Streptococcus mutans é o principal agente etiológico da cárie dental humana, uma doença infecciosa para a qual não há vacina disponível. A influência dos sistemas de transporte ABC na virulência bacteriana suporta seu uso como alvos vacinais. Em S. mutans a proteína ligadora de fosfato (PstS) influencia a expressão de fatores de virulência. A proposta deste trabalho foi caracterizar uma estratégia vacinal de mucosa anti-cárie baseada na proteína PstS como antígeno alvo. Inicialmente, a forma recombinante da proteína foi obtida em E. coli. A caracterização biofísica revelou uma estrutura secundária estável, semelhante a outras proteínas ligadoras e capaz de interagir com seu ligante. Epítopos antigênicos conservados foram identificados na proteína recombinante pela reatividade com soro anti-S. mutans. A proteína rPstS foi imunogênica por via sublingual, combinada ou não à LTK4R e anticorpos rPstS-específicos interferiram na colonização oral in vivo por S. mutans. Os resultados indicam que a proteína rPstS pode ser explorada em estratégias vacinais contra a cárie. / Streptococcus mutans is the main etiological agent of human dental caries, an infectious disease for which there is no vaccine available. The influence of ABC transport systems in bacterial virulence supports its use as vaccine targets. In S. mutans the phosphate binding protein (PstS) influences the expression of virulence traits. The purpose of this work was characterizing an anti-caries mucosal vaccine based on the PstS protein as target antigen. First, a recombinant form of the protein was obtained in E. coli. The biophysical characterization showed a stable secondary structure, similar to other binding proteins and able to interact with its ligand. Conserved antigenic epitopes were identified in the recombinant protein by reactivity with anti- S. mutans serum. The rPstS protein was immunogenic by the sublingual route in combination or not with LTK4R and rPstS-specific antibodies interfered with S. mutans oral colonization in vivo. The results indicated that the recombinant PstS protein can be exploited in vaccine strategies against dental caries.

Submitted by isabela.moljf@hotmail.com (isabela.moljf@hotmail.com) on 2017-05-12T14:04:16Z No. of bitstreams: 1 erikamariahenriquesmonteiro.pdf: 6797940 bytes, checksum: 8ec07139ac5eeaa9eb035c731131e662 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-05-12T15:43:44Z (GMT) No. of bitstreams: 1 erikamariahenriquesmonteiro.pdf: 6797940 bytes, checksum: 8ec07139ac5eeaa9eb035c731131e662 (MD5) / Made available in DSpace on 2017-05-12T15:43:44Z (GMT). No. of bitstreams: 1 erikamariahenriquesmonteiro.pdf: 6797940 bytes, checksum: 8ec07139ac5eeaa9eb035c731131e662 (MD5) Previous issue date: 2012-07-17 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A abrangência da utilização de fitoterápicos e de plantas medicinais é vasta e engloba fins variados, também em relação à saúde bucal. O controle do biofilme bacteriano dentro das diversas especialidades odontológicas é de grande importância, pois aponta tanto para a prevenção, como para o tratamento, da cárie e de doenças periodontais. Com a finalidade de auxiliar os métodos convencionais de higiene bucal, muitos agentes químicos vêm sendo estudados, entre produtos com ação antimicrobiana e antiinflamatória. O presente trabalho teve o objetivo de propor o desenvolvimento um enxaguatório bucal bifásico contendo extratos naturais. Fezse o screening fotoquímico dos extratos aquosos de Eucalyptus globulus, Mentha arvensis e a identificação por CG/EM dos compostos principais do óleo de gergelim. Realizou-se experimentos para testar a ação antimicrobiana dos extratos aquosos de Eucalyptus globulus, Mentha arvensis e o óleo de gergelim (teste de difusão em agar, concentração mínima inibitória (CIM) e concentração mínima de aderência (CIMA) frente a cepa padrão Streptococcus mutans ATCC 25175. Realizou-se também ensaios para avaliar a ação antiinflamatória e antinociceptiva do óleo de gergelim. Nos extratos aquosos de M. arvensis não foram encontrados taninos, flavonóides e saponinas, também não apresentaram atividade sobre a bactéria testada. O extrato aquoso de E. globlulus foi positivo para a presença de taninos, flavonóides e saponinas. Também apresentou atividade sobre Streptococcus mutans ATCC 25175 observando um halo de 20,0 ± 0,66 cm (p<0,05), a CIM e a CIMA foi determinada com a E4 (12,5 mg/mL), o óleo de gergelim não apresentou halo no teste de difusão em Agar, acredita-se que seja pelo fator de difusão do óleo, pois em meio líquido apresentou atividade,a CIM e CIMA foi a OG 3 (25%). O óleo de gergelim apresentou atividade antiinflamatória e antinociceptiva nos ensaios realizados e no CG/EM deste óleo mostrou a presença de ácido graxos insaturados que já possuem atividades antiinflamatórias comprovadas. Conclui-se que os extratos aquosos de E. globlulus e o óleo de gergelim possuem potencial para serem empregados como prováveis enxaguatórios bucais. / The extent of use of herbal and medicinal plants is vast and encompasses varied purposes, also in relation to oral health. The control of the biofilm within the various specialties dental is of great importance because it points for both the prevention and for the treatment of dental caries and periodontal disease. In order to assist the conventional oral hygiene methods, many chemical agents have been studied, among products with antimicrobial and anti-inflammatory action. This study aimed to develop a proposed two-phase mouthwash containing natural extracts. There was photochemical screening of aqueous extracts of Eucalyptus globulus, Mentha arvensis, identifying by GC / MS of the main compounds of sesame oil. We conducted experiments to test the antimicrobial activity of aqueous extracts of Eucalyptus globlulus, Mentha arvensis and sesame oil (agar diffusion test, minimum inhibitory concentration (MIC) and minimum concentration of adhesion (MICA) compared to standard strain Streptococcus mutans ATCC 25175. Also conducted tests to evaluate the antinociceptive and anti-inflammatory action of sesame oil. The aqueous extract of M. arvensis were not found tannins, flavonoids and saponins, this also did not show activity against the bacteria tested. The aqueous extract of E. globulus was positive for the presence of tannins, flavonoids and saponins. Also showed activity against Streptococcus mutans ATCC 25175 observed a halo of 20.0 ± 0.66 cm (p <0.05), MIC and MICA was determined with the E4 (12.5 mg/ml), the oil sesame not present in the test halo Agar diffusion is believed to be by a factor of oil dilution, as in liquid medium was active, the MICA and MIC was OG 3 (25%l). Sesame oil presented antinociceptive and antiinflammatory tests and GC / MS of this oil showed the presence of unsaturated fatty acid that already have proven antiinflammatory activities. It is concluded that the aqueous extract of E. globlulus and sesame oil are potentially likely to be mployed as mouthwashes.

Neste estudo focamos no papel de cinco transportadores da família ABC (ATP-binding cassete) envolvidos com a captação ativa de oligopeptídeos, poliaminas, fosfato inorgânico, glutamato e glutamina, em duas espécies bacterianas: Streptococcus mutans, que causa a cárie, e Escherichia coli enterohemorrágica (EHEC), responsável por diarreias e síndrome hemolítica urêmica em humanos. Com relação a inativação do sistema de transporte de oligopeptídeos de S. mutans não observamos alteração do crescimento bacteriano ou da aderência à superfícies abióticas. A deleção do sistema de captação de poliaminas não interferiu com o crescimento em meio rico, porém aumentou a resistência à ambiente ácidos. Inativação do sistema de transporte de fosfato inorgânico reduziu a aderência de S. mutans. Sobre os sistemas de transporte de glutamato e glutamina, mutantes de S. mutans apresentaram alterações nas taxas de crescimento e adesão à superfícies. Inativação da proteína OppA de EHEC não afetou a produção da toxina Stx bem como a patogenicidade in vitro e in vivo de EHEC. Em suma, o presente estudo demonstra que o papel dos transportadores ABC na fisiologia e patogenicidade de bactérias pode variar de acordo com a espécies bem como com o substrato transportado. / This study focuses on the role of five ABC (ATP-binding cassette) transport systems related to active uptake of oligopeptides, polyamines, inorganic phosphate, glutamate and glutamine. In this work we study two bacterial species: Streptococcus mutans, which causes caries, and enterohaemorrhagic Escherichia coli (EHEC), responsible for diarrhea and hemolytic uremic syndrome in humans. The inactivation of the S. mutans oligopeptide transport system did not change the bacterial growth and adherence to abiotic surfaces. Inactivation of the polyamine uptake system did not interfere with growth in rich medium, but increased the survival of bacteria in acid environments. Inactivation of the inorganic phosphate transport system reduced the adherence of S. mutans. Regarding the glutamate and glutamine transport systems, the S. mutans mutants showed changes in growth rates and adhesion to abiotic surfaces. Inactivation of the EHEC OppA protein, did not affect the production of the Stx toxin neither affected the in vitro and in vivo pathogenicity of the strain. Collectively, the present study shows that the roles of ABC transporters in the physiology and pathogenicity of bacteria may vary according to the species involved as well as the substrate transported.

Indiana University-Purdue University Indianapolis (IUPUI) / Objective: The aim of this study is to investigate the effects of cinnamon water extract on nicotine-induced Streptococcus mutans biofilm. This study utilized S. mutans biofilm assays with varying concentrations of nicotine/cinnamon water extract levels. Design: A preliminary experiment was carried out to confirm the most likely effective concentration of cinnamon water extract on S. mutans biofilm. Then a 24-hour culture of S. mutans UA159 in microtiter plates was treated with varying nicotine concentrations (0-32 mg/ml) in TSBS at the same time with or without the optimum cinnamon water extract concentration. A spectrophotometer was used to determine total growth absorbance and planktonic growth. The microtiter plate wells were washed, fixed and stained with crystal violet dye and the absorbance measured to determine biofilm formation. Results: The results indicated that cinnamon water extract was able to inhibit biofilm formation significantly (p<0.05) at 5 mg/ml cinnamon water extract, therefore, 5 mg/ml of cinnamon water extract was recognized as the MIC for S. mutans biofilm formation. When combined with nicotine, cinnamon water extract sub-MIC (2.5 mg/ml) demonstrated a significant inhibitory effect (p<0.05) in biofilm and total absorbance measures at high concentrations of nicotine (8 mg/ml and above). In addition, cinnamon water extract showed a significant effect (p<0.05) at very low concentrations of nicotine (0.25 and 0.5 mg/ml) in all measures (biofilm, planktonic and total absorbance). However, at low concentrations of nicotine (2 and 4 mg/ml), there was a significant increase (p<0.05) in biofilm growth, whereas planktonic growth was significantly (p<0.05) decreased at the same concentrations. Conclusion: These results provided more evidence regarding the negative effects of nicotine and also demonstrated the positive influence of cinnamon water extract in reducing nicotine-induced biofilm formation, which needs be confirmed by in-vivo studies.

Objectifs: Le but de cette étude clinique était de comparer un groupe d’adultes ayant un parodonte sain avec un groupe d’adultes atteints de parodontite chronique en terme de risque carieux et mesures cliniques et microbiologiques de la carie. Méthodes: Quatre-vingt-seize individus ont été divisés en deux groupes en fonction de leur état de santé parodontal et ont été appariés pour l'âge, le sexe et l'origine ethnique. Trente-huit sujets étaient atteints de parodontite chronique définie comme ayant au moins quatre dents avec ≥ 1 site avec une profondeur de sondage ≥ 4 mm et une perte d'attache clinique ≥ 2 mm, et 58 sujets présentaient un parodonte sain. Par la suite, les groupes ont été subdivisés en deux groupes en fonction de leur statut carieux : les participants ayant au moins une lésion carieuse non traitée sur une surface dentaire et ceux n’ayant pas de lésion carieuse non traitée. Les données ont été recueillies par le biais d’un questionnaire, un examen clinique et des échantillons de plaque supra- et sous-gingivale. L’évaluation de la charge buccale de Streptococcus mutans et de six agents pathogènes parodontaux a été réalisée par la technique d'amplification de la réaction en chaine de la polymérase (PCR). Les données ont été analysées à l'aide d’analyses statistiques descriptives et bivariées. Résultats: Les individus atteints de parodontite chronique étaient 3,5 fois plus susceptibles d'avoir des caries que les individus en bonne santé (OR 3,5 ; IC: 1,5 - 8,3 ; P = 0,006). Les sujets à la fois atteints de parodontite chronique et de caries dentaires ont eu un niveau d’éducation significativement plus faible que les sujets ayant un parodonte sain et sans caries dentaires (OR 6,0 ; IC: 1,7 à 21,7 ; P = 0,04). La proportion de sujets ayant une charge buccale élevée de Porphyromonas gingivalis (P. g.) et Treponema denticola (T. d.) était significativement plus élevée chez les patients atteints de parodontite chronique et de carie que chez les patients sains présentant des caries (P. g.: OR 8,6 ; IC: 2,4 - 30,3 ; P = 0,004 et T. d.: OR 10,0 ; CI: 2,6 - 38.1 ; P = 0,003). Conclusions: Les résultats de cette étude suggèrent que, chez les sujets adultes atteints de la parodontite chronique, la fréquence des caries est plus élevée que chez les sujets ayant un parodonte sain. De plus, le faible niveau d'éducation influence négativement le statut parodontal des individus. / Aim: The aim of this clinical study was to compare adults with a healthy periodontium and those with chronic periodontitis, in terms of caries’ risk and caries’ clinical and microbiological measures. Methods: Ninety-six healthy adults were divided into chronic periodontitis (n= 38) and healthy periodontium (n=58) based on their periodontal status, and matched for age, gender, and ethnic background. Chronic periodontitis was defined as having at least four teeth with ≥1 site with a pocket depth ≥4 mm and clinical attachment loss ≥2 mm. Each group were subsequently subdivided in 2 groups according to their caries status: participants having at least one untreated decayed surface and those with no untreated caries. Data were collected by means of self-administrated questionnaire, clinical examination, and supra- and subgingival plaque sampling. Assessments of oral levels of Streptococcus mutans and six periodontal pathogens were conducted by PCR amplification techniques. Data were analyzed using descriptive and bivariate statistical tests. Results: Individuals with chronic periodontitis were 3.5 times more likely to have caries than healthy individuals (OR 3.5; CI: 1.5 – 8.3; P = 0.006). Subjects with both chronic periodontitis and dental caries had a significantly lower level of education than periodontally healthy subjects without dental caries (OR 6.0; CI: 1.7 – 21.7; P = 0.04). A significant higher proportion of subjects with high oral levels of Porphyromonas gingivalis (P. g.) and Treponema denticola (T. d.) was found among subjects with chronic periodontitis and untreated caries compared to periodontally healthy subjects with untreated caries (P. g.: OR 8.6; CI: 2.4 – 30.3; P = 0.004 and T. d.: OR 10.0; CI: 2.6 – 38.1; P = 0.003). Conclusion: The results from this study suggest that, adults with chronic periodontitis are more prone to caries disease than those adults with a healthy periodontium. Furthermore, low educational level could have a negative impact on the periodontal status of individuals.

Le but de ce travail de mémoire était d'explorer des moyens pour augmenter la perméabilité des biofilms de Streptococcus mutans aux macromolécules en utilisant des agents potentiellement perturbateurs de la structure des biofilms. L’acide éthylènediamine tétraacétique (EDTA) ainsi que l’acide acétylsalicylique (aspirine) sont les agents perturbateurs choisis. Le changement de perméabilité des biofilms de S. mutans a été déterminé en mesurant les coefficients de diffusion globale du polyéthylène glycol (PEG) et de diffusion locale de dextrans. Les coefficients de diffusion globale ont été mesurés par spectroscopie infrarouge avec un échantillonnage par réflexion totale atténuée (ATR) alors que la spectroscopie par corrélation de fluorescence (SCF) a été utilisée pour la mesure des coefficients de diffusion locale. Les résultats ont démontré que l’incorporation de l’EDTA à une concentration de 7.5 (m/v) % dans la solution de diffusion permet d’améliorer les propriétés de transport du PEG dans les biofilms en augmentant sa pénétrabilité et son coefficient de diffusion globale. Par contre, aucune variation n’a été constatée dans la valeur du coefficient de diffusion locale de dextran fluorescent. Cette différence peut être expliquée, entre autres, par l'échelle des mesures et la nature différente des molécules diffusantes. L’aspirine n’a démontré aucun effet sur le transport du PEG à travers les biofilms de S. mutans. La pénétration accrue du PEG en présence de l’EDTA a été corrélée aux tests de viabilité des cellules bactériennes. En effet, la combinaison de la pénicilline G (PenG) avec l’EDTA 2 (m/v) % a eu comme effet l’augmentation du pouvoir biocide d’un facteur 3. De plus, les images de microscopie à épifluorescence et de microscopie confocale à balayage de laser ont démontré que les bactéries dans le cœur des microcolonies sont plus affectées par la PenG lorsque le milieu contient de l'EDTA. A la lumière des résultats obtenus, il s’avère que l’incorporation d'agents perturbateurs de la structure des biofilms est une option sérieuse à considérer dans l’éradication des biofilms microbiens. Plus d’études devront être effectuées afin d’investiguer l’effet d’autres molécules possédant les propriétés perturbatrices de la structure des biofilms sur la résistance de ces derniers aux agents antimicrobiens. / The aim of this study was to investigate the role of biofilm structure disrupting potential agents in the biofilm permeability to macromolecules. The effect of ethylenediamine tetraacetic (EDTA) and acetylsalicylic acid (aspirin), two potential disrupting agents, on the diffusion of poly (ethylene glycol) (PEG) and dextran in Streptococcus mutans biofilms was investigated. Attenuated total reflection (ATR) infrared spectroscopy was used to measure the global diffusion coefficients, while the local diffusion coefficients were measured using fluorescence correlation spectroscopy (FCS). On one hand, the results showed that EDTA, at a concentration of 7.5 (w/v) %, increased PEG penetrability and global diffusion coefficient in biofilms. No effect was noticed for the local diffusion coefficient of fluorescent dextran molecules. This difference was associated with, amongst others, the scale of the measurements and the different nature of the diffusing species. On the other hand, aspirin had no effect on the S. mutans biofilm permeability for PEG. The enhanced penetration of PEG in the presence of EDTA was correlated with bacterial cell viability. The bactericidal effect of penicillin G (PenG) alone and in combination with EDTA, was assessed for the biofilms. The results showed that antibiotic effect of PenG was improved by a factor of 3 when the antibiotic was used in combination with EDTA compared to the same antibiotic used alone. Moreover images obtained by epifluorescence microscopy and laser scanning confocal microscopy indicated that the bacteria in the core of the microcolonies were more affected by PenG when the milieu contained EDTA. These results revealed that the use of disrupting agents of biofilm structure is a promising approach to eradicate biofilms showing resistance to classical antimicrobial agent treatments. More studies should be done to investigate the effect of others potentials disrupting agents on the biofilm permeability to macromolecules.

Este estudo teve por objetivo analisar in vitro o efeito inibitório de materiais res tauradores contendo o monômero ant ibac ter iano brometo de metacriloiloxidodecilpiridínio (MDPB) no desenvolvimento de lesões de cárie secundária em raízes dentárias submetidas a um desafio cariogênico. Cavidades com características de cavidades classe V (2 mm x 2mm) foram preparadas em setenta e cinco fragmentos de raízes dentárias humanas. As amostras foram divididas aleatoriamente em 5 grupos (n=15 amostras/grupo) e restauradas como segue: (I) sistema adesivo sem MDPB + resina composta sem MDPB (controle negativo); (II) cimento de ionômero de vidro resino-modificado (CIV-RM, controle positivo); (III) sistema adesivo sem MDPB e resina composta com MDPB (2,83% MDPB); (IV) sistema adesivo contendo MDPB e resina composta sem MDPB; e (V) sistema adesivo contendo MDPB e resina composta contendo MDPB. Lesões de cárie secundária radicular foram produzidas utilizando-se um modelo bacteriológico de indução de cáries artificiais. As amostras restauradas foram imersas em um meio de cultura contendo Streptococcus mutans e sacarose por 15 dias. Fatias (80 ± 20 m) das amostras foram utilizadas para a mensuração da média das profundidades das lesões formadas adjacentes à ambas as margens das restaurações através de microscopia de luz polarizada. Os resultados foram expressos em porcentagem relativa à média de profundidade encontrada no grupo controle negativo (GI), que foi considerado 100%. Os dados foram comparados por ANOVA complementada pelo teste de Tukey (p0.05). As profundidades das lesões adjacentes às cavidades preenchidas com CIV-RM (GII; 85,17 ± 15,2%) foram significantemente (p < 0.01) menores que as profundidades das cavidades adjacentes às restaurações realizadas com resina composta sem MDPB (GI; 100,00 ± 10,04%). As profundidades das lesões adjacentes às cavidades restauradas com resina contendo MDPB (GIII; 82,68 ± 12,81% e GV; 85,65 ± 15,42%), independentemente do sistema adesivo utilizado, foram similares àquelas do CIV-RM (GII). A média das profundidades das lesões nestes grupos diminuiu 13% (GV) e 17% (GIII) em relação ao controle negativo (GI). Portanto, nas condições deste estudo, sugerimos que a resina composta contendo o monômero antibacteriano MDPB é capaz de inibir o avanço de lesões de cárie secundária em raízes dentárias. / This in vitro study aimed to analyze the potential of restorative materials containing the antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB) for controlling the advance of bacterial induced secondary root caries-like lesions. Class V cavities (2 mm x2 mm) were prepared in seventy-five human root fragments. Specimens were randomly divided in 5 groups (n=15 fragments per group) and restored as follows: (I) MDPB-free adhesive system + MDPB-free composite (negative control); (II) resin modified glass ionomer (RM-GIC; positive control); (III) MDPB-free adhesive system + MDPB-containing composite (2.83% MDPB); (IV) MDPB-containing adhesive system + MDPB-free composite; (V) MDPB-containing adhesive system + MDPB-containing composite. Artificial secondary root caries lesions were produced by a biological artificial caries challenge. The restored specimens were immersed into a culture medium containing Streptococcus mutans and sucrose for 15 days. Histological slices (80 ± 20 m) of the specimens were used for measuring the mean depths of the artificial lesions produced in both margins of the restorations using polarized light microscopy. Results were expressed in percentage related to the mean depth of the negative control, considered 100%. Data were compared by ANOVA followed by the Tukeys test (p 0.05). The depths of lesions adjacent to cavities filled with RM-GIC (GII; 85.17 ± 15.2%) were significantly (p < 0.01) shallower than those adjacent to restorations with MDPB-free composite (GI; 100.00 ± 10.04%), despite the presence of MDPB in the adhesive system (GIV; 101.95 ± 21.32%). The depths of lesions adjacent to cavities restored with MDPBcontaining 3 composite (GIII; 82.68 ± 12.81%and GV; 85.65 ± 15.42%), despite the adhesive system used, were similar to those of RM-GIC (GII). Mean lesions depths in these groups decreased 13% (GV) to 17% (GIII) in relation to the negative control (GI). Concluding, MDPB-containing composite inhibits the progression of artificial secondary root caries lesions regardless of adhesive systems.

A cárie dentária é uma doença bucal de alta prevalência e impactante em países em desenvolvimento. É causada pela presença de biofilme dentário rico em bactérias acidogênicas e acidúricas, como Streptococcus mutans e Lactobacillus casei. Neste sentido, a fitoterapia tem sido aplicada na odontologia devido ao seu conhecido efeito antimicrobiano, tendo potencial para prevenir doenças como a cárie dentária. Portanto, o presente estudo tem como objetivo testar o potencial antimicrobiano de extratos bruto e etanólico das folhas de Myracrodruon urundeuva (M. urundeuva.) e Qualea grandiflora (Q. grandiflora) sobre S.mutans e L. casei. Para tal, determinaram-se a Concentração Inibitória Mínima (CIM) e a Concentração Bactericida Mínima (CBM). A CIM foi definida como a menor concentração do agente antimicrobiano capaz de inibir 100% o crescimento microbiano (absorbância) em relação aos controles negativos. Para a CBM, alíquotas foram removidas dos poços que não apresentaram nenhuma absorbância (viabilidade, concentrações CIM) e semeadas em placas de ágar BHI, incubadas por 24 h a 37°C em estufa de CO2 5%. A CBM foi determinada considerando a menor concentração dos extratos capaz de impedir o crescimento bacteriano visível. Cepas de S. mutans (ATCC 21175) e L. casei (ATTC 334) foram ativadas em BHI e caldo Rogosa, respectivamente. A CIM foi determinada pela técnica de diluição em microplacas de 96 poços (100 l de extrato + 80 l BHI/Rogosa + 20 l da bactéria diluída em BHI/Rogosa equivalente a 5x105 UFC/mL), as quais foram incubadas por 24 h (S. mutans) e 48 h (L. casei) a 37°C em estufa de CO2 5%. Os extratos de M. urundeuva e Q. grandiflora inicialmente foram diluídos em BHI/Rogosa variando as concentrações entre 2 mg/ml a 0,00012207 mg/ml e os mesmos extratos diluídos em álcool foram avaliados nas concentrações entre 20 mg/ml a 0,00244 mg/ml para S. mutans e L. casei. Não foi possível determinar a CIM e a CBM para os extratos diluídos no BHI/Rogosa. Foram utilizados como controle positivo a clorexidina e como controles negativos BHI/Rogosa com e sem álcool a 5%. As CIMs (CBMs) da M. urundeuva e Q. grandiflora, diluídas em álcool, e clorexidina contra S. mutans foram 2,5 mg/ml (2,5 mg/ml), 5,0 mg/ml (--) e 0,00468 mg/ml (0,00937 mg/ml), respectivamente. Em relação ao L. casei, as CIMs (CBMs) da M. urundeuva e Q. grandiflora, diluídas em álcool, e da clorexidina foram 0,156 mg/ml (0,312 mg/ml), 0,156 0,625 mg/ml (0,312 0,625mg/ml) e 0,00468 mg/ml (0,3 mg/ml), respectivamente. Como conclusão, nosso estudo mostrou que L. casei (ATTC 334) é mais susceptível aos extratos que S. mutans (ATCC 21175) e o extrato M. urundeuva apresenta melhor efeito antimicrobiano que a Q. grandiflora em S. mutans (ATCC 21175), porém os dois extratos apresentam efeito similar sobre L. casei (ATTC 334) e ambos foram inferiores à CHX. / Dental caries is an oral disease of high prevalence and impact in developing countries. It is caused by the presence of a dental biofilm rich in acidogenic and aciduric bacteria, such as Streptococcus mutans and Lactobacillus casei. Accordingly, phytotherapy has been applied in dentistry due to its known antimicrobial effect, having potential to prevent diseases such as dental caries. Therefore, the present study aims to test the antimicrobial potential of crude and ethanolic extracts of Myracrodruon urundeuva (M. urundeuva) and Qualea grandiflora (Q. grandiflora) leaves on S. mutans and L. casei. For this, the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were determined. MIC was defined as the lowest concentration of the antimicrobial agent capable of inhibiting 100% the microbial growth in comparison to the negative controls. For MBC, aliquots were removed from the wells that did not show any absorbance (viability, concentrations than MIC) and seeded on BHI agar plates, incubated for 24 h at 37°C and 5% CO2. The MBC was determined considering the lowest concentration of extracts capable of preventing visible bacterial growth. Strains of S. mutans (ATCC 21175) and L. casei (ATTC 334) were activated in BHI and Rogosa broth. MIC was determined by the dilution technique in 96-wells microplates (100 l of extract + 80 l BHI/ Rogosa + 20 l of bacterium diluted in BHI/Rogosa equivalent to 5x105 CFU/ml), which were incubated for 24 h (S. mutans) and 48 h (L. casei) at 37°C and 5% CO2. The extracts were firstly diluted in BHI/Rogosa varying the concentrations between 2 mg/mL and 0.00012207 mg/ml; the same extracts were also diluted in alcohol at concentrations ranging from 20 mg/ml to 0.00244 mg/ml and both tested against S. mutans and L. casei. It was not possible to determine the MIC and MBC for the extracts diluted in BHI/Rogosa. Chlorhexidine was used as positive control, while BHI/Rogosa with or without 5% alcohol were used as negative controls. The MICs (MBCs) of M. urundeuva and Q. grandiflora diluted in alcohol, and clorexidine against S. mutans were 2.5 mg/ml (2.5 mg/ml), 5.0 mg/ml (--) and 0.00468 mg/ml (0.00937 mg/ml), respectively. In respect to L. casei, the MICs (MBCs) of M. urundeuva and Q. grandiflora, diluted in alcohol, and chlorhexidine were 0.156 mg/ml (0.312 mg/ml), 0.156 0.625 mg/ml (0.312 0.625 mg/ml) and 0.00468 mg/ml (0.3 mg/ml), respectively. In conclusion, our study showed that L. casei (ATTC 334) is more susceptible than S. mutans (ATCC 21175) to the extracts and the extract of M. urundeuva has a better antimicrobial effect than Q. grandiflora against S. mutans (ATCC 21175), but both extracts have similar effect on L. casei (ATTC 334) and they were inferior to CHX.

Objectifs: Le but de cette étude clinique était de comparer un groupe d’adultes ayant un parodonte sain avec un groupe d’adultes atteints de parodontite chronique en terme de risque carieux et mesures cliniques et microbiologiques de la carie. Méthodes: Quatre-vingt-seize individus ont été divisés en deux groupes en fonction de leur état de santé parodontal et ont été appariés pour l'âge, le sexe et l'origine ethnique. Trente-huit sujets étaient atteints de parodontite chronique définie comme ayant au moins quatre dents avec ≥ 1 site avec une profondeur de sondage ≥ 4 mm et une perte d'attache clinique ≥ 2 mm, et 58 sujets présentaient un parodonte sain. Par la suite, les groupes ont été subdivisés en deux groupes en fonction de leur statut carieux : les participants ayant au moins une lésion carieuse non traitée sur une surface dentaire et ceux n’ayant pas de lésion carieuse non traitée. Les données ont été recueillies par le biais d’un questionnaire, un examen clinique et des échantillons de plaque supra- et sous-gingivale. L’évaluation de la charge buccale de Streptococcus mutans et de six agents pathogènes parodontaux a été réalisée par la technique d'amplification de la réaction en chaine de la polymérase (PCR). Les données ont été analysées à l'aide d’analyses statistiques descriptives et bivariées. Résultats: Les individus atteints de parodontite chronique étaient 3,5 fois plus susceptibles d'avoir des caries que les individus en bonne santé (OR 3,5 ; IC: 1,5 - 8,3 ; P = 0,006). Les sujets à la fois atteints de parodontite chronique et de caries dentaires ont eu un niveau d’éducation significativement plus faible que les sujets ayant un parodonte sain et sans caries dentaires (OR 6,0 ; IC: 1,7 à 21,7 ; P = 0,04). La proportion de sujets ayant une charge buccale élevée de Porphyromonas gingivalis (P. g.) et Treponema denticola (T. d.) était significativement plus élevée chez les patients atteints de parodontite chronique et de carie que chez les patients sains présentant des caries (P. g.: OR 8,6 ; IC: 2,4 - 30,3 ; P = 0,004 et T. d.: OR 10,0 ; CI: 2,6 - 38.1 ; P = 0,003). Conclusions: Les résultats de cette étude suggèrent que, chez les sujets adultes atteints de la parodontite chronique, la fréquence des caries est plus élevée que chez les sujets ayant un parodonte sain. De plus, le faible niveau d'éducation influence négativement le statut parodontal des individus. / Aim: The aim of this clinical study was to compare adults with a healthy periodontium and those with chronic periodontitis, in terms of caries’ risk and caries’ clinical and microbiological measures. Methods: Ninety-six healthy adults were divided into chronic periodontitis (n= 38) and healthy periodontium (n=58) based on their periodontal status, and matched for age, gender, and ethnic background. Chronic periodontitis was defined as having at least four teeth with ≥1 site with a pocket depth ≥4 mm and clinical attachment loss ≥2 mm. Each group were subsequently subdivided in 2 groups according to their caries status: participants having at least one untreated decayed surface and those with no untreated caries. Data were collected by means of self-administrated questionnaire, clinical examination, and supra- and subgingival plaque sampling. Assessments of oral levels of Streptococcus mutans and six periodontal pathogens were conducted by PCR amplification techniques. Data were analyzed using descriptive and bivariate statistical tests. Results: Individuals with chronic periodontitis were 3.5 times more likely to have caries than healthy individuals (OR 3.5; CI: 1.5 – 8.3; P = 0.006). Subjects with both chronic periodontitis and dental caries had a significantly lower level of education than periodontally healthy subjects without dental caries (OR 6.0; CI: 1.7 – 21.7; P = 0.04). A significant higher proportion of subjects with high oral levels of Porphyromonas gingivalis (P. g.) and Treponema denticola (T. d.) was found among subjects with chronic periodontitis and untreated caries compared to periodontally healthy subjects with untreated caries (P. g.: OR 8.6; CI: 2.4 – 30.3; P = 0.004 and T. d.: OR 10.0; CI: 2.6 – 38.1; P = 0.003). Conclusion: The results from this study suggest that, adults with chronic periodontitis are more prone to caries disease than those adults with a healthy periodontium. Furthermore, low educational level could have a negative impact on the periodontal status of individuals.

In addition to occasional opportunistic colonization of the oral mucosa, Candida albicans is frequently found in carious dentin. The yeast’s potential to induce dental caries as a consequence of its pronounced ability to produce and tolerate acids was investigated. Eighty caries-active Osborne-Mendel rats were raised on an ampicillin-supplemented diet and exposed to C. albicans and/or Streptococcus mutans, except for controls. Throughout the 28-day test period, the animals were offered the modified cariogenic diet 2000a, containing 40% various sugars. Subsequently, maxillary molars were scored for plaque extent. After dissection, the mandibular molars were evaluated for smooth surface and fissure caries. Test animals exposed to C. albicans displayed considerably more advanced fissure lesions (p < 0.001) than non-exposed controls. While S. mutans yielded similar results, a combined association of C. albicans and S. mutans had no effect on occlusal caries incidence. Substituting dietary sucrose by glucose did not modify caries induction by C. albicans. However, animals fed a diet containing 20% of both sugars showed no differences to non-infected controls. Smooth surface caries was not generated by the yeast. This study provides experimental evidence that C. albicans is capable of causing occlusal caries in rats at a high rate. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.