Gostaríamos de agradecer o apoio de financiamento para este trabalho a partir do Canadian Institutes of Health Research (CIHR), a Fundação Krembil Família, o Presidente Halbert em Neural Repair and Regeneration, Phillip e Peggy DeZwirek, e Gordon Yao para a contribuição para a Figura 2 . Klaus Zweckberger foi financiado por uma doação da &quot;Deutsche Forschungsgesellschaft&quot; (DFG).

Nota: Os seguintes protocolo experimental foi aprovado pelo comitê de cuidados com os animais da University Health Network (Toronto, Canadá) e está em conformidade com as políticas estabelecidas no guia para o cuidado e uso de animais de experimentação preparado pelo Conselho Canadense de cuidados com os animais . 1. Cervical Aneurysm Clipe Contusão / Compression Modelo <ol><li> Antes de instrumentos e autoclave cirurgia manter condições estéreis durante todo o procedimento surgcial, colocando os instrumentos em um banho de álcool a 70%. </li><li> Anestesiar ratos machos Wistar (250-270 g) com uma combinação de oxigénio (O2), óxido nitroso (N2O) (1: 1), e 1,8-2,2% de isoflurano e suportar a respiração espontânea, através de uma máscara de anestesia de gás. Para a indução de anestesia começar com 5% de isoflurano durante 1 minuto e depois reduzir. Antes de iniciar a cirurgia, a profundidade da anestesia controle, dando um estímulo doloroso (ex. Nas patas). Aplicar pomadas gordasnos olhos para evitar a secura e a prevenção de infecções subsequentes. </li><li> Coloque os ratos sobre uma almofada de aquecimento (37 ° C) e fixar a cabeça em um quadro estereotáxico. </li><li> Raspar a região cirúrgica ao redor da coluna cervical e desinfectar com iodopovidona e 70% de álcool. </li><li> Faça uma incisão na linha média acima da espinha da vértebra cervical (C2) chegando ao processus proeminente do corpo vertebral torácica 2 (T2). </li><li> Corte através da camada externa dos músculos da coluna diretamente na linha média (para evitar o sangramento) no sentido crânio-caudal e mais dissecar as camadas musculares mais profundas sem rodeios até chegar ao spinosus processus e as lâminas. Insira afastadores. </li><li> Orientar o proeminente spinosus processus do T2 corpo vértebra para observar os níveis direcionados para laminectomy. Após a identificação e preparação de micro-cirúrgica das lâminas selecionado, cortar o flavae ligamenti para soltar a lâmina e do spinosus processus. Finalmente, cut através das lâminas com um cortador de osso de lateral para a medula espinhal e remover aqueles suave, evitando qualquer compressão da medula espinhal em si. NOTA: O mais comum são os níveis C5 / 6, C6 / 7, ou C7 / T1. A taxa de mortalidade peri-operatória aumenta o mais rostral do nível da lesão. Sangramento do seio venoso paravertebral é comum e pode ser abordada por compressão cuidado com esponja. </li><li> Antes de inserir o clipe para traumatizar a cabo, identificar emergente raízes nervosas para poupá-los a partir de recorte (especialmente a nível C5 / 6). </li><li> A fim de garantir a indução grampo liso, soltar a dura-máter do lado ventral dorsal dos corpos vertebrais com um gancho e preparar um corredor para o grampo. </li><li> Finalmente, insira o clipe aberto e deixá-lo tirar fechada (fechamento rápido) para alcançar uma contusão. Clipe força de fecho e duração de encerramento clipe determinar a intensidade do trauma e o grau de compressão. Comumente usado são forças clipe de entre 15-35 g e um clippiduração ng de por exemplo 1 min. (Figura 1). </li><li> Após a remoção do clipe, adaptar-se os músculos em duas camadas e fechar a ferida. </li><li> Pare de anestesia e deixar o rastro dos animais sob sua observação contínua até que ele recupere a consciência suficiente para decúbito esternal. Por último, coloque o rato em uma gaiola individual e siga as diretrizes de tratamento pós-operatório. </li><li> Dado que os animais lutam com a gravidade deste tipo de lesão, você deve prestar atenção especial para tratamentos pós-operatórios: <ol><li> Administrar analgésicos (buprenorfina e meloxicam durante 3 dias e 5 dias, respectivamente, e de acordo com os sintomas clínicos). </li><li> Dê subcutaneamente solução salina adicional durante 3 dias (2 vezes por dia, 5-10 mililitros (ml)). </li><li> Fornecer antibióticos em água potável 2 dias antes e até sete dias após a cirurgia (por exemplo, moxifloxacina) </li><li> Aperte a bexiga urinária 2-3 vezes ao dia até que a recuperação da função da bexiga é constantely aparente. </li><li> Observe déficits neurológicos e condição fisiológica dos animais operados pelo menos uma vez por dia. </li></ol></li></ol> 2. A injeção de SAPs e NPCs (14 dias após a lesão) <ol><li> Induzir a anestesia, conforme descrito no item 1.1. para 1,3, fixar a cabeça do rato em um quadro estereotáxico, retirar os pontos ou clipes de feridas, e desinfectar a ferida e a área cirúrgica com iodopovidona e 70% de álcool. </li><li> Dissecar cuidadosamente os músculos paravertebrais, insira afastadores, remover o tecido cicatricial microscopicamente da dura-máter, e re-expor o local da lesão. </li><li> Prepare SAPs em uma concentração de 1% (w / v) para executar um gel de matriz extracelular. SAPs QL6 tem um pH fisiologicamente compatível e não precisam de ser tamponada, antes da injecção. Para a visualização de PAE na medula espinhal, usar um derivado fluorescente de QL6 (QL6-FITC). </li><li> Injectar SAPs (5 microlitros (ul) para o centro da lesão, distribuídos em duas porções, cada2,5 ul bilateral da linha média. Usando uma seringa de Hamilton ligada à armação estereotáxica com um micro capilar de vidro (100 micrómetros (um) de diâmetro externo (OD)). Abra a dura-máter cuidadosamente com a ponta de uma agulha afiada, e inserir o tubo capilar de vidro estereotaxicamente 2 milímetros (mm) para a medula espinhal traumatizado. </li><li> Após a injecção de 1/3 do volume, remover a agulha a uma profundidade 1,5 mm, e após mais 1/3 a 1 mm. Após a injecção de todo o volume, e antes da remoção da seringa, esperar 5 minutos para estabilizar a formação de gel. </li><li> Para gerar NPCs, use camundongos adultos DsRed (ou YFP camundongos positivos, verde) e isolar e cultivar-los da zona paraventrical 4,18,21. </li><li> Avaliar a viabilidade de NPCs por coloração com azul de tripano, indicando a presença de ~ 90% de células vivas na suspensão de células. Diluir as células em meio de crescimento (50 x 103 células vivas / l) e, em seguida, usá-los para o transplante de células. </li><li> Faça quatro 2 ul (8 ul vo total delume, contendo 4 x 10 5 NPCs) injeções intra-espinhal bilateralmente a 2 mm rostral e caudal do local da lesão. Após a abertura da dura-máter, inserir o micro capilar de vidro Hamilton 1,5 milímetros abaixo da superfície dorsal da medula espinal e injectar 2 ul de suspensão de células. Escolha uma velocidade de injecção de cerca de 0,5 ul / min (min). </li><li> No final de cada injecção e antes da remoção do capilar para fora do cabo, para esperar pelo menos 1 min permitir que o tecido estica para acomodar o volume da célula nova. (Figura 2) </li></ol> 3. Implantação de subdural Bombas para Growth Factor Aplicação <ol><li> A fim de enriquecer o cerebral fluido espinhal (CSF) com fatores de crescimento apoiando a sobrevivência da célula, use bombas de micro-osmótica diluindo fatores de crescimento sub-durally ao longo de 7-14 dias, com uma taxa de diluição de 0,5 l / h, um diâmetro de 0,04 cateter cm OD, e um volume do reservatório de 100 ul. </li><li> Escolha factores de crescimento preferidos(Por exemplo, factor de crescimento derivado do cérebro (BDGF), o factor de crescimento epidérmico (EGF), factor de crescimento de fibroblastos (FGF)), encher as bombas de 6-10 horas antes da implantação, bombas e equilibrar num banho de água a 37 ° C. </li><li> Imediatamente após injeções NPC, preparar um recesso subcutânea para colocar a bomba. Locais preferidos são os flancos laterais da toracotomia região abdominal evitar maior desconforto local provocada pela própria bomba. </li><li> Para ganhar a maior concentração de fatores de crescimento, garantir que a ponta do cateter aberto termina próximo ao local da lesão. Portanto, executar um salto-laminectomy do nível superior ou inferior adjacente. Por exemplo, se o SCI é a C7 / T1, realizar uma pequena laminectomia de C5. </li><li> Coloque a bomba no recesso subcutânea, encurtar o cateter com o comprimento necessário, e prenda-o com várias suturas (6.0) nos músculos paravertebrais evitando qualquer luxação associada à circulação. </li><li> Após abertura da dura (por exemplo, em C5) com a ponta afiadade uma agulha, introduzir o cateter no espaço subdural e deixá-lo deslizar em direção caudal, sem qualquer resistência e sem ferir o cabo. A lâmina pulada de eg C6 serve como ponto adicional de fixação e estabilização do cateter. </li><li> Certifique-se de que o cateter corra bem e não se dobra. </li><li> Fechar músculos por camada, e pele, com suturas ou grampos. (Figura 3) </li><li> Pare de anestesia e deixar o rastro dos animais sob sua observação contínua até que ele recupere a consciência suficiente para decúbito esternal. Por fim, colocá-los de volta em uma única gaiola e siga as diretrizes de tratamento pós-operatório. </li><li> Fornecer tratamento pós-operatório especial, embora, os animais poderiam ter recuperado parcialmente a este ponto-time: <ol><li> Administrar analgésicos (Buprenorthine e meloxicam durante 3 dias e 5 dias, respectivamente, e de acordo com os sintomas clínicos). </li><li> Fornecer antibióticos em garrafas de água até 2 dias antes7 dias pós-cirurgia (por exemplo, moxifloxacina). </li><li> Continue apertando a bexiga urinária, se ainda for necessário. </li><li> Dê fluidos por via subcutânea adicionais, se os ratos aparecem desidratado. </li><li> Mantenha observando a função neurológica e estado fisiológico dos animais operados pelo menos uma vez por dia. </li><li> Administrar tratamento de imunossupressão dois dias antes da injecção NPC e até 7 dias após o transplante (minociclina) e até o sacrifício (Sandimmune), respectivamente. </li></ol></li></ol> Avaliação 4. Tissue <ol><li> Sacrificar os animais no final do período de observação (por exemplo. 4 semanas após a SCI) em anestesia profunda (5% de isoflurano durante 2-3 min) e perfundir-los transcardíaca com 50 ml de solução salina fria (4 ° C), seguido por 150 ml de 4 frias % de paraformaldeído em 0,1 M de solução salina tamponada com fosfato (PBS). </li><li> Remover a medula espinhal e colocá-lo em 4% de paraformaldeído em 0,1 M de solução salina tamponada com fosfato (PBS) durante 24 horas. </li><li&gt; Coloque as medulas espinais em solução de sacarose a 25% para crioprotecção durante 48 horas. </li><li> Fornecer criocortes longitudinais com uma espessura de &lt;30 um. </li><li> Para coloração imuno-histoquímica de todos os núcleos de células-proporcionando um uso do fundo DAPI (1: 1.000). DsRed NPCs positivos aparecem em vermelho, QL-6 FITC aparece verde, e ambos, portanto, não precisa de ser marcadas especialmente. (Figura 4). </li><li> Para a microscopia eletrônica de varredura (SEM) deixe amostras de molho em glutaraldeído a 4 ° C, durante 2 horas, desidratam-los lentamente em passos de 10% de incremento de etanol durante 5 minutos e coloque-os em um pressurizado líquido CO 2 sifão para a 1 hora. Scaffolds revestimento com ouro usando um revestidor por crepitação. Tire fotos com a Hitachi S-3400N microscópio eletrônico de varredura. (Figura 5) </li></ol>

<ol>
	<li>Sekhon, L. H., Fehlings, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidemiology,+demographics,+and+pathophysiology+of+acute+spinal+cord+injury.">Epidemiology, demographics, and pathophysiology of acute spinal cord injury.</a> Spine (Phila Pa 1976). 26, S2-S12 (2001).</li><li>Fehlings, M. G., Tator, C. H., Linden, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+relationships+among+the+severity+of+spinal+cord+injury,+motor+and+somatosensory+evoked+potentials+and+spinal+cord+blood+flow).">The relationships among the severity of spinal cord injury, motor and somatosensory evoked potentials and spinal cord blood flow).</a> Electroencephalogr Clin Neurophysiol. 74, 241-259 (1989).</li><li>Thuret, S., Moon, L. D., Gage, F. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Therapeutic+interventions+after+spinal+cord+injury.">Therapeutic interventions after spinal cord injury.</a> Nat Rev Neurosci. 7, 628-643 (2006).</li><li>Iwasaki, M., Wilcox, J. T., Nishimura, Y., Zweckberger, K., Suzuki, H., Wang, J., Liu, Y., Karadimas, S. K., Fehlings, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synergistic+effects+of+self-assembling+peptide+and+neural+stem/progenitor+cells+to+promote+tissue+repair+and+forelimb+functional+recovery+in+cervical+spinal+cord+injury.">Synergistic effects of self-assembling peptide and neural stem/progenitor cells to promote tissue repair and forelimb functional recovery in cervical spinal cord injury.</a> Biomaterials. 35, 2617-2629 (2014).</li><li>Holmes, T. C., de Lacalle, S., Su, X., Liu, G., Rich, A., Zhang, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extensive+neurite+outgrowth+and+active+synapse+formation+on+self-assembling+peptide+scaffolds.">Extensive neurite outgrowth and active synapse formation on self-assembling peptide scaffolds.</a> Proc Natl Acad Sci U S A. 97, 6728-6733 (2000).</li><li>Dong, H., Paramonov, S. E., Aulisa, L., Bakota, E. L., Hartgerink, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Self-assembly+of+multidomain+peptides:+balancing+molecular+frustration+controls+conformation+and+nanostructure.">Self-assembly of multidomain peptides: balancing molecular frustration controls conformation and nanostructure.</a> J Am Chem Soc. 129, 12468-12472 (2007).</li><li>Rivlin, A. S., Tator, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+duration+of+acute+spinal+cord+compression+in+a+new+acute+cord+injury+model+in+the+rat.">Effect of duration of acute spinal cord compression in a new acute cord injury model in the rat.</a> Surg Neurol. 10, 38-43 (1978).</li><li>Poon, P. C., Gupta, D., Shoichet, M. S., Tator, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clip+compression+model+is+useful+for+thoracic+spinal+cord+injuries:+histologic+and+functional+correlates.">Clip compression model is useful for thoracic spinal cord injuries: histologic and functional correlates.</a> Spine (Phila Pa 1976). 32, 2853-2859 (2007).</li><li>Fehlings, M. G., Tator, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+relationships+among+the+severity+of+spinal+cord+injury,+residual+neurological+function,+axon+counts,+and+counts+of+retrogradely+labeled+neurons+after+experimental+spinal+cord+injury.">The relationships among the severity of spinal cord injury, residual neurological function, axon counts, and counts of retrogradely labeled neurons after experimental spinal cord injury.</a> Exp Neurol. 132, 220-228 (1995).</li><li>Joshi, M., Fehlings, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+characterization+of+a+novel,+graded+model+of+clip+compressive+spinal+cord+injury+in+the+mouse:+Part+2.+Quantitative+neuroanatomical+assessment+and+analysis+of+the+relationships+between+axonal+tracts,+residual+tissue,+and+locomotor+recovery.">Development and characterization of a novel, graded model of clip compressive spinal cord injury in the mouse: Part 2. Quantitative neuroanatomical assessment and analysis of the relationships between axonal tracts, residual tissue, and locomotor recovery.</a> J Neurotrauma. 19, 191-203 (2002).</li><li>Joshi, M., Fehlings, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+characterization+of+a+novel,+graded+model+of+clip+compressive+spinal+cord+injury+in+the+mouse:+Part+1.+Clip+design,+behavioral+outcomes,+and+histopathology.">Development and characterization of a novel, graded model of clip compressive spinal cord injury in the mouse: Part 1. Clip design, behavioral outcomes, and histopathology.</a> J Neurotrauma. 19, 175-190 (2002).</li><li>Cigognini, D., Satta, A., Colleoni, B., Silva, D., Doneg&#224;, M., Antonini, S., Gelain, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+early+and+late+effects+into+the+acute+spinal+cord+injury+of+an+injectable+functionalized+self-assembling+scaffolds.">Evaluation of early and late effects into the acute spinal cord injury of an injectable functionalized self-assembling scaffolds.</a> PLoS One. 6 (5), e19782 (2011).</li><li>Hou, T., Wu, T., Wang, L., Liu, Y., Li, M., Long, Z., Chen, H., Li, Y., Wang, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+prostheses+fabricated+with+motor+neurons+seeded+in+self-assembling+peptides+promotes+partial+functional+recovery+afters+spinal+cord+injury+in+rats.">Cellular prostheses fabricated with motor neurons seeded in self-assembling peptides promotes partial functional recovery afters spinal cord injury in rats.</a> Tissue eng Part A. 18 (9-10), (2012).</li><li>Gelain, F., Cigognini, D., Caprini, A., Silva, D., Colleoni, B., Doneg&#224;, M., Antonini, S., Cohen, B. E., Vescovi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+bioactive+motifs+and+their+use+in+functionalized+self-assembling+peptides+for+NPC+differentiation+and+neural+tissue+engineering.">New bioactive motifs and their use in functionalized self-assembling peptides for NPC differentiation and neural tissue engineering.</a> Nanoscale. 4 (9), 2946-2957 (2012).</li><li>Liu, Y., Ye, H., Satkunendrarajah, K., Yao, G. S., Bayon, Y., Fehlings, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+self-assembling+peptide+reduces+glial+scarring,+attenuates+post-traumatic+inflammation+and+promotes+neurological+recovery+following+spinal+cord+injury.">A self-assembling peptide reduces glial scarring, attenuates post-traumatic inflammation and promotes neurological recovery following spinal cord injury.</a> Acta Biomater. 9, 8075-8088 (2013).</li><li>Rosner, J., Avalos, P., Axosta, F., Liu, J., Drazin, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+potential+for+cell+therapy+combined+with+growth+factors+in+spinal+cord+injury.">The potential for cell therapy combined with growth factors in spinal cord injury.</a> Stem Cell Int. , 826754 (2012).</li><li>Lu, P., Wang, Y., Graham, L., McHale, K., Gao, M., Wu, D., Brock, J., Blesch, A., Rosenzweig, E. S., Havton, L. A., Zheng, B., Conner, J. M., Marsala, M., Tuszynsky, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long+distance+growth+and+connectivity+of+neural+stem+cells+after+severe+spinal+cord+injury.">Long distance growth and connectivity of neural stem cells after severe spinal cord injury.</a> Cell. 150, 1265-1273 (2012).</li><li>Karimi-Abdolrezaee, S., Schut, D., Wang, J., Fehlings, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chondrioitinase+and+grwoth+factors+enhance+activation+and+oligodendrocyte+differentiation+of+endogenous+neural+precursor+cells+after+spinal+cord+injury.">Chondrioitinase and grwoth factors enhance activation and oligodendrocyte differentiation of endogenous neural precursor cells after spinal cord injury.</a> PLoS One. 7 (5), e37589 (2012).</li><li>Awad, B. I., Carmody, M. A., Steinmetz, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Potential+role+of+growth+factors+in+the+management+of+spinal+cord+injury.">Potential role of growth factors in the management of spinal cord injury.</a> World Neurosurg. (13), 1875-8750 (2013).</li><li>Kojima, A., Tator, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intrathecal+administration+of+epidermal+growth+factor+and+fibroblast+growth+factor+2+promotes+ependymal+proliferation+and+functional+recovery+after+spinal+cord+injury+in+adult+rats.">Intrathecal administration of epidermal growth factor and fibroblast growth factor 2 promotes ependymal proliferation and functional recovery after spinal cord injury in adult rats.</a> J Neurotrauma. 19 (2), 223-238 (2002).</li><li>Karimi-Abdolrezaee, S., Eftekharpour, E., Wang, J., Cindi, M. M., Fehlings, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Delayed+trasplantation+of+adult+neural+presursor+cells+promotes+remyelination+and+functional+neurological+recovery+after+spinal+cord+injury.">Delayed trasplantation of adult neural presursor cells promotes remyelination and functional neurological recovery after spinal cord injury.</a> J Neurosci. 26 (13), 3377-3389 (2006).</li><li>Burdick, J. A., Ward, M., Liang, E., Young, M. J., Langer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stimulation+of+neurite+outgrowth+by+neurotrophins+delivered+from+degradable+hydrogels.">Stimulation of neurite outgrowth by neurotrophins delivered from degradable hydrogels.</a> Biomaterials. 27, 452-459 (2006).</li></ol>

Covidien supplied the QL6 SAP for this study.

Este protocolo foi desenvolvido para permitir que o leitor para realizar um modelo de lesão cervical em ratos e usar uma abordagem de tratamento combinado com SAPs e NPCs que promovem uma melhor recuperação após SCI cervical. Particularmente em relação a outros modelos de trauma cervical, como o (hemi) -transection modelo ou os modelos de queda de peso e de concussão, o modelo clipe contusão / compressão representa tanto importantes mecanismos de trauma fisiopatológicos - portanto,...

Mais de 50% das lesões da medula espinal estão relacionadas com a coluna cervical. Na clínica definindo dois principais mecanismos patofisiológicos são descritos: a contusão inicial da espinal medula e, subsequentemente, a compressão permanente causada por fracturas ósseas, hemorragias ou inchaço dos tecidos. O clipe de aneurisma imita contusão modelo / compressão ambos os mecanismos fisiopatológicos: tirando o clipe cria uma contusão e a duração do recorte representa o componente de compressão, admitindo que a compressão em clínicas causadas por fraturas ósseas, hemorragias ou inchaço dos tecidos última significativamente maior. O clipe de aneurisma utilizado é modificado por uma mola anel garantindo força exata e reprodutível recorte. Especialmente em comparação com o hemi-transection ou o modelo de contusão, este clip de aneurisma imita modelo melhores ambientes clínicos. Enquanto pacientes com lesões torácicas sofrem de paraplegia, a maioria dos pacientes com lesão cervicaluries são tetraplégica e completamente dependente. A estrutura anatômica do cordão cervical, no entanto, apresenta diferenças significativas em relação à coluna vertebral torácica ou lombar e, portanto, é dirigida em particular neste protocolo. O desenvolvimento de cavidades intramedulares e tecido cicatricial são obstáculos para a recuperação e regeneração. Para superar estes obstáculos à utilização de material de andaime é uma abordagem promissora. Péptidos de auto-montagem pode ser injectado directamente no epicentro da lesão. Lá eles se reúnem em andaimes nano-fibras ponte da cavidade e melhorar o ambiente inibitório, reduzindo a inflamação e cicatrização de tecidos. Enquanto materiais rígidos causar danos consideráveis ​​da medula espinhal durante a implantação, os péptidos de fluido podem ser injectados com segurança e sem causar danos adicionais grave. Melhorar o ambiente inibitório com peptídeos de auto-montagem antes do transplante de células-tronco, assim, apoiar a célula integração, diferenciação e, finalmente, a recuperação funcional após a lesão da medula espinal cervical.

<table border="0" cellpadding="0" cellspacing="0" width="541">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:273px;">Name</td>
			<td style="width:268px;">Company</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Aneurysmal clip</td>
			<td>SharpTech</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgical microscope</td>
			<td>Leica</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micro injection system</td>
			<td>World Precision Instruments, Inc.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Small animal stereotaxic instrument</td>
			<td>David Kopf Instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hamilton syringe</td>
			<td>Hamilton company</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Subdural pumps</td>
			<td>Alzet osmotic micro pump 1007D</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgical instrument</td>
			<td>Fine Science tools</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Isoflurane USP</td>
			<td>Pharmaceutical Partners of Canada Inc.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">0.9% Sodium Chloride injection USP</td>
			<td>Baxter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">7.5% Povidone iodine</td>
			<td>Purdue Pharma</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">70% Isopropyl alcohol USP</td>
			<td>GreenField Ethanol Inc.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">QL6 SAP</td>
			<td>Covidien</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">0.4% Trypan blue</td>
			<td>Gibco</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Platelet-Derived Growth Factor (PDGF)</td>
			<td>Sigma</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Epidermal Growth Factor (EGF)</td>
			<td>Sigma</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fibroblast Growth Factor (FGF)</td>
			<td>Sigma</td>
		</tr>
	</tbody>
</table>

synergetic use of neural precursor cells and self-assembling peptides in experimental cervical spinal cord injury

Traumatismos da medula espinal (SCI) causar lesão neurológica grave e consequências psicológicas, econômicas e sociais para os pacientes e suas famílias. Clinicamente, mais de 50% da SCI afecta a coluna cervical 1. Como uma consequência da lesão primária, uma cascata de mecanismos secundários, incluindo a inflamação, a apoptose, e desmielinização ocorrem finalmente levando a cicatrização de tecidos e de desenvolvimento de cavidades intramedulares 2,3. Ambos representam barreiras físicas e químicas para o transplante de células, integração e regeneração. Portanto, moldar o ambiente inibitório e colmatar cavidades para criar um ambiente favorável para o transplante de células e regeneração é um alvo terapêutico promissor 4. Aqui, um modelo de contusão / compressão de SCI cervical usando um clipe de aneurisma é descrito. Este modelo é mais clinicamente relevantes do que outros modelos experimentais, uma vez que transection ou rupturas do cabo completo são raros. Também em COMPARAÇÃOn para o modelo de queda de peso, o que, em particular, as colunas danos no dorso, compressão circunferencial da medula espinal é vantajoso. Clipe força de fecho e tempo de exposição pode ser ajustada para atingir diferente gravidade da lesão. Uma mola anel facilita a calibração precisa e constância da força clip. Em condições fisiológicas, peptídeos auto-montagem sintéticos (SAP) de auto-montar em nanofibras e assim, estão apelando para aplicação em SCI 5. Eles podem ser injectados directamente na lesão minimizando os danos ao cabo. SAPs são estruturas biocompatíveis erguer andaimes para colmatar cavidades intramedulares e, portanto, equipar o cabo danificado por tratamentos regenerativos. K2 (QL) 6K2 (QL6) é um romance SAP introduzido por Dong et a l. 6 Em comparação com outros peptídeos, QL6 auto-monta em β-folhas de pH neutro 6 0,14 dias após a LM, após a fase aguda, SAPs são injectadas no centro da lesão e as células precursoras neurais (NPC) são injected em colunas dorsais adjacentes. A fim de suportar a sobrevivência de células, o transplante é combinado com a administração subdural contínua de factores de crescimento de micro-bombas osmóticas durante 7 dias.

<html> Ao executar o procedimento acima descrito, irá obter um andaime SAP colmatar a cavidade e que oferece uma melhoria do ambiente inibitório, menos tecido cicatricial, e um aumento na sobrevivência NPC. A Figura 4 mostra uma secção longitudinal de uma medula espinal de rato obtido no local da lesão 6 semanas após SCI e 4 semanas após a injeção QL6 SAP e transplante NPC. Peptídeos QL6 foram injetados com êxito ao cabo, agregadas no epicentro e difusa rostro-caudal na penumbra. Microscópio...

Watch this Scientific Journal Video about Synergetic Use of Neural Precursor Cells and Self-assembling Peptides in Experimental Cervical Spinal Cord Injury at JoVE.com

Synergetic Use of Neural Precursor Cells and Self-assembling Peptides in Experimental Cervical Spinal Cord Injury

Treating cervical spinal cord injury with both self-assembling peptides (SAP) and neural precursor cells (NPC), together with growth factors, is a promising approach to promote regeneration and recovery. A contusion/compression aneurysm clip rat model of cervical SCI and combined treatment involving SAP injection and NPC transplantation is established.

Use Synergetic de células neuronais precursoras e Peptídeos de auto-montagem em Experimental Cervical Spinal Cord Injury

Spinal cord injuries (SCI) cause serious neurological impairment and psychological, economic, and social consequences for patients and their families. ...

synergetic-use-neural-precursor-cells-self-assembling-peptides

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Medicine

9.1K visualizações.  University Health Network, Krembil Neuroscience Centre. Treating cervical spinal cord injury with both self-assembling peptides (SAP) and neural precursor cells (NPC), together with growth factors, is a promising approach to promote regeneration and recovery. A contusion/compression aneurysm clip rat model of cervical SCI and combined treatment involving SAP injection and NPC transplantation is established. 

Vídeo: Synergetic Use of Neural Precursor Cells and Self-assembling Peptides in Experimental Cervical Spinal Cord Injury

Intraspinal Cell Transplantation for Targeting Cervical Ventral Horn in Amyotrophic Lateral Sclerosis and Traumatic Spinal Cord Injury

Respiratory compromise due to phrenic motor neuron loss is a debilitating consequence of a large proportion of human traumatic spinal cord injury (SCI) cases 1 and is the ultimate cause of death in patients with the motor neuron disorder, amyotrophic laterals sclerosis (ALS) 2.
ALS is a devastating neurological disorder that is characterized by relatively rapid degeneration of upper and lower motor neurons. Patients ultimately succumb to the disease on average 2-5 years following diagnosis because of respiratory paralysis due to loss of phrenic motor neuron innnervation of the diaphragm 3. The vast majority of cases are sporadic, while 10% are of the familial form. Approximately twenty percent of familial cases are linked to various point mutations in the Cu/Zn superoxide dismutase 1 (SOD1) gene on chromosome 21 4. Transgenic mice 4,5 and rats 6 carrying mutant human SOD1 genes (G93A, G37R, G86R, G85R) have been generated, and, despite the existence of other animal models of motor neuron loss, are currently the most highly used models of the disease.
Spinal cord injury (SCI) is a heterogeneous set of conditions resulting from physical trauma to the spinal cord, with functional outcome varying according to the type, location and severity of the injury 7. Nevertheless, approximately half of human SCI cases affect cervical regions, resulting in debilitating respiratory dysfunction due to phrenic motor neuron loss and injury to descending bulbospinal respiratory axons 1. A number of animal models of SCI have been developed, with the most commonly used and clinically-relevant being the contusion 8.
Transplantation of various classes of neural precursor cells (NPCs) is a promising therapeutic strategy for treatment of traumatic CNS injuries and neurodegeneration, including ALS and SCI, because of the ability to replace lost or dysfunctional CNS cell types, provide neuroprotection, and deliver gene factors of interest 9.
Animal models of both ALS and SCI can model many clinically-relevant aspects of these diseases, including phrenic motor neuron loss and consequent respiratory compromise 10,11. In order to evaluate the efficacy of NPC-based strategies on respiratory function in these animal models of ALS and SCI, cellular interventions must be specifically directed to regions containing therapeutically relevant targets such as phrenic motor neurons. We provide a detailed protocol for multi-segmental, intraspinal transplantation of NPCs into the cervical spinal cord ventral gray matter of neurodegenerative models such as SOD1G93A mice and rats, as well as spinal cord injured rats and mice 11.

Neural precursor transplantation is a promising strategy for protecting and/or replacing lost/dysfunctional cervical phrenic motor neurons in spinal cord injury (SCI) and the motor neuron disorder, amyotrophic laterals sclerosis (ALS). We provide a protocol for cell delivery to cervical spinal cord ventral horn in rodent models of ALS and SCI.

Intra Transplante de Células de Segmentação Chifre Cervical Ventral em Esclerose Lateral Amiotrófica e lesão medular traumática

Respiratory compromise due to phrenic motor neuron loss is a debilitating consequence of a large proportion of human traumatic spinal cord injury (SCI) ...

Medicina

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory testing in rat SCI models is needed. However, until recently, no single testing approach had been validated for SCI so that standardized methods have not been implemented across labs. Additionally, available testing methods could not be implemented acutely or when severe motor impairments existed, preventing studies of the development of SCI-induced allodynia3. Here we present two validated sensory testing methods using von Frey Hair (VFH) monofilaments which quantify changes in tactile sensory thresholds after SCI4-5. One test is the well-established Up-Down test which demonstrates high sensitivity and specificity across different SCI severities when tested chronically5. The other test is a newly-developed dorsal VFH test that can be applied acutely after SCI when allodynia develops, prior to motor recovery4-5. Each VFH monofilament applies a calibrated force when touched to the skin of the hind paw until it bends. In the up-down method, alternating VFHs of higher or lower forces are used on the plantar L5 dermatome to delineate flexor withdrawal thresholds. Successively higher forces are applied until withdrawal occurs then lower force VFHs are used until withdrawal ceases. The tactile threshold reflects the force required to elicit withdrawal in 50% of the stimuli. For the new test, each VFH is applied to the dorsal L5 dermatome of the paw while the rat is supported by the examiner. The VFH stimulation occurs in ascending order of force until at least 2 of 3 applications at a given force produces paw withdrawal. Tactile sensory threshold is the lowest force to elicit withdrawal 66% of the time. Acclimation, testing and scoring procedures are described. Aberrant trials that require a retest and typical trials are defined. Animal use was approved by Ohio State University Animal Care and Use Committee.

We describe two tactile sensory testing methods for acute or chronic periods of spinal cord injury in rats. These validated procedures can detect the development and maintenance of allodynia-like sensations.

Aguda e crônica teste sensorial tátil após a lesão medular em ratos

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

A Contusive Model of Unilateral Cervical Spinal Cord Injury Using the Infinite Horizon Impactor

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of spinal cord injury (SCI) in which the thoracic spinal cord is injured. Additionally, because most human cord injuries occur as the result of blunt, non-penetrating trauma (e.g. motor vehicle accident, sporting injury) where the spinal cord is violently struck by displaced bone or soft tissues, the majority of SCI researchers are of the opinion that the most clinically relevant injury models are those in which the spinal cord is rapidly contused.1 Therefore, an important step in the preclinical evaluation of novel treatments on their way to human translation is an assessment of their efficacy in a model of contusion SCI within the cervical spinal cord. Here, we describe the technical aspects and resultant anatomical and behavioral outcomes of an unilateral contusive model of cervical SCI that employs the Infinite Horizon spinal cord injury impactor.
Sprague Dawley rats underwent a left-sided unilateral laminectomy at C5. To optimize the reproducibility of the biomechanical, functional, and histological outcomes of the injury model, we contused the spinal cords using an impact force of 150 kdyn, an impact trajectory of 22.5&deg; (animals rotated at 22.5&deg;), and an impact location off of midline of 1.4 mm. Functional recovery was assessed using the cylinder rearing test, horizontal ladder test, grooming test and modified Montoya's staircase test for up to 6 weeks, after which the spinal cords were evaluated histologically for white and grey matter sparing.
The injury model presented here imparts consistent and reproducible biomechanical forces to the spinal cord, an important feature of any experimental SCI model. This results in discrete histological damage to the lateral half of the spinal cord which is largely contained to the ipsilateral side of injury. The injury is well tolerated by the animals, but does result in functional deficits of the forelimb that are significant and sustained in the weeks following injury. The cervical unilateral injury model presented here may be a resource to researchers who wish to evaluate potentially promising therapies prior to human translation.

A reliable and repeatable way to produce a cervical unilateral spinal cord injury using the Infinite Horizon impactor is described. The method takes advantage of a custom designed frame and clamp to stabilize the spine. The standardized procedure and biomechanical injury parameters result in sufficient and sustained injuries.

Um Modelo contundente de lesão da medula espinhal cervical unilateral Usando o Impactor horizonte infinito

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of ...

Controlled Cervical Laceration Injury in Mice

Use of genetically modified mice enhances our understanding of molecular mechanisms underlying several neurological disorders such as a spinal cord injury (SCI). Freehand manual control used to produce a laceration model of SCI creates inconsistent injuries often associated with a crush or contusion component and, therefore, a novel technique was developed. Our model of cervical laceration SCI has resolved inherent difficulties with the freehand method by incorporating 1) cervical vertebral stabilization by vertebral facet fixation, 2) enhanced spinal cord exposure, and 3) creation of a reproducible laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm in depth without associated contusion. Compared to the standard methods of creating a SCI laceration such as freehand use of a scalpel or scissors, our method has produced a consistent lesion. This method is useful for studies on axonal regeneration of corticospinal, rubrospinal, and dorsal ascending tracts.

A novel technique to create a reproducible in vivo model of cervical spinal cord laceration injury in the mouse is described. This technique is based on spine stabilization by fixation of the cervical facets and laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm.

Controlada por laceração do colo do útero em camundongos

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment (RMCA) in Individuals with Chronic Spinal Cord Injury

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination is disrupted by spinal cord injury (SCI), control of respiratory muscles innervated below the injury level is compromised1,2 leading to respiratory muscle dysfunction and pulmonary complications. These conditions are among the leading causes of death in patients with SCI3. Standard pulmonary function tests that assess respiratory motor function include spirometrical and maximum airway pressure outcomes: Forced Vital Capacity (FVC), Forced Expiratory Volume in one second (FEV1), Maximal Inspiratory Pressure (PImax) and Maximal Expiratory Pressure (PEmax)4,5. These values provide indirect measurements of respiratory muscle performance6. In clinical practice and research, a surface electromyography (sEMG) recorded from respiratory muscles can be used to assess respiratory motor function and help to diagnose neuromuscular pathology. However, variability in the sEMG amplitude inhibits efforts to develop objective and direct measures of respiratory motor function6. Based on a multi-muscle sEMG approach to characterize motor control of limb muscles7, known as the voluntary response index (VRI)8, we developed an analytical tool to characterize respiratory motor control directly from sEMG data recorded from multiple respiratory muscles during the voluntary respiratory tasks. We have termed this the Respiratory Motor Control Assessment (RMCA)9. This vector analysis method quantifies the amount and distribution of activity across muscles and presents it in the form of an index that relates the degree to which sEMG output within a test-subject resembles that from a group of healthy (non-injured) controls. The resulting index value has been shown to have high face validity, sensitivity and specificity9-11. We showed previously9 that the RMCA outcomes significantly correlate with levels of SCI and pulmonary function measures. We are presenting here the method to quantitatively compare post-spinal cord injury respiratory multi-muscle activation patterns to those of healthy individuals.

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment RMCA in Individuals with Chronic Spinal Cord Injury

The purpose of this publication is to present our original work on a multi-muscle surface electromyographic approach to quantitatively characterize respiratory muscle activation patterns in individuals with chronic spinal cord injury using vector-based analysis.

Avaliação da ativação muscular respiratória Usando respiratória Motor Control Assessment (ACMD) em indivíduos com lesão medular crônica

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination ...

A Murine Model of Cervical Spinal Cord Injury to Study Post-lesional Respiratory Neuroplasticity

A cervical spinal cord injury induces permanent paralysis, and often leads to respiratory distress. To date, no efficient therapeutics have been developed to improve/ameliorate the respiratory failure following high cervical spinal cord injury (SCI). Here we propose a murine pre-clinical model of high SCI at the cervical 2 (C2) metameric level to study diverse post-lesional respiratory neuroplasticity. The technique consists of a surgical partial injury at the C2 level, which will induce a hemiparalysis of the diaphragm due to a deafferentation of the phrenic motoneurons from the respiratory centers located in the brainstem. The contralateral side of the injury remains intact and allows the animal recovery. Unlike other SCIs which affect the locomotor function (at the thoracic and lumbar level), the respiratory function does not require animal motivation and the quantification of the deficit/recovery can be easily performed (diaphragm and phrenic nerve recordings, whole body ventilation). This pre-clinical C2 SCI model is a powerful, useful, and reliable pre-clinical model to study various respiratory and non-respiratory neuroplasticity events at different levels (molecular to physiology) and to test diverse putative therapeutic strategies which might improve the respiration in SCI patients.

Respiratory failure is the leading cause of death following a cervical spinal cord injury. Having a reproducible, quantifiable, and reliable pre-clinical animal model of respiratory failure induced by a partial cervical injury will help to understand the subsequent respiratory and non-respiratory neuroplasticity and allow testing putative repair strategies.

Um modelo murino de Spinal Cord Injury do colo do útero para o Estudo de pós-lesional respiratória Neuroplasticidade

A cervical spinal cord injury induces permanent paralysis, and often leads to respiratory distress. To date, no efficient therapeutics have been developed ...

Controlled Cortical Impact Model for Traumatic Brain Injury

Every year over a million Americans suffer a traumatic brain injury (TBI). Combined with the incidence of TBIs worldwide, the physical, emotional, social, and economical effects are staggering. Therefore, further research into the effects of TBI and effective treatments is necessary. The controlled cortical impact (CCI) model induces traumatic brain injuries ranging from mild to severe. This method uses a rigid impactor to deliver mechanical energy to an intact dura exposed following a craniectomy. Impact is made under precise parameters at a set velocity to achieve a pre-determined deformation depth. Although other TBI models, such as weight drop and fluid percussion, exist, CCI is more accurate, easier to control, and most importantly, produces traumatic brain injuries similar to those seen in humans. However, no TBI model is currently able to reproduce pathological changes identical to those seen in human patients. The CCI model allows investigation into the short-term and long-term effects of TBI, such as neuronal death, memory deficits, and cerebral edema, as well as potential therapeutic treatments for TBI.

Traumatic brain injuries (TBIs) remain a serious health problem. Using the controlled cortical impact surgery model, research on the effects of TBI and possible treatment methods may be performed.

Controlado Modelo Impact Cortical para lesão cerebral traumática

Every year over a million Americans suffer a traumatic brain injury (TBI).  Combined with the incidence of TBIs worldwide, the physical, emotional, ...

A Radio-telemetric System to Monitor Cardiovascular Function in Rats with Spinal Cord Transection and Embryonic Neural Stem Cell Grafts

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter connected to a radio transmitter into the femoral artery of rats that underwent a T4 spinal cord transection with or without grafting of embryonic brainstem-derived neural stem cells expressing green fluorescent protein. Compared to other methods such as cannula insertion or tail-cuff, telemetry is advantageous to continuously monitor blood pressure and heart rate in freely moving animals. It is also capable of long term multiple data acquisitions. In spinal cord injured rats, basal cardiovascular data under unrestrained condition and autonomic dysreflexia in response to colorectal distension were successfully recorded. In addition, cardiovascular parameters before and after SCI can be compared in the same rat if a transmitter is implanted before a spinal cord transection. One limitation of the described telemetry procedure is that implantation in the femoral artery may influence the blood supply to the ipsilateral hindlimb.

We present a protocol for using a radio-telemetric system to record cardiovascular parameters in T4 spinal cord transected rats eight weeks after embryonic brainstem neural stem cell grafting into the lesion site. Telemetry is an advanced technique to accurately evaluate cardiovascular function in conscious freely moving spinal cord injured rats.

Um sistema de rádio-telemetria para monitorar a função cardiovascular em ratos com Medula Espinhal transecção e Embryonic Stem Neural enxertos de células-

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter ...

Neural Stem Cell Transplantation in Experimental Contusive Model of Spinal Cord Injury

Spinal cord injury is a devastating clinical condition, characterized by a complex of neurological dysfunctions. Animal models of spinal cord injury can be used both to investigate the biological responses to injury and to test potential therapies. Contusion or compression injury delivered to the surgically exposed spinal cord are the most widely used models of the pathology. In this report the experimental contusion is performed by using the Infinite Horizon (IH) Impactor device, which allows the creation of a reproducible injury animal model through definition of specific injury parameters. Stem cell transplantation is commonly considered a potentially useful strategy for curing this debilitating condition. Numerous studies have evaluated the effects of transplanting a variety of stem cells. Here we demonstrate an adapted method for spinal cord injury followed by tail vein injection of cells in CD1 mice. In short, we provide procedures for: i) cell labeling with a vital tracer, ii) pre-operative care of mice, iii) execution of a contusive spinal cord injury, and iv) intravenous administration of post mortem neural precursors. This contusion model can be utilized to evaluate the efficacy and safety of stem cell transplantation in a regenerative medicine approach.

Spinal cord injury is a traumatic condition that causes severe morbidity and high mortality. In this work we describe in detail a contusion model of spinal cord injury in mice followed by a transplantation of neural stem cells.

Neural Stem Cell Transplantation em Experimental contusão Model of Spinal Cord Injury

Spinal cord injury is a devastating clinical condition, characterized by a complex of neurological dysfunctions. Animal models of spinal cord injury can ...

Calibrated Forceps Model of Spinal Cord Compression Injury

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The calibrated forceps model of compression injury is a convenient, low cost, and very reproducible animal model for SCI. We used a pair of modified forceps in accordance with the method published by Plemel et al. (2008) to laterally compress the spinal cord to a distance of 0.35 mm. In this video, we will demonstrate a dorsal laminectomy to expose the spinal cord, followed by compression of the spinal cord with the modified forceps. In the video, we will also address issues related to the care of paraplegic laboratory animals. This injury model produces mice that exhibit impairment in sensation, as well as impaired hindlimb locomotor function. Furthermore, this method of injury produces consistent aberrations in the pathology of the SCI, as determined by immunohistochemical methods. After watching this video, viewers should be able to determine the necessary supplies and methods for producing SCI of various severities in the mouse for studies on SCI and/or treatments designed to mitigate impairment after injury.

Spinal cord injury models should be highly reproducible. We demonstrate that the calibrated forceps compression model of spinal cord injury is an easy to use surgical method for generating reproducible injuries to the murine spinal cord.

Calibrado Fórceps Modelo de Lesão Medular Compression

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The ...