The Clinical Pattern and Prevalence of Streptococcus mutans and Streptococcus sobrinus among Adult and Children Patients with Dental Caries

Abstract Objective: To explore the clinical pattern, host factors, and presentation of Streptococcus mutans related to caries incidence among children and adults visiting Universitas Airlangga dental clinic. Material and Methods: This was an observational study with a cross-sectional approach with 50 patients in each group of carious children (6-12 years) and adults (18-35 years). Dental decay samples were taken by sterile excavator, put in a BHI's transport medium, and directly incubated overnight at 37 ºC. The next day, they were sub-cultured microbiologically in Tryptone Yeast Cystine Sucrose Bacitracin (TYCSB) selective medium. Bacterial species and serogroups were examined by PCR. All patient's data were collected from medical records and direct observation. Results: Caries were mostly media type in both children and adults. Oral hygiene (OHIS) in children was higher than in adults but not significantly different according to their DMFT. The highest scores for decay, missed and filled teeth were 16, 8 and 7, with an average of 6.82, 1.22 and 0.63, considered quite high. Conclusion: The prevalence of S. mutans was higher in children's caries than in adults, but among the adult patients the co-incidence of S. mutans and S. sobrinus was associated with higher DMFT. The mutans serotypes e, f, and d were more prevalent among children than adults.

Creating Antibacterial Properties in Flowable Dental Composites by Incorporation of 3, 4-dihydropyrimidin-2(1H)-ones

ABSTRACT Objective: To investigate the antibacterial, mechanical, physical properties and water sorption of flowable dental composites containing 3,4-dihydropyrimidin-2(1H)-ones. Material and Methods: 3,4-dihydropyrimidin-2(1H)-ones was synthesized and the antibacterial activity of flowable dental composites containing 0-5 wt% 3,4-dihydropyrimidin-2(1H)-ones and also of their mechanical and physical properties on flowable dental composites were investigated. Flexural strength was measured by a three-point bending test. Compressive strength (CS), Water sorption (WS) and depth of cure (DOC) were investigated. The data were analyzed by One-way ANOVA test. The level of significance was determined as p<0.01. Results: The direct contact test demonstrates that by increasing the 3,4-dihydropyrimidin-2(1H)-ones content, the bacterial growth is significantly diminished (p<0.001). The average flexural strength results show that with increasing 3,4-dihydropyrimidin-2(1H)-ones until 3% in the composite, no significant difference was observed in flexural strength (p>0.001) and the mean of compressive strength results show no significant difference between 0-4% groups (p>0.001). The mean of water sorption and depth of cure results shows no significant difference between groups (p>0.001). Conclusion: Incorporation of 3,4-dihydropyrimidin-2(1H)-ones into flowable resin composites in 3% wt can reduce the activity of Streptococcus mutans.

Sucrose, Lactose, and Xylitol Exposures Affect Biofilm Formation of Streptococcus mutans

Abstract Objective: To determine the level of biofilm formation of S. mutans after being exposed to 5% sucrose, 8% lactose, or 1% xylitol. Material and Methods: This research was a laboratory-based experimental study with post-test only control group design. S. mutans was grown in test tubes containing tryptose soy broth (TSB) medium supplemented with 1% glucose. They were incubated at 37° C for 24 hours to grow the biofilms. The culture was then exposed to 5% sucrose, 8% lactose or 1% xylitol, incubated for 24 hours at 37° C, and examined using ELISA at a wavelength of 625 nm. The statistical analysis was performed using a one-way analysis of variance followed by the least significant difference test (a=0.05). Results: There were some differences in the biofilm formation of S. mutans after exposure to 5% sucrose, 8% lactose, or 1% xylitol (p<0.05). An LSD test indicated significant differences among the biofilm formations after exposure to 5% sucrose and 8% lactose and between 5% sucrose and 1% xylitol. In comparison, there were no significant differences (p>0.05) between 8% lactose and 1% xylitol. Conclusion: Sucrose, lactose and xylitol can form biofilms and the formation of lactose biofilms is the same as xylitol.

Type-2 Diabetes Mellitus Individuals Carry Different Periodontal Bacteria

ABSTRACT Objective: To identify etiologic microbiota associated periodontal diseases among diabetes patients and the factors related to the most commonly identified bacteria species. Material and Methods: Periodontal plaque samples from 11 diabetic participants and 13 non-diabetic controls were collected to assess their aerobic and anaerobic bacterial growth. Different distinct colonies were identified by microscopic and 16srDNA sequencing. Pearson's chi-square tests were conducted to examine any association between categorical variables. Results: The diabetic subjects revealed a more intense plaque formation with a mean plaque index of 2.4 compared to 1.8 in non-diabetics. A total of 86 bacteria were isolated from 24 plaque samples, 44 were aerobic, and 42 were anaerobic. Only aerobic isolates, 22 from diabetic patients and 22 from non-diabetic patients, were evaluated in these analyses. Bacillus spp. (B. cereus mainly) and Klebsiella spp. (K. pneumoniae, K. aerogenes, K. oxytoca) were detected markedly higher in non-diabetic individuals than in diabetic subjects (p=0.026 and p=0.021, respectively). Some bacteria were only identified in the dental plaque of diabetic individuals, namely, Bacillus mojavensis, Enterobacter cloacae, Proteus mirabilis, Staphylococcus epidermidis, Staphylococcus hominis, Staphylococcus pasteuri, Streptococcus mutans, and Streptococcus pasteurianus. The presence of acid reflux and jaundice were significantly associated with the most common bacterial isolate, namely Bacillus spp., with the p-values of 0.007 and 0.001, respectively. Conclusion: Type-2 diabetes mellitus is associated with a higher amount of dental plaques. Periodontal plaque samples from diabetic and non-diabetic subjects possess differential microbial communities. Diabetic plaques contain more versatile microbes predominated by gram-positive streptococci and staphylococci.

Sucrose and Xylitol-Induced Streptococcus mutans Biofilm Adherence

Abstract Objective: To study the adherence of Streptococcus mutans biofilm after induction with sucrose and xylitol. Material and Methods: Laboratory experimental study incorporating posttest-only control group design. S. mutans biofilm was generated for 24 hours at a temperature of 37°C using BHIB with 5% sucrose and BHIB with 1% xylitol. An adherence assay was conducted in accordance with the method applied previously. The quantity of adhered bacteria was measured by means of a spectrophotometer at 570 nm. The data were presented as mean and standard deviation. Results: A biofilm induced with sucrose has a higher adherence level (0.9294 ± 0.0431) compared with one induced with xylitol (0.5095 ± 0.0392). Sucrose induces adherence levels by increasing glucan binding protein and glucosyltransferase of the bacteria, whereas xylitol will inhibit the glycolysis process of the bacteria. Conclusion: The adherence of sucrose-induced S. mutans biofilm is higher than that of xylitol-induced S. mutans biofilm.

Atividade antifúngica de metabólitos produzidos por Streptococcus mutans sobre Candida albicans / Antifungal activity of metabolites produced by Streptococcus mutans on Candida albicans

RESUMO A formação de biofilmes e produção de hifas por Candida albicans são importantes fatores de virulência, principalmente, para a aderência à mucosa e invasão tecidual. A busca por metabólitos secundários produzidos por S. mutans é de suma importância, pois poderá fornecer novas estratégias terapêuticas no combate às infecções por Candida, possibilitando o desenvolvimento de medicamentos capazes de inibir os mecanismos de patogenicidade das espécies desse gênero. Desse modo, o objetivo desse estudo foi obter o extrato bruto e frações a partir do sobrenadante da cultura de 4 h de S. mutans (UA159) e avaliar seus efeitos sobre os mecanismos de patogenicidade de C. albicans (ATCC18804) por meio de estudos in vitro. Após o cultivo de S. mutans, o extrato bruto foi obtido via partição líquido-líquido com acetato de etila (3x) e posteriormente fracionado em coluna de sílica derivatizada C-18 eluída com gradiente MeOH:H2O, fornecendo cinco frações (SM-F1, SM-F2, SM-F3, SM-F4 e SM-F5). A identificação das substâncias contidas no extrato bruto e frações foi realizada utilizando cromatografia gasosa acoplada a espectrometria de massas (CGEM), sendo encontradas as seguintes substâncias: ácido octanóico e uridina no extrato bruto, ácido propanóico, (3R)-3-Methyl-1,4-bis(trimethylsilyl)piperazine-2,5- dione, pirimidina, gulose e oleamida na SM-F1 e ácido nicotínico e triptofano na SM- F2. O extrato bruto e as frações foram submetidos aos ensaios de bioatividade sobre a formação de hifas de C. albicans e analisados por microscopia óptica e microscopia eletrônica de varredura (MEV). O extrato bruto e a fração SM-F2, que resultou em maior inibição das hifas, foram investigados na expressão de genes de C. albicans envolvidos no mecanismo de filamentação (CPH1, EFG1, HWP1, UME6 e YWP1) por PCR quantitativo em tempo real (RT-qPCR). Além disso, o potencial de inibição do extrato bruto, frações SM-F1 e SM-F2 foi avaliado sobre a formação de biofilmes de C. albicans, analisados pela quantificação de células viáveis (UFC/mL) e MEV Os resultados demonstraram inibição significativa na formação de hifas de C. albicans pelo extrato bruto e principalmente, pela SM-F2, com alterações significativas na expressão de todos os genes analisados. O extrato bruto e SM-F2 regularam negativamente a expressão dos genes CPH1, EFG1, HWP1 e UME6, em contrapartida, regularam positivamente a expressão do gene YWP1, envolvido no mecanismo de dispersão das leveduras. Nas análises de UFC/mL, os resultados demonstraram redução nas contagens de C. albicans nos biofilmes formados quando em contato com o extrato bruto, frações SM-F1 e SM-F2 em todas as concentrações testadas, sendo que o extrato bruto reduziu totalmente as células de C. albicans na concentração 15 mg/mL. Os resultados do nosso estudo demonstraram que o extrato bruto e frações de S. mutans apresentaram efeitos inibitórios sobre importantes mecanismos de virulência de C. albicans, fornecendo evidências que S. mutans produz substâncias com ação antifúngica, tornando-o promissor na busca de novos compostos antimicrobianos para prevenção e tratamento das candidoses humanas(AU)

The biofilm formation and hyphae production by Candida albicans are important virulence factors, mainly for mucosal adhesion and tissue invasion. The search for secondary metabolites produced by S. mutans is of great importance, since it may provide new therapeutic strategies against Candida infections, enabling the development of drugs that inhibit the mechanisms of pathogenicity of the species of this genus. Thus, the objective of this study was to obtain the crude extract and fractions from the supernatant of the 4 h culture of S. mutans (UA159) and evaluate their effects on the mechanisms of pathogenicity of C. albicans (ATCC18804) through in vitro studies. After culture of S. mutans, the crude extract was obtained via liquid-liquid partition with ethyl acetate (3x) and then fractionated on a C-18 derivatized silica column eluted with MeOH:H2O gradient, providing five fractions (SM-F1, SM-F2, SM-F3, SM-F4 and SM-F5). The identification of the substances contained in the crude extract and fractions was performed by gas chromatography coupled to mass spectrometry (GC-MS). The following substances were found: octanoic acid and uridine in crude extract, propanoic acid, (3R) -3-methyl -1,4-bis (trimethylsilyl) piperazine-2,5-dione, pyrimidine, gulose and oleamide in SM-F1 and nicotinic acid and tryptophan in SM-F2. The crude extract and fractions were submitted to bioactivity assays on hyphae formation by C. albicans and analyzed by optical microscopy and scanning electron microscopy (SEM). The crude extract and SM-F2 fraction, which resulted in highest inhibition of hyphae, were investigated in the expression of C. albicans genes involved in the filamentation mechanism (CPH1, EFG1, HWP1, UME6 and YWP1) by quantitative real-time PCR (RT-qPCR). In addition, the potential of inhibition of the crude extract, SM-F1 and SM-F2 fractions in biofilms of C. albicans were evaluated by the quantification of viable cells (CFU/mL) and SEM. The data obtained were statistically analyzed by Graph Pad Prism 5.0, with a significance level of 5%. The results demonstrated a significant inhibition on C. albicans hyphae formation by the crude extract and mainly by SMF2, with significant alterations in the expression of all genes analyzed. The crude extract and SM-F2 negatively regulated the expression of the CPH1, EFG1, HWP1 and UME6 genes, in contrast, positively regulated the expression of the YWP1 gene, involved in the mechanism of yeast dispersion. In the CFU/mL analyzes, the results showed a reduction in the counts of C. albicans in the biofilms formed when in contact with the crude extract, SM-F1 and SM-F2 fractions in all tested concentrations and the crude extract totally reduced C. albicans cells at 15 mg/mL concentration. The results of our study demonstrated that the crude extract and fractions of S. mutans presented inhibitory effects on important mechanisms of virulence of C. albicans, providing evidence that S. mutans produces substances with antifungal action, making it promising in the search for new compounds antimicrobials for the prevention and treatment of human candidoses(AU)

Estudio in vitro del Efecto Antibacteriano de la Oleorresina de Copaifera reticulata y el Aceite Esencial de Origanum majoricum Frente a Streptococcus mutans y Enterococcus Faecalis Bacterias de Importancia en Patologías Orales / In vitro Study of the Antibacterial Effect of Oleoresin of Copaifera reticulata and the Essential Oil of Origanum majoricum against Streptococcus mutans and Enterococcus faecalis Bacteria of Importance in Oral Pathologies

RESUMEN: El objetivo del estudio fue determinar el efecto antibacteriano in vitro de la oleorresina de Copaifera reticulata (C. reticulata) "copaiba" y del aceite esencial de Oreganum majoricum (O. majoricum) "orégano" frente a Streptococcus mutans (S. mutans) y Enterococcus faecalis (E. faecalis). Se desarrollaron pruebas de sensibilidad activando primero las cepas bacterias a enfrentar. La oleorresina de copaiba fue diluida con dimetilsulfósido (DMSO), obteniéndose al final concentraciones a probar de 100 %, 50 %, 25 %, y 12,5 %. En relación al aceite esencial de orégano este se probó solamente al 100 %. Para la prueba de difusión en agar con discos, se tomaron inóculos 100 µL de cada cepa bacteriana a una turbidez de 0,5 de Mc Farlam, para ser sembrados por diseminación en placas de tripticasa soya agar, para luego colocar los discos de forma equidistante cargados con las diferentes concentraciones de los productos naturales, se utilizaron como control positivo a la clorhexidina al 0,12 % y al DMSO como control negativo. Se incubaron las placas por el método de la vela en extinción a 37 °C, por un periodo de 24 horas, pasado el tiempo se realizó la lectura de los halos de inhibición. Los resultados obtenidos por la copaiba, determinaron un efecto antibacteriano en sus cuatro concentraciones, siendo los mayores halos de inhibición a la concentración del 100 %, copaiba genero mayores halos promedios para S, mutans de 30,00 ± 0,00 mm y para E. faecalis de 8,3 ± 0,50 mm. Para el caso del orégano se producen halos a la concentración del 100 % con un promedio de 25,3 ± 0,96 mm para S. mutans y para E. faecalis de 9,5 ± 1,29 mm. Se concluye del estudio que tanto copaiba como el orégano presentan un efecto antibacteriano para ambas bacterias, siendo su mayor efecto antibacteriano para ambos productos naturales sobre S. mutans.

ABSTRACT: The objective of the study was to determine the in vitro antibacterial effect of the oleoresin of Copaifera reticulata (C. reticulata) "copaiba" and of the essential oil of Oreganum majoricum (O. majoricum) "oregano" against Streptococcus mutans (S. mutans) and Enterococcus faecalis (E. faecalis). Sensitivity tests were developed by first activating the bacteria strains to be confronted. The oleoresin of copaiba was diluted with dimethylsulphoside (DMSO), obtaining final concentrations to be tested of 100 %, 50 %, 25 %, and 12.5 %. In relation to the essential oil of oregano, it was only 100 % tested. For the disk agar diffusion test, 100 mL of each bacterial strain was taken at a turbidity of 0.5 of Mc Farlam, to be planted by dissecting trypticase soy agar plates, and then placing the disks equidistantly loaded with the different concentrations of natural products; 0.12 % chlorhexidine was used as a positive control and DMSO as negative control. The plates were incubated by the candle method in extinction at 37 °C, for a period of 24 hours, after which time the inhibition halos were read. The results obtained by the copaiba, determined an antibacterial effect in its four concentrations, being the biggest halos of inhibition at the concentration of 100 %, copaiba genus higher average halos for S. mutans of 30.00 ± 0.00 mm and for E. faecalis of 8.3 ± 0.50 mm. In the case of oregano, haloes are produced at a concentration of 100 % with an average of 25.3 ± 0.96 mm for S. mutans and for E. faecalis 9.5 ± 1.29 mm. It is concluded from the study that both copaiba and oregano present an antibacterial effect for both bacteria, being its greater antibacterial effect for both natural products on S. mutans.

Comparisons of IgA response in saliva and colostrum against oral streptococci species

Abstract The present study compared IgA specificity against oral streptococci in colostrum and saliva samples. Sixty-two mother-and-child pairs were included; samples of colostrum (C) and saliva (MS) were collected from the mothers and saliva samples were collected from babies (BS). The specificity of IgA against Streptococcus mutans and S. mitis were analyzed by western blot. Only 30% of babies’ samples presented IgA reactivity to S. mutans, while 74 and 80% of MS and C, respectively, presented this response. IgA reactivity to S. mutans virulence antigens (Ag I/II, Gtf and GbpB) in positive samples showed differences between samples for Gtf and especially for GbpB (p < 0.05), but responses to Ag I/II were similar (p > 0.05). The positive response of Gtf-reactive IgA was different between C (90%) and MS (58%) samples (p < 0.05), but did not differ from BS (p > 0.05). GbpB was the least detected, with 48 and 26% of C and MS, and only 5% of BS samples presenting reactivity (p > 0.05). Eight percent of MS and C samples presented identical bands to SM in the same time-point. In conclusion, the differences of IgA response found between C and MS can be due to the different ways of stimulation, proliferation and transportation of IgA in those secretions. The colostrum has high levels of IgA against S. mutans virulence antigens, which could affect the installation and accumulation process of S. mutans, mainly by supplying anti-GbpB IgA to the neonate.

Validade e confiabilidade de kits para detecção dos níveis de estreptococos do grupo mutans e lactobacilos na saliva de crianças e adultos / Validity and reliability of kits for salivary detection of mutans streptococci and lactobacilli in infants and adults

Objetivo: Avaliar a validade e a confiabilidade dos resultados obtidos por meio de kits pré-fabricados disponíveis no mercado brasileiro para a detecção e quantificação dos níveis de estreptococos do grupo mutans e lactobacilos na saliva de crianças e adultos. Método: Amostras salivares de 15 crianças (12-24meses) e 14 adultos (17-35 anos) foram coletadas e analisadas quanto aos níveis salivares de estreptococos do grupo mutans e lactobacilos pelo uso dos kits Dentacult I e II (Laborclin) e por método microbiológico convencional, baseado em cultura em meios seletivos. Os resultados de ambos os métodos foram comparados pelo uso dos coeficientes Kappa ponderado (Kp) e de correlação de Spearman. Para a determinação da concordância inter e intraexaminador da leitura dos resultados obtidos com o uso dos kits, os dados foram comparados por meio do Kp. As análises estatísticas foram realizadas utilizando o software Minitab 15 (versão 15.1.0.0.). Foi adotado o nível de significância de 5%. Resultados: Os resultados apresentados pelo kit Dentalcult I apresentaram concordância regular e correlação não-significante com os obtidos com o método microbiológico convencional (rs=0,53; p maior que 0,05). Por outro lado, os resultados obtidos com o kit Dentalcult II apresentaram boa concordância e correlação estatisticamente significante com os do método convencional (rs=0,77; p menor que 0,05). Os kits Dentalcult I e II tiveram confiabilidade de leitura perfeita (Kp=1) em momentos distintos, independentes do examinador. A concordância interexaminadores foi perfeita para o kit Dentalcult II (Kp=1) e ótima para o Dentalcult I (Kp=0,94). Conclusão: O kit Dentalcult II pode ser indicado para a estimativa dos níveis salivares de estreptococos do grupo mutans em substituição aos métodos convencionais.

Objective: To compare the results of commercially available kits for determination of mutans streptococci (MS) and lactobacilli (LB) salivary levels in infants and adults. Method: Salivary samples of 15 children (aged 12-24 months-old) and 14 adults (aged 17-35 years-old) were collected and evaluated for salivary levels of MS and LB by using the Dentacult I and II kits, respectively (Laborclin) and conventional methods based on culture in selective media. The results of both methods were compared to those obtained by the weighted kappa coefficients (Kp) and Spearman's correlation. Statistical analysis was performed using Minitab software 15 (version 15.1.0.0). It was adopted a significance level of 5%. Results: The data obtained with the Dentalcult I kit showed regular agreement and no significant correlation with the conventional microbiological method (rs=0.53, p greater than 0.05). On the other hand, results provided by the Dentalcult II kit produced good agreement and were statistical significant correlated with those of the conventional method (rs=0.77, p less than 0.05). The kits Dentalcult I and II presented perfect reading reliability (Kp=1) in distinct moments, not -dependent on the examiner. The inter-examiner agreement was considered perfect for the kit Dentalcult II (Kp=1) and satisfactory for Dentalcult I (Kp=0.94). Conclusion: The kit Dentalcult II proved validity and reliability for detecting salivary MS.

Dental caries vaccine.

Dental caries is one of the most common diseases in humans. In modern times, it has reached epidemic proportions. Dental caries is an infectious microbiologic disease of the teeth that results in localized dissolution and destruction of the calcified tissue. Dental caries is a mulitifactorial disease, which is caused by host, agent, and environmental factors. The time factor is important for the development and progression of dental caries. A wide group of microorganisms are identified from carious lesions of which S. mutans , Lactobacillus acidophilus , and Actinomyces viscosus are the main pathogenic species involved in the initiation and development of dental caries. In India, surveys done on school children showed caries prevalence of approximately 58%. Surveys among the U.S. population showed an incidence of 45.3% in children and 93.8% in adults with either past or present coronal caries. Huge amounts of money and time are spent in treating dental caries. Hence, the prevention and control of dental caries is the main aim of public health, eventually the ultimate objective of public health is the elimination of the disease itself. Recently, dental caries vaccines have been developed for the prevention of dental caries. These dental caries vaccines are still in the early stages.

Production of monoclonal antibodies against Streptococcus mutans antigens

Several studies have been conducted in the last decades aiming to obtain an anti-caries vaccine, however some studies have demonstrated cross reactivity between Streptococcus mutans surface antigens and the human cardiac tissue. In this work, the reactivity of five anti-Streptococcus mutans monoclonal antibodies (MoAb) (24A, 56G, C8, E8 and F6) was tested against oral streptococci, cardiac antigens and skeletal and cardiac myosins, aiming to evaluate the specificity of these MoAb. The hybrid producers of immunoglobulins of the IgG2b class were cloned by limit dilution and expanded in vivo. MoAb were tested by ELISA. The hybrid 24A reacted with S. mutans CCT 1910, S. salivarius CCT 0365 and S. pyogenes T23. No reactivity difference was observed among the tested species. Cross reactivity with heart and cardiac myosin was not confirmed and only reaction with myosin of skeletal muscle was observed (p = 0.0381). The hybrid 56G reacted with all the tested microorganisms and there was statistically significant difference between S. mutans and S. pyogenes T23 (p < 0.001). This hybrid also reacted with myosin of skeletal muscle (p = 0.0095). C8, E8 and F6 presented low reactivity against oral streptococci strains and no reactivity against cardiac antigens. The data of this study showed that the 24A and 56G anti-S. mutans MoAb presented reactivity with S. pyogenes and S. salivarius, reinforcing the occurrence of common antigens between these species. The tested MoAb presented low cross-reactivity with myosin of skeletal muscle, but anti-heart activity could not be confirmed.

Diversos estudos foram realizados nas últimas décadas com o intuito de se obter uma vacina anticárie dentária, mas alguns trabalhos têm demonstrado reatividade cruzada entre antígenos de superfície de Streptococcus mutans e tecido cardíaco humano. Neste trabalho, foi testada a reatividade de cinco anticorpos monoclonais (AcMo) anti-Streptococcus mutans (24A, 56G, C8, E8 e F6) contra estreptococos orais, antígenos cardíacos e miosinas esquelética e cardíaca, no intuito de avaliar a especificidade desses AcMo. Os híbridos produtores de imunoglobulinas da classe IgG2b foram clonados por diluição limite e expandidos in vivo. Os AcMo foram testados por ELISA. O híbrido 24A reagiu com S. mutans CCT 1910, S. salivarius CCT 0365 e S. pyogenes T23. Nenhuma diferença de reatividade foi detectada entre as espécies analisadas. Reatividade cruzada com coração e miosina cardíaca não foi confirmada, existindo somente reação com miosina de músculo esquelético (p = 0,0381). O híbrido 56G reagiu com todos os microrganismos testados e houve diferença estatisticamente significante entre S. mutans e S. pyogenes T23 (p < 0,001). Este híbrido também reagiu com miosina de músculo esquelético (p = 0,0095). C8, E8 e F6 apresentaram baixa reatividade contra cepas de estreptococos orais e nenhuma reatividade com antígenos cardíacos. Os dados deste trabalho demonstraram que os AcMo 24A e 56G anti-S. mutans reagiram com S. pyogenes e S. salivarius, confirmando a existência de antígenos comuns entre essas espécies. Esses AcMo avaliados apresentaram baixa reatividade cruzada com miosina de músculo esquelético, porém a atividade anticoração não foi confirmada.

Correlation among mutans streptococci counts, dental caries, and IgA to Streptococcus mutans in saliva

Duzentos e quarenta indivíduos divididos em cinco grupos foram estudados: crianças livres de cáries, crianças com cáries, crianças com cáries rampantes, adultos jovens com e sem cáries. Saliva total estimulada foi coletada, e todos os indivíduos foram investigados para CPO-D/ceo-d seguindo-se os critérios da OMS e o índice de higiene oral simplificado (IHOS). Foi também realizada a quantificação da microbiota aeróbica total e de estreptococos do grupo mutans na saliva. Além disso, o nível de IgA anti-S. mutans foi determinado por ELISA. Crianças com cárie rampante apresentaram maior valor de IHOS. A contagem total de microrganismos mais elevada foi encontrada no grupo de crianças com cárie. Nenhuma diferença estatisticamente significativa foi observada em relação a fluxo salivar, IHOS e contagens de microrganismos entre os grupos de adultos jovens. Nenhuma correlação entre contagem de estreptococos do grupo mutans e níveis de IgA anti-Streptococcus mutans foi observada em nenhum dos grupos estudados. Foi encontrada uma correlação entre níveis mais altos de IgA anti-S. mutans e ausência de cáries entre os adultos jovens, mas não entre as crianças.

Efeitos da imunização com antígenos de superfície de Streptococcus mutans na auto-reatividade / Effeets of immunization with Streptococcus mutans' surface antigens in auto-reaetivity of BALB/c mice

Camundongos BALB/c foram imunizados com antígenos de superfície de Streptococcus mutans, cepa GS5. Observamos que a imunização elevou significativamente os níveis de IgGs anticoração e antimiosina livres e presentes em imunocomplexos. No Westem-blotting estes auto-anticorpos reagiram com miosina cardíaca e principalmente com uma banda de 35kDa do extrato cardíaco. A análise histológica dos corações demonstrou ausência de alterações significativas nas válvulas ou miocárdio. Também não foram encontrados anticorpos ligados ao tecido cardíaco. O tratamento com ciclofosfamida foi capaz de reduzir os níveis de auto-anticorpos. não alterando porém o aspecto histológico do coração. Os resultados deste trabalho mostraram que os antígenos de superfície da cepa GS5 de S. mutans não foram capazes de iniciar uma lesão cardíaca em camundongos BALB/c, embora elevassem consideravelmente o nível de anticorpos livres e presentes em imunocomplexos reativos com coração e miosina.

Detecçäo de anticorpo anticoraçäo em camundongos Balb/c imunizados com Streptococcus mutans / Detection of heart-reactive antibody in Balb/c mice immunized with Streptococcus mutans

Anticorpos para antígenos cardíacos foram analisados por ELISA em 14 soros de camundongos Balb/c hiperimunizados com Streptococcus mutans, inativado pelo formaldéido. Os níveis de anticorpos da classe IgG anticoraçäo e antimiosina elevaram-se significativamente nos animais imunizados quando comparados com os controles, especialmente no grupo A, imunizado e reestimulado com antígenos solúveis de S. mutans. Neste grupo, os resultados do "Western Blot" mostraram reatividade com miosina cardíaca e uma banda de 35 kDa. A análise histológica dos coraçöes dos animais do grupo B, imunizado e reestimulado com antígenos de superfície do microrganismo, demonstrou a presença de degeneraçäo celular, tipo hidrópica e hialina e focos inflamatórios constituídos de linfócitos e macrófagos no miocárdio e pericárdio. Os resultados deste trabalho reforçam a hipótese da existência de mimetismo antigênico entre tecido cardíaco e S. mutans e chamam a atençäo para o risco de desenvolvimento de anticorpos reativos com antígenos próprios induzidos por vacina anticárie com componentes estreptocócicos

Vacunas anticaries nuevas estrategias para la inmunizacion contra la caries dental / Dental caries vaccine new strategies for immunization against dental caries

La vacunacion contra la caries, tiene por objeto la estimulacion del sistema inmunitario, para aumentar la secrecion de anticuerpos a nivel salival, y asi evitar la acumulacion y proliferacion de bacterias cariogenicas sobre las superficies dentarias. La evidencia cientifica acumulada en los ultimos treinta años presenta a los Streptococcus mutans, como los principales causantes de las caries en el ser humano. Este microorganismo ha sido el objetivo en la mayoria de las investigaciones en el campo de la inmunidad. La estrategia tradicional es la inmunizacion activa, mediante la inoculacion o la ingestion de la vacuna. Recientemente, en alimentos de consumo masivo se incluyo el uso de Streptococcus lactis modificados geneticamente con una proteina de los Streptococcus mutans, iniciando la estimulacion de anticuerpos. La inmunizacion pasiva, mediante la aplicacion local de anticuerpos es una alternativa en evaluacion, con resultados muy alentadores. Existen diversas vacunas anticaries que resultan efectivas e inocuas en animales de experimentacion. Algunas de estas formas de inmunizacion estan sujetas a pruebas clinicas en voluntarios y se espera que en pocos años puedan ser utilizadas ampliamente.

Análise dos efeitos da soluçäo de fluoreto estanhoso associado ao flúor fosfato acidulado sobre a placa bacteriana e na quantidade de Streptococcus mutans em indivíduos de 21 a 23 anos de idade / Evaluation of the effects of a stannous fluoride mouthrinse associated to the acidulated phosphate fluoride on dental plaque and on salivary Streptococcus mutans in patients aged 21 to 23 years

O presente trabalho teve a finalidade de avaliar os efeitos de uma soluçäo de enxágüe contendo fluoreto estanhoso (SnF2) a l,64 por cento e flúor fosfato acidulado a 0,3l por cento sobre a placa bacteriana e na quantidade de Streptococcus mutans na saliva em indivíduos com idade variando entre 21 a 23 anos. Quinze voluntários utilizaram a soluçäo experimental uma vez ao dia, durante vinte três dias. Tanto o índice de placa como a análise salivar foram realizados antes e após a fase experimental, quando se observou que a utilizaçäo da soluçäo experimental propociou uma diminuiçäo significativa no índice de placa e na quantidade de Streptococcus mutans na saliva, levando os autores a concluir que este pode ser um método efetivo na prevençäo da cárie dentária, bem como de problemas periodontais

Detecçäo e caracterizaçäo das classes de anticorpos anti-Streptococcus mutans presentes na saliva de crianças em diferentes fases de instalaçäo da dentiçäo decídua / Detection and characterization of salivary antibodies against S. mutans in edentulous and dentate children

Um total de 52 crianças, pertecentes à faixa etária de 3 a 34 meses e em diferentes fases de dentiçäo, foram avaliadas quanto à presença de anticorpos salivares dirigidos contra antígenos protéicos de um extrato de Streptococcus mutans. Crianças edêntulas (G1), com dentes anteriores erupcionados (G2), ou com molares decíduos erupcionados (G3) foram selecionadas para o presente estudo. Extrato da bactéria (S. mutans ATCC 25175) foi obtido, e a detecçäo dos anticorpos salivares foi realizada através da técnica ELISA. Apenas 5 por cento das amostras dos G2 ou G3 apresentaram níveis baixos ou indetectáveis de IgA anti-S. mutans. Nas crianças edêntulas essa porcentagem também foi baixa, em torno de 8 por cento (1/12). Com relaçäo à IgG e IgM, respectivamente 83 por cento (10/12) e 67 por cento (8/12) amostras do G1 apresentou valores baixos ou indetectáveis de anticorpos. Nas amostras do G2, 70 por cento apresentaram títulos baixos de IgM e IgG, e nas amostras do G3, 57 por cento apresentaram valores baixos para IgG e 63 por cento para IgM, respectivamente. A separaçäo eletroforética do extrato revelou que a maioria das amostras apresentaram IgA, IgG e IgM dirigidos contra determinadas proteínas, tais como as de 150 kDa, 74 kDa e 39 kDa. Outras amostras apresentaram anticorpos que reconheceram apenas algumas das bandas reveladas por SDS-PAGE. Apenas 1 amostra do G1, reconheceu a banda de 185 kDa com conjugado anti-IgM e duas amostras do G3 apresentaram IgA anti-antígeno I/II. Nenhuma das amostras apresentou IgG anti-antígeno I/II. É possível que, a exposiçäo freqüente da bactéria seja necessária para haver resposta específica a adesina I/II

Humoral immune response to mutans streptococci associated with dental caries.

BACKGROUND. Mutans streptococci are important aetiological agents in dental caries and their prolonged contact with oral tissues evokes a variety of immune responses through local secretory and systemic antibodies. Patterns of such humoral responses in Indian children have not been reported and we undertook the present study to examine these. METHODS. One hundred and twenty-six children with dental caries and 55 matched controls were studied and saliva and sera collected from them. The tests on these specimens included total salivary and systemic immunoglobulins of different classes using radial immunodiffusion and Streptococcus mutans specific IgA, IgG and IgM using specifically standardized enzyme immunoassays. RESULTS. Children with caries had higher levels of IgG (1350 +/- 9.9 mg/dl; controls 1110 +/- 6.7 mg/dl) and IgA (260 +/- 1.8 mg/dl; controls 190 +/- 1 mg/dl) in the serum but their saliva had lower levels of total IgG (160 +/- 0.7 mg/dl; controls 340 +/- 2.9 mg/dl) and IgA (130 +/- 0.5 mg/dl; controls 410 +/- 3 mg/dl). IgM levels in caries children and controls were not significantly different. Higher levels of Streptococcus mutans specific IgA were detected in the saliva of 95 out of 126 (75%) children with caries compared to 13 out of 55 (22%) controls. Specific serum IgG and IgA levels were also increased in 105 and 114 children with caries, although the levels were not as high as those in saliva. Total and specific salivary and serum IgM antibodies were similar in children with caries and control subjects. CONCLUSION. The nature of the humoral immune response in Indian children with dental caries suggests that Streptococcus mutans specific salivary and serum antibodies may play a major role in pathogenesis. Our findings may have importance when devising methods for follow up and prognosis as well as for vaccination strategies.

Differential cell-mediated immune response to S. mutans in children with low and high dental caries.

Role of cell-mediated immune response (CMI) in dental caries was studied in 171 subjects, comprising of 86 children with low caries (LC), 31 with high caries (HC), and 54 age matched controls. [3H]thymidine mediated lymphoblast transformation test (LTT) using mutans streptococci antigen as stimulant was used to study the stimulation index (SI) of in vitro cultured lymphocytes from these children. The analysis revealed low stimulation index in high caries children whereas low caries children exhibited high stimulation index normally ranging between 2 to 6. The findings indicated that low caries children had strong CMI response as compared to high caries children. Although, the findings are based on limited number of samples, it certainly lays emphasis on protective or regulatory role of CMI in different phases of dental caries.