Desequilíbrios no desenvolvimento da microbiota bucal podem gerar algumas patologias e, entre elas, está a cárie dental, uma doença crônica - contagiosa, de etiologia multifatorial, mas extremamente relacionada à bactéria Streptococcus mutans, que através de seu metabolismo fermentativo destrói estruturas mineralizadas do dente. Além disso, o aumento da resistência à antibioticoterapia, devido à existência do biofilme (também formado pelo S. mutans) e aos tratamentos bactericidas mal sucedidos, faz do desenvolvimento de técnicas antimicrobianas alternativas um importante foco nesta área de pesquisa. Assim, na busca de novas metodologias contra microrganismos, estudos usando a Terapia fotodinâmica (TFD) têm sido empregados para se obter a inativação de bactérias como o S. mutans. A TFD é uma modalidade terapêutica em que há a integração de uma luz visível, um corante e oxigênio que quando interagem, levam à produção de diferentes espécies reativas de oxigênio (ERO?s) que irão desencadear uma seqüência de eventos biológicos, resultando numa possível morte ou inativação celular, inclusive de microrganismos como o S. mutans. O objetivo deste trabalho foi avaliar a ação da TFD sobre a bactéria S. mutans em meio planctônico ou formando biofilme. Assim, a metodologia padronizada possibilitou o cultivo da bactéria em meio líquido (planctônico) ou na forma de biofilme, tornando-se possível a realização de testes com a aplicação da terapia fotodinâmica. Uma análise das curvas de crescimento possibilitou avaliar o metabolismo anaeróbico de S. mutans que consome açúcares fermentáveis (glicose, frutose, sacarose e maltose), mostrando-se hábil na produção de ácidos. Conforme proposto, um fotopolimerizador de resina odontológica foi empregado como fonte de luz na TFD. Esta luz é emitida em comprimentos de onda variáveis (possível graças à troca do filtro de luz do equipamento) que fotoativam os diferentes corantes empregados tais como: Rose bengal, a Protoporfirina IX (PPIX) e a Zinco ftalocianina. Estes agentes fotossensíveis, depois de irradiados, produzem espécies reativas de oxigênio, principalmente o oxigênio singlete, que leva à inativação bacteriana observada nos experimentos realizados. Em contrapartida, as concentrações de luz e corantes empregados na inativação bacteriana apresentaram valores de baixa toxicidade para culturas de fibroblastos. Além disso, a grande diferença entre as concentrações de corante que causa a morte da bactéria na presença e na ausência de luz, indica que os compostos produzidos pela fotoativação são os responsáveis pela morte bacteriana em meio planctônico. Quanto ao biofilme formado por S. mutans, é possível promover uma diminuição da sua formação tanto com a TFD quanto com o emprego de flúor. Células irradiadas imediatamente após a adição do corante foram semelhantemente inativadas após TFD, podendo ser desconsiderada uma incubação prévia para que agente fotossensível fosse homogeneamente distribuído. Aparentemente a TFD não acarretou em mudanças na atividade ATPase total extraída da membrana da bactéria. Os experimentos para a determinação da localização dos corantes na célula ou para a avaliação de sua capacidade acidogênica não foram conclusivos em relação à possibilidade das espécies reativas estarem causando danos à parede, membrana plasmática ou ainda em alguma biomolécula citoplasmática da bactéria. Em adição, os estudos da peroxidação lipídica, de análise do DNA e de proteínas de stress após a TFD não trouxeram nenhuma informação adicional conclusiva sobre o alvo específico das ERO?s que causam a inativação bacteriana. Esta metodologia, apesar de ainda não poder ser aplicada na clínica, poderá complementar as técnicas antimicrobianas convencionais, por ser uma metodologia simples, de fácil emprego e baixo custo, podendo inclusive ser associada à possibilidade do uso de diferentes agentes fotossensíveis e adequada às necessidades da prática odontológica. / Unbalance in the development of the mouth macrobiota can originate some pathologies, dental cavity being among them - a chronic disease - contagious, of multifactorial etiology, but extremely related to the Streptococcus mutans bacteria, which, through its fermentative metabolism, destroys tooth mineralized structures. Besides that, the increase in resistance to antibiotic therapy, due to the existence of biofilm (also formed by S. Mutans) and to non successful bactericide treatments, makes alternative antimicrobial techniques development an important focus in this research area. So, in search of new methodologies against microorganisms studies using the Photodynamic Therapy (TFD) have been employed to achieve inactivation of bacteria such as S. mutans. TFD is a therapeutical modality in which there is an integration of a visible light, a dye and oxygen and, when they interact, there is the production of different reactive oxygen species (ERO?s) which will lead to a sequence of biological events, resulting in a possible cellular death or inactivation, including microorganisms such as S. mutans. The aim of this work was to evaluate the action of TFD on S. mutans bacteria in planktonic medium of forming biofilm. So, the standardized methodology allowed the cultivation of bacteria in liquid media (planktonic) or in the biofilm form, making it possible to do tests with the application of the photodynamic therapy. The analysis of the growth curves allowed the evaluation of the anaerobic metabolism of S. mutans, which consumes fermentable sugars (glucose, fructose, sucrose and maltose), and is apt in acid production. According to what is was proposed, a odontological resin photopolymerizer was used as light source in TFD. This light is sent in variable wave lengths (made possible thanks to the change of the light filter in the equipment) which photoactivate the different dyes used, such as Rose Bengal, Protoporfirine IX (PPIX) and the Zinc Phtalocyanine. These photosensitive agents, after being irradiated, produce reactive oxygen species, specially the singlet oxygen, which leads to the bacterial inactivation observed in the experiments done. Nonetheless, the light concentrations and dyes used in the bacterial inactivation have shown low toxicity values for fibroblasts cultures. Besides that, the high difference between the dye concentrations which causes the death of bacteria in the presence and absence of light indicates that the compounds produced by the photoactivation are the responsible for the bacterial death in planktonic media. As for the biofilm formed by S. mutans, it is possible to have a decrease in its formation with TFD as much as with the use of fluoride. Cells irradiated immediately after the dye addition were similarly inactivated after TFD, so a previous incubation for the homogeneous distribution of the photosensitive agent can be discarded. Apparently, the TFD did not cause changes in the total APTase activity extracted from bacteria membrane. The experiments for the determination of dye location in the cell or for the evaluation of its acidogenic capacity were not conclusive in relation to the possibility of reactive species to be causing damage to the wall, plasmatic membrane or still in some cytoplasmatic biomolecule of the bacteria. In addition, the studies of lipidic peroxidation, DNA analysis and stress proteins after TFD did not bring any conclusive additional information about the specific target of ERO?s that cause bacterial inactivation. This methodology, although can?t be applied in clinic yet, can complement the conventional antimicrobial techniques, being a simple methodology, of easy used and low cost, which can also possibly be associated to the use of different photosensitive agents and adjusted to the needs of odontological practice.

Made available in DSpace on 2014-06-11T19:30:59Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-07-28Bitstream added on 2014-06-13T21:01:51Z : No. of bitstreams: 1 paradella_tc_dr_sjc.pdf: 4215723 bytes, checksum: f39144211c5619098d46e5c06f9aa776 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Objetivo deste trabalho foi avaliar diferentes materiais restauradores após indução de cárie secundária in vitro, in situ e aderência de S. mutans sobre estes materiais. Oitenta e quatro molares humanos foram restaurados com diferentes materiais (Z250, Freedom, Vitremer e Fuji IX) e subdivididos em: desafio cariogênico com S. mutans ou ciclagem de pH (n=12). Para o estudo in situ, aparelhos removíveis contendo um espécime restaurado com cada material foram utilizados por 12 voluntários por 14 dias (dia e noite). Uma gota de sacarose a 20% foi pingada nos espécimes 8X/dia. O grupo controle teve espécimes restaurados e não submetidos aos desafios cariogênicos. Os espécimes foram submetidos à microscopia de luz polarizada e EDS. Para a aderência, espécimes dos materiais foram imersos em cultura de S. mutans ATCC 35688 em caldo sacarosado. Após 48h (37°C/5% CO2), foi realizada contagem das unidades formadoras de colônias (log UFC/mL). Os resultados foram submetidos à análise estatística Kruskall-Wallis e Student-Newman-Keuls ou Comparação de Dunn e demonstraram diferenças entre as metodologias in vitro e in situ (p<0,05), porém sem diferenças entre as metodologias in vitro. Fuji IX demonstrou maior concentração de cálcio e menor desmineralização do esmalte adjacente, diferindo dos demais materiais. Os valores médios de aderência (±desvio padrão) foram: Z250 6,41 (0,15)a; Freedom 4,90 (0,95)b; Vitremer 5,13 (0,27)b; Fuji IX 4,89 (0,62)b. Letras diferentes significam diferença estatística significante. Concluiu-se que conforme a metodologia de desafio cariogênico, os materiais restauradores apresentaram diferentes resultados e que materiais fluoretados apresentaram menor aderência de S. mutans. / The purpose of this study was to different restorative materials under in vitro and in situ secondary caries-inducing methodologies and the adherence of S. mutans to restorative materials. Eighty-four extracted human third molars were restored with different materials (Z250, Freedom, Vitremer and Fuji IX) and divided into: S. mutans cariogenic challenge or pH-cycling cariogenic challenge (n=12). For the in situ study, removable appliances having enamel specimens restored with each material were used by 12 volunteers for 14 days (day and night). One drop of 20% sucrose solution was dripped on the specimens 8X/day. Control group had restored specimens that were not submitted to any cariogenic challenge. Specimens were submitted to polarized light microscopy, SEM and EDS. For the adherence testing, specimens of each restorative material were immersed in S. mutans ATCC 35688 culture in sucrose broth. After 48h (37°C/ 5% CO2), the countage of colony forming units was performed (log cfu/mL). Results were submitted to Kruskall-Wallis and Student-Newman-Keuls or Dunn Comparison statistical analysis and showed differences between in vitro and in situ methodologies (p<0.05), however no differences were found between in vitro methodologies. Enamel adjacent to Fuji IX showed higher calcium concentration and less demineralization, differing from the other materials. Mean adherence values (±standard deviation) were: Z250 6.41 (0.15)a; Freedom 4.90 (0.95)b; Vitremer 5.13 (0.27)b; Fuji IX 4.89 (0.62)b. Different letters indicate significant statistical difference. It was concluded that according to secondary-caries inducing methodology, restorative materials presented different results and fluoridated materials presented less S. mutans adherence.

Made available in DSpace on 2017-03-14T14:10:06Z (GMT). No. of bitstreams: 0 Previous issue date: 2015. Added 1 bitstream(s) on 2017-03-14T14:42:48Z : No. of bitstreams: 1 000874703_20170916.pdf: 127430 bytes, checksum: 6c7f9d658a64027218edeedd8022d757 (MD5) Bitstreams deleted on 2017-09-22T12:16:27Z: 000874703_20170916.pdf,. Added 1 bitstream(s) on 2017-09-22T12:17:18Z : No. of bitstreams: 1 000874703.pdf: 1294999 bytes, checksum: 1c6b907cb99545ab91c73127d7a207c0 (MD5) / As restaurações e selantes podem sofrer cárie como qualquer material, devido à possibilidade das bactérias de aderir-se a suas superfícies. Objetivo principal foi colher e quantificar (contagem) das unidades formadoras de colônias de S. mutans de forma direta pela técnica da moldeira (TR) em restaurações de resina composta (RC), amálgama (AM) e em selantes de fóssulas e fissura. Material e método: Recrutaram-se 40 pacientes para o estudo de avaliação da TR, 65 para a comparação das restaurações de AM e RC e 38 para o selante. Critérios inclusão: pacientes 18-50 anos, mínimo 28 dentes na boca, pelo menos uma restauração oclusal de RC e de AM avaliadas alfa pelo critério Ryge (Kappa 0,8), e um dente com indicação de selante. Numa primeira parte, no mesmo paciente foram realizadas amostragem de saliva, impressões de RC ou AM com a TR e a técnica toothpick (TT). Na segunda e terceira etapa num mesmo paciente foram realizadas impressões de RC ou AM com a TR e no caso de selante num mesmo dente foi realizado antes e após da aplicação. TR foi feita numa moldeira de fluoreto com ágar tripticase, extrato levedura, L-cisteína, sacarose e bacitracina (TYCSB) preparada previamente. As amostras foram incubadas a 37ºC durante 48 horas, após uma contagem de unidades formadoras de colônias (UFC) foi realizado por um operador calibrado. Os resultados foram analisados com um nível de confiança do 95%. Resultados: TR foi capaz de colher e isolar S. mutans, e ter uma correlação positiva significativa (r = 0,95) com TT. Não houve correlação significativa entre TR e grupo saliva (r = 0,47). RC tive diferença estatística significativa com maior número de UFC de S. mutans do que AM (p< 0,05). A mediana de UFC/cm2 foi de 111,00 para antes aplicação de selante e 22,00 após aplicação(p< 0,05). Conclusões : TR pode colher UFC de S. mutans desde as superfícies ... (Resumo completo, clicar acesso eletrônico abaixo) / The restorations suffer secondary decay as anything material, because of the possibility of bacteria from adhering to theirs surfaces. Therefore the aim of the study is quantified (count) the colonies forming units (CFU) of S. mutans by tray technique in resin composite (RC), amalgam (AM) restorations and sealants of pits and fissures. Material and Method: 40 patients were selected to assess the tray technique, 65 patients to compare AM restorations and RC and 38 patients for sealant. Inclusion criteria: Patients 18 - 50 years old, with 28 teeth, at least one occlusal restorations with RC and AM, evaluated alfa Ryge criteria (Kappa 0.8), and at least with one teeth with indication of occlusal sealant. First study, int the same patient was taken saliva samples, for RC or AM took the impression by TR and toothpick technique (TT). The second and third parts in the same patient were taken RC and AM impression by TR and the sealant of study in the same tooth was performed before and after the sealant application. This method was performed with a fluoride tray previously loaded with a solid tripticase yeast extract cysteine sucrose and bacitracin agar. The samples were incubated at 37°C for 48 hours, after the colonies forming units (CFU) were quantified by a calibrated operator (Kappa 0.8). The results were analyzed with a confidence level of 95%. Results: TR was able to recover and isolated S. mutans, and had a positive correlation (r= 0.95) with TT. There was no significant correlation between TR and saliva (r= 0.47). RC restorations, there was statistical significant difference, with higher number of CFU of S. mutans than AM (p< 0.05). The median CFU/cm2 was 111,00 before the sealant application and 22,00 after sealant application (p< 0.05). Conclusion: The TR was be able recover CFU of S. mutans from occlusal restorations with positive correlation and significant with TT...(Complete abstract electronic access below)

Resumo: O objetivo deste estudo foi avaliar a contaminação de escovas dentais por Streptococcus grupo mutans e a ocorrência de infecção cruzada por Streptococcus mutans após armazenagem conjunta das mesmas. A amostra foi composta de 46 crianças de 6 a 7 anos de idade divididas em Grupo A com 22 e Grupo B com 24 alunos, de acordo com a sala de aula que freqüentavam. As amostras de biofilme obtidas da cavidade bucal das crianças no início do estudo e o biofilme das escovas dentais das crianças que compartilhavam o mesmo ambiente, foram recolhidas após 21 dias e cultivadas sob condições adequadas em meio de cultura SB20. Em cada escova foram isoladas UFC com morfologia típica, as quais foram identificadas por meio de testes bioquímicos e o polimorfismo genético foi detectado pela técnica de AP-PCR. Pode-se observar 94 UFC de Streptococcus mutans isoladas a partir das escovas dentais. O crescimento bacteriano ocorreu em 14 escovas dentais, sendo que 6 escovas pertenciam às crianças do Grupo A, e 8 às crianças do Grupo B. Dos microrganismos encontrados nas escovas dentais, 83,92% eram Streptococcus mutans. Todos os isolados apresentaram perfil genético distinto, com exceção de 4 e 3 amostras dos Grupo A e B respectivamente, os quais pertenciam a mesma escova dental. Pode-se concluir que dos microrganismos encontrados, ocorreu maior prevalência de colonização por Streptococcus mutans nas crianças e escovas analisadas, não havendo evidência de que a armazenagem conjunta de escovas dentais pode ser um veículo para infecção cruzada por este microrganismo. / Abstract: The aim of this study was to evaluate the contamination of toothbrushes by mutans streptococci and cross-section infection of Streptococcus mutans. Forty-six children aged from 6 and 7 years were divided into two groups, Group A with 22 children and Group B with 24, according to attending class. A dental plaque sample from each participant was collected at the beginning of the study and the plaque present in toothbrushes was collected after 21 days. The samples were processed and cultivated under adequate conditions in SB20 medium. Typical morphotype mutans streptococci colonies were isolated and submitted to bioquimical tests for identification. An arbitrarily primed polymerase chain reaction (AP-PCR) method was performed for genotypic characterization of Streptococcus mutans. Fourteen toothbrushes were contaminated by mutans streptococci, 6 in Group A and 8 in Group B. Streptococcus mutans represented 83,92% of the mutans streptococci identified. The presence of matching genotypes of Streptococcus mutans has occurred in 4 strains of Group A and in 3 strains of Group B. However, for both groups, the matching genotypes belonged to the same toothbrush. In conclusion, Streptococcus mutans was the predominant specie in both dental plaque and toothbrush. The latter did not represent a source of cross-sectional infection of such microorganism. / Orientador: Angela Cristina Cilense Zuanon / Coorientador: Denise Madalena Palomari Spolidorio / Banca: Josimeri Hebling / Banca: Marcelo Fabiano Gomes Boriollo / Mestre

Resumo: O objetivo do presente estudo foi avaliar a microbiota bucal de mães, e pares de crianças, atendidas no programa educativo-preventivo da Bebê Clínica da Faculdade de Odontologia de Araçatuba-UNESP, nas faixas etárias de 6, 12, 18 e 24 meses, relacionando-a com a dieta, higiene bucal, presença ou não de dentes, prevalência de cárie e condição gengival. Após exame clínico e levantamento da dieta alimentar das crianças, foi realizada coleta dos espécimes de saliva, biofilme nas faixas etárias determinadas, que foram transferidos para frascos contendo meio de transporte VMGA III, submetidos a diluições seriadas em VMGA I, seguido de inoculação em meios de cultura adequados e incubação em diferentes condições redox e períodos de tempo. Além disso, a identificação dos isolados foi também realizada através de amplificação do DNA por PCR, após armazenamento dos espécimes em água ultra pura, e os níveis salivares de estreptococos do grupo mutans foram avaliados por real-time PCR. Nas faixas etárias avaliadas, não se observou correlação entre prevalência de patógenos orais e tipo de hábitos de dieta e higiene oral, sendo esta relacionada ao número de dentes irrompidos, principalmente com relação à prevalência de cocos cariogênicos e suas populações. Os microrganismos mais freqüentemente detectados aos 6 meses, pertenciam ao gênero Actinomyces, Campylobacter rectus, e à espécie Fusobacterium nucleatum. De maneira geral, aos 6 meses, a colonização da cavidade bucal dos bebês, por esses microrganismos, era transitória, por vezes não sendo observados em coletas subseqüentes. Aos 12 meses, observou-se uma elevação na prevalência de Actinomyces sp., Fusobacterium nucleatum e Streptococcus sobrinus. Nesta faixa etária, a presença de Eikenella corrodens foi... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The purpose of this study was to evaluate oral microorganisms of 50 infants and their mothers, in the ages of 6, 12, 18 and 24 months, integrants of Baby-Clinic's preventive program (Dental School of Aracatuba- UNESP). This factor was related to diet, oral hygiene habits, presence of teeth, prevalence of caries and periodontal conditions. After clinical exam, saliva and oral biofilm were collected and stored for culture and PCR process. Salivary levels of Streptococcus mutans were measured by Real time PCR. In the ages of the infants examined, after statistical analysis, it could be observed that no correlation was found between presence of oral pathogens and diet and hygiene habits, otherwise, this correlation could be seen with increase of erupted teeth. Most frequent microorganisms found in oral flora of infants at six months were Actinomyces, Campylobacter rectus and Fusobacterium nucleatum. Microorganisms observed at one period, could not be found in the other, suggesting that it was not a resident flora. At 12 months, data revealed increase in prevalence of Actinomyces sp., Fusobacterium nucleatum e S. sobrinus. In this age, the presence of Eikenella corrodens was significant. At 18 months, significant increase in the population of Actinomyces sp., Eikenella corrodens, Fusobacterium nucleatum, Streptococcus mutans e S. sobrinus was found. At 24 months, oral microflora was similar to the anterior but Streptococcus mutans and S. sobrinus, had an increase of population. Positive correlation between microorganisms found in mouth of mother and children in all periods was observed. For this reason, educative and preventive measures, which aim control of oral biofilm, must be priority, and made associated with attendance of the infants, as mothers are main transmissive ways to their children. Besides, the control of oral biofilm, as eruption... (Complete abstract click electronic access below) / Orientador: Robson Frederico Cunha / Coorientador: Elerson Gaetti Jardim Júnior / Banca: Sandra Maria Herondina Coelho Ávila de Aguiar / Banca: Renato Moreira Arcieri / Banca: Leila Maria Cesário Pereira Pinto / Banca: Edo Hirata / Doutor

Submitted by Rosivalda Pereira (mrs.pereira@ufma.br) on 2017-05-10T19:02:24Z No. of bitstreams: 1 ElizabethCosta.pdf: 1115019 bytes, checksum: 7bb115996a69101584c50ccc1ee319ae (MD5) / Made available in DSpace on 2017-05-10T19:02:24Z (GMT). No. of bitstreams: 1 ElizabethCosta.pdf: 1115019 bytes, checksum: 7bb115996a69101584c50ccc1ee319ae (MD5) Previous issue date: 2016-02-29 / Fundação de Amparo à Pesquisa e ao Desenvolvimento Científico e Tecnológico do Maranhão (FAPEMA) / Introduction: Several risk factors are associated with the ECC, including the S. mutans, identified as responsible for the onset of the disease; maternal saliva, the main source of primary infection to the child, socioeconomic factors, dietary habits and oral hygiene shared in the family environment. Thus, this thesis is divided into two chapters: Chapter I aimed at analyzing the contamination of S. mutans in the mother mediated by the contamination of S. mutans in the child together with the ECC. Chapter II aimed at analyzing the association between the consumption of added sugar by the mother and the frequency of the child's added sugar consumption. Methods: The sample consisted of 697 preschool children between 24 and 71 months old and their mothers. The mothers answered a questionnaire on their general and oral health as well as the child and a food frequency questionnaire (FFQ) to assess the diet of the binomial and the consumption of added sugar by the mother and the child. Mothers and children were subjected to an oral clinical examination to measure the caries index (DMF/ceo), IPV, ISG, anthropometric analysis of the mother's waist circumference and microbiological examination of saliva in 400 mothers / children pairs. A theoretical model based on causal diagrams (DAGs) that allows control of confounding factors in the most appropriate manner and at the same time avoids unnecessary adjustments to mothers / children mediators has been prepared. For Article 1, two models have been suggested, adjusted and analyzed: a total effect model and a mediated effect model of the association between maternal contamination S. mutans and ECC. The Paramed test was used to quantify the proportion of maternal S. mutans contamination effects mediated by contamination by S. mutans in the child in the ECC. To Article 2, two models have been suggested to study the association between the consumption of added sugar by the mother and child: an Effect 1 model and an Effect 2 model. The multinomial regression analysis was used to test the models, based on the DAGs. The marginal structural model was estimated by weighting by the inverse of the probability of consumption exposure in order to assess the association between consumption of added sugar by the mother and child. We estimated the prevalence ratio (PR) and confidence intervals (95% CI), in a 5% significance level. Results: In the total effect model, the presence of high colonization of S. mutans in the mother, the largest measurement of the waist circumference of the mother were associated with the ECC; Maternal IPV; child's age ≥ 4 years old and higher frequency of sugar consumption by the child. In the mediated effect model, the high maternal colonization remained linked to the ECC. Paramed test showed the proportion of mediated effect in 33% of maternal contamination of S. mutans by contamination in the child of S. mutans in the ECC. In effect model 1, the second and third tertile of the mother added sugar consumption was associated with the second tertile of added sugar consumption by the child; likewise the second and third tertile of consumption by the mother were also associated with the third tertile of the child's added sugar consumption. In effect model 2, all these variables of maternal consumption were also associated with the greatest child's consumption tertiles; yet maternal CW ≥88cm was also associated with the highest tertile of sugar consumption by the child. In marginal structural model, the consumption of added sugar by the mother was associated with added sugar consumption by the child. Conclusions: The contamination of S. mutans in the child only partially measured the association of the mother's S. mutans contamination in ECC. Other factors must be considered in the mother/child binomial, such as diet, oral hygiene practices and history of caries in the family environment. Sugar consumption by the mother and the highest maternal metabolic risk seem to explain the child's added sugar consumption in early childhood. / Introdução: Diversos fatores de risco estão associados à CPI, dentre eles os S.mutans, apontados como responsável pelo início da doença; a saliva materna, como principal fonte de infecção primária para o filho, os fatores socioeconômicos, os hábitos de dieta e de higiene bucal compartilhados no ambiente familiar. Assim sendo, esta tese foi dividida em dois capítulos: O capítulo I teve como objetivo, analisar a contaminação de S.mutans na mãe mediado pela contaminação de S.mutans no filho em associação com a CPI. O capítulo II teve como objetivo analisar a associação entre o consumo de açúcar de adição pela mãe e a frequência do consumo de açúcar de adição pelo filho. Métodos: A amostra foi composta por 697 pré-escolares de 24 a 71 meses de idade e suas mães. As mães responderam a um questionário sobre saúde geral e bucal da mãe e do filho e questionário de frequência alimentar (QFA), para avaliar a dieta do binômio e o consumo de açúcar de adição pela mãe e consumo de açúcar de adição pelo filho. Mães e filhos foram submetidos a um exame clínico bucal para aferição do índice de cárie (CPOD/ceo), IPV, ISG, análise antropométrica da circunferência da cintura da mãe e exame microbiológico da saliva em 400 pares mães/filhos. Foi elaborado um modelo teórico baseado em diagramas causais (DAGs) que permite o controle de fatores de confusão da forma mais adequada e ao mesmo tempo evita ajustes desnecessários para mediadores mães/filhos. Para o Artigo 1, dois modelos foram sugeridos, ajustados e analisados: um modelo de efeito total e outro modelo de efeito mediado da associação entre contaminação de S. mutans materna e CPI. O teste paramed foi usado para quantificação da proporção do efeito da contaminação de S. mutans materna mediada pela contaminação por S. mutans no filho na CPI. Para o artigo 2, foram sugeridos dois modelos para o estudo da associação entre o consumo de açúcar de adição pela mãe e o consumo de açúcar de adição pelo filho: um modelo de Efeito 1 e um modelo de Efeito 2. A análise de regressão multinomial foi usada para testar os modelos, baseados nos DAGs. Modelo estrutural marginal foi estimado pela ponderação pelo inverso da probabilidade do consumo da exposição, para avaliar a associação entre consumo de açúcar de adição pela mãe e consumo de açúcar de adição pelo filho. Estimou-se o razão de prevalência (RP) e os respectivos intervalos de confiança (IC 95%), a um nível de significância de 5%. Resultados: No modelo de efeito total, foram associadas à CPI, a presença de alta colonização de S. mutans na mãe, as maiores medidas da circunferência da cintura materna; IPV materno; idade da criança ≥ 4 anos e maior frequência de consumo de açúcar pelo filho. No modelo de efeito mediado, a alta colonização materna permaneceu associada à CPI. O teste paramed mostrou a proporção do efeito mediado de 33% da contaminação materna de S. mutans pela contaminação no filho de S. mutans na CPI. No modelo de efeito 1, o segundo e terceiro tercil do consumo de açúcar de adição pela mãe foram associados o segundo tercil do consumo de açúcar de adição pela filho; da mesma forma o segundo e terceiro tercil de consumo pela mãe foram também associados ao terceiro tercil do consumo de açúcar de adição pelo filho. No modelo de efeito 2, todas essas variáveis de consumo maternas também foram associadas aos maiores tercis de consumo do filho; ainda a CC materna ≥88cm também foi associada ao maior tercil de consumo de açúcar pelo filho. No modelo estrutural marginal, o consumo de açúcar de adição pela mãe foi associado ao consumo de açúcar de adição pelo filho. Conclusões: A contaminação de S. mutans no filho media apenas parcialmente a associação da contaminação de S. mutans da mãe na CPI. Outros fatores devem ser considerados no binômio mãe-filho, como hábitos alimentares, práticas de higiene bucal e a história da cárie no ambiente familiar. O consumo de açúcar pela mãe e o maior risco metabólico materno parecem explicar o consumo de açúcar de adição pelo filho na primeira infância.

En el presente estudio se buscó determinar el efecto de la aplicación de la combinación de los barnices de Fluoruro de sodio 5 % y Diacetato de clorhexidina 1 % en los niveles de Streptococcus mutans en saliva, en niños que asistieron al Servicio de Odontopediatría del Hospital Nacional Docente Madre Niño San Bartolomé y cumplieron con los criterios de inclusión. Método: Estudio experimental, prospectivo y longitudinal. Los 45 niños con edades fluctuantes entre 3 a 5 años, con caries de esmalte en fórmula temporal completa fueron asignados aleatoriamente en 3 grupos de 15 cada uno. Se aplicaron los agentes quimioterapéuticos aleatoriamente, el primer grupo recibió la aplicación de la combinación de Fluoruro de sodio al 5 % y Diacetato de clorhexidina 1 %, el segundo grupo recibió solo la aplicación de Fluoruro de sodio al 5 % y el grupo control sólo un barniz placebo. Se aplicaron los agentes, siguiendo los instructivos, tres dosis en un lapso de 10 días; después de 8 semanas se realizó una evaluación microbiológica de S. mutans. Resultados: La combinación Fluoruro de sodio al 5 % y Diacetato de clorhexidina al 1 % redujo el nivel S. mutans con una media representativa de 15,267 ± 9,816 (p: 0.000), el grupo que recibió solo Fluoruro de sodio al 5 %, también redujo los valores de S. mutans con una media relativamente mayor de 16,267 ± 7,146 (p: 0,000) y finalmente el grupo control que recibió el barniz placebo, generó reducción microbiológica pero en menor proporción con una media de 9,467 ± 9,326 (p: 0,001), demostrando todos diferencias estadísticamente significativas. Conclusión: La aplicación combinada de Fluoruro de sodio al 5 % y Diacetato de clorhexidina al 1 % es efectiva para reducir los niveles de S. mutans que se encuentran en la saliva de los niños; sin embargo al comparar los grupos entre sí no existió diferencia significativa. / In the present study sought to determine the effect of the application of the combination of varnishes Sodium fluoride 5% Diacetate chlorhexidine 1% in the levels of Streptococcus mutans in saliva, on children who attended the Service of Pediatric Dentistry at the National Hospital Docente Madre Niño San Bartolome and met the inclusion criteria. Method: Experimental, prospective, longitudinal study. The 45 children with fluctuating aged 3-5 years with enamel caries full temporal formula were randomized into 3 groups of 15 each. Randomly chemotherapeutic agents were applied, the first group received the application of the combination of Sodium fluoride and 5 % Diacetate Chlorhexidine 1 %, the second group received only the application of Sodium fluoride 5 % and the control group only placebo varnish . Agents were applied, following the instructions, three doses within 10 days after 8 weeks microbiological evaluation of S. mutans was performed. Results: The combination Sodium Fluoride 5 % and Diacetate Chlorhexidine 1 % reduced the level S. mutans with a representative average of 15,267 ± 9,816 (p: 0.000 ), the group that received Sodium fluoride 5 % also reduced the values of S. mutans with a relatively higher average of 16,267 ± 7,146 (p: 0.000) and finally the control group receiving the placebo varnish, generated microbiological reduction but to a lesser extent with an average of 9,467 ± 9,326 (p:0.001), showing statistically significant differences across. Conclusion: The combined application of Sodium fluoride 5 % and Diacetate chlorhexidine 1 % is effective in reducing the levels of S. mutans found in the saliva of children, however when comparing the groups with each other, no significant difference existed. / Tesis

Tooth decay is a serious health risk and a significant contributor to health care costs in both industrialized and developing nations. Tooth decay is the end result of a change in the balance of plaque ecology towards more acidogenic and aciduric bacterial species. The primary force facilitating this change is an increase in the amount and frequency of simple carbohydrate, in particular, sucrose ingestion by the host. The acidity of plaque increases after host ingestion of fermentable carbohydrates and this promotes demineralization of the tooth enamel which is restored by salivary buffering and mineral deposition. Frequent and prolonged periods of low plaque pH drive cycles of enamel homeostasis towards demineralization, which ultimately leads to the formation of dental caries. Streptococcus mutans is the main etiologic agent in the development of dental caries. The cariogenic potential of S. mutans is based on their ability to produce and tolerate large amounts of acid and to adhere to and accumulate large numbers on the surface of a tooth. They are capable of efficiently fermenting a variety of simple carbohydrates and can produce high concentrations of acid, even in a low pH environment. However, it is the ability of S. mutans to rapidly synthesize copious amounts of water-insoluble and water-soluble glucan from dietary sucrose, which allow the bacteria to accumulate large enough numbers to dominate the dental plaque and significantly lower the plaque pH. Synthesis of glucan is mediated by glucosyltransferase enzymes and is crucial to sucrose-dependent adherence and to the cariogenicity of S. mutans. S. mutans also makes four non-GTF glucan-binding proteins: GbpA and GbpD are secreted and released proteins that contain a region that is homologous to the glucan-binding domains of the Gtf enzymes, and GbpC is a cell wall bound protein that confers the property of dextran-dependent aggregation during stressful conditions, and GbpB whose glucan-binding properties appear secondary to its role in cell-wall metabolism. It was hypothesized that Gbps A, C, and D primarily function to shape the architecture of S. mutans biofilms which in turn affects the cariogenicity of S. mutans. To test this hypothesis, a panel of Gbp mutants was constructed from S. mutans strain UA130 that encompasses all deletions of Gbps (GbpA, GbpC and GbpD) individually and in combination. Specific pathogen-free rats were infected with the WT S. mutans UA130 strain along with each of the Gbp mutants, were fed a high sucrose diet for seventy days, and were then scored for caries. Significant attenuation of caries was observed in some but not all gbp mutants. Biofilms were also grown and analyzed via confocal microscopy and COMSTAT image analysis software. Architectural differences were found with all of the gbp mutants when compared to the wild-type, most notably the mutant strains lost significant biofilm depth. Several of the architectural parameters correlated with caries attenuation. It was concluded that deletion of one or more Gbps resulted in a partial loss of the cohesive properties of S. mutans biofilms and changes in biofilm architecture. In several cases this resulted in significant attenuation of cariogenicity but not a complete loss. The architectural changes that resulted from this loss of biofilm cohesiveness and the specific combinations of Gbp deletions that lead to significant attenuation suggested specific roles for each Gbp in biofilm formation. Furthermore, the attenuation of Gbp mutant strains could not be explained by differences in acidogenicity or aciduricity among the mutants. Therefore it was concluded that Gbps A, C and D make profound contributions to biofilm architecture and changes in biofilm architecture, as a result of loss of Gbp-mediated cohesion, affects S. mutans cariogenicity.

Infectious diseases e.g. gastric ulcer, caries and perodontitis, are caused by bacteria in a biofilm. Adhesion of bacteria to host ligands e.g. proteins, polypeptides and glycoproteins, is a key event in biofilm formation and colonization of surfaces such as mucosa and tooth tissues. Thus, host ligands could contribute to the susceptibility to infectious diseases. The general aim of this doctoral thesis was to study the effect of phosphorylated polypeptides and gp-340 in saliva and milk on oral bacterial adhesion and aggregation. Statherin is a non-glycosylated, phosphorylated polypeptide in saliva. The polypeptide inhibits precipitation and crystal growth of calcium phosphate and mediates adhesion of microorganisms. By using a hybrid peptide construct, the domain for adhesion of Actinomyces isolated from human infections and from rodents was found to reside in the C-terminal end, and the adhesion was inhibitable. With alanine substitution the peptide recognition epitope in the C-terminal end was delineated to Q and TF, where QAATF was an optimal inhibitory peptide. In contrast, human commensal Actinomyces bound to the middle region in a non-inhibitable fashion. Gp-340 is another protein in saliva, and it is a large, multifunctional glycoprotein. Four novel size variants (I-IV) of salivary gp-340 were distinguished within individuals, and their glycoforms were characterized. All four size variants were identical in the N-terminal amino acid sequence and shared core carbohydrates. Low-glyco lung gp-340, high-glyco saliva gp-340, and size variants I-III aggregated bacteria differently. Human milk, which shares many traits with saliva, could inhibit adhesion of Streptococcus mutans to saliva-coated hydroxyapatite (s-HA), a model for teeth, in an individually varying fashion. Human milk caseins, lactoferrin, secretory IgA, and IgG inhibited the binding avidly. By using synthetic peptides the inhibitory epitope in b-casein was mapped to a C-terminal stretch of 30 amino acids. Inhibition by human milk, secretory IgA and the b-casein-derived inhibitory peptide was universal among a panel of mutans streptococci. The main conclusions are: (i) statherin mediates differential binding of commensal versus infectious Actinomyces strains with small conformation-dependent binding epitopes, (ii) salivary gp-340 has individual polymorphisms that at least affect binding of bacteria, (iii) human milk inhibits S. mutans adhesion to s-HA in an individually varying fashion, and the C-terminal end of human milk β-casein is one inhibitory component. Together these results suggest that the studied host ligands can influence the composition of the oral biofilm. Statherin may protect the host from colonization of bacteria associated with infections. Gp-340 size variants may affect functions related to host innate immune defences such as interactions with a wide array of bacteria, and human milk may have a protective effect in infants from colonization of mutans streptococci.

Este estudio se efectuó con el propósito de: conecer la prevalencia con que se presenta la caries dental, determinar el pH salival y los niveles salivales de Streptococcus mutans en adolescentes con síndrome Down comparándolos con adolescentes normales en el Perú. Para ello se examinaron 33 adolescentes con síndrome Down y 33 adolescentes normales con edades comprendidas entre 13 y 19 años de edad de 3 centros de educación especial, y 1 escuela de educación secundaria de Lima respectivamente. Los resultados de la investigación revelan que : La prevalencia de caries dental en adolescentes con síndrome Down fue menor que la de los adolescentes normales. El índice de CPO-S de los adolescentes con síndrome Down fue 11.06 y el de los adolescentes normales fue de 12.45, sin existir diferencia estadística significativa entre estos valores. El valor del promedio del pH salival encontrado en los adolescentes con síndrome Down fue de 7,27 mientras que los adolescentes normales fue de 6,91, existiendo diferencia estadística significativa entre estos valores. En cuanto a los niveles salivales de Streptococcus mutans se encontró que el mayor porcentaje de los adolescentes con síndrome Down y adolescentes normales tuvieron un recuento medio. Haciendo una comparación de promedios de unidades formadoras de colonias se encontró los siguientes valores, para los adolescentes con síndrome Down 335015.76 y para los adolescentes normales 383695.76 no existiendo diferencia estadística significativa entre estos valores. / This study was done with the purpose of: to know the prevalency of dental caries and to determine the salivary pH and the salivary levels of Streptococcus mutans in teenagers with Down Síndrome, comparing them with normal teenagers, in Perú. For this purpose, 33 teenagers with Down Syndrome and 33 normal teenagers between 13 and 19 years old of 3 special education centers and 1 secundary school of Lima respectively were examinated. The results of the research reveal that: The dental caries prevalency in teenagers with Down Syndrome was lower than the prevalency in normal teenagers. The CPO-S indicator in teenagers with Down Syndrome was 11.06 and in normal teenagers was 12.45. There is no statistically significant difference between those values. The average of the salivary pH found in teenagers with Down Syndrome was 7.27, while in normal teenagers was 6.91. Statistically significant difference exist between those values. About the salivary levels of Streptococcus mutans, the result was that the majority of teenagers with Down Syndrome and normal teenagers had an intermediate recount. Doing a comparisson of the UFC of Streptococcus mutans average we found the following values: for the teenagers with Down Syndrome the average was 335015.76 and for the normal teenagers the average was 383695.76. There is no statistically significant difference between those values.