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Doctoral Thesis
DOI
https://doi.org/10.11606/T.5.2023.tde-01122023-165917
Document
Author
Full name
Thiego Pedro Freitas Araújo
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2023
Supervisor
Committee
Barros Filho, Tarcisio Eloy Pessoa de (President)
Torelli, Alessandro Gonzalez
Cristante, Alexandre Fogaça
Meves, Robert
Title in Portuguese
Injeção em camundongos de células-tronco mononucleares de cordão umbilical e placenta humana após 3 a 6 semanas de lesão medular espinhal experimental
Keywords in Portuguese
Avaliação histopatológica
Avaliação motora
Células-tronco mononucleares
Cordão umbilical
Traumatismos da medula espinal
Abstract in Portuguese
O trauma raquimedular é dano catastrófico na vida do doente e até o momento nenhuma medida se mostrou francamente eficaz para o tratamento. O paciente passa da fase aguda para a fase subaguda e por último a fase crônica. E mesmo na última fase, alterações neurofisiológicas continuam a acontecer. A descoberta das células-tronco e o desenvolvimento da manipulação delas abriram uma nova esperança para o lesado medular. As células-tronco retiradas do sangue de cordão umbilical e placenta humana apresentam várias vantagens tanto para o uso em pesquisa quanto clínico. Na literatura, a maioria dos estudos com animais é realizada na fase aguda e subaguda, tendo uma lacuna de conhecimento na fase crônica do trauma raquimedular. Devido ao exposto acima, criou-se interesse nesse estudo para a pesquisa em células-tronco em animais com lesão medular na fase crônica. O objetivo deste estudo foi avaliar, em modelo padronizado, a regeneração nervosa e a recuperação funcional motora de camundongos Balb C com paraplegia cirurgicamente provocada em resposta ao uso de células-tronco, na fase crônica, do sangue do cordão umbilical e placentário de humanos após 3 semanas e 6 semanas do trauma raquimedular. No total, 48 camundongos foram sorteados em 6 grupos de 8 animais. O grupo 1 recebeu as células-tronco 3 semanas após o trauma, o grupo 2, 6 semanas após o trauma. No grupo 3 foi injetada solução fisiológica no sítio da lesão 3 semanas após o trauma e no grupo 4, 6 semanas após o trauma. O grupo 5 foi submetido apenas a lesão medular e o grupo 6 a laminectomia sem lesão medular. As escalas utilizadas para a avaliação motora foram BMS, MFS e escada horizontal por 12 semanas. Os grupos de intervenção apresentaram melhora motora nas escalas analisadas, principalmente na BMS e na MFS, em relação aos controles, com significância estatística. Na análise histopatológica os grupos intervenção apresentaram menor grau de lesão em relação aos grupos controle com significância estatística. Em relação ao brotamento axonal, os grupos intervenção apresentaram aumento do brotamento axonal em relação aos controles, sendo que na porção caudal, teve significância estatística. Conclui-se que o uso de células-tronco em camundongos, na fase crônica em 3 e 6 semanas do trauma raquimedular, traz benefícios funcionais e histopatológicos para os mesmos
Title in English
Injection of mononuclear stem cells from human umbilical cord and placenta into mice after 3 to 6 weeks of experimental spinal cord injury
Keywords in English
Histopathological assessment
Mononuclear stem cells
Motor assessment
Spinal cord injury
Umbilical cord
Abstract in English
Spinal cord trauma is a catastrophic event to the patient's life and so far no measure has been shown to be frankly effective for the treatment. The patient passes from acute phase to subacute phase and finally to chronic phase. And even in the last phase, neurophysiological changes continue to occur. The discovery of stem cells and the development of their manipulation opened up new hope for the spinal cord injured person. Stem cells taken from human umbilical cord and placental blood have several advantages for both research and clinical use. In the literature, most studies with animals are carried out in the acute and sub-acutephases, with a lack of knowledge in the chronic phase of spinal cord injury. Due to the above, interest was created in this study for research on stem cells in animals with spinal cord injury. The aim of this study was to evaluate, in a standardized model, nerve regeneration and motor functional recovery in Balb C mice with surgically provoked paraplegia in response to the use of stem cells, in chronic phase, from human umbilical cord and placental blood after 3 weeks and 6 weeks of spinal cord trauma. In total, 48 mice were randomly assigned to 6 groups of 8 animals. Group 1 received stem cells 3 weeks after the trauma, group 2, six weeks after the trauma. In group 3, saline solution was injected at the injury site 3 weeks after the trauma and in group 4, six weeks after the trauma. Group 5 was submitted to spinal cord injury only and group 6 to laminectomy without spinal cord injury. The scales used for motor assessment were BMS, MFS and horizontal ladder 12 weeks. The intervention groups showed motor improvement in the analyzed scales, mainly in the BMS and MFS, in relation to the controls, with statistical significance. In the histopathological analysis, the intervention groups showed a lower degree of injury compared to the control groups, with statistical significance. Regarding axonal sprouting, the intervention groups showed an increase in axonal sprouting in relation to the controls, and in the caudal portion, with statistical significance. It is concluded that the use of stem cells in mice in the chronic phase at 3 and 6 weeks of spinal cord trauma, brings functional and histopathological benefits for them
 
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Publishing Date
2023-12-11
 
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