Avaliação histológica e imunofluorescente da lesão compressiva na medula espinhal de ratos wistar tratados com células-tronco mesenquimais
Autor(a) principal: | |
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Data de Publicação: | 2013 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | LOCUS Repositório Institucional da UFV |
Texto Completo: | http://locus.ufv.br/handle/123456789/2358 |
Resumo: | The traumatic spinal cord injury often results in severe neurological deficits, and mostly irreversible. The cell therapy represents a new strategy for the treatment of these lesions, particularly with the use of stem cells (SC), with satisfactory results in several experimental models. This study compared the spinal cord injury in animals treated and not treated with mesenchymal stem cells (MSC) from bone marrow. MSCs were obtained from bone marrow of rats, cultured, characterized, labeled with Qtracker 655® and transplanted in the sixth passage in animals with spinal cord injury.Spinal cord injury (SCI) was performed with 2-Fr Fogarty catheter after T9-T10 laminectomy. The animals were randomly divided into six experimental groups of 5 animals each and submitted to different treatments: control group (CG), with only spinal cord injury and treated groups (GT) that suffered the SCI and received transplants of cells 3 hours after injury. The animals were euthanized 24 hours, 7 and 21 days postinjury. For light microscopy, was measured the percentage of lesion area in the fragment of macroscopic lesion. We also investigated, by fluorescence microscopy, the effect of MSC on inflammation, astrocytic reactivity and activation of endogenous stem cells. There was no statistical difference between GC and MSC as the size of the lesion area. Although MSC were not found at the site of the lesion, in the evaluation immunofluorescent was seen in the animals treated with MSC was significant attenuation of the inflammatory response with a reduction in the number of macrophages labeled with anti-CD68 antibody (ED1), decreased immunoreactivity of astrocytes (GFAP+) and greater activation of endogenous stem cells (nestin+). Our results indicate that cell transplantation has a beneficial effect on recovery from traumatic spinal cord injury. As MSC transplanted nanolabeling were not observed at the lesion site, we suggest that the therapeutic effect of these cells can occur by some telecrine mechanism. |
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Araújo, Marta Rochahttp://lattes.cnpq.br/7226988137025989Neves, Clóvis Andradehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4785611E1Monteiro, Betânia Souzahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4771761J3Carlo, Ricardo Junqueira Delhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4788143A92015-03-26T13:04:17Z2013-07-182015-03-26T13:04:17Z2013-02-28ARAÚJO, Marta Rocha. Histological and immunofluorescent evaluation of compressive lesion in the spinal cord of Wistar rats treated with mesenchymal stem cells. 2013. 47 f. Dissertação (Mestrado em Análises quantitativas e moleculares do Genoma; Biologia das células e dos tecidos) - Universidade Federal de Viçosa, Viçosa, 2013.http://locus.ufv.br/handle/123456789/2358The traumatic spinal cord injury often results in severe neurological deficits, and mostly irreversible. The cell therapy represents a new strategy for the treatment of these lesions, particularly with the use of stem cells (SC), with satisfactory results in several experimental models. This study compared the spinal cord injury in animals treated and not treated with mesenchymal stem cells (MSC) from bone marrow. MSCs were obtained from bone marrow of rats, cultured, characterized, labeled with Qtracker 655® and transplanted in the sixth passage in animals with spinal cord injury.Spinal cord injury (SCI) was performed with 2-Fr Fogarty catheter after T9-T10 laminectomy. The animals were randomly divided into six experimental groups of 5 animals each and submitted to different treatments: control group (CG), with only spinal cord injury and treated groups (GT) that suffered the SCI and received transplants of cells 3 hours after injury. The animals were euthanized 24 hours, 7 and 21 days postinjury. For light microscopy, was measured the percentage of lesion area in the fragment of macroscopic lesion. We also investigated, by fluorescence microscopy, the effect of MSC on inflammation, astrocytic reactivity and activation of endogenous stem cells. There was no statistical difference between GC and MSC as the size of the lesion area. Although MSC were not found at the site of the lesion, in the evaluation immunofluorescent was seen in the animals treated with MSC was significant attenuation of the inflammatory response with a reduction in the number of macrophages labeled with anti-CD68 antibody (ED1), decreased immunoreactivity of astrocytes (GFAP+) and greater activation of endogenous stem cells (nestin+). Our results indicate that cell transplantation has a beneficial effect on recovery from traumatic spinal cord injury. As MSC transplanted nanolabeling were not observed at the lesion site, we suggest that the therapeutic effect of these cells can occur by some telecrine mechanism.A lesão traumática da medula espinhal geralmente resulta em déficits neurológicos graves e, na maioria, irreversíveis. As terapias celulares representam uma nova estratégia para o tratamento destas lesões, sobretudo, com a utilização de células-tronco (CT); que experimentalmente vem exibindo resultados satisfatórios em vários modelos animais. O presente trabalho comparou a lesão da medula espinhal em animais tratados e não-tratados com células-tronco mesenquimais (CTM) da medula óssea. As CTM foram obtidas da medula óssea de ratos Wistar, cultivadas, caracterizadas, nanomarcadas com Qtracker 655® e transplantadas na sexta passagem para animais com lesão medular. A lesão espinhal compressiva foi realizada com insuflação de 50μL, de solução salina, do cuff do cateter de Forgat n.º 2 Fr. no espaço epidural T8, durante 5 minutos. Os animais foram distribuídos aleatoriamente em dois grupos experimentais com 15animais em cada e submetidos a tratamentos distintos: grupos controles (GC), que sofreram somente a lesão medular e grupos tratados (GT), que sofreram a lesão medular e receberam o transplante de células na concentração de 1,0 x 107células/mL de tampão fosfato salino(PBS), 3 horas após a lesão. Realizou-se a eutanásia dos animais com 24 horas, 7 e 21 dias pós-lesão. Por microscopia de luz, foi mensurado o percentual de área lesionada no fragmento da lesão macroscópica. Também foi investigado, por microscopia de fluorescência, o efeito das CTM sobre a inflamação, reatividade astrocítica e ativação de células-tronco endógenas. Embora as CTM não tenham sido encontradas no local da lesão, na avaliação imunofluorescente foi visto que nos animais tratados com CTM houve atenuação significativa da resposta inflamatória com redução no número de macrófagos/micróglia marcados com anti-CD68 (ED1); diminuição da imunorreatividade dos astrócitos (GFAP+) e maior ativação de célulasix tronco endógenas (nestin+). Nossos resultados indicam que o transplante de CTM tem ação benéfica sobre a recuperação da lesão traumática da medula espinhal. Como as CTM transplantadas nanomarcadas não foram observadas no local da lesão, sugerimos que o efeito terapêutico dessas células pode ocorrer por algum tipo de mecanismo telécrino.Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorapplication/pdfporUniversidade Federal de ViçosaMestrado em Biologia Celular e EstruturalUFVBRAnálises quantitativas e moleculares do Genoma; Biologia das células e dos tecidosLesão medularTerapia celularCélula-tronco mesenquimalSpinal cord injuryCell therapyMesenchymal stem cellCNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERALAvaliação histológica e imunofluorescente da lesão compressiva na medula espinhal de ratos wistar tratados com células-tronco mesenquimaisHistological and immunofluorescent evaluation of compressive lesion in the spinal cord of Wistar rats treated with mesenchymal stem cellsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf1254312https://locus.ufv.br//bitstream/123456789/2358/1/texto%20completo.pdfe5b413aa6bd065b30e242d2961064291MD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain78295https://locus.ufv.br//bitstream/123456789/2358/2/texto%20completo.pdf.txtd12669a376d26d902d43e557ec698d8fMD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3722https://locus.ufv.br//bitstream/123456789/2358/3/texto%20completo.pdf.jpg9244d87bea43f989748d4e27d3ad55afMD53123456789/23582016-04-08 23:05:37.643oai:locus.ufv.br:123456789/2358Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-09T02:05:37LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false |
dc.title.por.fl_str_mv |
Avaliação histológica e imunofluorescente da lesão compressiva na medula espinhal de ratos wistar tratados com células-tronco mesenquimais |
dc.title.alternative.eng.fl_str_mv |
Histological and immunofluorescent evaluation of compressive lesion in the spinal cord of Wistar rats treated with mesenchymal stem cells |
title |
Avaliação histológica e imunofluorescente da lesão compressiva na medula espinhal de ratos wistar tratados com células-tronco mesenquimais |
spellingShingle |
Avaliação histológica e imunofluorescente da lesão compressiva na medula espinhal de ratos wistar tratados com células-tronco mesenquimais Araújo, Marta Rocha Lesão medular Terapia celular Célula-tronco mesenquimal Spinal cord injury Cell therapy Mesenchymal stem cell CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL |
title_short |
Avaliação histológica e imunofluorescente da lesão compressiva na medula espinhal de ratos wistar tratados com células-tronco mesenquimais |
title_full |
Avaliação histológica e imunofluorescente da lesão compressiva na medula espinhal de ratos wistar tratados com células-tronco mesenquimais |
title_fullStr |
Avaliação histológica e imunofluorescente da lesão compressiva na medula espinhal de ratos wistar tratados com células-tronco mesenquimais |
title_full_unstemmed |
Avaliação histológica e imunofluorescente da lesão compressiva na medula espinhal de ratos wistar tratados com células-tronco mesenquimais |
title_sort |
Avaliação histológica e imunofluorescente da lesão compressiva na medula espinhal de ratos wistar tratados com células-tronco mesenquimais |
author |
Araújo, Marta Rocha |
author_facet |
Araújo, Marta Rocha |
author_role |
author |
dc.contributor.authorLattes.por.fl_str_mv |
http://lattes.cnpq.br/7226988137025989 |
dc.contributor.author.fl_str_mv |
Araújo, Marta Rocha |
dc.contributor.advisor1.fl_str_mv |
Neves, Clóvis Andrade |
dc.contributor.advisor1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4785611E1 |
dc.contributor.referee1.fl_str_mv |
Monteiro, Betânia Souza |
dc.contributor.referee1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4771761J3 |
dc.contributor.referee2.fl_str_mv |
Carlo, Ricardo Junqueira Del |
dc.contributor.referee2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4788143A9 |
contributor_str_mv |
Neves, Clóvis Andrade Monteiro, Betânia Souza Carlo, Ricardo Junqueira Del |
dc.subject.por.fl_str_mv |
Lesão medular Terapia celular Célula-tronco mesenquimal |
topic |
Lesão medular Terapia celular Célula-tronco mesenquimal Spinal cord injury Cell therapy Mesenchymal stem cell CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL |
dc.subject.eng.fl_str_mv |
Spinal cord injury Cell therapy Mesenchymal stem cell |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL |
description |
The traumatic spinal cord injury often results in severe neurological deficits, and mostly irreversible. The cell therapy represents a new strategy for the treatment of these lesions, particularly with the use of stem cells (SC), with satisfactory results in several experimental models. This study compared the spinal cord injury in animals treated and not treated with mesenchymal stem cells (MSC) from bone marrow. MSCs were obtained from bone marrow of rats, cultured, characterized, labeled with Qtracker 655® and transplanted in the sixth passage in animals with spinal cord injury.Spinal cord injury (SCI) was performed with 2-Fr Fogarty catheter after T9-T10 laminectomy. The animals were randomly divided into six experimental groups of 5 animals each and submitted to different treatments: control group (CG), with only spinal cord injury and treated groups (GT) that suffered the SCI and received transplants of cells 3 hours after injury. The animals were euthanized 24 hours, 7 and 21 days postinjury. For light microscopy, was measured the percentage of lesion area in the fragment of macroscopic lesion. We also investigated, by fluorescence microscopy, the effect of MSC on inflammation, astrocytic reactivity and activation of endogenous stem cells. There was no statistical difference between GC and MSC as the size of the lesion area. Although MSC were not found at the site of the lesion, in the evaluation immunofluorescent was seen in the animals treated with MSC was significant attenuation of the inflammatory response with a reduction in the number of macrophages labeled with anti-CD68 antibody (ED1), decreased immunoreactivity of astrocytes (GFAP+) and greater activation of endogenous stem cells (nestin+). Our results indicate that cell transplantation has a beneficial effect on recovery from traumatic spinal cord injury. As MSC transplanted nanolabeling were not observed at the lesion site, we suggest that the therapeutic effect of these cells can occur by some telecrine mechanism. |
publishDate |
2013 |
dc.date.available.fl_str_mv |
2013-07-18 2015-03-26T13:04:17Z |
dc.date.issued.fl_str_mv |
2013-02-28 |
dc.date.accessioned.fl_str_mv |
2015-03-26T13:04:17Z |
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.citation.fl_str_mv |
ARAÚJO, Marta Rocha. Histological and immunofluorescent evaluation of compressive lesion in the spinal cord of Wistar rats treated with mesenchymal stem cells. 2013. 47 f. Dissertação (Mestrado em Análises quantitativas e moleculares do Genoma; Biologia das células e dos tecidos) - Universidade Federal de Viçosa, Viçosa, 2013. |
dc.identifier.uri.fl_str_mv |
http://locus.ufv.br/handle/123456789/2358 |
identifier_str_mv |
ARAÚJO, Marta Rocha. Histological and immunofluorescent evaluation of compressive lesion in the spinal cord of Wistar rats treated with mesenchymal stem cells. 2013. 47 f. Dissertação (Mestrado em Análises quantitativas e moleculares do Genoma; Biologia das células e dos tecidos) - Universidade Federal de Viçosa, Viçosa, 2013. |
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http://locus.ufv.br/handle/123456789/2358 |
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por |
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Universidade Federal de Viçosa |
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Mestrado em Biologia Celular e Estrutural |
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UFV |
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BR |
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Análises quantitativas e moleculares do Genoma; Biologia das células e dos tecidos |
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Universidade Federal de Viçosa |
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