Biomateriais xenogênicos ósseos não interferem na viabilidade e proliferação de células-tronco mesenquimais: estudo piloto

Detalhes bibliográficos
Autor(a) principal: Stroparo, Jeferson Luis de Oliveira
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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spelling 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
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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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