Effect of different irrigation protocols on the removal of mono and multispecies biofilms, depth of penetration and changes in the dentin surface

Detalhes bibliográficos
Ano de defesa: 2021
Autor(a) principal: Costa, Vanessa Abreu Sanches Marques
Orientador(a): Não Informado pela instituição
Banca de defesa: Não Informado pela instituição
Tipo de documento: Tese
Tipo de acesso: Acesso aberto
Idioma: eng
Instituição de defesa: Biblioteca Digitais de Teses e Dissertações da USP
Programa de Pós-Graduação: Não Informado pela instituição
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
Palavras-chave em Português:
Link de acesso: https://www.teses.usp.br/teses/disponiveis/25/25147/tde-03122021-171536/
Resumo: The aim of the present study was to evaluate the antimicrobial action against mono and multispecies biofilms, the chemical changes in dentine surface, by Attenuated Total Reflectance in Fourier Transform Infrared Spectroscopy (ATR-FTIR), microhardness, Picrosirius Red staining and the influence of contact time and activation of irrigants on dentinal penetration by Optical Coherence Tomography (OCT), after submission to different irrigation protocols. Dentine samples from bovine teeth (N=20) were submitted to the irrigation protocols after in vitro induction of monospecies and dual-species biofilms. Live/dead dye and a confocal laser scanning microscope were used to measure the percentage of living cells. The samples for ATRFTIR (N=10) were analyzed before and after the treatments and the proportions of amide III / phosphate and carbonate / phosphate were determined. For the OCT, crosssectional images were obtained from the samples (N=5), before and after the irrigation protocols, and the mean depth of penetration () was obtained. Microhardness (N=12) was measured with Knoop indenter under a 25 g load for 15 seconds, before and after treatments. Samples passed through a histological processing to Sirius Red staining (n=12) after irrigation schemes. Data were statistically compared (P <.05). For the E. faecalis biofilm, the PUI groups had the best antimicrobial action regardless of chemical action (P <.05). Easy Clean worked best with chlorhexidine, regardless of time or NaOCl in longer contact time (P <.05). Against E. faecalis and S. oralis, only Easy Clean with saline solution, regardless of time, did not have an antimicrobial action (P >.05), as well as PUI saline solution, for the biofilm of E. faecalis with C.albicans, and Easy Clean in less agitation time with saline solution (P >.05).The irrigant contact time intensified the dissolution of the dentin collagen reducing the amide III/phosphate ratio when in contact for 30 minutes with NaOCl (P <.05); In the carbonate/phosphate ratio, there were no intergroup differences (P >.05), only intra-group changes in conventional irrigation, PUI and EasyClean treatments with saline solution (P <.05). The diffusion values of irrigants in dentin reveal in the intragroup analysis that, except for conventional irrigation with saline, there was a significant diffusion of irrigants, through the tested protocols (P <.05). In the intergroup analysis, no significant differences were observed (P >.05). Except for the saline group, immersed in 30 minutes, there were no statistically significant differences in relation to their initial microhardness values (P >.05) and there were no intergroup differences (P >.05). All groups showed birefringence for greenish, yellow and red fibers, demonstrating the maturation dynamics of the matrix. The agitation of irrigants by the PUI technique and continuous rotation with Easy clean, as well as the association with the factors of time and chemical action, favors the microbial reduction on the dentin surface among the studied biofilms. The use of NaOCl for prolonged time or with continuous agitation causes greater deproteination of the organic matrix of dentin compared to saline solution. Some carbonate ions are removed in the inorganic phase of the dentine by the saline solution. The agitation and contact time of the sodium hypochlorite, chlorhexidine or saline irrigating solutions and the longer contact time favor the penetration of these irrigants in the root dentin. The irrigation regimes applied in this study are not capable of altering dentinal microhardness. The solutions of chlorhexidine and NaOCl, when in contact with the dentin for a prolonged period, can cause a disorganization in the fibrillar network or modify the morphological structure of the dentin substrate.
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spelling Effect of different irrigation protocols on the removal of mono and multispecies biofilms, depth of penetration and changes in the dentin surfaceEfeito de diferentes protocolos de irrigação na remoção de biofilmes mono e multiespécies, profundidade de penetração e alterações da superfície dentináriaBiofilmesBiofilmsEspectroscopia de infravermelho com transformada de FourierSpectroscopy fourier transform infraredTomografia de coerência ópticaTomography optical coherenceThe aim of the present study was to evaluate the antimicrobial action against mono and multispecies biofilms, the chemical changes in dentine surface, by Attenuated Total Reflectance in Fourier Transform Infrared Spectroscopy (ATR-FTIR), microhardness, Picrosirius Red staining and the influence of contact time and activation of irrigants on dentinal penetration by Optical Coherence Tomography (OCT), after submission to different irrigation protocols. Dentine samples from bovine teeth (N=20) were submitted to the irrigation protocols after in vitro induction of monospecies and dual-species biofilms. Live/dead dye and a confocal laser scanning microscope were used to measure the percentage of living cells. The samples for ATRFTIR (N=10) were analyzed before and after the treatments and the proportions of amide III / phosphate and carbonate / phosphate were determined. For the OCT, crosssectional images were obtained from the samples (N=5), before and after the irrigation protocols, and the mean depth of penetration () was obtained. Microhardness (N=12) was measured with Knoop indenter under a 25 g load for 15 seconds, before and after treatments. Samples passed through a histological processing to Sirius Red staining (n=12) after irrigation schemes. Data were statistically compared (P <.05). For the E. faecalis biofilm, the PUI groups had the best antimicrobial action regardless of chemical action (P <.05). Easy Clean worked best with chlorhexidine, regardless of time or NaOCl in longer contact time (P <.05). Against E. faecalis and S. oralis, only Easy Clean with saline solution, regardless of time, did not have an antimicrobial action (P >.05), as well as PUI saline solution, for the biofilm of E. faecalis with C.albicans, and Easy Clean in less agitation time with saline solution (P >.05).The irrigant contact time intensified the dissolution of the dentin collagen reducing the amide III/phosphate ratio when in contact for 30 minutes with NaOCl (P <.05); In the carbonate/phosphate ratio, there were no intergroup differences (P >.05), only intra-group changes in conventional irrigation, PUI and EasyClean treatments with saline solution (P <.05). The diffusion values of irrigants in dentin reveal in the intragroup analysis that, except for conventional irrigation with saline, there was a significant diffusion of irrigants, through the tested protocols (P <.05). In the intergroup analysis, no significant differences were observed (P >.05). Except for the saline group, immersed in 30 minutes, there were no statistically significant differences in relation to their initial microhardness values (P >.05) and there were no intergroup differences (P >.05). All groups showed birefringence for greenish, yellow and red fibers, demonstrating the maturation dynamics of the matrix. The agitation of irrigants by the PUI technique and continuous rotation with Easy clean, as well as the association with the factors of time and chemical action, favors the microbial reduction on the dentin surface among the studied biofilms. The use of NaOCl for prolonged time or with continuous agitation causes greater deproteination of the organic matrix of dentin compared to saline solution. Some carbonate ions are removed in the inorganic phase of the dentine by the saline solution. The agitation and contact time of the sodium hypochlorite, chlorhexidine or saline irrigating solutions and the longer contact time favor the penetration of these irrigants in the root dentin. The irrigation regimes applied in this study are not capable of altering dentinal microhardness. The solutions of chlorhexidine and NaOCl, when in contact with the dentin for a prolonged period, can cause a disorganization in the fibrillar network or modify the morphological structure of the dentin substrate.O objetivo do presente estudo foi avaliar a ação antimicrobiana contra biofilmes mono e multiespécies, as alterações químicas na superfície da dentina, por reflexão totalmente atenuada em espectroscopia por transformada de Fourier no infravermelho (ATR-FTIR), microdureza, coloração com Picrosirius Red e a influência do tempo de contato e ativação de irrigantes na penetração dentinária por tomografia de coerência óptica (OCT), após diferentes protocolos de irrigação. Amostras de dentina bovina (N=20) foram submetidas aos protocolos de irrigação após indução in vitro de biofilmes. Corante Live/Dead e um microscópio confocal de varredura a laser foram usados para medir a porcentagem de células vivas. As amostras para ATR-FTIR (N=10) foram analisadas antes e após os tratamentos e as proporções de amida III/fosfato e carbonato/fosfato foram determinadas. Para a OCT, foram obtidas imagens transversais das amostras (N=5), antes e após os protocolos de irrigação, e obtida a profundidade média de penetração (). A microdureza (N=12) foi medida com indentador Knoop sob carga de 25 g por 15 segundos, antes e após os tratamentos. As amostras passaram por um processamento histológico, para coloração Picrosirius Red (N=12), após os regimes de irrigação. Os dados foram comparados estatisticamente (P <.05). Para o biofilme de E. faecalis, os grupos PUI tiveram a melhor efeito antimicrobiano independente da ação química (P <.05). Easy Clean atuou melhor com clorexidina, independente do tempo ou NaOCl em tempo de contato maior (P <0,05). Contra E. faecalis e S. oralis, apenas Easy Clean com solução salina, independente do tempo, não teve ação antimicrobiana (P> .05), assim como PUI ou Easy Clean em menor tempo de agitação com solução salina (P> .05), para o biofilme de E. faecalis com C. albicans. O tempo de contato do NaOCl (30 minutos) intensificou a dissolução do colágeno dentinário reduzindo a relação amida III/fosfato (P< .05 ). Na relação carbonato/fosfato, não houve diferenças intergrupos (P> .05), apenas alterações intragrupo na irrigação convencional, PUI e EasyClean com solução salina (P <.05). Os valores de difusão dos irrigantes na dentina revelam na análise intragrupo que, exceto para a irrigação convencional com soro fisiológico, houve uma difusão significativa dos irrigantes, através dos protocolos testados (P <.05). Na análise intergrupo, não foram observadas diferenças significativas (P > .05). Exceto para o grupo solução salina, imerso em 30 minutos, não houve diferenças estatisticamente significativas em relação aos seus valores iniciais de microdureza (P> .05) e não apresentaram diferenças intergrupos (P> .05). Todos os grupos apresentaram birrefringência para fibras esverdeadas,amarelas e vermelhas, demonstrando a dinâmica de maturação da matriz. A agitação dos irrigantes pela técnica PUI e rotação contínua, assim como a associação com os fatores tempo e ação química, favorecem a redução microbiana na superfície dentinária entre os biofilmes estudados. O uso de NaOCl por tempo prolongado ou com agitação contínua causa maior desproteinação da matriz orgânica da dentina em relação ao soro fisiológico. Alguns íons carbonato são removidos na fase inorgânica da dentina pela solução salina. A agitação e tempo de contato das soluções irrigantes de hipoclorito de sódio, clorexidina ou salina e o maior tempo de contato favorecem a penetração desses irrigantes na dentina radicular. Os regimes de irrigação aplicados neste estudo não são capazes de alterar a microdureza dentinária. As soluções de clorexidina e NaOCl, quando em contato com a dentina por um período prolongado, podem causar uma desorganização na rede fibrilar ou modificar a estrutura morfológica do substrato dentinário.Biblioteca Digitais de Teses e Dissertações da USPVivan, Rodrigo RicciCosta, Vanessa Abreu Sanches Marques2021-04-07info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttps://www.teses.usp.br/teses/disponiveis/25/25147/tde-03122021-171536/reponame:Biblioteca Digital de Teses e Dissertações da USPinstname:Universidade de São Paulo (USP)instacron:USPLiberar o conteúdo para acesso público.info:eu-repo/semantics/openAccesseng2023-12-03T13:00:27Zoai:teses.usp.br:tde-03122021-171536Biblioteca Digital de Teses e Dissertaçõeshttp://www.teses.usp.br/PUBhttp://www.teses.usp.br/cgi-bin/mtd2br.plvirginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.bropendoar:27212023-12-03T13:00:27Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)false
dc.title.none.fl_str_mv Effect of different irrigation protocols on the removal of mono and multispecies biofilms, depth of penetration and changes in the dentin surface
Efeito de diferentes protocolos de irrigação na remoção de biofilmes mono e multiespécies, profundidade de penetração e alterações da superfície dentinária
title Effect of different irrigation protocols on the removal of mono and multispecies biofilms, depth of penetration and changes in the dentin surface
spellingShingle Effect of different irrigation protocols on the removal of mono and multispecies biofilms, depth of penetration and changes in the dentin surface
Costa, Vanessa Abreu Sanches Marques
Biofilmes
Biofilms
Espectroscopia de infravermelho com transformada de Fourier
Spectroscopy fourier transform infrared
Tomografia de coerência óptica
Tomography optical coherence
title_short Effect of different irrigation protocols on the removal of mono and multispecies biofilms, depth of penetration and changes in the dentin surface
title_full Effect of different irrigation protocols on the removal of mono and multispecies biofilms, depth of penetration and changes in the dentin surface
title_fullStr Effect of different irrigation protocols on the removal of mono and multispecies biofilms, depth of penetration and changes in the dentin surface
title_full_unstemmed Effect of different irrigation protocols on the removal of mono and multispecies biofilms, depth of penetration and changes in the dentin surface
title_sort Effect of different irrigation protocols on the removal of mono and multispecies biofilms, depth of penetration and changes in the dentin surface
author Costa, Vanessa Abreu Sanches Marques
author_facet Costa, Vanessa Abreu Sanches Marques
author_role author
dc.contributor.none.fl_str_mv Vivan, Rodrigo Ricci
dc.contributor.author.fl_str_mv Costa, Vanessa Abreu Sanches Marques
dc.subject.por.fl_str_mv Biofilmes
Biofilms
Espectroscopia de infravermelho com transformada de Fourier
Spectroscopy fourier transform infrared
Tomografia de coerência óptica
Tomography optical coherence
topic Biofilmes
Biofilms
Espectroscopia de infravermelho com transformada de Fourier
Spectroscopy fourier transform infrared
Tomografia de coerência óptica
Tomography optical coherence
description The aim of the present study was to evaluate the antimicrobial action against mono and multispecies biofilms, the chemical changes in dentine surface, by Attenuated Total Reflectance in Fourier Transform Infrared Spectroscopy (ATR-FTIR), microhardness, Picrosirius Red staining and the influence of contact time and activation of irrigants on dentinal penetration by Optical Coherence Tomography (OCT), after submission to different irrigation protocols. Dentine samples from bovine teeth (N=20) were submitted to the irrigation protocols after in vitro induction of monospecies and dual-species biofilms. Live/dead dye and a confocal laser scanning microscope were used to measure the percentage of living cells. The samples for ATRFTIR (N=10) were analyzed before and after the treatments and the proportions of amide III / phosphate and carbonate / phosphate were determined. For the OCT, crosssectional images were obtained from the samples (N=5), before and after the irrigation protocols, and the mean depth of penetration () was obtained. Microhardness (N=12) was measured with Knoop indenter under a 25 g load for 15 seconds, before and after treatments. Samples passed through a histological processing to Sirius Red staining (n=12) after irrigation schemes. Data were statistically compared (P <.05). For the E. faecalis biofilm, the PUI groups had the best antimicrobial action regardless of chemical action (P <.05). Easy Clean worked best with chlorhexidine, regardless of time or NaOCl in longer contact time (P <.05). Against E. faecalis and S. oralis, only Easy Clean with saline solution, regardless of time, did not have an antimicrobial action (P >.05), as well as PUI saline solution, for the biofilm of E. faecalis with C.albicans, and Easy Clean in less agitation time with saline solution (P >.05).The irrigant contact time intensified the dissolution of the dentin collagen reducing the amide III/phosphate ratio when in contact for 30 minutes with NaOCl (P <.05); In the carbonate/phosphate ratio, there were no intergroup differences (P >.05), only intra-group changes in conventional irrigation, PUI and EasyClean treatments with saline solution (P <.05). The diffusion values of irrigants in dentin reveal in the intragroup analysis that, except for conventional irrigation with saline, there was a significant diffusion of irrigants, through the tested protocols (P <.05). In the intergroup analysis, no significant differences were observed (P >.05). Except for the saline group, immersed in 30 minutes, there were no statistically significant differences in relation to their initial microhardness values (P >.05) and there were no intergroup differences (P >.05). All groups showed birefringence for greenish, yellow and red fibers, demonstrating the maturation dynamics of the matrix. The agitation of irrigants by the PUI technique and continuous rotation with Easy clean, as well as the association with the factors of time and chemical action, favors the microbial reduction on the dentin surface among the studied biofilms. The use of NaOCl for prolonged time or with continuous agitation causes greater deproteination of the organic matrix of dentin compared to saline solution. Some carbonate ions are removed in the inorganic phase of the dentine by the saline solution. The agitation and contact time of the sodium hypochlorite, chlorhexidine or saline irrigating solutions and the longer contact time favor the penetration of these irrigants in the root dentin. The irrigation regimes applied in this study are not capable of altering dentinal microhardness. The solutions of chlorhexidine and NaOCl, when in contact with the dentin for a prolonged period, can cause a disorganization in the fibrillar network or modify the morphological structure of the dentin substrate.
publishDate 2021
dc.date.none.fl_str_mv 2021-04-07
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
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url https://www.teses.usp.br/teses/disponiveis/25/25147/tde-03122021-171536/
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language eng
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dc.rights.driver.fl_str_mv Liberar o conteúdo para acesso público.
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Liberar o conteúdo para acesso público.
eu_rights_str_mv openAccess
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dc.publisher.none.fl_str_mv Biblioteca Digitais de Teses e Dissertações da USP
publisher.none.fl_str_mv Biblioteca Digitais de Teses e Dissertações da USP
dc.source.none.fl_str_mv
reponame:Biblioteca Digital de Teses e Dissertações da USP
instname:Universidade de São Paulo (USP)
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instname_str Universidade de São Paulo (USP)
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reponame_str Biblioteca Digital de Teses e Dissertações da USP
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repository.name.fl_str_mv Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)
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