Efeitos da fototerapia com energia fracionada do laser de baixa intensidade em células-tronco da polpa de dentes decíduos esfoliados humanos

Detalhes bibliográficos
Ano de defesa: 2019
Autor(a) principal: Almeida Junior, Luciano Aparecido De lattes
Orientador(a): Sakai, Vivien Thiemy lattes
Banca de defesa: Moretti, Ana Beatriz Da Silveira, Santos, Gérsika Bitencourt
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal de Alfenas
Programa de Pós-Graduação: Programa de Pós-Graduação em Ciências Odontológicas
Departamento: Faculdade de Odontologia
País: Brasil
Palavras-chave em Português:
Área do conhecimento CNPq:
Link de acesso: https://repositorio.unifal-mg.edu.br/handle/123456789/1335
Resumo: Low-level laser irradiation (LLL) therapy may be able to induce proliferation and differentiation into several cellular types, with remarkable effects in the process of tissue repair, inflammation decrease and analgesia. For pulp repair after conservative procedures, pulp stem cells have the potential to differentiate into odontoblasts and to produce dentine, requiring a specific stimulus to trigger this process. The present study aimed to evaluate in vitro whether the LLL delivering fractionated total energy (multiple irradiation) or single irradiation stimulates regeneration-associated events (viability and proliferation) in stem cells from human exfoliated deciduous teeth (SHED). Cells received LLL irradiation (InGaAlP – 660 nm), according to the following experimental groups: GI (single irradiation: 2.5 J/cm², 10 s, 0.10 J), GII (single irradiation: 5.0 J/cm², 20 s, 0.20 J), GIII (single irradiation: 7.5 J/cm², 30 s, 0.30 J), GIV (2 irradiations: 2.5 J/cm², 10 s, 0.20 J), GV (3 irradiations: 2.5 J/cm², 10 s, 0.30 J) and GVI (non-irradiated). Cell viability was assessed by MTT and trypan blue exclusion (TBE) methods, while cell proliferation was evaluated by crystal violet (CV) and sulforhodamine B (SRB) assays after 24, 48 and 72 hours after the first irradiation. By MTT, there was no difference between groups at 24 and 72 hours. At 48 hours, the groups with multiple irradiation (GIV and GV) presented higher cell viability rates. The mean percentages of viable cells for all groups by TBE method were 91.04%, 96.63% and 97.48% at 24, 48 and 72 hours, respectively. By CV, there was no significant difference between groups at 24 and 48 hours; at 72 hours, GII, GIII and GIV presented higher cell proliferation. By SRB, GI and GIV presented lower proliferation rates in all the periods. When the groups presenting the same total energy were compared, GII (0.20 J) presented lower cell viability rates and higher cell proliferation rates in comparison to GIV; GIII (0.30 J) presented similar results to those of GV, with higher cell viability and proliferation. In conclusion, the application of laser delivering fractionated total energy (2 or 3 applications of 2,5 J/cm²) induced higher cell viability at 48 hours, while the 2.5 J/cm² single irradiation decreased cell viability in such period and proliferation over time. The 5.0 and 7.5 J/cm² single doses and the 3 applications of 2.5 J/cm² maintained cell viability and stimulated proliferation of SHED at 72 hours.
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spelling Almeida Junior, Luciano Aparecido Dehttp://lattes.cnpq.br/7146420151581867Marques, Nádia Carolina TeixeiraMoretti, Ana Beatriz Da SilveiraSantos, Gérsika BitencourtSakai, Vivien Thiemyhttp://lattes.cnpq.br/47690510923830702019-04-03T14:00:53Z2019-01-23ALMEIDA JUNIOR, Luciano Aparecido de. Efeitos da fototerapia com energia fracionada do laser de baixa intensidade em células-tronco da polpa de dentes decíduos esfoliados humanos. 2019. 62 f. Dissertação (Mestrado em Ciências Odontológicas) - Universidade Federal de Alfenas, Alfenas, MG, 2019.https://repositorio.unifal-mg.edu.br/handle/123456789/1335Low-level laser irradiation (LLL) therapy may be able to induce proliferation and differentiation into several cellular types, with remarkable effects in the process of tissue repair, inflammation decrease and analgesia. For pulp repair after conservative procedures, pulp stem cells have the potential to differentiate into odontoblasts and to produce dentine, requiring a specific stimulus to trigger this process. The present study aimed to evaluate in vitro whether the LLL delivering fractionated total energy (multiple irradiation) or single irradiation stimulates regeneration-associated events (viability and proliferation) in stem cells from human exfoliated deciduous teeth (SHED). Cells received LLL irradiation (InGaAlP – 660 nm), according to the following experimental groups: GI (single irradiation: 2.5 J/cm², 10 s, 0.10 J), GII (single irradiation: 5.0 J/cm², 20 s, 0.20 J), GIII (single irradiation: 7.5 J/cm², 30 s, 0.30 J), GIV (2 irradiations: 2.5 J/cm², 10 s, 0.20 J), GV (3 irradiations: 2.5 J/cm², 10 s, 0.30 J) and GVI (non-irradiated). Cell viability was assessed by MTT and trypan blue exclusion (TBE) methods, while cell proliferation was evaluated by crystal violet (CV) and sulforhodamine B (SRB) assays after 24, 48 and 72 hours after the first irradiation. By MTT, there was no difference between groups at 24 and 72 hours. At 48 hours, the groups with multiple irradiation (GIV and GV) presented higher cell viability rates. The mean percentages of viable cells for all groups by TBE method were 91.04%, 96.63% and 97.48% at 24, 48 and 72 hours, respectively. By CV, there was no significant difference between groups at 24 and 48 hours; at 72 hours, GII, GIII and GIV presented higher cell proliferation. By SRB, GI and GIV presented lower proliferation rates in all the periods. When the groups presenting the same total energy were compared, GII (0.20 J) presented lower cell viability rates and higher cell proliferation rates in comparison to GIV; GIII (0.30 J) presented similar results to those of GV, with higher cell viability and proliferation. In conclusion, the application of laser delivering fractionated total energy (2 or 3 applications of 2,5 J/cm²) induced higher cell viability at 48 hours, while the 2.5 J/cm² single irradiation decreased cell viability in such period and proliferation over time. The 5.0 and 7.5 J/cm² single doses and the 3 applications of 2.5 J/cm² maintained cell viability and stimulated proliferation of SHED at 72 hours.terapia de irradiação com laser de baixa intensidade (LBI) pode ser capaz de induzir a proliferação e a diferenciação de diversos tipos celulares, com efeitos consideráveis no processo de reparo tecidual, diminuição da inflamação e analgesia. Para o reparo pulpar após procedimentos conservadores, as células-tronco pulpares apresentam potencial de diferenciação em odontoblastos e de produção de dentina, necessitando de um estímulo específico para iniciar este processo. O presente estudo teve como objetivo avaliar in vitro se o LBI aplicado com energia total fracionada (múltiplas irradiações) ou com irradiação única estimula eventos associados à regeneração (viabilidade e proliferação) em células-tronco de dentes decíduos esfoliados humanos (SHED). As células foram irradiadas com LBI (InGaAlP – 660 nm) de acordo com os seguintes grupos experimentais: GI (irradiação única: 2,5 J/cm², 10 s, 0,10 J), GII (irradiação única: 5,0 J/cm², 20 s, 0,20 J), GIII (irradiação única: 7,5 J/cm², 30 s, 0,30 J), GIV (2 irradiações: 2,5 J/cm², 10 s, 0,20 J), GV (3 irradiações: 2,5 J/cm², 10 s, 0,30 J) e GVI (não irradiado). A viabilidade celular foi analisada pelos métodos MTT e de exclusão do azul de tripan (EAT), enquanto que a proliferação celular foi avaliada pelos ensaios cristal violeta (CV) e sulforrodamina B (SRB) após 24, 48 e 72 horas da primeira irradiação. Pelo ensaio MTT, não houve diferença entre os grupos às 24 e 72 horas. Às 48 horas, os grupos de múltiplas irradiações (GIV e GV) apresentaram maiores taxas de viabilidade celular. As porcentagens médias de células viáveis para todos os grupos pelo método EAT foram de 91,04%, 96,63% e 97,48% às 24, 48 e 72 horas, respectivamente. Pelo ensaio CV, às 24 e 48 horas não houve diferença significativa entre os grupos; já às 72 horas, GII, GIII e GV obtiveram maior proliferação celular. Pelo SRB, GI e GIV apresentaram menores índices de proliferação celular em todos os períodos. Quando comparados os grupos de mesma energia total, GII (0,20 J) apresentou menores taxas de viabilidade celular e maiores níveis de proliferação celular em relação a GIV; já GIII (0,30 J) demonstrou resultados semelhantes aos de GV, com maior viabilidade e proliferação celular. Conclui-se que a aplicação do laser com energia total fracionada (2 ou 3 aplicações de 2,5 J/cm²) induziu maior viabilidade celular às 48 horas, enquanto que a irradiação única de 2,5 J/cm² reduziu a viabilidade celular neste período e a proliferação celular ao longo do tempo. Já as doses únicas de 5,0 e 7,5 J/cm² e 2,5 J/cm² em 3 aplicações mantiveram a viabilidade e estimularam a proliferação das SHED às 72 horas.Fundação de Amparo à Pesquisa do Estado de Minas Gerais - FAPEMIGapplication/pdfporUniversidade Federal de AlfenasPrograma de Pós-Graduação em Ciências OdontológicasUNIFAL-MGBrasilFaculdade de Odontologiainfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-nd/4.0/Células-TroncoProliferação de CélulasTerapia a Laser de Baixa IntensidadeCIENCIAS DA SAUDE::ODONTOLOGIAEfeitos da fototerapia com energia fracionada do laser de baixa intensidade em células-tronco da polpa de dentes decíduos esfoliados humanosinfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/publishedVersion4673435736271820140600600600-2070498469879244349-1527361517405938873reponame:Repositório Institucional da Universidade Federal de Alfenas - RiUnifalinstname:Universidade Federal de Alfenas (UNIFAL)instacron:UNIFALAlmeida Junior, Luciano Aparecido DeLICENSElicense.txtlicense.txttext/plain; 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dc.title.pt-BR.fl_str_mv Efeitos da fototerapia com energia fracionada do laser de baixa intensidade em células-tronco da polpa de dentes decíduos esfoliados humanos
title Efeitos da fototerapia com energia fracionada do laser de baixa intensidade em células-tronco da polpa de dentes decíduos esfoliados humanos
spellingShingle Efeitos da fototerapia com energia fracionada do laser de baixa intensidade em células-tronco da polpa de dentes decíduos esfoliados humanos
Almeida Junior, Luciano Aparecido De
Células-Tronco
Proliferação de Células
Terapia a Laser de Baixa Intensidade
CIENCIAS DA SAUDE::ODONTOLOGIA
title_short Efeitos da fototerapia com energia fracionada do laser de baixa intensidade em células-tronco da polpa de dentes decíduos esfoliados humanos
title_full Efeitos da fototerapia com energia fracionada do laser de baixa intensidade em células-tronco da polpa de dentes decíduos esfoliados humanos
title_fullStr Efeitos da fototerapia com energia fracionada do laser de baixa intensidade em células-tronco da polpa de dentes decíduos esfoliados humanos
title_full_unstemmed Efeitos da fototerapia com energia fracionada do laser de baixa intensidade em células-tronco da polpa de dentes decíduos esfoliados humanos
title_sort Efeitos da fototerapia com energia fracionada do laser de baixa intensidade em células-tronco da polpa de dentes decíduos esfoliados humanos
author Almeida Junior, Luciano Aparecido De
author_facet Almeida Junior, Luciano Aparecido De
author_role author
dc.contributor.author.fl_str_mv Almeida Junior, Luciano Aparecido De
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/7146420151581867
dc.contributor.advisor-co1.fl_str_mv Marques, Nádia Carolina Teixeira
dc.contributor.referee1.fl_str_mv Moretti, Ana Beatriz Da Silveira
dc.contributor.referee2.fl_str_mv Santos, Gérsika Bitencourt
dc.contributor.advisor1.fl_str_mv Sakai, Vivien Thiemy
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/4769051092383070
contributor_str_mv Marques, Nádia Carolina Teixeira
Moretti, Ana Beatriz Da Silveira
Santos, Gérsika Bitencourt
Sakai, Vivien Thiemy
dc.subject.por.fl_str_mv Células-Tronco
Proliferação de Células
Terapia a Laser de Baixa Intensidade
topic Células-Tronco
Proliferação de Células
Terapia a Laser de Baixa Intensidade
CIENCIAS DA SAUDE::ODONTOLOGIA
dc.subject.cnpq.fl_str_mv CIENCIAS DA SAUDE::ODONTOLOGIA
description Low-level laser irradiation (LLL) therapy may be able to induce proliferation and differentiation into several cellular types, with remarkable effects in the process of tissue repair, inflammation decrease and analgesia. For pulp repair after conservative procedures, pulp stem cells have the potential to differentiate into odontoblasts and to produce dentine, requiring a specific stimulus to trigger this process. The present study aimed to evaluate in vitro whether the LLL delivering fractionated total energy (multiple irradiation) or single irradiation stimulates regeneration-associated events (viability and proliferation) in stem cells from human exfoliated deciduous teeth (SHED). Cells received LLL irradiation (InGaAlP – 660 nm), according to the following experimental groups: GI (single irradiation: 2.5 J/cm², 10 s, 0.10 J), GII (single irradiation: 5.0 J/cm², 20 s, 0.20 J), GIII (single irradiation: 7.5 J/cm², 30 s, 0.30 J), GIV (2 irradiations: 2.5 J/cm², 10 s, 0.20 J), GV (3 irradiations: 2.5 J/cm², 10 s, 0.30 J) and GVI (non-irradiated). Cell viability was assessed by MTT and trypan blue exclusion (TBE) methods, while cell proliferation was evaluated by crystal violet (CV) and sulforhodamine B (SRB) assays after 24, 48 and 72 hours after the first irradiation. By MTT, there was no difference between groups at 24 and 72 hours. At 48 hours, the groups with multiple irradiation (GIV and GV) presented higher cell viability rates. The mean percentages of viable cells for all groups by TBE method were 91.04%, 96.63% and 97.48% at 24, 48 and 72 hours, respectively. By CV, there was no significant difference between groups at 24 and 48 hours; at 72 hours, GII, GIII and GIV presented higher cell proliferation. By SRB, GI and GIV presented lower proliferation rates in all the periods. When the groups presenting the same total energy were compared, GII (0.20 J) presented lower cell viability rates and higher cell proliferation rates in comparison to GIV; GIII (0.30 J) presented similar results to those of GV, with higher cell viability and proliferation. In conclusion, the application of laser delivering fractionated total energy (2 or 3 applications of 2,5 J/cm²) induced higher cell viability at 48 hours, while the 2.5 J/cm² single irradiation decreased cell viability in such period and proliferation over time. The 5.0 and 7.5 J/cm² single doses and the 3 applications of 2.5 J/cm² maintained cell viability and stimulated proliferation of SHED at 72 hours.
publishDate 2019
dc.date.accessioned.fl_str_mv 2019-04-03T14:00:53Z
dc.date.issued.fl_str_mv 2019-01-23
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format masterThesis
status_str publishedVersion
dc.identifier.citation.fl_str_mv ALMEIDA JUNIOR, Luciano Aparecido de. Efeitos da fototerapia com energia fracionada do laser de baixa intensidade em células-tronco da polpa de dentes decíduos esfoliados humanos. 2019. 62 f. Dissertação (Mestrado em Ciências Odontológicas) - Universidade Federal de Alfenas, Alfenas, MG, 2019.
dc.identifier.uri.fl_str_mv https://repositorio.unifal-mg.edu.br/handle/123456789/1335
identifier_str_mv ALMEIDA JUNIOR, Luciano Aparecido de. Efeitos da fototerapia com energia fracionada do laser de baixa intensidade em células-tronco da polpa de dentes decíduos esfoliados humanos. 2019. 62 f. Dissertação (Mestrado em Ciências Odontológicas) - Universidade Federal de Alfenas, Alfenas, MG, 2019.
url https://repositorio.unifal-mg.edu.br/handle/123456789/1335
dc.language.iso.fl_str_mv por
language por
dc.relation.department.fl_str_mv 4673435736271820140
dc.relation.confidence.fl_str_mv 600
600
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dc.relation.cnpq.fl_str_mv -2070498469879244349
dc.relation.sponsorship.fl_str_mv -1527361517405938873
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Alfenas
dc.publisher.program.fl_str_mv Programa de Pós-Graduação em Ciências Odontológicas
dc.publisher.initials.fl_str_mv UNIFAL-MG
dc.publisher.country.fl_str_mv Brasil
dc.publisher.department.fl_str_mv Faculdade de Odontologia
publisher.none.fl_str_mv Universidade Federal de Alfenas
dc.source.none.fl_str_mv reponame:Repositório Institucional da Universidade Federal de Alfenas - RiUnifal
instname:Universidade Federal de Alfenas (UNIFAL)
instacron:UNIFAL
instname_str Universidade Federal de Alfenas (UNIFAL)
instacron_str UNIFAL
institution UNIFAL
reponame_str Repositório Institucional da Universidade Federal de Alfenas - RiUnifal
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repository.name.fl_str_mv Repositório Institucional da Universidade Federal de Alfenas - RiUnifal - Universidade Federal de Alfenas (UNIFAL)
repository.mail.fl_str_mv repositorio@unifal-mg.edu.br
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