Detecção da microbiota endodôntica de dentes sem vitalidade pulpar com ou sem lesão periapical radiográfica, por meio das técnicas de checkerboard DNA-DNA hybridization e cultura microbiológica
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Data
2006-09-26
Autores
Rodrigues, Vivian Maria Tellaroli [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
A proposta do estudo foi avaliar a microbiota endodôntica de dentes de humanos sem vitalidade pulpar com ou sem lesão periapical visível radiograficamente por meio das técnicas de Checkerboard DNA-DNA Hybridization e cultura microbiológica. Foram utilizados 20 dentes unirradiculados, divididos em casos de necrose pulpar sem lesão periapical (Grupo I) e dentes com necrose pulpar e lesão periapical (Grupo II). A colheita do material do canal radicular foi realizada com lima K e cones de papel absorvente. Para a técnica de cultura, as amostras foram semeadas em meios As, Ask, Ms e Tio's, para avaliação da presença de microrganismos anaeróbios, aeróbios e facultativos. Para a técnica de Checkerboard DNA-DNA Hybridization, foi realizado o processamento do DNA extraído das amostras e das sondas de DNA de 33 microrganismos. Os resultados mostraram nas duas técnicas empregadas a presença de microrganismos. A técnica de hibridação DNA - DNA checkerboard permitiu a identificação de 31 espécies diferentes no Grupo I e de 32 espécies diferentes no Grupo II. Fusobacterium nucleatum spp polymorphum (80%), Porphyromonas gingivalis (80%), Prevotella melaninogenica (90%) predominaram no Grupo I. As sondas Streptococcus mitis e Selenomonas noxia não foram detectadas em nenhuma amostra deste grupo. No Grupo II, Fusobacterium nucleatum spp polymorphum, Neisseria mucosa e Porphyromonas gingivalis foram detectadas em todos os casos e Selenomonas noxia não foi encontrada em nenhuma das amostras deste grupo. Houve tendência para maior presença de bactérias anaeróbias obrigatórias moderadas no Grupo II, embora sem diferença significante (p>0,05), em relação ao Grupo I, de acordo com a cultura microbiológica. Foi observada maior freqüência de espécies bacterianas anaeróbias obrigatórias moderadas nos canais radiculares, tanto para o Grupo I como para o Grupo II, de acordo com a técnica de hibridação DNA-DNA checkerboard.
The purpose of this study was to evaluate the endodontic microbiota from human's teeth, with no pulp vitality, with or without radiographic chronic periapical lesion, using a culture microbiologic method and checkerboard DNA-DNA hybridization technique. The whole sample, with 20 single root's teeth, was collected and divided into: teeth with no pulp vitality without periapical lesion (Group I) and teeth with no pulp vitality with periapical lesion (Group II). The samples were obtained with K file and absorbent paper points. To the culture technique, the samples were grown by using media like As, Ask, Ms and Tio's. To checkerboard DNA-DNA hybridization technique, the samples of DNA extracted crossed with microorganisms' DNA obtained from ATCC. The results demonstrated in two techniques used that in all the samples identified microorganisms, at least 20 ufc/ml. The checkerboard DNA-DNA hybridization technique demonstrated the identification of 31 different species of microorganisms in Group I and 32 different species of microorganisms in Group II. Fusobacterium nucleatum spp polymorphum (80%), Porphyromonas gingivalis (80%), Prevotella melaninogenica (90%) predominated in Group I. The probes Streptococcus mitis and Selenomonas noxia were not detected in no sample in this group. At Group II, Fusobacterium nucleatum spp polymorphum, Neisseria mucosa and Porphyromonas gingivalis were detected in all cases, and Selenomonas noxia was not present in sample of this group. It was tendency to greater number strict anaerobe microorganisms in Group II, however without statistical significance (p>0.05), in relation Group I, in accord with culture microbiologic method. It was detected a greater frequency of anaerobe strict microorganisms species in root canal, in both Groups I and II, in accord with the checkerboard DNA-DNA hybridization technique.
The purpose of this study was to evaluate the endodontic microbiota from human's teeth, with no pulp vitality, with or without radiographic chronic periapical lesion, using a culture microbiologic method and checkerboard DNA-DNA hybridization technique. The whole sample, with 20 single root's teeth, was collected and divided into: teeth with no pulp vitality without periapical lesion (Group I) and teeth with no pulp vitality with periapical lesion (Group II). The samples were obtained with K file and absorbent paper points. To the culture technique, the samples were grown by using media like As, Ask, Ms and Tio's. To checkerboard DNA-DNA hybridization technique, the samples of DNA extracted crossed with microorganisms' DNA obtained from ATCC. The results demonstrated in two techniques used that in all the samples identified microorganisms, at least 20 ufc/ml. The checkerboard DNA-DNA hybridization technique demonstrated the identification of 31 different species of microorganisms in Group I and 32 different species of microorganisms in Group II. Fusobacterium nucleatum spp polymorphum (80%), Porphyromonas gingivalis (80%), Prevotella melaninogenica (90%) predominated in Group I. The probes Streptococcus mitis and Selenomonas noxia were not detected in no sample in this group. At Group II, Fusobacterium nucleatum spp polymorphum, Neisseria mucosa and Porphyromonas gingivalis were detected in all cases, and Selenomonas noxia was not present in sample of this group. It was tendency to greater number strict anaerobe microorganisms in Group II, however without statistical significance (p>0.05), in relation Group I, in accord with culture microbiologic method. It was detected a greater frequency of anaerobe strict microorganisms species in root canal, in both Groups I and II, in accord with the checkerboard DNA-DNA hybridization technique.
Descrição
Palavras-chave
Endodontia, Bactéria, Microbiologia, Biologia molecular, Canais radiculares, Endodontic, Microbiologic technique, Molecular biology, Dental pulp cavity
Como citar
RODRIGUES, Vivian Maria Tellaroli. Detecção da microbiota endodôntica de dentes sem vitalidade pulpar com ou sem lesão periapical radiográfica, por meio das técnicas de checkerboard DNA-DNA hybridization e cultura microbiológica. 2006. 109 f. Dissertação (mestrado) - Universidade Estadual Paulista, Faculdade de Odontologia de Araraquara, 2006.