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Master's Dissertation
DOI
https://doi.org/10.11606/D.47.2008.tde-15012009-160608
Document
Author
Full name
Beatriz de Freitas Azevedo Levy
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2008
Supervisor
Committee
Chadi, Gerson (President)
Cristante, Alexandre Fogaça
Taricco, Mario Augusto
Title in Portuguese
Efeitos das células de schwann cultivadas e tratadas com o pedf no trofismo de neurônios medulares cultivados e na recuperação motora de ratos submetidos ao trauma contuso da medula espinal pelo impactor
Keywords in Portuguese
Células de Schwann
Fator derivado do epitélio pigmentado
Lesão medular
Neurônios
Neuroplasticidade
Trofismo
Abstract in Portuguese
O presente estudo objetivou a avaliação do potencial regenerativo e trófico das CS e do fator neurotrófico PEDF sobre os neurônios da medula espinal através da cultura destas células in vitro assim como no modelo experimental da lesão medular contusa in vivo. CS foram cultivadas a partir de fragmentos de nervo ciático de ratos adultos jovens. As culturas primárias foram purificadas e utilizadas em experimentos in vitro e in vivo. Para avaliar se a cultura de CS tratadas com o PEDF proporciona o trofismo dos neurônios da medula espinal, estes foram cultivados a partir de fragmentos da medula espinal de ratos Wistar neonatos e tratados com o meio condicionado das CS (MCCS) pré-tratadas ou não com formas específicas do PEDF. A resposta trófica foi estudada através da quantificação do número de células bem como da área dos prolongamentos e dos corpos celulares dos neurônios cultivados sobreviventes no decurso temporal da cultura, utilizando-se método estereológico específico. Em outra série de experimentos, Ratos Wistar adultos foram submetidos à lesão medular contusa no 10º-11° segmento torácico pelo sistema NYU Impactor e tratados com inoculações locais de CS e do seu meio condicionado na presença ou não do PEDF. Ao término do procedimento cirúrgico, os ratos foram submetidos aos testes do comportamento BBB, CBS durante 6 semanas. Os tratamentos aplicados à cultura de neurônios da medula espinal demonstraram que o MCCS, pré-tratado ou não com diferentes formas moleculares do PEDF e os fatores por si só possuem ações tróficas e plásticas em relação aos neurônios da medula espinal. Foi observado que o MCCS exerceu um efeito de arborização e de trofismo sobre o corpo neuronal dos neurônios medulares, sendo que este efeito foi potencializado quando o MCCS foi obtido a partir de CS pré-tratadas com os fatores nPEDF ou coPEDF, e de intensidade ainda maior quando as formas puras dos fatores em questão foram adicionadas à cultura. Os animais submetidos ao trauma contuso da medula espinal que foram tratados com a inoculação das CS, pré-tratadas ou não com o nPEDF, do meio condicionado destas células e dos fatores nPEDF e coPEDF apresentaram melhor recuperação funcional em relação ao grupo controle. A partir dos dados obtidos neste trabalho pode-se dizer que as CS, seu meio condicionado e o PEDF possuem ações importantes sobre os neurônios da medula espinal, possibilitando a modulação do processo de recuperação funcional deste órgão após o trauma.
Title in English
Effect of the cells of Schwann cultivated and treated with the PEDF in the trophism to spinal cord neurons cultivated and in the motor recovery of rats submitted to the trauma contused of the spinal cords for the Impactor
Keywords in English
Neuronal plasticity
Neurons
Pigment epitelium-derived factor
Schwann cells
Spinal cord (wounds and injuries)
Trophism
Abstract in English
The present study aimed the assessment of the trophic and regenerative potential of the Schwann cells (SC) and of the neurotrophic factor PEDF on the spinal cord neurons by means of cultures of those cells in vitro as well as in an experimental model of a contused spinal cord lesion in vivo. SC were cultivated from sciatic nerves fragments of young adult rats. The primary cultures were purified and used in in vitro and in vivo experiments. To evaluate if the SC cultures treated with PEDF provide the trophism of the spinal cord neurons, these were cultivated from spinal cords fragments of new born rats and treated with SC conditional medium (SCCM), of cells pre-treated or not with specifics isoforms of PEDF. The trophic responses were analyzed by the quantification of the cell number, and the area of sprounting and perycaria of the survived neuronal cells in the culture time course using a specific stereological method. In another series of experiment, adults Wistar rats were submitted to a contuse spinal cord lesion in the 10º-11º thoracic level by means of the NYU Impactor and treated with local injection of SC and your CM with the presence or not of the PEDF. In the end of the surgical procedure, the rats were submitted to behaviors testes as the BBB and the CBS during the 6 follow weeks .The spinal cord neuron treatments show that the SCCM, prétreated or not with specifics molecular isoforms of PEDF and this factors by themselves demonstrated trophic and plastic actions with the spinal cord neurons. It was observed that the SCCM exerted a sprouting and trophic effect on the neuronal body of the spinal cord neurons, and that this effect was enhanced when the SCCM was achieved from SC pre-treated with PEDF, and even better when the pures forms of the PEDF was added to the culture. The animals that received the spinal cord lesion and were treated with injection of SC, pre-treated or not with nPEDF, or with the conditioned medium of those cells and the factors nPEDF or coPEDF showed a better functional recovery when compared to the control groups. The results of this study showed that the SC, your conditioned medium and the PEDF play an important role in the spinal cord neurons, allowing the modulation of the functional recovery process of this organ after the trauma.
 
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Publishing Date
2009-02-11
 
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