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Use of fibrin sealant combined with mesenchymal stem cells for anterior funiculus injury repair: effects on inflammation, axonal regeneration and neuroprotection.

Grant number: 12/22750-9
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): May 01, 2013
Effective date (End): August 31, 2017
Field of knowledge:Biological Sciences - Morphology - Cytology and Cell Biology
Principal researcher:Alexandre Leite Rodrigues de Oliveira
Grantee:Aline Barroso Spejo
Home Institution: Instituto de Biologia (IB). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
Associated scholarship(s):16/19954-2 - Targeted dorsal root regeneration towards spinal motoneurons by intramuscular Neurotrophin-3 gene therapy, BE.EP.DR


The limited regenerative capacity of neurons in the central nervous system and the formation of an inhibitory environment at the site of injury represent a major challenge for regeneration and repair of injuries to the spinal cord. In this sense, the influence of mesenchymal stem cells (MSC) in the spinal Cord microenvironment, glial scar formation and its effects on inflammation, control of reactive gliosis, neuronal survival and synaptic plasticity in injured motoneurons, becomes an important object of study. Additionally, the use of fibrin sealant as a substrate for the MSC may contribuit positively to the successful regeneration in the CNS. MSC modify the inflammatory environment in the acute phase and reduce inhibitory effects of glial scar in the subacute and chronic phase, providing a permissive environment for axonal extension. It can also provide growth factors that promote survival and axonal elongation through injury to the spinal cord. To evaluate the neuroprotective, regenerative, modulatory of inflammation and glial scar, the treatment with fibrin sealant and / or therapy with MSC after a central lesion, Will be performed a perforating injury in the region of the right anterior funiculus in spinal levels L4 and L5 ( n = 5 per experimental group). After survival rate 7 days after the lesion will be performed (1) RT-PCR in real time to VEGF, IL4, IL3, TNF, IL6, arginase-1, iNOS, and TGF² and (2) with double labeling immunohistochemistry for IBA1 (marker of microglia and macrophages) and Arginase-1 (macrophage marker M2) in situ to demonstrate the presence / reactivity of such cells. After 14 days survival will be performed (1) by counting neuronal Nissl staining, and (2) immunohistochemistry for synaptophysin (a marker of synapses). With survival of 8 weeks, will be held (1) behavioral test for gait analysis of animals (Catwalk), (2) analysis of the glial scar by FISH (fluorescence in situ hybridization) and IH. In this procedure will be doubly labeled GFAP (a marker for astrocytes - IH) and chondroitin sulfate (FISH), (3) analysis of neuronal regeneration by glial scar by FISH and IH. In this procedure will be doubly labeled neurofilaments (IH) and GAP-43 (marker of neuron regeneration - FISH) and (4) qualitative analysis of the production of collagen and laminin by MSC through IH.

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
SPEJO, A. B.; CHIAROTTO, G. B.; FERREIRA, A. D. F.; GOMES, D. A.; FERREIRA JR, R. S.; BARRAVIERA, B.; OLIVEIRA, A. L. R.. Neuroprotection and immunomodulation following intraspinal axotomy of motoneurons by treatment with adult mesenchymal stem cells. JOURNAL OF NEUROINFLAMMATION, v. 15, . (13/16168-8, 14/06892-3, 12/22750-9)
BISCOLA, NATALIA PERUSSI; CARTAROZZI, LUCIANA POLITTI; ULIAN-BENITEZ, SUZANA; BARBIZAN, ROBERTA; CASTRO, MATEUS VIDIGAL; SPEJO, ALINE BARROSO; FERREIRA, JR., RUI SEABRA; BARRAVIERA, BENEDITO; RODRIGUES OLIVEIRA, ALEXANDRE LEITE. Multiple uses of fibrin sealant for nervous system treatment following injury and disease. Journal of Venomous Animals and Toxins including Tropical Diseases, v. 23, n. 1, . (13/23030-2, 12/08101-8, 11/23377-7, 13/04409-0, 14/06892-3, 12/13514-0, 12/20456-6, 14/11405-4, 12/22750-9, 10/00729-2, 09/53846-9, 12/02689-3, 11/02889-0)
SPEJO, ALINE BARROSO; TELES, CAROLINE BRANDAO; ZUCCOLI, GIULIANA DA SILVA; RODRIGUES DE OLIVEIRA, ALEXANDRE LEITE. Synapse preservation and decreased glial reactions following ventral root crush (VRC) and treatment with 4-hydroxy-tempo (TEMPOL). Journal of Neuroscience Research, v. 97, n. 4, p. 520-534, . (14/12707-4, 12/22750-9, 14/12671-0, 14/06892-3)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
SPEJO, Aline Barroso. Use of fibrin sealant combined with mesenchymal stem cells for anterior funiculus injury repair: effects on inflammation, axonal regeneration and neuroprotection. 2017. Doctoral Thesis - Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia Campinas, SP.

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