Publicación a texto completo no autorizada por el autor / Evalúa la actividad inhibitoria de los aceites esenciales de Mentha spicata (hierba buena) y Eucalyptus globulus (eucalipto) de origen comercial sobre el desarrollo de biopelículas de Streptococcus mutans ATCC 25175 en un modelo in vitro, emulando las condiciones de la placa dental. La composición de los aceites esenciales (AE) se evaluó por Cromatografía de Gases acoplada a Espectrómetro de Masas (CG/EM) siendo los principales metabolitos el R-(–)-carvona (57,93%) y Llimoneno (12,907%) para Mentha spicata y el 1,8-cineol (eucaliptol) (65,83%) para Eucalyptus globulus. Se evaluó la actividad inhibitoria de los AE por los métodos de difusión en pozo de agar y microdilución colorimétrica. Los halos de inhibición fueron de 18,3 ± 0,47 mm y 27,0 ± 0,82 mm y los CMI de 1,8484 x 10-3 mg/mL y 1,9168 x 10-3 mg/mL, para los AE de Mentha spicata y Eucalyptus globulus respectivamente. La actividad frente a biopelículas se evaluó en un sustrato de piezas de esmalte dental bovino, empleando medio basal de mucina (BMM), en condiciones de anaerobiosis y ciclos diarios de exposición a la sacarosa, para emular las condiciones de la cavidad oral. Los AE se aplicarón a una concentración de 0,5% en un vehículo salino estéril con polisorbato 20 al 1%. Tras 72 horas de cultivo, se observó una reducción significativa (P <0,001%) en la biomasa de la biopelícula, evaluada por su turbidez en suspensión, y en el recuento de organismos recuperables respecto al control. Los efectos de ambos AE no resultaron significativamente distintos entre sí. Ambos AE presentan actividad antimicrobiana frente a S. mutans en cultivos planctónicos y de biopelícula, por lo que podría tener gran potencial para el desarrollo de productos farmacéuticos y sanitarios en el área de la salud oral. / Tesis

Made available in DSpace on 2016-06-02T19:02:44Z (GMT). No. of bitstreams: 1 6426.pdf: 2868015 bytes, checksum: e3bf13cc85e7606dc2d0f7dbb387ebaa (MD5) Previous issue date: 2014-11-24 / Financiadora de Estudos e Projetos / The evaluation of new materials and biomaterials modified with silver nanoparticles is a line of research that has been gaining strength in various knowledge areas including dentistry. Even considering international scientific publications, however, there are still few studies evaluating dental restoration materials that are modified with silver nanoparticles. Proposal: this study aimed to compare a photopolymerizable composite resin modified with an antimicrobial silver nanoparticle additive (an experimental resin), with the photopolymerizable composite resin in its unmodified form (a commercially available dental restoration material), by evaluating the in vitro antibacterial properties and also some of the primary physical properties of the resin. Methodology: composite resin disks were made for three experimental groups: unmodified resin (control group) and modified resin with different silver nanoparticle concentrations: 0.3% and 0.6% wt (groups 1 and 2). The antibacterial activity on the surface of the samples was evaluated by in vitro growth of of Streptococcus mutans and Lactobacillus acidophilus biofilms in a 20% sucrose medium, which is followed by counting the viable cells from the biofilms through serial dilutions after three incubation periods - 1, 4 and 7 days (n = 9). Optical transmittance was tested to measure the percentage of transmittance in the samples (n = 9). The surface roughness was evaluated using Atomic Force Microscopy (n = 9). The colour of the samples and the colour difference between the groups was measured in a colorimetry test according to the CIELab system. The presence of silver in the nanoparticle additive was analysed using X-Ray Fluorescence Spectroscopy. The quality of the dispersion and distribution of silver on the samples was analysed using Transmission Microscopy. The liberation of silver in artificial saliva was also evaluated in vitro after three incubation periods (60, 120 and 210 days) with agitation of the samples, using the Inductively Coupled Plasma Optical Emission Spectroscopy technique. Results: for the three incubation times, the number of viable cells was statistically lower in groups 1 and 2 than in the control group (p < 0.05), and groups 1 and 2 were not statistically different (p > 0.05) for the two bacterial species (ANOVA Two way/Tukey). There was no significant difference in optical transmittance between group 1 and the control (p > 0.05), and group 2 had a lower transmittance value than the other groups (p < 0.05) (ANOVA/Tukey). The analysis of the values of the arithmetic roughness, which was obtained using the NanoScope software, showed that there was no significant difference in surface roughness between the three groups (p > 0.05) (ANOVA/Tukey). From the measurements of the colours, the total colour difference (&#916;E) between all of the groups was higher than 1 (the minimum value that represents a colour alteration perceptible to human vision) and lower than 3,3 (critical &#916;E). The presence of silver in the nanoparticle additive was confirmed in the X-Ray Fluorescence test. Microscopic analysis showed good distribution of silver for groups 1 and 2 (with silver in the four quadrants of the regions analysed). The dispersion was better for group 1 than for group 2 (which showed a higher number of particle clusters). After the three incubation times, silver was not detected in the artificial saliva for the three groups or; if it was present, the concentration was below the limit of detection for this technique (< 0.02 mg/L). Conclusions: the antibacterial property on the surfaces of the experimental resins was verified for the two cariogenic bacteria considered. Furthermore, the addition of silver did not cause negative alterations to the resin modified with the lower concentration of nanoparticles, which makes the continuation of this study possible and valuable. / A avaliação de novos materiais e biomateriais modificados com nanopartículas de prata é uma linha de pesquisa que vem ganhando força em várias áreas do conhecimento, inclusive na área odontológica. Porém, mesmo considerando a publicação científica internacional, ainda há poucas pesquisas que avaliam materiais restauradores dentários modificados desta forma. Proposição: o propósito deste estudo foi comparar a resina composta fotopolimerizável modificada com um aditivo antimicrobiano nanoparticulado de prata (uma resina experimental), com a resina composta fotopolimerizável em sua forma não modificada, um material restaurador dentário disponível comercialmente, avaliando a propriedade antibacteriana, in vitro, e também algumas de suas principais propriedades físicas. Metodologia: foram confeccionados discos de resina composta para 3 grupos experimentais: resina não modificada (grupo controle), e resina modificada com diferentes concentrações de nanopartículas de prata, 0,3 e 0,6% (grupos 1 e 2). A atividade antibacteriana na superfície das amostras foi avaliada pelo crescimento in vitro de biofilme de Streptococcus mutans e Lactobacillus acidophilus em meio de cultura contendo 20% de sacarose, seguido de contagem de células viáveis provenientes dos biofilmes após 3 períodos de incubação, 1, 4 e 7 dias, através de diluições seriadas (n=9). Um ensaio de transmitância óptica foi realizado para medir a porcentagem de transmitância das amostras (n=9). A rugosidade superficial foi avaliada por Microscopia de Força Atômica (n=9). A cor das amostras e a diferença de cor entre os grupos foi medida em um ensaio de colorimetria considerando o sistema CIELab. A presença de prata no aditivo nanoparticulado foi analisada por meio da Espectrometria de Fluorescência de Raios X. A qualidade da dispersão e distribuição da prata nas amostras foi analisada através da Microscopia de Transmissão. Foi também avaliada a liberação de prata em saliva artificial, in vitro, após 3 períodos de incubação das amostras, sob agitação, 60, 120 e 210 dias, através da técnica de Espectrometria de Emissão Óptica por Plasma Acoplado Indutivamente. Resultados: para os 3 tempos de incubação, o número de células viáveis foi estatisticamente mais baixo nos grupos 1 e 2 do que no grupo controle (p < 0,05), e os grupos 1 e 2 não mostraram entre si diferença estatisticamente significante (p > 0,05), para as 2 espécies bacterianas (ANOVA Two way/Tukey). Não houve diferença significante na transmitância óptica entre o grupo 1 e o controle (p > 0,05), já o grupo 2 apresentou um valor de transmitância menor que os demais grupos (p < 0,05) (ANOVA/Tukey). A análise dos valores de rugosidade aritmética, obtidos através do software NanoScope, mostrou que não houve diferença significante na rugosidade superficial entre os 3 grupos (p > 0,05) (ANOVA/Tukey). A partir das medições de cores, a diferença total de cor (&#916;E) entre todos os grupos foi superior a 1 (valor mínimo que representa uma alteração de cor perceptível à visão humana) e inferior a 3,3 (&#916;E crítico). A presença de prata no aditivo nanoparticulado foi confirmada no ensaio de Fluorescência de Raios X. A análise microscópica demonstrou boa distribuição de prata para o grupo 1 e 2 (com prata nos 4 quadrantes das regiões analisadas), e a dispersão foi melhor para o grupo 1 do que para o grupo 2 (que apresentou maior número de aglomerados de partículas). Após os 3 tempos de incubação, não foi detectada a presença de prata na saliva artificial, para os 3 grupos, sendo que, se presente, ela encontrava-se em uma concentração abaixo do limite de detecção da técnica (< 0,02 mg/L). Conclusões: a propriedade antibacteriana foi verificada nas superfícies das resinas experimentais para as duas espécies bacterianas cariogênicas consideradas, e a adição de prata não provocou alterações negativas na resina modificada com a menor concentração de nanopartículas, o que viabiliza a continuidade do estudo.

Submitted by Izabel Franco (izabel-franco@ufscar.br) on 2016-09-14T14:14:33Z No. of bitstreams: 1 TeseVHP.pdf: 3273273 bytes, checksum: afe82ad5c25bb61d489c9fcaec24a835 (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-09-16T19:20:25Z (GMT) No. of bitstreams: 1 TeseVHP.pdf: 3273273 bytes, checksum: afe82ad5c25bb61d489c9fcaec24a835 (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-09-16T19:20:33Z (GMT) No. of bitstreams: 1 TeseVHP.pdf: 3273273 bytes, checksum: afe82ad5c25bb61d489c9fcaec24a835 (MD5) / Made available in DSpace on 2016-09-16T19:20:46Z (GMT). No. of bitstreams: 1 TeseVHP.pdf: 3273273 bytes, checksum: afe82ad5c25bb61d489c9fcaec24a835 (MD5) Previous issue date: 2015-05-07 / Não recebi financiamento / Demineralization and caries are often present in patients with orthodontic braces due to the increased accumulation of dental biofilm containing Streptococcus mutans, which produces acids that attack the tooth surface. The Antimicrobial Photodynamic Therapy (APDT) has been shown as an alternative in microbial inactivation in dentistry. This work, carried out both in vitro and in vivo, aims to evaluate the antimicrobial effect of APDT on Streptococcus mutans present in the dental biofilm accumulated on the metal surface of orthodontic appliances and in patients undergoing orthodontic treatment. The in vitro study evaluated the susceptibility of biofilm formed by S. mutans on the metal surface under the application of orthodontic appliances APDT induced Curcumin diluted surfactant (SDS) and blue light-emitting diode (LED). The samples were treated with the photosensitizer (PS) at a concentration of 1 g/L, 0.1% surfactant and exposed to LED light (455 ± 10 nm, 46mW/cm²), fluence of 30 J/cm². The experimental groups studied were: control group (P-L-S-); light (P-L+S-), surfactant (P-L-S+) and photosensitizer (P+L-S-) groups; surfactant-light (P-L+S+), photosensitizer-surfactant (P+L-S+) and PDT (P+L+S-) groups; and the PDT-surfactant (P+L+S+) group. Each group was named accordingly to the addiction or not of the photosensitizer (P+ or P-), surfactant (S+ or S-) and application or not of light (L+ or L-), respectively. The colonies grown on plates of brain heart infusion (BHI) were counted (CFU / mL) and the results were analyzed by ANOVA and the post hoc Tukey test (p <0.05). The results showed that in this research protocol, the group P+L+S+ was able to promote significant decrease (p<0.001) on the population of S. mutans of 99.999% compared to the control group. In the in vivo study, 24 volunteers were randomly selected in order to evaluate the clinical utility of APDT and the APDT association with the surfactant (SDS) as oral decontamination agents in orthodontic patients. The selected patients (n = 24) were randomly divided into seven groups: G1 - Negative control, G2 - Curcumin mouthwash, G3 - Curcumin mouthwash+SDS, G4 - light irradiation, G5 - APDT, G6 - APDT+SDS and G7 - Positive Control (chlorhexidine 0.12% moutwash). Saliva samples were collected from unstimulated way of each patient in three steps (S) as follows: S1 - initial condition S2 - Treatment with rinsing (water, curcumin or chlorhexidine) and S3 - After APDT. The photosensitiser used was curcumin with concentration of 1 g/L. Two types of blue LED light source emitting in 450±10nm were used in the present study: one emitting with an intensity of 33.54 mW/cm² and other with 212 mW/cm². Evaluation of microbial reduction, and survival fraction in each of the studied levels, was performed using the Kruskal-Wallis ANOVA test. Statistical analysis showed that only G6 and G7 presented results of bacterial inactivation that showed statistically significant differences (p<0.05) compared to those observed in G1. These results indicated that APDT, when combined with SDS surfactant, may be used as an adjunct agent for convenient oral decontamination promoter in vivo. / Desmineralizações e cáries dentais estão comumente presentes em indivíduos portadores de aparelho fixo ortodôntico, isto ocorre devido ao aumento de acúmulo de biofilme dental contendo Streptococcus mutans, os quais produzem ácidos que atacam a superfície dental. A terapia fotodinâmica antimicrobial (TFDA) tem sido mostrada como alternativa em inativação microbiana na Odontologia. Este trabalho, realizado in vitro e in vivo, tem o objetivo de avaliar o efeito antimicrobiano da TFDA sobre Streptococcus mutans em biofilme dental sobre superfície metálica de acessórios ortodônticos e em indivíduos sob tratamento ortodôntico. O estudo in vitro avaliou a susceptilidade de biofilme formado por S. mutans sobre superfície metálica de acessórios ortodônticos sob aplicação de TFDA induzida por Curcumina diluída em surfactante (SDS) e por diodo emissor de luz azul (LED). Nas amostras obtidas de lâminas de bandas ortodônticas foram formados biofilmes e tratadas com fotossensibilizador (FS) com concentração de 1g/L, surfactante a 0,1% e exposto a luz LED (455±10 nm, 46mW/cm²), com fluência de30J/cm2. Os grupos experimentais estudados foram: C- (somente lavagem com solução fisiológica); L+ (aplicação apenas de luz); S+L- (aplicação de surfactante); S+L+ (aplicação de surfactante e luz); F+L- (aplicação de fotossensibilizador sem luz); F+L+ (aplicação de fotossensibilizador e luz); F+L-S+ (aplicação de fotossensibilizador sem luz + surfactante) e F+L+S+ (aplicação de fotossensibilizador e luz + surfactante). As colônias cultivadas em placas com Brain Heart Infusion (BHI) foram contadas (UFC/mL) e os resultados foram analisados por ANOVA e teste Tukey post hoc (p<0.05). Os resultados obtidos demonstraram que no protocolo deste estudo o grupo F+S+L+ foi capaz de promover diminuição significativa(p<0,001) da população de S. mutans em 99,999% comparado ao grupo controle. No estudo in vivo, 24 voluntários foram selecionados aleatoriamente, com o objetivo de avaliar a eficiência clínica da TFDA e da associação da TFDA com o surfactante SDS como agentes de descontaminação bucal em indivíduos ortodônticos. Os indivíduos selecionados foram distribuídos aleatoriamente em sete grupos experimentais: G1 - controle negativo, G2 - bochechos com curcumina, G3 – bochechos com curcumina + SDS, G4 - irradiação com luz apenas, G5 - TFDA, G6 - TFDA + SDS e G7 - Controle Positivo (clorexidina 0,12%). Amostras de saliva foram coletadas de maneira não estimulada de cada um dos indivíduos em três etapas (s), como segue: S1 - Condição inicial, S2 - Tratamento com bochechos (água, clorexidina ou curcumina) e S3 - Após TFDA. O fotossensibilizador usado foi a Curcumina em concentração de 1g/L. Foi utilizado no estudo dois tipos de equipamentos de luz com LED azul (450 ± 10nm, 33.54 mW/cm2 e 212 mW/cm2). A avaliação da redução microbiana, e da fração de sobrevivência em cada um dos níveis estudados, foi realizada utilizando-se o teste ANOVA Kruskal-Wallis. A análise estatística mostrou que apenas os grupos 6 e 7 apresentaram resultados de inativação bacteriana que apresentavam diferenças estatisticamente significantes (p <0,05) em relação aos resultados observados no grupo 1. Estes resultados indicam que a TFDA associada com o surfactante SDS pode ser utilizado como agente auxiliar conveniente para promover a descontaminação bucal.

FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Over the past few years, there has been increasing evidence of the effect of the oral health over the general health of individuals, supported by a series of biological and epidemiological studies that show a relation between the mouth and many diseases, including cardiovascular diseases. Structural deficiencies and functional abnormalities of heart valves represent an important cause of cardiovascular morbidity and mortality in Brazil, and a few defects have been recently associated with infectious agents. The aim of this study was to identify cariogenic and periodontopathogenic bacteria in dental plaque, saliva and heart valves, without clinical endocarditis, of patients with heart valve diseases, and correlate these findings with the oral health status of the patients. Oral exams using the DMTF (decayed, missing and filled teeth) and PSR (Periodontal Screening and Recording) indexes to evaluate caries and periodontal disease, respectively, were performed. Samples of supragingival and subgingival dental plaque, saliva and cardiac valves were evaluated, through Real Time Polymerase Chain Reaction, for the presence of DNA of Streptococcus mutans (S. mutans), Prevotella intermedia (P. intermedia), Porphyromonas gingivalis (P. gingivalis) and Treponema denticola (T. denticola). A total of 114 samples were collected from 42 patients with a mean age of 55.6  13.8 years. The average number of missing teeth due to caries was 23.52  9.41 teeth per patient, and according to the highest score of periodontal disease observed for each patient, excluding edentulous patients (44.0%), periodontal pockets over 4mm (43.4%) and dental calculus (34.7%) were detected in a higher number of patients. The molecular analysis of the oral samples revealed high frequency of S. mutans and P. intermedia in supragingival dental plaques, subgingival dental plaques and saliva of dentate and edentulous patients (variation 60.0% - 100.0%), while P. gingivalis and T. denticola were detected in a smaller number of oral samples (variation 17.6% - 64.0%). The microorganism most frequently detected in heart valve samples was the S. mutans (89.3%), followed by P. intermedia (19.1%), P. gingivalis (4.2%) e T. denticola (2.1%). Significant difference was observed between the frequency of P. intermedia, P. gingivalis and T. denticola in the heart valve and dental plaque, as oposed to S. mutans. The identification of oral bacteria, especially S. mutans, in heart valves of patients with a previous history of dental caries and gingivitis/periodontitis suggests the possible involvement of these pathogens in the etiopathogenesis of heart valve diseases. / Atualmente, cada vez mais se tem evidÃncias do efeito da condiÃÃo oral na saÃde geral dos indivÃduos, atravÃs de uma sÃrie de estudos epidemiolÃgicos e biolÃgicos que mostram uma relaÃÃo entre a boca e diversas doenÃas, incluindo as doenÃas cardiovasculares. Desordens estruturais e nas funÃÃes das vÃlvulas cardÃacas representam uma importante causa de morbidade e mortalidade cardiovascular no Brasil, sendo alguns processos, como a estenose aÃrtica degenerativa, mais recentemente associados a agentes infecciosos. O objetivo desta pesquisa foi identificar bactÃrias cariogÃnicas e periodontopatogÃnicas na placa dental, saliva e vÃlvulas cardÃacas, sem endocardite clÃnica, de pacientes com doenÃa valvar, correlacionando esses achados à condiÃÃo bucal dos indivÃduos. AvaliaÃÃo, quanto Ãs doenÃas cÃrie e periodontal, foi realizada, atravÃs dos Ãndices CPO-D (Dentes Permanentes Cariados, Perdidos e Obturados) e PSR (Registro Periodontal Simplificado), respectivamente. Amostras de placa dental supragengival, subgengival, saliva e vÃlvula cardÃaca foram coletadas para investigaÃÃo da presenÃa de DNA, atravÃs de PCR (ReaÃÃo em Cadeia de Polimerase) em tempo real, de Streptococcus mutans (S. mutans), Prevotella intermedia (P. intermedia), Porphyromonas gingivalis (P. gingivalis) e Treponema denticola (T. denticola). Um total de 114 amostras foi coletado de 42 pacientes com mÃdia de idade de 55.6  13.8 anos. A mÃdia de dentes perdidos devido à cÃrie, por paciente, foi em torno de 23.52  9.41 e, segundo o maior grau de doenÃa periodontal observado no indivÃduo, excluindo-se os pacientes desdentados totais (44.0%), bolsa superior a 4 mm (43.4%) e o cÃlculo dental (34.7%) esteve presente em um maior nÃmero de pacientes. A anÃlise molecular das amostras bucais revelou alta frequÃncia de S. mutans e P. intermedia nas placas supragengival, subgengival e saliva de pacientes dentados e desdentados (variando entre 60.0% e 100.0%), enquanto que P. gingivalis e T. denticola estiveram presentes em menor nÃmero de amostras bucais (variando entre 17.6% e 64.0%). O micro-organismo mais frequentemente encontrado nas amostras valvares foi o S. mutans (89.3%), seguido da P. intermedia (19.1%), P. gingivalis (4.2%) e T. denticola (2.1%). DiferenÃa significativa foi encontrada entre a presenÃa de P. intermedia, P. gingivalis e T. denticola na vÃlvula e na placa dental, diferentemente do S. mutans. A identificaÃÃo de bactÃrias orais, principalmente S. mutans, em vÃlvulas cardÃacas de pacientes com elevada experiÃncia prÃvia de cÃrie e ocorrÃncia de gengivite/periodontite, sugere o possÃvel envolvimento desses patÃgenos nas doenÃas valvares.

Objetivo: Evaluar in vitro el efecto antibacteriano y citotóxico del extracto metanólico del café peruano y colombiano frente a cepas de Streptococcus mutans (ATCC® 25175) y Streptococcus sanguinis (ATCC® 10556). Métodos: El extracto metanólico de café peruano y colombiano fue preparado utilizando una proporción 1:2 (P/V). Para determinar las propiedades antibacterianas de cada extracto se utilizó el método de difusión en pozo y la concentración mínima inhibitoria (CMI) se determinó mediante el método de microdilución. Como control positivo se utilizo la clorhexidina al 2%. La viabilidad celular y la citotoxicidad de los extractos fueron realizados utilizando el ensayo de reducción de MTT en la línea celular MDCK. Resultados: El extracto metanólico del café colombiano (Coffea) tuvo un mayor efecto antibacteriano (18.9 ± 2.8 mm) en comparación al café peruano (15+2.54mm), mientras que para el grupo de Streptoccocus mutans y para el grupo de Streptoccocus sanguinis el café colombiano y peruano mostraron halos de 18.8 ± 2.77 mm y 14.7± 3.8 mm, respectivamente. La concentración mínima inhibitoria (CMI) para los extractos del café colombiano y peruano fue de 3.1 μg/ml y 1.6 μg/ml para Streptoccocu mutans y 6.3 μg/ml para Streptoccocu sanguinis, respectivamente. El ensayo de MTT muestras que ambos extractos no fueron citotóxicos a altas concentraciones. Conclusiones: Los hallazgos que se encontraron en este estudio muestran que los extractos del colombiano y peruano (Coffea) tienen propiedades antibacterianas efectivas contra cepas de Streptococcus mutans y Streptococcus sanguinis. / Objective: To evaluate in vitro the antibacterial and cytotoxic effect of the methanolic extract of peruvian and colombian coffee against strains of Streptococcus mutans (ATCC® 25175) and Streptococcus sanguinis (ATCC® 10556). Methods: An extract was prepared for each type of coffee, carrying out nine tests independently, chlorhexidine 2% was used as a positive control. The well diffusion method was performed to determine the antibacterial properties of each extract. The minimum inhibitory concentration (MIC) was determined by the microdilusion method and the cytotoxicity was analyzed by the MTT reduction test using a MDCK cell line. Results: The methanolic extract of colombian coffee (Coffea) had a greater antibacterial effect (18.9 ± 2.8 mm) compared to peruvian coffee (15 + 2.54mm) for the group of Streptoccocus mutans and for the group of Streptoccocus sanguinis, colombian coffee and peruvian obtained 18.8 ± 2.77 mm and 14.7 ± 3.8 mm, respectively. The minimum inhibitory concentration (MIC) for colombian and peruvian coffee extracts was 3.1 μg / ml and 1.6 μg / ml for S. mutans and 6.3 μg / ml for S. sanguinis, respectively. These extracts were not cytotoxic at high concentrations. Conclusions: The findings found in this study show that colombian and peruvian extracts (Coffea) have effective antibacterial properties against Streptococcus Mutans and Sanguinis strains. Further study with Coffea is recommended as there is very little research on this. / Tesis

Indiana University-Purdue University Indianapolis (IUPUI) / Both quaternary ammonium bromide (QAB) and furanone derivatives were synthesized, characterized and formulated into dental resin composites for improved antibacterial properties. Compressive strength (CS) and S. mutans viability were used to evaluate the mechanical strength and antibacterial activity of the restoratives. The effects of chain length, loading, saliva and aging on CS and S. mutans viability were investigated. Chapter 2 describes how we studied and evaluated the formulated antibacterial resin composites by incorporating the synthesized QAB-containing oligomers into the formulation. The results show that all the QAB-modified resin composites showed significant antibacterial activity and mechanical strength reduction. Increasing chain length and loading significantly enhanced the antibacterial activity but dramatically reduced the CS as well. The 30-day aging study showed that the incorporation of the QAB accelerated the degradation of the composite, suggesting that the QAB may not be well suitable for development of antibacterial dental resin composites or at least the QAB loading should be well controlled. Chapter 3 describes how we studied and evaluated the formulated antibacterial resin composite by incorporating the synthesized furanone derivative into the formulation. The results show that the modified resin composites showed a significant antibacterial activity without substantially decreasing the mechanical strengths. With 5 to 30% addition of the furanone derivative, the composite kept its original CS unchanged but showed a significant antibacterial activity with a 16-68% reduction in the S. mutans viability. Further, the antibacterial function of the new composite was found not to be affected by human saliva. The aging study indicates that the composite may have a long-lasting antibacterial function. In summary, we have developed a novel QAB- and furanone-containing antibacterial system for dental restoratives. Both QAB- and furanone-modified resin composites have demonstrated significant antibacterial activities. The QAS-modified experimental resin composite may not be well suitable for development of antibacterial dental resin composites due to its accelerated degradation in water unless the QAB loading is well controlled. The furanone-modified resin composite shows nearly no reduction in mechanical strength after incorporation of the antibacterial furanone derivative. It appears that the furanone-modified resin composite is a clinically attractive dental restorative that can be potentially used for long-lasting restorations due to its high mechanical strength and permanent antibacterial function.

Diss. (sammanfattning) Umeå : Umeå universitet, 2010.

Indiana University-Purdue University Indianapolis (IUPUI) / The impact of tobacco smoking on human health is well documented. The influence of smoking on tooth loss and cardiovascular diseases was investigated in the current study via both epidemiology and in vitro studies. From analyzing the 2006 Behavioral Risk Factor Surveillance System (2006 BRFSS) database, we confirmed that smoking was significantly associated with the number of teeth lost in a dose-dependent manner and smoking cessation reduced the risk when compared to those subjects continuing to smoke. In addition, the virulence factors related to caries were compared between Streptococcus mutans and Streptococcus gordonii in response to cigarette smoking condensate (CSC) treatment. We observed that S. gordonii was more susceptible to CSC treatment than S. mutans. CSC significantly enhanced S. mutans sucrose-dependent and independent adherence. Western blot assays revealed that several bacterial surface proteins including glucosyltransferase (GTF), glucan-binding proteins and antigen I/II, were significantly upregulated for the treated S. mutans. These findings suggested that the oral environment with CSC may favor a cariogenic dominant composition, which may increase the risk for smokers to develop caries. We also found that smoking and oral health status modified each other and synergistically increased the risk of CVD and this joint effect was more pronounced among the youngest age group using the 2006 BRFSS database. To further understand the joint effect, we conducted an in vitro study to investigate bacterial attachment to fibronectin and endothelial cells in response to smoking condensate treatment. Our study clearly demonstrated CSC significantly enhanced S. mutans attachment to both soluble and immobilized fibronectin as well as endothelial cells. Furthermore, our data suggested that bacteria possessed several adhesins that bound to host tissues and endothelial cells also had multiple receptors for bacterial attachment. Among these adhesins, antigen I/II seemed essential for bacterial attachment to endothelial cells without CSC. The knowledge of bacterial attachment to host tissues in the presence of CSC may help in developing different preventive or therapeutic strategies against attachment and colonization of the host by S. mutans.

Guatemalan medicinal plants were collected and screened for the presence of essential oils using steam distillation. Oil was found in 63 species from 24 families and was tested in tube dilution assays for activity against Escherichia coli, Staphylococcus aureus, Streptococcus mutans, Lactobacillus acidophilus and Candida albicans. Several essential oils were highly active with 20 instances of oils inhibiting the microbes at an MIC of 0.31 µl/ml. Oils were also tested against cancerous and established cell lines using a 15% (v/v) agar-media which was developed to improve essential oil solubility. Assays were performed against three cancer lines: Stomach (AGS: CRL-1739), Skin (A375: CRL-1619), Tongue (CAL27: CRL-2095) and an established Monkey Kidney cell line (Vero C 1008: CRL-1586). Assessment of viability was performed using the Neutral Red assay with results indicating that many of the oils significantly inhibited cancer cell lines in vitro with 24 individual instances producing an IC50 of 0.20 µl/ml or less. Therapeutic indices indicated that many of the highly inhibitory oils were more cytotoxic to cancerous cell lines than to the established cell line.

Introduktion och syfte: Kariesutveckling influeras av faktorer hos både värd och bakterier. Men när olika individer exponeras för samma nivåer av externa riskfaktorer, är en del individer mer mottagliga för karies jämfört med andra. En förklaring skulle kunna vara olika glykosylering av glykoprotein i saliven. I denna pilotstudie undersökte vi skillnaden i nivåer av monosackariderna sialinsyra, fukos och galaktos hos personer som aldrig haft karies och personer som har/har haft karies. Syftet var även att undersöka om plackenzym kan vara en modifierare av nämnda glykoprotein.Material och metod: Två grupper, med 10 individer i varje, inkluderades i studien. Ena gruppen hade DMFT = 0 och den andra DMFT ≥ 1. Saliv och plack samlades och innehållet av bundna monosackarider (sialinsyra, fukos och galaktos) samt glykosidaser (sialidas, β-fukosidas, β-galaktosidas, α-glukosidas och N-acetylglukosaminidas) analyserades med en fluorometer. Även salivflödet kalkylerades.Resultat: Innehållet av både sialinsyra och galaktos var signifikant högre i gruppen med DMFT = 0, medan innehållet av fukos inte skilde sig åt signifikant mellan grupperna. Ingen signifikant skillnad kunde ses mellan de två grupperna avseende enzymaktivitet och salivflöde.Konklusion: Högre nivåer av bunden sialinsyra och galaktos fanns hos gruppen med DMFT = 0. Resultaten indikerar att dessa monosackarider kan vara en möjlig markör för oral hälsa. Större longitudinella studier behövs för att verifiera sambandet. / Introduction and aim: Caries development is affected by factors within bacteria and host. But when different individuals are exposed to same levels of external risk factors, some individuals are still more susceptible to caries. One explanation could be different glycosylation of salivary glycoproteins. In this pilot study, we investigated the difference in levels of the monosaccharides sialic acid, fucose and galactose between individuals with or without previous caries experience. We also aimed to investigate if plaque glycosidases could be a modifier of these glycoproteins.Material and method: Two groups, with 10 subject in each, were included in this study. One group had DMFT = 0 and the other DMFT ≥ 1. Saliva and plaque were collected and content of bound monosaccharides (sialic acid, fucose and galactose) and glycosidases (sialidase, α-fucosidase, β-galactosidase, α-glucosidase and N-acetylglucosaminidase) were detected using absorbance and fluoroscens respectively. Salivary flow rate was also measured.Results: Content of both sialic acid and galactose were significantly higher in the group with DMFT = 0, while the content of fucose did not differ significantly between the groups. No significant differences could be seen between the two groups (DMFT = 0 and DMFT ≥ 1) regarding any of the investigated glycosidases and salivary flow rate. Conclusion: Higher levels of bound sialic acid and galactose were found in the group with DMFT = 0 and the results indicate that these monosaccharides could be a possible marker for oral health. Larger longitudinal studies are needed to verify this correlation.