Biomateriais xenogênicos ósseos não interferem na viabilidade e proliferação de células-tronco mesenquimais: estudo piloto
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório do Centro Universitário Braz Cubas |
Texto Completo: | https://repositorio.cruzeirodosul.edu.br/handle/123456789/2135 |
Resumo: | The use of cells, especially mesenchymal stem cells (MSCs), is the subject of numerous researches, including tissue engineering. These cells are undifferentiated, presenting capacity for differentiation and self-renewal. They have the potential to transform into multiple cell types of the body, such as adipocytes, chondrocytes and osteoblasts, depending on the stimulus they receive. The main objective of this work was to evaluate, in an in vitro model, the influence of bovine xenogenic biomaterials on mesenchymal stem cells from human exfoliated deciduous teeth, in order to identify a biomaterial with higher potential cell carrier, for subsequent in vivo and/or clinical application. The following groups were used: 1) Control C (control), containing only MSCs; 2) Group BP, containing MSCs and Bonefill Porous® (0.60-1.50 mm); 3) Group BO, containing MSCs and Bios-Oss® (0.25-1 mm). The MSCs (P3) used came from a deciduous tooth in exfoliation from a 7-year-old male donor. A cell aliquot was submitted to immunophenotyping by flow cytometry. Cell viability (neutral red), cytotoxicity (TMT), and cell proliferation (violet crystal) tests were performed; all groups were submitted to morphological analysis by light microscopy (ML). Due to the results of previous tests, the biomaterial considered with superior performance was submitted to ultrastructural evaluation by scanning electron microscopy (SEM). The times of 24, 48 and 72 h of cultivation were used. Mesenchymal stem cell validation demonstrated positive surface markers for CD105, CD73, CD90, and negative for CD45, CD34, CD14, CD19, and HLA-DR. The results showed that both biomaterials maintained cell viability and cytotoxicity similar to group C. As for proliferation, there was a difference for less in the BO group compared to the other groups. To ML, the BP group presented more spread and adhered cells than the BO group. Thus, at SEM, the cells of the BP group presented characteristics of cells more active than those of group C, with secretion of vesicles to the extracellular matrix. Therefore, it can be concluded that BP presented a higher potential stem cell carrier for future studies with in vivo and/or clinical application. |
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Biomateriais xenogênicos ósseos não interferem na viabilidade e proliferação de células-tronco mesenquimais: estudo pilotoOdontologia clínicaCélulas-tronco mesenquimaisMateriais biocompatíveisDente decíduoPolpa dentáriaCNPQ::CIENCIAS DA SAUDE::ODONTOLOGIAThe use of cells, especially mesenchymal stem cells (MSCs), is the subject of numerous researches, including tissue engineering. These cells are undifferentiated, presenting capacity for differentiation and self-renewal. They have the potential to transform into multiple cell types of the body, such as adipocytes, chondrocytes and osteoblasts, depending on the stimulus they receive. The main objective of this work was to evaluate, in an in vitro model, the influence of bovine xenogenic biomaterials on mesenchymal stem cells from human exfoliated deciduous teeth, in order to identify a biomaterial with higher potential cell carrier, for subsequent in vivo and/or clinical application. The following groups were used: 1) Control C (control), containing only MSCs; 2) Group BP, containing MSCs and Bonefill Porous® (0.60-1.50 mm); 3) Group BO, containing MSCs and Bios-Oss® (0.25-1 mm). The MSCs (P3) used came from a deciduous tooth in exfoliation from a 7-year-old male donor. A cell aliquot was submitted to immunophenotyping by flow cytometry. Cell viability (neutral red), cytotoxicity (TMT), and cell proliferation (violet crystal) tests were performed; all groups were submitted to morphological analysis by light microscopy (ML). Due to the results of previous tests, the biomaterial considered with superior performance was submitted to ultrastructural evaluation by scanning electron microscopy (SEM). The times of 24, 48 and 72 h of cultivation were used. Mesenchymal stem cell validation demonstrated positive surface markers for CD105, CD73, CD90, and negative for CD45, CD34, CD14, CD19, and HLA-DR. The results showed that both biomaterials maintained cell viability and cytotoxicity similar to group C. As for proliferation, there was a difference for less in the BO group compared to the other groups. To ML, the BP group presented more spread and adhered cells than the BO group. Thus, at SEM, the cells of the BP group presented characteristics of cells more active than those of group C, with secretion of vesicles to the extracellular matrix. Therefore, it can be concluded that BP presented a higher potential stem cell carrier for future studies with in vivo and/or clinical application.O uso de células, em especial as células-tronco mesenquimais (CTMs), é objeto de inúmeras pesquisas, incluindo a engenharia tecidual. Estas células são indiferenciadas, apresentando capacidade de diferenciação e auto-renovação. Tem o potencial de se transformar em múltiplos tipos celulares do organismo, podem se diferenciar em adipócitos, condrócitos e osteoblastos, dependendo do estímulo que recebem. O objetivo principal do presente trabalho foi avaliar em modelo in vitro a influência de biomateriais xenogênicos bovinos sobre células-tronco mesenquimais da polpa de dente decíduo, no intuito de identificar um biomaterial com maior potencial carreador de células, para posterior aplicação in vivo e/ou clínica. Os seguintes grupos foram usados: 1) Grupo C (controle), contendo apenas CTMs; 2) Grupo BP, contendo CTMs e Bonefill Porous® (0,60-1,50 mm); 3) Grupo BO, contendo CTMs e Bio-Oss® (0,25-1 mm). As CTMs (P3) utilizadas foram provenientes de um dente decíduo em exfoliação, de um doador do sexo masculino de 7 anos de idade. Uma alíquota de células foi submetida à imunofenotipagem por citometria de fluxo. Foram realizados os ensaios de viabilidade celular (vermelho neutro), citotoxicidade (MTT), e proliferação celular (cristal violeta); todos os grupos foram submetidos a uma análise morfológica por microscopia de luz (ML). Devido aos resultados dos testes anteriores, o biomaterial considerado com desempenho superior foi submetido à avaliação ultraestrutural por microscopia eletrônica de varredura (MEV). Foram utilizados os tempos de 24, 48 e 72 h de cultivo. A validação das células-tronco mesenquimais demonstrou marcadores de superficie positivos para CD105, CD73, CD90, e negativos para CD45, CD34, CD14, CD19, e HLADR. Os resultados evidenciaram que ambos os biomateriais mantiveram viabilidade celular e citotoxicidade semelhantes ao grupo C. Quanto à proliferação, houve diferença para menor no grupo BO em relação aos outros grupos. À ML, o grupo BP apresentou células mais espraiadas e aderidas do que o grupo BO. Assim, ao MEV, as células do grupo BP apresentaram características de células mais ativas do que as do grupo C, com secreção de vesículas para a matrix extracelular. Logo, pode-se concluir que o BP apresentou maior potencial carreador de células-tronco para futuros trabalhos com aplicação in vivo e/ou clínica.Universidade PositivoBrasilPós-GraduaçãoPrograma de Pós-Graduação em Odontologia ClínicaUPZielak, João Césarhttp://lattes.cnpq.br/5991949730989569Leão, Moira Pedrosohttp://lattes.cnpq.br/5243777898770664Stroparo, Jeferson Luis de Oliveira2021-05-11T22:27:58Z20202021-05-11T22:27:58Z2020info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttps://repositorio.cruzeirodosul.edu.br/handle/123456789/2135porinfo:eu-repo/semantics/openAccessreponame:Repositório do Centro Universitário Braz Cubasinstname:Centro Universitário Braz Cubas (CUB)instacron:CUB2021-09-09T19:50:41Zoai:repositorio.cruzeirodosul.edu.br:123456789/2135Repositório InstitucionalPUBhttps://repositorio.brazcubas.edu.br/oai/requestbibli@brazcubas.edu.bropendoar:2021-09-09T19:50:41Repositório do Centro Universitário Braz Cubas - Centro Universitário Braz Cubas (CUB)false |
dc.title.none.fl_str_mv |
Biomateriais xenogênicos ósseos não interferem na viabilidade e proliferação de células-tronco mesenquimais: estudo piloto |
title |
Biomateriais xenogênicos ósseos não interferem na viabilidade e proliferação de células-tronco mesenquimais: estudo piloto |
spellingShingle |
Biomateriais xenogênicos ósseos não interferem na viabilidade e proliferação de células-tronco mesenquimais: estudo piloto Stroparo, Jeferson Luis de Oliveira Odontologia clínica Células-tronco mesenquimais Materiais biocompatíveis Dente decíduo Polpa dentária CNPQ::CIENCIAS DA SAUDE::ODONTOLOGIA |
title_short |
Biomateriais xenogênicos ósseos não interferem na viabilidade e proliferação de células-tronco mesenquimais: estudo piloto |
title_full |
Biomateriais xenogênicos ósseos não interferem na viabilidade e proliferação de células-tronco mesenquimais: estudo piloto |
title_fullStr |
Biomateriais xenogênicos ósseos não interferem na viabilidade e proliferação de células-tronco mesenquimais: estudo piloto |
title_full_unstemmed |
Biomateriais xenogênicos ósseos não interferem na viabilidade e proliferação de células-tronco mesenquimais: estudo piloto |
title_sort |
Biomateriais xenogênicos ósseos não interferem na viabilidade e proliferação de células-tronco mesenquimais: estudo piloto |
author |
Stroparo, Jeferson Luis de Oliveira |
author_facet |
Stroparo, Jeferson Luis de Oliveira |
author_role |
author |
dc.contributor.none.fl_str_mv |
Zielak, João César http://lattes.cnpq.br/5991949730989569 Leão, Moira Pedroso http://lattes.cnpq.br/5243777898770664 |
dc.contributor.author.fl_str_mv |
Stroparo, Jeferson Luis de Oliveira |
dc.subject.por.fl_str_mv |
Odontologia clínica Células-tronco mesenquimais Materiais biocompatíveis Dente decíduo Polpa dentária CNPQ::CIENCIAS DA SAUDE::ODONTOLOGIA |
topic |
Odontologia clínica Células-tronco mesenquimais Materiais biocompatíveis Dente decíduo Polpa dentária CNPQ::CIENCIAS DA SAUDE::ODONTOLOGIA |
description |
The use of cells, especially mesenchymal stem cells (MSCs), is the subject of numerous researches, including tissue engineering. These cells are undifferentiated, presenting capacity for differentiation and self-renewal. They have the potential to transform into multiple cell types of the body, such as adipocytes, chondrocytes and osteoblasts, depending on the stimulus they receive. The main objective of this work was to evaluate, in an in vitro model, the influence of bovine xenogenic biomaterials on mesenchymal stem cells from human exfoliated deciduous teeth, in order to identify a biomaterial with higher potential cell carrier, for subsequent in vivo and/or clinical application. The following groups were used: 1) Control C (control), containing only MSCs; 2) Group BP, containing MSCs and Bonefill Porous® (0.60-1.50 mm); 3) Group BO, containing MSCs and Bios-Oss® (0.25-1 mm). The MSCs (P3) used came from a deciduous tooth in exfoliation from a 7-year-old male donor. A cell aliquot was submitted to immunophenotyping by flow cytometry. Cell viability (neutral red), cytotoxicity (TMT), and cell proliferation (violet crystal) tests were performed; all groups were submitted to morphological analysis by light microscopy (ML). Due to the results of previous tests, the biomaterial considered with superior performance was submitted to ultrastructural evaluation by scanning electron microscopy (SEM). The times of 24, 48 and 72 h of cultivation were used. Mesenchymal stem cell validation demonstrated positive surface markers for CD105, CD73, CD90, and negative for CD45, CD34, CD14, CD19, and HLA-DR. The results showed that both biomaterials maintained cell viability and cytotoxicity similar to group C. As for proliferation, there was a difference for less in the BO group compared to the other groups. To ML, the BP group presented more spread and adhered cells than the BO group. Thus, at SEM, the cells of the BP group presented characteristics of cells more active than those of group C, with secretion of vesicles to the extracellular matrix. Therefore, it can be concluded that BP presented a higher potential stem cell carrier for future studies with in vivo and/or clinical application. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020 2020 2021-05-11T22:27:58Z 2021-05-11T22:27:58Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://repositorio.cruzeirodosul.edu.br/handle/123456789/2135 |
url |
https://repositorio.cruzeirodosul.edu.br/handle/123456789/2135 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Positivo Brasil Pós-Graduação Programa de Pós-Graduação em Odontologia Clínica UP |
publisher.none.fl_str_mv |
Universidade Positivo Brasil Pós-Graduação Programa de Pós-Graduação em Odontologia Clínica UP |
dc.source.none.fl_str_mv |
reponame:Repositório do Centro Universitário Braz Cubas instname:Centro Universitário Braz Cubas (CUB) instacron:CUB |
instname_str |
Centro Universitário Braz Cubas (CUB) |
instacron_str |
CUB |
institution |
CUB |
reponame_str |
Repositório do Centro Universitário Braz Cubas |
collection |
Repositório do Centro Universitário Braz Cubas |
repository.name.fl_str_mv |
Repositório do Centro Universitário Braz Cubas - Centro Universitário Braz Cubas (CUB) |
repository.mail.fl_str_mv |
bibli@brazcubas.edu.br |
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1798311340814630912 |