1. Configure: Frame e Design Pinça para Segurar o Animal <ol><li> A moldura e grampo para prender o animal foi customizado para acomodar o horizonte infinito (IH) Impactor de lesão medular. </li><li> A base do quadro é uma plataforma de alumínio cortado em as seguintes dimensões (30,2 cm x 20,3 cm x 1,3 cm), a fim de se encaixar no suporte de guia tabela que vem padrão com o dispositivo IH (Figura 1A). </li><li> Quatro Flexaframe placas de apoio do pé (Fisher Scientific, Toronto, ON) estão ligados à plataforma e oito hastes Flexaframe de apoio (Fisher Scientific, Toronto, ON, 30,5 cm) são montados utilizando oito conectores Flexaframe de apoio (Figura 1B). </li><li> Dois conectores Flexaframe de suporte adicionais, inerentes aos valores médios duas varas, a casa feita sob encomenda vale para o grampo (Figura 1B). </li><li> O ângulo pelo qual medula espinal do animal é rodado em relação à vertical é estabelecida por keeping uma da haste horizontal no lugar e variando a altura da haste de outro horizontal (Figura 2). </li><li> O grampo é 35,6 milímetros de comprimento, 25,4 milímetros de altura e com uma mandíbula milímetros 7,6 concebido para agarrar rigidamente sob o processo transverso de C4 a C6 (Figura 1C). Detalhes adicionais sobre o desenho grampo têm sido descritos anteriormente por Choo et ai., 2009. </li></ol> 2. Cirurgia <ol><li> Macho Sprague Dawley (Charles River Laboratories) pesando 300-350 g foram anestesiados com isofluorano (4% para a indução e 2% para manutenção) em oxigénio (1 L / min). </li><li> Uma vez que os animais estão sob o plano da anestesia, o animal é colocado em um quadro estereotáxico (Kopf, Tujunga, CA). </li><li> Para minimizar a hemorragia durante a cirurgia mL procedimento de lidocaína 0,4 (20 mg / mL; Bimeda - MTC Animal Health Inc., Cambridge, Ontario, Canadá) com epinefrina é injectado por via intramuscular em torno do local cirúrgico no dorsoregião do pescoço l. </li><li> Um 4-5 centímetros incisão na linha média dorsal é feita usando um bisturi estéril (# 15), a partir da base do crânio e estendendo caudalmente. </li><li> Pinças esterilizadas Adson sem rodeios são utilizados para dissecar através da musculatura dorsal para alcançar a coluna, e um afastador Alm estéril (Ferramentas Ciência Belas, North Vancouver, BC) é inserido para manter os músculos separados. </li><li> Usando o bisturi # 15, os músculos sobreposição de lâminas de C4-C7 são raspadas, iniciando na linha média e varrendo-os lateralmente. </li><li> Uma incisão com um bisturi estéril (# 15) é feita para o músculo ligado aos processos transversais em ambos os lados da espinha, a fim de encaixar a braçadeira por baixo dos processos transversais de C4 a C6. </li><li> Com uma multa estéril ponta Friedman-Pearson Rongeur (Ferramentas Ciência Belas, North Vancouver, BC), a lâmina C5 esquerda é cuidadosamente removida para visualizar a dura ea medula espinhal. </li><li> Uma haste com um diâmetro de 1,5 mm é deslizado por baixo dos braçospara sustentar o animal para cima, elevando assim a coluna um pouco e facilitar a inserção de fixação. </li><li> Monte a mandíbula da pinça estéril para laterais processos transversos de C4 a C6 e aperte os parafusos. </li><li> Retire o afastador Alm. </li></ol> 3. Lesão Medular <ol><li> Depois de a braçadeira é montado sobre o animal, o animal é movido para o pêndulo IH. </li><li> O grampo é inserida nos suportes de metal sobre as duas hastes meio da armação que foram protegidos com um ângulo de 22,5 ° fora de horizontal (Figura 2). </li><li> A tomada de tesoura que oferece ajustes de altura estável, (VWR, Mississauga, ON) é levantada até que o animal está plano. </li><li> Assegure-se que o grampo é horizontal pela colocação de um pequeno nível de cilindro na parte superior do grampo e apertando os parafusos. É importante que todos os parafusos estão apertados ea configuração é rígida, sem qualquer movimento. </li><li> O resto do procedimento é realizado sob uma microscope (Leica MZ8). </li><li> Abaixar e têm como objectivo a ponta do pêndulo (15 mm de diâmetro, com arestas arredondadas), utilizando o botão de ajuste vertical e os dois botões de ajuste horizontal no pêndulo IH até que o centro da ponta do pêndulo paira acima do vértice do C6 processo espinal. </li><li> Uma vez que a ponta do pêndulo está centrada, girar o botão de ajuste do eixo y um e dois por sua vez, quinto (1,4 mm) para mover a ponta lateralmente para o lado esquerdo e do eixo x botão de ajuste horizontal para mover a ponta do pêndulo para o centro do C5. </li><li> Abaixar a ponta até que é um pouco acima da dura a fim de verificar que a ponta do pêndulo é destinada a metade lateral da matéria cinzenta. </li><li> Gire o botão de ajuste vertical de duas voltas para levantar a ponta 4 mm acima da dura-máter. </li><li> Certifique-se que a área de impacto é seca utilizando um cotonete ou vara. </li><li> Definir a força desejada a 150 kdyn sobre o programa e clique em &quot;Iniciar Experiment&quot; para acionar o pêndulo. </li><li> Após a lesão, a ferida é fechada em layers com Vicryl 5-0. A buprenorfina (0,03 mg / kg SC, Temgesic; Schering-Plough Corporation, Kenilworth, NJ) e solução salina (10 mL) é administrada por via subcutânea duas vezes ao dia antes e durante dois dias após a cirurgia. Os animais são cuidadosamente monitorizados duas vezes por dia durante 2 semanas e uma vez por semana durante 6 semanas pós-lesão. </li></ol> 4. Os resultados representativos Vinte e nove macho Sprague Dawley (Charles River Laboratories) pesando 300-350 g foram feridas com uma definição de força de 150 kdyn. A ponta do pêndulo foi destinado 1,4 milímetros lateral à linha mediana, a um ângulo de 22,5 ° fora da vertical. A força média real foi de 155,55 ± 0,73 kdyn. O deslocamento média foi de 1512,72 ± 27,86 uM ea velocidade foi 120,24 ± 0,52 mm / s (Figura 3). Desfechos Comportamento A recuperação funcional foi avaliada utilizando a horizontal escada, teste de criação do cilindro, da preparação de teste,e modificado Montoya teste escada 1. Os animais foram treinados antes da lesão e avaliada em 2, 4 e 6 semanas pós-lesão. Houve prejuízos forelimb significativas sofridas ao longo do período experimental. Teste de Escada Horizontal. Antes da lesão, os animais feita apenas 4,75 ± 0,73% erros no membro anterior ipsilateral ao percorrer toda a escada espaçadas irregularmente horizontal. Após a lesão, os animais demonstraram aumento significativo na porcentagem de erros membro anterior. Os erros por cento ipsilaterais forelimb foram 26,97 ± 2,92%, 26,23 ± 2,84% e 22,06 ± 2,05% a 2, 4 e 6 semanas após a lesão, respectivamente (Figura 4A). Importante, a insuficiência forelimb sobre este teste foi mantida durante as 6 semanas. Criação de teste do cilindro. O percentual de forelimb ipsilateral (esquerdo + ambos) o uso durante a exploração diminuiu significativamente após a SCI.Antes da lesão, os animais utilizados no membro anterior ipsilateral 75,12 ± 2,25%. Após a lesão, os animais utilizados no membro anterior ipsilateral 8,59 ± 1,80% em 2 semanas, 14,25 ± 2,65% a 4 semanas e 11,76 ± 2,66% em 6 semanas (Figura 4B). Modificado Montoya teste Staircase. O número de pelotas recuperados com o membro anterior ipsilateral diminuiu drasticamente após a lesão. Antes da lesão, os animais recolhidos 84,85 ± 2,88% das recompensas alimentares. No entanto, aos 2, 4 e 6 semanas após a lesão, os animais recuperados somente 30,91 ± 4,03%, 28,94 ± 4,38% e 25,86 ± 3,09% das pelotas (Figura 4C). Higiene teste. Houve quedas dramáticas nos escores de cuidados especiais após a lesão. Depois de SCI, as pontuações ipsilaterais coçar foram 2 semanas: 2,00 ± 0,17, 4 semanas: 1,83 ± 0,17 e 6 semanas: 1,79 ± 0,11 (FIGUre 4D). Resultados histológicos A massa branca e poupadores matéria Grey. Um exemplo de uma lesão da medula espinal, a uma força de 150 kdyn, o ângulo de 22,5 ° e têm como objectivo lateral de 1,4 milímetros é apresentada na Figura 5. A lesão resultou em danos substanciais à substância branca e cinzenta no lado ipsilateral. Ambos corticoespinhal e vias rubroespinal ficaram feridas e 23 dos 29 animais apresentaram lesão parenquimatosa contido no lado ipsilateral. A extensão longitudinal do dano da matéria branca e cinzenta foi 2400 e 2400 rostralmente caudalmente (Figura 6). Ao adicionar as seções para fornecer uma estimativa grosseira do &quot;spread cumulativo&quot; de branco e cinza poupador matéria (2000 mM rostral e caudal do epicentro), o lado ipsilateral teve apenas 51,8% da substância branca poupado e 39,7% da massa cinzenta comparados remanescente para o lado contralateral (Figura 6). &lt;img alt = &quot;Figura 1&quot; src = &quot;files/ftp_upload/3313/3313fig1.jpg /&quot; /&gt; Figura 1. Horizonte Infinito impactor lesão medular. A. Impactor horizonte infinito e global definida. B. Quadro configurar. C. imagem Close-up do grampo para a realização de processos transversos cervicais. D. Especificação (unidade: polegada) do grampo. Tolerância sugerida &lt;0,002 em (Choo et al. De 2009). <a href="http://www.jove.com/files/ftp_upload/3313/3313fig1large.jpg" target="_blank">Clique aqui para ver maior figura</a> . <img alt="A Figura 2" src="/files/ftp_upload/3313/3313fig2.jpg" /> Figura 2. A. Moldura ilustração e correspondente configurado para atingir a rotação da medula espinal de 0 ° (neutro) ou B. 22,5 °, com o objectivo lateral de 1,4 mm. <img alt="A Figura 3" src="/files/ftp_upload/3313/3313fig3.jpg" /> Figura 3. Força representativa e gráficos de deslocamento para impactor horizonte infinito. A seta indicates o tempo em que a ponta do pêndulo atingiu 20 kdyn eo ponto quando a gravação de deslocamento é iniciada. A força real é lido a partir do pico da curva de força versus tempo eo deslocamento correspondente é calculado. Os gráficos abaixo mostram uma típica 150 contusão kdyn, Deslocamento A. vs gráfico de tempo, B. Força vs gráfico Time. Estes gráficos mostram que a força real obtido foi de 152 kdyn, eo deslocamento do pêndulo para a medula foi medido como sendo de 1287 uM. <a href="/files/ftp_upload/3313/3313fig3large.jpg" target="_blank">para ver figura maior</a> . <img alt="A Figura 4" src="/files/ftp_upload/3313/3313fig4.jpg" /> Figura 4. Avaliações comportamentais para a força de 150 kdyn, angulação do pêndulo de 22,5 ° e têm como objectivo de 1,4 mm à esquerda da linha média. A. Teste de escada horizontal. B. teste criação do cilindro. C. Modificado teste escadaria Montoya. D. Higiene teste. O membro anterior ipsilateral resultou em significativa e SusNed deficiências em comparação com o membro anterior contralateral. <img alt="A Figura 5" src="/files/ftp_upload/3313/3313fig5.jpg" /> Figura 5. Imagens representativas da medula lesionada de rostral caudal de 1600 mm a 1600 uM. <img alt="A Figura 6" src="/files/ftp_upload/3313/3313fig6.jpg" /> Figura 6. Avaliações histológicas. A. Matéria por cento branco e cinza poupado. Importa cerca de 2000 mM de o epicentro da lesão foi em grande parte poupada. B. cumulativa branco e cinza assunto poupado com 2.000 mM de o epicentro da lesão. O lado ipsilateral tinham significativamente menos poupado substâncias branca e cinzenta em relação ao lado contralateral. <a href="/files/ftp_upload/3313/3313fig6large.jpg" target="_blank">Clique aqui para ver maior figura</a> .

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Neste artigo, descrevemos um modelo de contusão cervical unilateral com o horizonte infinito (IH) impactor, a uma força de 150 kdyn, um ângulo de 22,5 ° fora da vertical, e um objectivo lateral, de 1,4 mm da linha média. Com essas configurações, fomos capazes de produzir sofridos déficits comportamentais no membro anterior ipsilateral com o rompimento do parênquima contido em grande parte para o lado ipsilateral, onde verificou-se que ocorreu um dano considerável para as regiões onde as vias rubroespinal, ret...

<table border="1"><tbody><tr><td> Nome do equipamento </td><td> Companhia </td><td> Número de catálogo </td><td> Comentários </td></tr><tr><td> Impactor horizonte infinito </td><td> Sistemas de precisão e Instrumentação </td><td> IH-0400 </td><td></td></tr><tr><td> Folha de metal de alumínio </td><td> Metalsupermarlets.com </td><td> APT6061/500 </td><td></td></tr><tr><td> Pés Flexaframe placas de suporte </td><td> Fishers Científico </td><td> 1466625Q </td><td></td></tr><tr><td> Apoio hastes Flexaframe </td><td> Fishers Científico </td><td> 1466610GQ </td><td></td></tr><tr><td> Flexaframe Suporte Conectores </td><td> Fishers Científico </td><td> 1466620Q </td><td></td></tr><tr><td> Grampo 1 </td><td> Custom Made </td><td></td><td> Choo et al., 2009 </td></tr><tr><td> Titulares de metal </td><td> Custom Made </td><td></td><td> Veja acima </td></tr><tr><td> Impactor ponta </td><td> Custom Made </td><td></td><td> Diâmetro: 1,15 milímetros </td></tr><tr><td> Estereotáxica </td><td> David Kopf Instruments </td><td> Modelo 900 </td><td></td></tr><tr><td> Nível Cilindro </td><td> Yijia FERRAMENTAS </td><td> YJ-SL0620 </td><td></td></tr><tr><td> Microscópio </td><td> Leica </td><td></td><td> Modelo #: MZ8 </td></tr><tr><td> Laboratório tesoura jack </td><td> VWR </td><td> 12620-902 </td><td></td></tr></tbody></table>

a contusive model of unilateral cervical spinal cord injury using the infinite horizon impactor

Embora a maioria dos humanos lesões da medula espinhal ocorrer na medula cervical, a grande maioria da pesquisa de laboratório emprega modelos animais de lesão da medula espinal (SCI), na qual o cabo vertebral torácica é ferido. Além disso, porque a maioria dos humanos lesões da medula ocorrer como resultado de romba, trauma não penetrante (acidente de veículo automóvel, por exemplo, lesões desportivas) onde a medula espinal é violentamente atingido por osso deslocado ou tecidos moles, a maioria dos investigadores SCI são da opinião que a maioria dos modelos de lesão clinicamente relevantes são aqueles em que a medula espinal é rapidamente contundido. 1 Portanto, um passo importante para a avaliação pré-clínica de novos tratamentos no seu caminho para a tradução humana é uma avaliação da sua eficácia num modelo de contusão SCI dentro A medula cervical. Aqui, descrevemos os aspectos técnicos e consequentes resultados anatômicos e comportamentais de um modelo unilateral contundente da cervical SCI que emprega oHorizonte Infinito impactor lesão medular. Ratos Sprague Dawley foram submetidos a uma laminectomia unilateral do lado esquerdo no C5. Para optimizar a reprodutibilidade dos resultados biomecânicos, funcional e histológica do modelo de lesão, contundido as medulas espinais, utilizando uma força de impacto de 150 kdyn, uma trajectória de impacto de 22,5 ° (animais rodado a 22,5 °), e um local de impacto fora da linha média de 1,4 mm. A recuperação funcional foi avaliada através do cilindro de criação, teste escada horizontal, preparação e teste modificado Montoya escadaria até 6 semanas, após o que as medulas espinhais foram avaliados histologicamente para poupador substâncias branca e cinzenta. O modelo de lesão aqui apresentado transmite consistentes e reprodutíveis forças biomecânicas para a medula espinal, uma característica importante de qualquer modelo experimental SCI. Isto resulta em danos histológica discreta para a metade lateral da medula espinal, que é em grande parte contido em tele lado ipsilateral da lesão. A lesão é bem tolerado pelos animais, mas não resulta em déficits funcionais do membro anterior, que são significativa e sustentada nas últimas semanas após a lesão. O modelo de lesão cervical unilateral apresentado aqui pode ser um recurso para pesquisadores que desejam avaliar terapias potencialmente promissores antes da tradução humana.

Watch this Scientific Journal Video about A Contusive Model of Unilateral Cervical Spinal Cord Injury Using the Infinite Horizon Impactor at JoVE.com

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

Uma maneira confiável e repetível para produzir uma lesão da medula espinhal cervical unilateral utilizando o pêndulo horizonte infinito é descrito. O método leva vantagem de um quadro projetado e grampo para estabilizar a coluna. O procedimento padronizado e biomecânica lesão os parâmetros resultam em lesões suficientes e sustentados.

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 ...

a-contusive-model-unilateral-cervical-spinal-cord-injury-using

Research

JoVE Journal

Medicine

19.3K visualizações.  University of British Columbia. Uma maneira confiável e repetível para produzir uma lesão da medula espinhal cervical unilateral utilizando o pêndulo horizonte infinito é descrito. O método leva vantagem de um quadro projetado e grampo para estabilizar a coluna. O procedimento padronizado e biomecânica lesão os parâmetros resultam em lesões suficientes e sustentados.

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

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 ...

Medicina

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 ...

A Contusion Model of Severe Spinal Cord Injury in Rats

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is described to obtain a consistent model of severe SCI. Use of a stereotactic frame and computer controlled impactor allows for creation of reproducible injury. Hypothermia and urinary tract infection pose significant challenges in the post-operative period. Careful monitoring of animals with daily weight recording and bladder expression allows for early detection of post-operative complications. The functional results of this contusion model are equivalent to transection models. The contusion model can be utilized to evaluate the efficacy of both neuroprotective and neuroregenerative approaches.

A contusion model of severe spinal cord injury is described. Detailed pre-operative, operative and post-operative steps are described to obtain a consistent model.

Um modelo de contusão grave de lesão medular em ratos

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is ...

A Novel Vertebral Stabilization Method for Producing  Contusive Spinal Cord Injury

Clinically-relevant animal cervical spinal cord injury (SCI) models are essential for developing and testing potential therapies; however, producing reliable cervical SCI is difficult due to lack of satisfactory methods of vertebral stabilization. The conventional method to stabilize the spine is to suspend the rostral and caudal cervical spine via clamps attached to cervical spinous processes. &#160;However, this method of stabilization fails to prevent tissue yielding during the contusion as the cervical spinal processes are too short to be effectively secured by the clamps (Figure 1). &#160;Here we introduce a new method to completely stabilize the cervical vertebra at the same level of the impact injury. &#160;This method effectively minimizes movement of the spinal column at the site of impact, which greatly improves the production of consistent SCIs. &#160;We provide visual description of the equipment (Figure 2-4), methods, and a step-by-step protocol for the stabilization of the cervical 5 vertebra (C5) of adult rats, to perform laminectomy (Figure 5) and produce a contusive SCI thereafter. &#160;Although we only demonstrate a cervical hemi-contusion using the NYU/MASCIS impactor device, this vertebral stabilization technique can be applied to other regions of the spinal cord, or be adapted to other SCI devices. &#160;Improving spinal cord exposure and fixation through vertebral stabilization may be valuable for producing consistent and reliable injuries to the spinal cord. &#160;This vertebral stabilization method can also be used for stereotactic injections of cells and tracers, and for imaging using two-photon microscopy in various neurobiological studies.

Vertebral stabilization is necessary for minimizing variability, and for producing consistent experimental spinal cord injuries.&#160; Using a customized stabilizing apparatus in conjunction with the NYU/MASCIS impactor device, we have demonstrated here the proper equipment and procedure for generating reproducible hemi-contusive cervical (C5) spinal cord injuries in adult rats.

Um método de estabilização Novel Vertebral para a Produção de contusão Lesão Medular

Clinically-relevant animal cervical spinal cord injury (SCI) models are essential for developing and testing potential therapies; however, producing ...

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 ...

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

Spinal cord injuries (SCI) cause serious neurological impairment and psychological, economic, and social consequences for patients and their families. Clinically, more than 50% of SCI affect the cervical spine1. As a consequence of the primary injury, a cascade of secondary mechanisms including inflammation, apoptosis, and demyelination occur finally leading to tissue scarring and development of intramedullary cavities2,3. Both represent physical and chemical barriers to cell transplantation, integration, and regeneration. Therefore, shaping the inhibitory environment and bridging cavities to create a supportive milieu for cell transplantation and regeneration is a promising therapeutic target4. Here, a contusion/compression model of cervical SCI using an aneurysm clip is described. This model is more clinically relevant than other experimental models, since complete transection or ruptures of the cord are rare. Also in comparison to the weight drop model, which in particular damage the dorsum columns, circumferential compression of the spinal cord appears advantageous. Clip closing force and duration can be adjusted to achieve different injury severity. A ring spring facilitates precise calibration and constancy of clip force. Under physiological conditions, synthetic self-assembling peptides (SAP) self-assemble into nanofibers and thus, are appealing for application in SCI5. They can be injected directly into the lesion minimizing damage to the cord. SAPs are biocompatible structures erecting scaffolds to bridge intramedullary cavities and thus, equip the damaged cord for regenerative treatments. K2(QL)6K2 (QL6) is a novel SAP introduced by Dong et al.6 In comparison to other peptides, QL6 self-assembles into &#946;-sheets at neutral pH6.14 days after SCI, after the acute stage, SAPs are injected into the center of the lesion and neural precursor cells (NPC) are injected into adjacent dorsal columns. In order to support cell survival, transplantation is combined with continuous subdural administration of growth factors by osmotic micro pumps for 7 days.

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. ...

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 ...