Efeitos da fototerapia com energia fracionada do laser de baixa intensidade em células-tronco da polpa de dentes decíduos esfoliados humanos
| Ano de defesa: | 2019 |
|---|---|
| Autor(a) principal: | |
| Orientador(a): | |
| Banca de defesa: | , |
| 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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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 |
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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 |
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4673435736271820140 |
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600 600 600 |
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info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-nd/4.0/ |
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openAccess |
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http://creativecommons.org/licenses/by-nc-nd/4.0/ |
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application/pdf |
| dc.publisher.none.fl_str_mv |
Universidade Federal de Alfenas |
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Programa de Pós-Graduação em Ciências Odontológicas |
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UNIFAL-MG |
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Brasil |
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Faculdade de Odontologia |
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Universidade Federal de Alfenas |
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Universidade Federal de Alfenas (UNIFAL) |
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UNIFAL |
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UNIFAL |
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Repositório Institucional da Universidade Federal de Alfenas - RiUnifal |
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Repositório Institucional da Universidade Federal de Alfenas - RiUnifal - Universidade Federal de Alfenas (UNIFAL) |
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repositorio@unifal-mg.edu.br |
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