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Doctoral Thesis
DOI
https://doi.org/10.11606/T.25.2021.tde-01122021-132815
Document
Author
Full name
Denise Ferracioli Oda
Institute/School/College
Knowledge Area
Date of Defense
Published
Bauru, 2020
Supervisor
Committee
Andrade, Flaviana Bombarda de (President)
Garcia, Roberto Brandao
Midena, Raquel Zanin
Oliveira, Luciane Dias de
Title in English
Effect of PDT, and new chelating solutions on inactivation of bacterial LPS
Keywords in English
EDTA
Endotoxin
Etidronic acid
Photochemotherapy
Abstract in English
INTRODUCTION: In the necropulpectomies, in addition to the elimination of microorganisms and to neutralization of lipopolysaccharides (LPS), present in root canal system, it is fundamental, since that endotoxin is highly related to unsuccessful cases with painful symptomatology. OBJECTIVE: The present study evaluated the capacity of Photodynamic Therapy (PDT) and the different irrigants, to inactivate the bacterial LPS. MATERIALS AND METHODS: Eighty single-rooted human teeth were used, previously analyzed by radiographic images to confirm the single canal. The coronary portion was removed to obtain 14mm segment of the root, where the canals were properly prepared. The samples were placed vertically in 24-well plates and fixed with acrylic resin. The samples and all the materials used in the study were previously sterilized with Cobalt-60 Gamma radiation. Inside the laminar flow cabinet, 10L of LPS from Escherichia coli were inoculated inside in root canal of each sample and kept for 24 hours in a bacteriological oven. The inoculation procedure was repeated twice more and then the samples were divided into the following groups (n = 10): [G1] sodium hypochlorite at 2.5%; [G2] pyrogen-free water; [G3] trisodium EDTA at 17%; [G4] tetrasodium EDTA at 10%; [G5] Etidronate at 18%; [G6] Methylene blue at 5%; [G7] diode LASER; [G8] PDT (methylene blue + diode LASER). The time for action of the irrigating solutions, dye and irradiation, was 5 minutes for all groups. After the treatment, the canals were cleaned and filled with 5mL of pyrogen-free water, where it was agitated and aspirated by a pyrogen-free needle/syringe. The aspirated content of each specimen was kept under refrigeration at -80ºC until posterior analysis of Limulus Amebocyte Lysate. The obtained data did not pass the Shapiro Wilks normality test, so they were submitted to the Kruskal-Wallis test followed by the Dunns test. RESULTS: The obtained data presented relevant reduction only in the groups G3 (p = 0.0452) and G5 (p = 0.0097), in comparison to the control group G2. The medium percentage (Minimum/Maximum) of remaining endotoxin was: [G1] = 1.005 (0.3100/5.810); [G2] = 2.010 (0.8700/10.33); [G3] = 0.3800 (0.060 /2.150); [G4] = 1.195 (0.110/3,420); [G5] = 0.9000 (0.0/1.540); [G6] = 1.320 (0.1800/9.410); [G7] = 1.210 (0.4700/9.400); [G8] = 3.560 (0.4800/5.460).CONCLUSION: The PDT did not present efficacy in neutralizing the LPS and, among the evaluated solutions, trisodium EDTA at 17% and etidronate at 18% presented efficacy in inactivating the bacterial endotoxin.
Title in Portuguese
Efeito da PDT, e de novas soluções quelantes na inativação do LPS bacteriano
Keywords in Portuguese
EDTA
Endotoxinas
Etidronato
Fotoquimioterapia
Abstract in Portuguese
OBJETIVO: Nas necropulpectomias, além da eliminação de micro-organismos e neutralização de lipopolissacarídeos (LPS), presentes no sistema de canais radiculares, é fundamental, uma vez que esta endotoxina está altamente relacionada aos casos de insucesso e de sintomatologia dolorosa. Este trabalho avaliou a capacidade da Terapia fotodinâmica (PDT) e de diferentes irrigantes, para inativar o LPS bacteriano. MATERIAIS E MÉTODOS: Foram utilizados 80 dentes humanos unirradiculados, analisados previamente por imagens radiográficas para confirmação de canal único. A porção coronária foi removida para a obtenção de 14mm de segmento radicular, cujos canais foram devidamente preparados. As amostras foram posicionadas verticalmente em placas de cultura de 24 poços e fixados com resina acrílica. As amostras e todos os materiais a serem utilizados na pesquisa foram esterilizados, previamente, em radiação Gama cobalto 60. No interior da cabine de fluxo laminar, 10L de LPS de Escherichia coli foram inoculados no canal de cada amostra, e mantidas durante 24 horas em estufa bacteriológica. O procedimento de inoculação foi repetido mais 2 vezes e então as amostras foram divididas nos seguintes grupos (n=10): [G1] hipoclorito de sódio a 2,5%; [G2] água apirogênica; [G3] EDTA trissódico a 17%; [G4] EDTA tetrassódico a 10%; [G5] Etidronato a 18%; [G6] Azul de metileno a 5%; [G7] LASER de diodo e [G8] PDT (azul de metileno + LASER de diodo). O tempo de ação das soluções irrigadoras, do corante e da irradiação, foi de 5 minutos para todos os grupos. Após o tratamento, os canais foram lavados e preenchidos com 5mL de água apirogênica, onde a mesma foi agitada e aspirada por uma agulha/seringa apirogênica. O conteúdo aspirado de cada espécime foi mantido em refrigeração a -80ºC até posterior análise pelo método Limulus Amebocyte Lysate. Os dados obtidos não passaram no teste de normalidade Shapiro Wilks, sendo então submetido ao teste de Kruskal Wallis seguido de Dunn. RESULTADOS: Os dados obtidos apresentaram redução significativa apenas nos grupos G3 (p = 0,0452) e G 5 (p = 0,0097) em comparação ao grupo controle G2. A mediana (Mínima/Máxima) da porcentagem de endotoxina remanescente foram: [G1] = 1.005 (0.3100/5.810); [G2] = 2.010 (0.8700/10.33); [G3] = 0.3800 (0.060 /2.150); [G4] = 1.195 (0.110/3,420); [G5] = 0.9000 (0.0/1.540); [G6] = 1.320 (0.1800/9.410); [G7] = 1.210 (0.4700/9.400); [G8] = 3.560 (0.4800/5.460). CONCLUSÃO: A PDT não apresentou eficácia em neutralizar o LPS, e dentre as soluções avaliadas, o EDTA trissódico a 17% e o etidronato a 18, foram eficazes em inativar a endotoxina bacteriana.
 
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Publishing Date
2021-12-01
 
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