root canals in dogs.

Avaliou-se a presença de microrganismos nas estruturas dentais com necrose pulpar induzida experimentalmente, testando a eficácia de materiais com relação à atividade antibacteriana e influência no reparo tecidual. Utilizaram-se quatro cães, totalizando 64 raízes, provenientes de pré-molares, divididas em grupos. O Grupo I foi obturado com guta percha e cimento à base de óxido de zinco e eugenol, o Grupo II, com hidróxido de cálcio e o Grupo III, não foram obturados. Todos tiveram controle clínico e radiográfico quinzenal e após 120 dias, foram extraídos em bloco para análises. A histopatologia evidenciou infiltrado inflamatório, reabsorção cementária e óssea e tecido necrótico no delta apical, em diferentes proporções. A histomicrobiologia revelou presença de microrganismos nas estruturas estudadas, variando de acordo com o tratamento utilizado. Os resultados foram submetidos à análise estatística e foram significantes entre si (p>0,05). A guta-percha com óxido de zinco e eugenol apresentou-se eficaz com relação à atividade antibacteriana. O hidróxido de cálcio não se mostrou eficaz. Os resultados obtidos através da utilização destes materiais deixaram evidente que a guta-percha associada ao óxido de zinco e eugenol é o melhor protocolo a ser utilizado no tratamento endodôntico em cães.

The effect of timing temporary cements to treat induced pulp necrosis in the teeth of dogs

During endodontic therapy (pulpectomy, root canal debridement and root canal filling) microbiological management is a major concern. Bacteria present in dentine tubules, apical foramina and apical delta are causally related to failure of the procedure. Studies have shown that during single session endodontic treatment bacteria remain within dental structures. The aim of the present study was to evaluate endodontic treatment performed as two sessions, using temporary endodontic dressing materials for different periods in four groups of experimental dogs. A total of 80 roots of second and third upper premolar teeth and second, third and fourth lower premolar teeth were divided into four groups. The pulp chamber was opened with burrs and the pulp exposed for 60 days to induce pulpal inflammation and necrosis. Groups II, III and IV were treated with calcium hydroxide plus camphorated paramono-chlorophenol (PMCC) for 7, 15 and 30 days, respectively. In all groups, the root canals were filled with zinc oxide-eugenol and gutta-percha cones. Clinical and radiographical measurements were performed every 2 weeks. After 60 days a small block section containing the teeth, surrounding periapical tissues and the periodontium was removed for histological and microbiological study. Histological analysis revealed intense inflammatory response in all groups. Microbiological analysis showed microbial reduction inversely proportional to the period of time that the intracanal temporary medicament was left in place.

Em um tratamento endodôntico, a microbiota é o ponto primordial a ser levado em consideração, pois as bactérias presentes nos túbulos dentinários, nas foraminas e no delta apical em cães estão relacionadas aos insucessos do procedimento. Estudos revelam que tratamentos realizados em uma única sessão ainda permitem a permanência de bactérias nas estruturas dentárias, portanto propõe-se a execução em duas sessões, com diferentes tempos de "curativo de demora", para verificação da eficácia do protocolo em cães. Foram utilizados no total quatro cães, num total de 80 raízes, provenientes dos 2os e 3os pré-molares superiores e 2os, 3os e 4os pré-molares inferiores, divididas em quatro grupos de estudo. Nestes foi realizada abertura coronária, onde a polpa ficou exposta por 60 dias. Os Grupos II, III e IV foram tratados. O hidróxido de cálcio com p-monoclorofenol canforado, isto é, a pasta Calen/PMCC (S.S. White Artigos Dentários Ltda, RJ), foi utilizada como "curativo de demora". O Grupo II permaneceu com "curativo de demora" por sete dias, o Grupo III permaneceu por 15 dias e o Grupo IV por 30 dias, todos os grupos foram obturados com cimento à base de óxido de zinco e eugenol - Herodent (Vigodent S.A. Indústria e Comércio, SP) e guta-percha. Todos os grupos tiveram controle clínico e radiográfico quinzenal e após 60 dias os dentes foram extraídos em bloco (dente e periodonto) e passaram por processamento laboratorial para realização das análises histopatológica e histomicrobiológica. A análise histopatológica de forma geral mostrou intensa reação inflamatória em todos os grupos estudados, já na análise histomicrobiológica pode-se observar uma diminuição no número de microrganismos proporcional ao tempo de "curativo de demora" deixado intra-canal, portanto a permanência do "curativo" por 30 dias foi a mais eficaz.

Sterilisation of extracted human teeth for educational use.

Sixty intact, non-carious and unrestored teeth extracted due to periodontal disease were used to determine the most effective method of sterilisation. The teeth were divided into six groups, each containing 10 teeth. Group 1 teeth were immersed in 10% formalin for seven days, group 2 teeth were immersed in 3% hydrogen peroxide for seven days, group 3 teeth were immersed in 2.6% sodium hypochlorite for seven days, group 4 teeth were boiled in water at 100 degrees C for 20 minutes, group 5 teeth were autoclaved at 121 degrees C at 15 lbs psi for 30 minutes, and group 6 teeth were immersed in normal saline for seven days. After the treatment, the teeth were individually inoculated into trypticase soy broth and incubated for 48 hours. A questionnaire survey was also conducted to determine the awareness of dental students regarding infection due to extracted human teeth and the common disinfection/sterilisation methods used. Autoclaving at 121 degrees C, 15 lbs psi for 30 minutes and immersion in 10% formalin for seven days were effective in disinfecting/sterilising extracted human teeth. Chemicals such as 2.6% sodium hypochlorite, 3% hydrogen peroxide and boiling in water were not effective. The results indicate that autoclaving for 30 minutes or immersion in 10% formalin for seven days could be effectively used for disinfection/sterilisation of extracted human teeth.

Comparison of levels of mutans streptococci and lactobacilli in children with nursing bottle caries, rampant caries, healthy children with 3-5 dmft/DMFT and healthy caries free children.

Ten children from each group of nursing caries, rampant caries, 3-5 dmft/DMFT and caries free children were selected and plaque samples from carious lesion, early carious lesion, sound tooth surface as well as salivary sample were collected. The levels of mutans streptococci and lactobacilli were estimated at all the sample sites. The result further supported the role of mutans streptococci in the initiation and of lactobacilli in the progression of the dental caries.

Apical barrier formation time comparison of clinically infected with un-infected immature non-vital anterior teeth

This study included 60 non-vital immature anterior teeth to determine the barrier formation time. The teeth were divided into two categories: uninfected and clinically infected teeth. Calcium hydroxide paste was made using three different vehicles in both categories to compare the barrier formation time between clinically infected and non-infected non-vital immature anterior teeth

Sangramiento gingival y flora bacteriana en la gingivitis y la periodontitis / Gingival bleeding and bacterial microflora in gingivitis and periodontitis

Se estudiaron 30 sitios o áreas periodontales que presentaban gingivitis y 30 con periodontitis, con el objetivo de determinar la relación existente entre el sangramiento gingival y la flora microbiana presente en la gingivitis y la periodontitis. Los pacientes seleccionados no presentaban antecedentes de enfermedad general y no habían recibido medicación antimicrobiana ni tratamiento periodontal en los últimos 6 meses; en el caso de las mujeres, no podían estar embarazadas. En los dientes seleccionados se procedió a tomar la muestra cumpliendo con los requisitos establecidos; luego se examinó inmediatamente en el microscopio de campo oscuro. Los resultados obtenidos indica que no hubo relación entre los morfotipos microbianos y los diferentes valores del índice de sangramiento gingival

A comparative, qualitative and quantitative antimicrobial efficacies of mouthrinses containing chlorhexidine gluconate and essential oils.

A project was launched to evaluate and compare antimicrobial efficacy of Hexidine and Listerine over placebo in 10 day human experimental gingivitis study. A rigid study schedule for subject compliance spanning well over 3 months was sorted out and volunteers recruited for study rinsed all the 3 assigned mouthrinses containing (a) chlorhexidine gluconate, (b) "essential oils" and (c) flavoured distilled water one after the other at certain prefixed intervals. After 10 days of assigned mouthrinse regimen, where the assigned mouthrinse was the only method practiced by the volunteers for oral hygiene, Supragingival Plaque was quantitatively and qualitatively assayed. Qualitatively supragingival plaque was assayed by Gram staining (Direct smear and Thioglycollate Broth) and growth characteristics i.e; Aerobic, Microaerophilic and Anaerobic growth was noted. Quantitatively plaque was assayed by calculating total microbial load per tooth by preparing Mafarland Nephelometer standard and Spectrophotometric analysis. It is concluded that Hexidine and Listerine exert similar antimicrobial efficacy in 10 days experimental gingivitis study. Hexidine and Listerine exert their antimicrobial influence through reduction of total Aerobes and Anaerobes reducing total microbial load per tooth by 58% and 53% respectively as compared to placebo.