Cultivo de células da medula óssea humana sobre membranas de colágeno bovino e arcabouços de ácido poliglicóico polilático (PLGA)
Ano de defesa: | 2007 |
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Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | |
Tipo de documento: | Dissertação |
Tipo de acesso: | Acesso aberto |
Idioma: | por |
Instituição de defesa: |
Pontifícia Universidade Católica do Rio Grande do Sul
Porto Alegre |
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: | http://hdl.handle.net/10923/366 |
Resumo: | The bone marrow besides having a high number of heterogenic cells, like connective, hematopoietics, and neuronal cells, also presents a great number of stem cells. These cells are pluripotent, so can differentiate in any connective cell when a stimulus is given. The aim of this study was to evaluate bone marrow stem cells adhesion, proliferation, and differentiation to osteoblast in vitro, over a collagen membrane and a poli-lactic (glycolic) acid scaffold (PLGA), using either recombinant human bone morphogenetic protein (rhBMP-4) and platelet rich plasma (PRP). Bone marrow cells were aspirated from iliac crest and purify using specifics techniques. These cells were separated in main big groups cultured over collagen membrane and PLGA scaffold. Each culture had 4 distinct treatments: (1) with rhBMP-4; (2) with rhBMP-4 and PRP; (3) with PRP; and (4) control (only standard medium). The groups were evaluated after 6, 9 14 and 21 days. Microscopic analysis of collagen membrane and PLGA scaffold were performed using either Propidium Iodide, and Eosin and Hematoxilin. Real time PCR was performed to identify the expression level of osteopontin and osteocalcin, and standard PCR to osteopontin. The results showed cellular adhesion in both biomaterials, and suggest a higher cell proliferation and differentiation in rhBMP-4 and PRP, and PRP groups. The groups with rhBMP-4 had more differentiate cells that control group. The data obtained here indicate that PRP and/ or rhBMP-4 induce adhesion, proliferation and differentiation of bone marrow stem cells into osteogenic lineage. |
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Fritscher, Guilherme GenehrMachado, Denise Cantarelli2011-12-27T14:14:01Z2011-12-27T14:14:01Z2007http://hdl.handle.net/10923/366The bone marrow besides having a high number of heterogenic cells, like connective, hematopoietics, and neuronal cells, also presents a great number of stem cells. These cells are pluripotent, so can differentiate in any connective cell when a stimulus is given. The aim of this study was to evaluate bone marrow stem cells adhesion, proliferation, and differentiation to osteoblast in vitro, over a collagen membrane and a poli-lactic (glycolic) acid scaffold (PLGA), using either recombinant human bone morphogenetic protein (rhBMP-4) and platelet rich plasma (PRP). Bone marrow cells were aspirated from iliac crest and purify using specifics techniques. These cells were separated in main big groups cultured over collagen membrane and PLGA scaffold. Each culture had 4 distinct treatments: (1) with rhBMP-4; (2) with rhBMP-4 and PRP; (3) with PRP; and (4) control (only standard medium). The groups were evaluated after 6, 9 14 and 21 days. Microscopic analysis of collagen membrane and PLGA scaffold were performed using either Propidium Iodide, and Eosin and Hematoxilin. Real time PCR was performed to identify the expression level of osteopontin and osteocalcin, and standard PCR to osteopontin. The results showed cellular adhesion in both biomaterials, and suggest a higher cell proliferation and differentiation in rhBMP-4 and PRP, and PRP groups. The groups with rhBMP-4 had more differentiate cells that control group. The data obtained here indicate that PRP and/ or rhBMP-4 induce adhesion, proliferation and differentiation of bone marrow stem cells into osteogenic lineage.O estroma da medula óssea, além de conter uma população heterogênea de células conjuntivas, nervosas e hematopoiéticas, mantém uma quantidade de células com características de células tronco (células mesenquimais indiferenciadas). Estas possuem capacidade de se diferenciarem em qualquer célula de natureza conjuntiva quando estimuladas pelo organismo. Esta pesquisa teve como objetivo avaliar a adesão, a proliferação e a diferenciação de células mesenquimais da medula óssea cultivadas sobre membrana de colágeno bovino e sobre arcabouço de ácido poliglicólico polilático (PLGA) com a adição do gel de plasma rico em plaquetas (PRP) e/ ou da proteína morfogenética óssea recombinante humana (rhBMP-4). As células foram coletadas da medula óssea da crista do osso ilíaco humana e separadas por técnica específica. As células foram cultivadas em dois suportes: membrana de colágeno e arcabouço de PLGA, e receberam 4 tratamentos distintos: (1) com rhBMP-4, (2) com rhBMP-4 e PRP, (3) com PRP e (4) controle (somente meio de cultura). Os subgrupos foram avaliados em 6, 9, 14 e 21 dias. Foram realizadas análises microscópicas das membranas e arcabouços com as colorações de Iodeto de Propídio e de Hematoxilina e Eosina. Foi feito PCR em tempo real a fim de se avaliar a expressão de osteopontina e osteocalcina e PCR convencional para osteopontina. Os resultados mostraram adesão celular tanto na membrana de colágeno como no arcabouço de PLGA. Sugere-se haver uma maior proliferação e diferenciação celular nos grupos que foram adicionados PRP, e rhBMP-4 e PRP juntos. Nos grupos com rhBMP-4 parece haver uma maior diferenciação celular que o grupo controle. Os dados obtidos indicam que o PRP e/ ou a presença de rhBMP-4 induzem adesão, proliferação e diferenciação de células da medula óssea em células de linhagem osteogênica.Made available in DSpace on 2011-12-27T14:14:01Z (GMT). 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dc.title.pt_BR.fl_str_mv |
Cultivo de células da medula óssea humana sobre membranas de colágeno bovino e arcabouços de ácido poliglicóico polilático (PLGA) |
title |
Cultivo de células da medula óssea humana sobre membranas de colágeno bovino e arcabouços de ácido poliglicóico polilático (PLGA) |
spellingShingle |
Cultivo de células da medula óssea humana sobre membranas de colágeno bovino e arcabouços de ácido poliglicóico polilático (PLGA) Fritscher, Guilherme Genehr CIRURGIA BUCOMAXILOFACIAL MEDULA ÓSSEA TECIDO ÓSSEO - REGENERAÇÃO CULTURA DE CÉLULAS |
title_short |
Cultivo de células da medula óssea humana sobre membranas de colágeno bovino e arcabouços de ácido poliglicóico polilático (PLGA) |
title_full |
Cultivo de células da medula óssea humana sobre membranas de colágeno bovino e arcabouços de ácido poliglicóico polilático (PLGA) |
title_fullStr |
Cultivo de células da medula óssea humana sobre membranas de colágeno bovino e arcabouços de ácido poliglicóico polilático (PLGA) |
title_full_unstemmed |
Cultivo de células da medula óssea humana sobre membranas de colágeno bovino e arcabouços de ácido poliglicóico polilático (PLGA) |
title_sort |
Cultivo de células da medula óssea humana sobre membranas de colágeno bovino e arcabouços de ácido poliglicóico polilático (PLGA) |
author |
Fritscher, Guilherme Genehr |
author_facet |
Fritscher, Guilherme Genehr |
author_role |
author |
dc.contributor.author.fl_str_mv |
Fritscher, Guilherme Genehr |
dc.contributor.advisor1.fl_str_mv |
Machado, Denise Cantarelli |
contributor_str_mv |
Machado, Denise Cantarelli |
dc.subject.por.fl_str_mv |
CIRURGIA BUCOMAXILOFACIAL MEDULA ÓSSEA TECIDO ÓSSEO - REGENERAÇÃO CULTURA DE CÉLULAS |
topic |
CIRURGIA BUCOMAXILOFACIAL MEDULA ÓSSEA TECIDO ÓSSEO - REGENERAÇÃO CULTURA DE CÉLULAS |
description |
The bone marrow besides having a high number of heterogenic cells, like connective, hematopoietics, and neuronal cells, also presents a great number of stem cells. These cells are pluripotent, so can differentiate in any connective cell when a stimulus is given. The aim of this study was to evaluate bone marrow stem cells adhesion, proliferation, and differentiation to osteoblast in vitro, over a collagen membrane and a poli-lactic (glycolic) acid scaffold (PLGA), using either recombinant human bone morphogenetic protein (rhBMP-4) and platelet rich plasma (PRP). Bone marrow cells were aspirated from iliac crest and purify using specifics techniques. These cells were separated in main big groups cultured over collagen membrane and PLGA scaffold. Each culture had 4 distinct treatments: (1) with rhBMP-4; (2) with rhBMP-4 and PRP; (3) with PRP; and (4) control (only standard medium). The groups were evaluated after 6, 9 14 and 21 days. Microscopic analysis of collagen membrane and PLGA scaffold were performed using either Propidium Iodide, and Eosin and Hematoxilin. Real time PCR was performed to identify the expression level of osteopontin and osteocalcin, and standard PCR to osteopontin. The results showed cellular adhesion in both biomaterials, and suggest a higher cell proliferation and differentiation in rhBMP-4 and PRP, and PRP groups. The groups with rhBMP-4 had more differentiate cells that control group. The data obtained here indicate that PRP and/ or rhBMP-4 induce adhesion, proliferation and differentiation of bone marrow stem cells into osteogenic lineage. |
publishDate |
2007 |
dc.date.issued.fl_str_mv |
2007 |
dc.date.accessioned.fl_str_mv |
2011-12-27T14:14:01Z |
dc.date.available.fl_str_mv |
2011-12-27T14:14:01Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/masterThesis |
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masterThesis |
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publishedVersion |
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http://hdl.handle.net/10923/366 |
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http://hdl.handle.net/10923/366 |
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por |
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openAccess |
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Pontifícia Universidade Católica do Rio Grande do Sul Porto Alegre |
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Pontifícia Universidade Católica do Rio Grande do Sul Porto Alegre |
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