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Master's Dissertation
DOI
https://doi.org/10.11606/D.25.2021.tde-01122021-121035
Document
Author
Full name
Evelyn Giuliana Velásquez Espedilla
Institute/School/College
Knowledge Area
Date of Defense
Published
Bauru, 2021
Supervisor
Committee
Andrade, Flaviana Bombarda de (President)
Bramante, Clovis Monteiro
Junqueira, Juliana Campos
Ribeiro, Allan Victor
Title in Portuguese
Análise topográfica de biofilmes tratados com diferentes irrigantes sobre substratos também tratados, em microscopia de força atômica
Keywords in Portuguese
Adesão celular
Enterococcus faecalis
Irrigantes do canal radicular
Microscopia de força atômica
Abstract in Portuguese
Avaliou-se por meio da Microscopia de força atômica (AFM) a topografia de dentina radicular e biofilmes de Enterococcus faecalis, irrigados com soluções de uso endodôntico. Foram obtidos blocos de dentina da raiz de caninos humanos com o uso de uma trefina para osso e polidos até a espessura de 1 mm e esterilizados em autoclave. Foi reativada a cepa de E. faecalis (ATCC 29212) em culturas de 24h para atingir a concentração adequada de 12x108 UFC/mL. Os blocos de dentina foram divididos em dois experimentos diferenciados e com quatro grupos cada um quanto aos irrigantes (n=6): G1 (EDTA 17%), G2 (NaOCl 2,5%); G3 (NaOCl 2,5% + EDTA 17%), G4 (NaOCl 5% + HEBP 18%). As avaliações em AFM foram dispostas em duas etapas: Etapa I: primeira leitura da dentina, segunda leitura com o inoculo bacteriano e terceira leitura com o tratamento do biofilme pelas soluções. Etapa II: primeira leitura da dentina, segunda leitura do tratamento da superfície com as soluções e terceira leitura com o inóculo sobre a dentina radicular. As imagens foram avaliadas após padronização pelo teste Kappa por três examinadores. A análise estatística entre os grupos foi realizada com o teste de Kruskal-Wallis seguido de Dunn, entre as etapas o teste de Mann-Whitney e na avaliação da rugosidade foi empregado o teste ANOVA seguido de Tukey. Na etapa I os grupos mostraram diferenças entre si, sendo que o G2 e G3 demonstraram maior remoção de biofilme quando comparados ao G1 e na etapa II, o G3 mostrou menor adesão de E. faecalis após tratamento de superfície. A rugosidade do substrato também foi avaliada, demonstrando-se maior no grupo G3, diferente estatisticamente de G2, denotando a influência do agente quelante EDTA nesta propriedade. Concluiu-se que o tipo de solução irrigadora e a mistura delas é um fator importante para remover uma quantidade significativa de micro-organismos, como também podem alterar a superfície da dentina tratada dificultando a adesão dos mesmos. Dentre as soluções testadas, a associação NaOCl e EDTA eliminou maior quantidade de biofilmes formados assim como dificultou a adesão das bactérias sobre a dentina radicular.
Title in English
Topographic analysis of biofilms treated with different irrigants on substrates also treated in Atomic Force Microscopy
Keywords in English
Atomic force microscopy
Cell adhesion
Enterococcus faecalis
Root canal irrigants
Abstract in English
The topography of root dentin and Enterococcus faecalis biofilms, irrigated with endodontic solutions, were evaluated using atomic force microscopy (AFM). Dentin blocks were obtained from the root of human canines using a bone trephine and polished to a thickness of 1 mm and sterilized in an autoclave. The strain of E. faecalis (ATCC 29212) was reactivated in 24h cultures to reach the appropriate concentration of 12x108 CFU/mL. Dentin blocks were divided into two different experiments with four groups each with regard to irrigants (n=6): G1 (17% EDTA), G2 (2.5% NaOCl); G3 (2.5% NaOCl + 17% EDTA), G4 (5% NaOCl + 18% HEBP). The evaluations in AFM were arranged in two stages. Stage I: first reading of the dentin, second reading with the bacterial inoculum and third reading with the treatment of the biofilm by the solutions. Stage II: first reading of the dentin, second reading of the surface treatment with the solutions and third reading with the inoculum over the treated substrate. The images were evaluated after standardization by the Kappa test by three examiners. The statistical analysis between the groups was performed with the Kruskal-Wallis test followed by Dunn, between the experiments the Mann- Whitney test and in the roughness the ANOVA test was used followed by Tukey test. In stage I the groups showed differences between them, with G2 and G3 showing greater removal of biofilm when compared to G1 and in stage II, G3 showed less adhesion of E. faecalis after surface treatment. The substrate roughness was also evaluated, being higher in the G3 group, statistically different from G2, showing the influence of the EDTA chelating agent in this property. It was concluded that the type of irrigation solution and the mixture of them is an important factor to remove a significant amount of microorganisms as it can also change the surface of the treated dentin making them difficult to adhere. Among the tested solutions, the association of NaOCl and EDTA eliminated greater amount of formed biofilms, as difficult the adhesion of bacteria on the root dentin.
 
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Publishing Date
2021-12-01
 
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