Influência de diferentes tratamentos físico-químicos na adesão e viabilidade de fibroblastos de ligamento periodontal à superfície radicular simulando dentes avulsionados

Detalhes bibliográficos
Autor(a) principal: Moreira, Camila Said [UNESP]
Data de Publicação: 2016
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://hdl.handle.net/11449/137914
Resumo: The delayed reimplantation of avulsed teeth is a reality in dental offices. In an attempt to restore architecture and function of the periodontal ligament, avoiding ankylosis, preventing or modulating the replacement resorption, avulsed teeth root surface may be treated with various substances including EDTA (ethylenediaminetetraacetic acid) to 17%, always seeking a lower chance of occurring inflammatory or replacement resorption. Thus, the aim of this study was to evaluate the effectiveness of the treatment of discs cutted of the vestibular surface of bovine teeth roots with EDTA 17% with Emdogain® with hyaluronic acid and collagen through cell viability tests, quantification of cytokines IL -1ß, IL-6, IL-8, IL-10 and TNF-α by ELISA assay and, in addition, it was taken Illustrations by scanning electron microscopy of the accession of fibroblasts on the dentin disks. 155 dentin discs with 4,5 mm in diameter will be cut from the vestibular surface of the root surface of bovine teeth, previously scraped or not with periodontal curette. The discs were regularized, cleaned and autoclaved. The specimens were treated with the substances advocated and placed in 96 well plates are seeded in which primary cell cultures of human periodontal ligament fibroblasts that were in contact with the discs for 48 hours. The survival and cell viability on the surface of the disks are evaluated by XTT assay. The scanning electron microscopy was used to verify adherence of fibroblasts to the surface of the discs. Detection and quantification of cytokines were done by ELISA assay. The data were statistically analyzed by ANOVA and Tukey (p < 0.05). The results showed that the use of 17% EDTA for 5 minutes without stirring was more effective than the other protocols (p <0.05) and combined with hyaluronic acid was less cytotoxic promoting better cell viability. It was not possible to ensure the fibroblasts display in scanning electron microscopy. As for the ELISA assay, there was expression of IL- 1β, IL -6 and IL -8 (pro- inflammatory cytokines), but there was just statistically significant difference between IL -6 and IL -8. Thus, we can conclude that 17% EDTA for 5 minutes without stirring and hyaluronic acid showed less cytotoxicity index fibroblasts of the periodontal ligament, but there was inflammatory response by IL -6 and IL-8 expression .
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spelling Influência de diferentes tratamentos físico-químicos na adesão e viabilidade de fibroblastos de ligamento periodontal à superfície radicular simulando dentes avulsionadosInfluence of different physical-chemical treatment on periodontal ligament fibroblast adhesion to root surface of simulated avulsed teethTooth avulsionEdetic acidEnzyme-linked immunosorbent assayAvulsão dentáriaÁcido etilenodiaminotetracéticoEnsaio de imunoadsorção enzimáticaThe delayed reimplantation of avulsed teeth is a reality in dental offices. In an attempt to restore architecture and function of the periodontal ligament, avoiding ankylosis, preventing or modulating the replacement resorption, avulsed teeth root surface may be treated with various substances including EDTA (ethylenediaminetetraacetic acid) to 17%, always seeking a lower chance of occurring inflammatory or replacement resorption. Thus, the aim of this study was to evaluate the effectiveness of the treatment of discs cutted of the vestibular surface of bovine teeth roots with EDTA 17% with Emdogain® with hyaluronic acid and collagen through cell viability tests, quantification of cytokines IL -1ß, IL-6, IL-8, IL-10 and TNF-α by ELISA assay and, in addition, it was taken Illustrations by scanning electron microscopy of the accession of fibroblasts on the dentin disks. 155 dentin discs with 4,5 mm in diameter will be cut from the vestibular surface of the root surface of bovine teeth, previously scraped or not with periodontal curette. The discs were regularized, cleaned and autoclaved. The specimens were treated with the substances advocated and placed in 96 well plates are seeded in which primary cell cultures of human periodontal ligament fibroblasts that were in contact with the discs for 48 hours. The survival and cell viability on the surface of the disks are evaluated by XTT assay. The scanning electron microscopy was used to verify adherence of fibroblasts to the surface of the discs. Detection and quantification of cytokines were done by ELISA assay. The data were statistically analyzed by ANOVA and Tukey (p < 0.05). The results showed that the use of 17% EDTA for 5 minutes without stirring was more effective than the other protocols (p <0.05) and combined with hyaluronic acid was less cytotoxic promoting better cell viability. It was not possible to ensure the fibroblasts display in scanning electron microscopy. As for the ELISA assay, there was expression of IL- 1β, IL -6 and IL -8 (pro- inflammatory cytokines), but there was just statistically significant difference between IL -6 and IL -8. Thus, we can conclude that 17% EDTA for 5 minutes without stirring and hyaluronic acid showed less cytotoxicity index fibroblasts of the periodontal ligament, but there was inflammatory response by IL -6 and IL-8 expression .O reimplante tardio de dentes avulsionados é uma realidade dentro dos consultórios odontológicos. Na tentativa de se restabelecer a arquitetura e a função do ligamento periodontal, além de se evitar a anquilose, evitando ou modulando a reabsorção por substituição, a superfície radicular dos dentes avulsionados pode ser tratada com diversas substâncias, dentre elas o EDTA (ácido etilenodiamino tetra-acético) a 17%, sempre visando uma menor chance de ocorrer reabsorção inflamatória e por substituição. Sendo assim, o objetivo deste estudo foi avaliar a eficácia do tratamento de discos da superfície vestibular de raízes de dentes bovinos, com EDTA 17%, com Emdogain® , com ácido hialurônico e com colágeno, através de testes de viabilidade celular, quantificação das citocinas IL-1ß, IL-6, IL-8, IL-10 e TNF-α por ensaio ELISA, além da ilustração por microscopia eletrônica de varredura da adesão dos fibroblastos sobre os discos de dentina. Foram cortados 155 discos de dentina, com 4,5 mm de diâmetro, da face vestibular da superfície radicular de dentes bovinos, previamente raspados ou não com cureta periodontal. Os discos foram regularizados, limpos e autoclavados. Os espécimes foram tratados com as substâncias preconizadas e colocados em placas de 96 poços onde foram semeadas células de culturas primárias de fibroblastos humanos de ligamento periodontal, que ficaram em contato com os discos por 48 horas. A sobrevivência e a viabilidade celular na superfície dos discos foram avaliadas através do ensaio de XTT. A microscopia eletrônica de varredura foi utilizada para verificar a adesão de fibroblastos à superfície dos discos. A detecção e quantificação das citocinas foram realizadas pelo teste ELISA. Os dados foram submetidos à análise estatística pelo teste ANOVA e Tukey (p < 0,05). Os resultados obtidos mostraram que o uso de EDTA 17% por 5 minutos sem agitação foi mais eficaz que os outros protocolos (p < 0,05) e aliado ao ácido hialurônico, foi menos citotóxico promovendo melhor viabilidade celular. Não foi possível assegurar a visualização dos fibroblastos em microscopia eletrônica de varredura. Quanto ao ensaio de ELISA, houve expressão das citocinas IL-1β, IL-6 e IL-8 (citocinas pró-inflamatórias), porém houve diferença estatisticamente significante entre a IL-6 e a IL-8. Sendo assim, podemos concluir que o EDTA 17% por 5 minutos sem agitação e o ácido hialurônico apresentaram menor índice de citotoxicidade aos fibroblastos do ligamento periodontal, porém houve resposta inflamatória pela expressão de IL-6 e IL-8.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Universidade Estadual Paulista (Unesp)Camargo, Carlos Henrique Ribeiro Camargo [UNESP]Universidade Estadual Paulista (Unesp)Moreira, Camila Said [UNESP]2016-04-13T13:26:27Z2016-04-13T13:26:27Z2016-02-17info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttp://hdl.handle.net/11449/13791400086680233004145070P8porinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESP2024-01-24T06:35:07Zoai:repositorio.unesp.br:11449/137914Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T23:52:22.310357Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Influência de diferentes tratamentos físico-químicos na adesão e viabilidade de fibroblastos de ligamento periodontal à superfície radicular simulando dentes avulsionados
Influence of different physical-chemical treatment on periodontal ligament fibroblast adhesion to root surface of simulated avulsed teeth
title Influência de diferentes tratamentos físico-químicos na adesão e viabilidade de fibroblastos de ligamento periodontal à superfície radicular simulando dentes avulsionados
spellingShingle Influência de diferentes tratamentos físico-químicos na adesão e viabilidade de fibroblastos de ligamento periodontal à superfície radicular simulando dentes avulsionados
Moreira, Camila Said [UNESP]
Tooth avulsion
Edetic acid
Enzyme-linked immunosorbent assay
Avulsão dentária
Ácido etilenodiaminotetracético
Ensaio de imunoadsorção enzimática
title_short Influência de diferentes tratamentos físico-químicos na adesão e viabilidade de fibroblastos de ligamento periodontal à superfície radicular simulando dentes avulsionados
title_full Influência de diferentes tratamentos físico-químicos na adesão e viabilidade de fibroblastos de ligamento periodontal à superfície radicular simulando dentes avulsionados
title_fullStr Influência de diferentes tratamentos físico-químicos na adesão e viabilidade de fibroblastos de ligamento periodontal à superfície radicular simulando dentes avulsionados
title_full_unstemmed Influência de diferentes tratamentos físico-químicos na adesão e viabilidade de fibroblastos de ligamento periodontal à superfície radicular simulando dentes avulsionados
title_sort Influência de diferentes tratamentos físico-químicos na adesão e viabilidade de fibroblastos de ligamento periodontal à superfície radicular simulando dentes avulsionados
author Moreira, Camila Said [UNESP]
author_facet Moreira, Camila Said [UNESP]
author_role author
dc.contributor.none.fl_str_mv Camargo, Carlos Henrique Ribeiro Camargo [UNESP]
Universidade Estadual Paulista (Unesp)
dc.contributor.author.fl_str_mv Moreira, Camila Said [UNESP]
dc.subject.por.fl_str_mv Tooth avulsion
Edetic acid
Enzyme-linked immunosorbent assay
Avulsão dentária
Ácido etilenodiaminotetracético
Ensaio de imunoadsorção enzimática
topic Tooth avulsion
Edetic acid
Enzyme-linked immunosorbent assay
Avulsão dentária
Ácido etilenodiaminotetracético
Ensaio de imunoadsorção enzimática
description The delayed reimplantation of avulsed teeth is a reality in dental offices. In an attempt to restore architecture and function of the periodontal ligament, avoiding ankylosis, preventing or modulating the replacement resorption, avulsed teeth root surface may be treated with various substances including EDTA (ethylenediaminetetraacetic acid) to 17%, always seeking a lower chance of occurring inflammatory or replacement resorption. Thus, the aim of this study was to evaluate the effectiveness of the treatment of discs cutted of the vestibular surface of bovine teeth roots with EDTA 17% with Emdogain® with hyaluronic acid and collagen through cell viability tests, quantification of cytokines IL -1ß, IL-6, IL-8, IL-10 and TNF-α by ELISA assay and, in addition, it was taken Illustrations by scanning electron microscopy of the accession of fibroblasts on the dentin disks. 155 dentin discs with 4,5 mm in diameter will be cut from the vestibular surface of the root surface of bovine teeth, previously scraped or not with periodontal curette. The discs were regularized, cleaned and autoclaved. The specimens were treated with the substances advocated and placed in 96 well plates are seeded in which primary cell cultures of human periodontal ligament fibroblasts that were in contact with the discs for 48 hours. The survival and cell viability on the surface of the disks are evaluated by XTT assay. The scanning electron microscopy was used to verify adherence of fibroblasts to the surface of the discs. Detection and quantification of cytokines were done by ELISA assay. The data were statistically analyzed by ANOVA and Tukey (p < 0.05). The results showed that the use of 17% EDTA for 5 minutes without stirring was more effective than the other protocols (p <0.05) and combined with hyaluronic acid was less cytotoxic promoting better cell viability. It was not possible to ensure the fibroblasts display in scanning electron microscopy. As for the ELISA assay, there was expression of IL- 1β, IL -6 and IL -8 (pro- inflammatory cytokines), but there was just statistically significant difference between IL -6 and IL -8. Thus, we can conclude that 17% EDTA for 5 minutes without stirring and hyaluronic acid showed less cytotoxicity index fibroblasts of the periodontal ligament, but there was inflammatory response by IL -6 and IL-8 expression .
publishDate 2016
dc.date.none.fl_str_mv 2016-04-13T13:26:27Z
2016-04-13T13:26:27Z
2016-02-17
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
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dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
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dc.publisher.none.fl_str_mv Universidade Estadual Paulista (Unesp)
publisher.none.fl_str_mv Universidade Estadual Paulista (Unesp)
dc.source.none.fl_str_mv reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
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instname_str Universidade Estadual Paulista (UNESP)
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institution UNESP
reponame_str Repositório Institucional da UNESP
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repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
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