Este trabajo es apoyado por fondos del Programa marco de la Unión Europea de la OIT (FP7/2007-2013) en virtud de acuerdo de subvención n º 246556 (proyecto RBUCE-UP Europea), HandiMedEx asignado por la Junta de Inversión Pública francés. Marcel Bonay fue apoyada por la Chancellerie des las Universidades de Paris (Piernas Poix), el Fonds de Dotation Recherche en Santé respiratoire, y el Centro de Asistencia respiratoire un domicilio d&#39;Île de France (CARDIF)

Este protocolo fue aprobado por el comité de ética de la silla RBUCE-UP de Excelencia (Universidad de Paris Sud, acuerdo de subvención n º 246556) y la Universidad de Versailles Saint-Quentin-en-Yvelines. 1. Preparación de instrumentos quirúrgicos esterilizados <ol><li> Limpiar los instrumentos quirúrgicos con detergente de laboratorio. </li><li> Autoclave los instrumentos antes de la cirugía. </li><li> En una sesión quirúrgica, esterilizar las herramientas mediante la colocación de las puntas en un esterilizador de cuentas caliente durante 10 min a 180 ° C entre 2 cirugías. </li></ol> 2. Preparación de Drogas <ol><li> Prepare 2 x 1 ml jeringas para drogas drogas preanestésica cóctel y después de la operación. </li><li> De acuerdo con el peso de la rata, preparar una jeringa con drogas preanestésica: carprofeno (5 mg / kg), buprenorfina (50 mg / kg), Baytril (5 mg / kg) y dexmedetomidina (0,5 mg / kg). Completar el volumen a 1 ml con Ringer lactato. </li><li> Preparar enotra jeringa la reversión de los medicamentos pre-anestésicos: atipamezol (500 mg / kg). </li></ol> 3. La anestesia de la rata <ol><li> Administrar por vía subcutánea en el animal la solución de drogas pre anestésicas que se describen en el paso 2.2. Luego, se coloca el animal de nuevo en la jaula y espere hasta que aparezca el efecto sedante. </li><li> Coloque la rata en una cámara cerrada llena con 5% de isoflurano en el 100% de O 2, y esperar hasta que el ritmo respiratorio se ralentiza (alrededor de 30 segundos). A continuación, retire la rata de la cámara y colóquela en la mesa de la intubación. </li></ol> 4. Intubación orotraqueal <ol><li> Acuéstese al animal en la espalda, a continuación, fije la cabeza colocando una correa unida a los dientes delanteros a la mesa. </li><li> Con una luz de fibra óptica, iluminar el espacio torácico. Luego, coloque un laringoscopio (o un encargo a uno, Jou et al. 20 para más detalles) en la boca del animal. Visualizar las cuerdas vocales. </li><li> DeslizóE y colocar una guía orotraqueal en la tráquea (entre las cuerdas vocales). Deslice el tubo orotraqueal (16 g Tamaño del catéter) en la guía. </li><li> Retire la guía y comprobar con un espejo laríngeo colocado en el extremo del tubo orotraqueal para la presencia de humedad, lo que confirma la posición apropiada del tubo en la tráquea y no en el esófago. </li><li> Conectar el tubo a un ventilador de roedor (683 roedor ventilador, Harvard Apparatus) y ajustar la concentración de isoflurano al 2% (en 100% de O 2). </li><li> Asegure el tubo orotraqueal con cinta quirúrgica. </li></ol> 5. Cirugía Espinal <ol><li> Colocar el animal en posición decúbito ventral sobre una placa quirúrgica climatizada, con la nariz apuntando al ángulo de 90 ° para el cirujano. Mantener la temperatura del cuerpo en torno a 37,5 ° C durante toda la cirugía. </li><li> Afeitar el cabello con tijeras entre las escápulas y quitar el pelo con una gasa. </li><li> Limpie la piel con betadine, y luego con 70% de alcohol. Repita este paso 3x. </li><li> Un pellizco del dedo del pie se realiza antes de inicio de la cirugía para asegurar la profundidad anestésica adecuada. &#160; A continuación, realice una incisión en la piel rostro-caudal lateral con las tijeras entre los omóplatos. </li><li> Cortar el músculo acromiotrapezius rostro-caudal siguiendo el tendón para evitar el sangrado. Entonces disociar el músculo romboide para acceder a los músculos espinales (que rodean la vértebra). </li><li> Repliegue el músculo spinalis de C1 a C3 vértebra. La vértebra C2 es el uno con una apófisis prominente. </li><li> Limpie el músculo alrededor de la parte dorsal de la vértebra utilizando hisopos de algodón estériles. </li><li> Comience a retirar con cuidado la apófisis de C2 con una gubia. Luego, seguir meticulosamente hasta que se vea la parte dorsal de la médula espinal. Asegúrese de que la laminectomía es una laminectomía hemi dorsal. Preste especial atención a la duramadre que rodea la médula espinal, y las arterias en las proximidades de esta área. </li><li> Con # 55 pinzas, diseccionar rostro-caudalmente a lo largo de la duración C2, junto continuar lateralmente a cada lado rostral y caudal. </li><li> Esponja el líquido cefalorraquídeo. </li><li> Hacer una sección lateral bajo la raíz dorsal número cervical 2 con los micro tijeras. Compruebe con un bisturí micro que la extensión de la lesión es lo suficientemente cerca para llegar a la línea media de la médula espinal (véase la figura 2A para una vista dorsal de la lesión). Si no, entonces otro corte podría hacerse para completar la lesión. En caso de sangrado, use hisopos de algodón estériles. Tenga cuidado de no ir al lado contralateral, de lo contrario, el animal no se recupere de la lesión y tendrá una insuficiencia respiratoria. </li><li> La sutura de los músculos como una capa protectora y suturar de nuevo la piel. Limpie la herida con una gasa estéril Betadine saturado. </li><li> Turn-off del vaporizador isoflurano e inyectar los fármacos de reversión (atipamezol [500 mg / kg, im]), verifique la temperatura del cuerpo. </li><li> Cuando el animal comienza a respirar contra el ventilador,desconecte el tubo traqueal del ventilador y retire el tubo orotraqueal. Colocar el animal en una jaula climatizada para la recuperación. </li></ol> 6. El cuidado post-quirúrgico Después de la cirugía, los animales son controlados continuamente para garantizar el mejor entorno posible para la recuperación. Los antibióticos (Baytril, 5 mg / kg), anti-inflamatorio (Carprofeno, 5 mg / kg) y la buprenorfina (50 mg / kg) medicamentos se administran cada 12 horas durante los primeros 2 días después de la cirugía para prevenir infecciones y reducir la ocurrencia del dolor post-operatorio. Las ratas tienen acceso ad libitum al alimento suave y agua (o agua gelificada para el 1 º día postoperatorio). Fluidos subcutáneos se pueden usar para prevenir la deshidratación en los primeros pocos días después de la operación. El peso corporal y la ingesta de alimentos son monitoreados diariamente. Su entorno se enriquece a través del experimento y el tiempo después de la lesión (doble carcasa, tubos en sus jaulas).

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Dificultades técnicas de hacer el modelo de lesión C2 El modelo murino lesión C2 es una interesante herramienta para estudiar la neuroplasticidad respiratoria post-lesional. Sin embargo, los pasos necesarios para producir un modelo reproducible y fiable son numerosos y cada uno podría tener un impacto en el resultado del estudio. Por ejemplo, durante el proceso de intubación, un cuidado extremo debe ser tomada desde el tubo oro-traqueal puede producir una inflamación d...

Traumatismo de la médula espinal es una lesión común que se observa en la población humana con incidencias dramáticas, como la parálisis permanente. Sin embargo, la gravedad de la lesión depende del nivel y la extensión de la trauma inicial. La insuficiencia respiratoria es la causa principal de mortalidad después de una lesión de la médula espinal cervical superior (SCI) 1. Actualmente, el único tratamiento terapéutico es para colocar el paciente bajo asistencia ventilatoria. Dado que algunos pacientes pueden ser destetados de la asistencia ventilatoria 2, debido a la recuperación espontánea que se produce con un retraso post-lesional, la necesidad de desarrollar nuevas terapias no invasivas innovadoras es urgente 3. Tener un buen modelo de pre-clínico estandarizado para investigar el efecto de un SCI cervical en insuficiencia respiratoria y por lo tanto, para estudiar la aplicación de estrategias terapéuticas putativas, es esencial. En este artículo técnico, se describe un modelo murino pre-clínica o específicadeterioro respiratorio f inducida por una lesión medular cervical parcial en el nivel C2. Este modelo es utilizado actualmente por varios laboratorios de todo el mundo (para revisiones: 4-13). Sin embargo, ligeras diferencias en el procedimiento quirúrgico se pueden observar entre los diferentes investigadores para generar este modelo murino lesión cervical en particular. El efecto de un C2 SCI en la salida respiratoria fue descrita por primera vez en 1895 por Porter 14. Una hemisección de cuello uterino induce una desaferenciación de las motoneuronas frénicos de su unidad central (situado en el rVRG en el tronco cerebral, la figura 1A) en el lado ipsilateral de la lesión, que conduce a una actividad del nervio frénico en silencio y la parálisis posterior de diafragma. El lado contralateral se mantiene intacto y permite que el animal para sobrevivir. A diferencia de diferente SCI situado en un segmento inferior de la columna (por ejemplo una lesión contusa a nivel C4 15), se conserva la integridad del núcleo de las neuronas motoras frénico en ambos lados. Después de un CERVlesiones C2 ical, alguna actividad espontánea se puede observar en el lado ipsilateral (frénico y diafragma) debido a una activación de las vías sinápticas silenciosos contralateral que cruzan la línea media de la médula a nivel segmental C3-C6 (vías frénicos cruzados, CPP, Figura 1B) . La activación de la CPP, que es, por definición, una hemisección C2 combinado con un phrenicotomy contralateral que inducen una recuperación parcial del nervio frénico ipsilateral, puede ocurrir a partir de horas a semanas después de la lesión 16-18. El efecto beneficioso real de esta vía de CPP en la recuperación respiratoria se limita 19 y de la investigación y el tratamiento debe ser desarrollado para mejorar la magnitud de la restauración espontánea 3. Este protocolo proporciona un tipo poderoso de modelo murino pre-clínicos para estudiar la plasticidad respiratoria post-lesional en los distintos niveles (fisiología respiratoria de las motoneuronas pre y frénicos, interneuronas, molecular y cellular, la locomoción de la extremidad delantera, por ejemplo), así como un modelo para probar estrategias terapéuticas invasivas y no invasivas destinadas a mejorar la recuperación del aparato locomotor y respiratorio después de una lesión espinal C2 parcial cervical espinal.

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a murine model of cervical spinal cord injury to study post-lesional respiratory neuroplasticity

Una lesión de la médula espinal cervical induce parálisis permanente, y, a menudo conduce a la dificultad respiratoria. Hasta la fecha, no hay terapias eficaces se han desarrollado para mejorar / mejorar la insuficiencia respiratoria después de una lesión de la médula espinal cervical alta (SCI). Aquí se propone un modelo preclínico murino de alta SCI en la cervical 2 (C2) nivel metameric estudiar diversa neuroplasticidad respiratoria post-lesional. La técnica consiste en una lesión parcial quirúrgica en el nivel C2, que inducirá un hemiparalysis del diafragma debido a una desaferenciación de los frénico motoneuronas de los centros respiratorios situados en el tronco cerebral. El lado contralateral de la lesión permanece intacta y permite la recuperación de los animales. A diferencia de otros LIC que afectan la función locomotora (en la columna torácica y lumbar de nivel), la función respiratoria no requiere la motivación de los animales y la cuantificación del déficit / recuperación se puede realizar fácilmente (diafragma y la grabación del nervio frénicos, la ventilación de todo el cuerpo). Este modelo C2 SCI preclínica es un potente, útil y fiable modelo preclínico para estudiar diversos eventos de neuroplasticidad respiratorias y no respiratorias en diferentes niveles (molecular a la fisiología) y para probar diversas estrategias terapéuticas supuestos que podrían mejorar la respiración en pacientes con lesión medular.

Extensión de la lesión El éxito y la reproducibilidad de este modelo experimental particular, dependen de la experiencia de cada manipulador / cirujano. La cantidad subsiguiente de recuperación de las vías respiratorias (la actividad del nervio frénico y la actividad de diafragma) después de una lesión C2 está correlacionada con la ventrolateral restante salvó la materia blanca 21. Dado que la lesión es &quot;hecho a mano&quot; y requiere un poco de pr?...

Watch this Scientific Journal Video about A Murine Model of Cervical Spinal Cord Injury to Study Post-lesional Respiratory Neuroplasticity at JoVE.com

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

La insuficiencia respiratoria es la principal causa de muerte a raíz de una lesión de la médula espinal cervical. Tener un modelo animal preclínico reproducible, cuantificable y confiable de la insuficiencia respiratoria inducida por una lesión cervical parcial le ayudará a comprender la neuroplasticidad respiratoria y no respiratoria posterior y permiten probar estrategias de reparación putativos.

Un modelo murino de la médula espinal cervical Lesión en estudio post-lesional Respiratorio Neuroplasticidad

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

a-murine-model-cervical-spinal-cord-injury-to-study-post-lesional

Research

JoVE Journal

Medicine

13.4K vistas.  Université de Versailles Saint-Quentin-en-Yvelines.  La insuficiencia respiratoria es la principal causa de muerte a raíz de una lesión de la médula espinal cervical. Tener un modelo animal preclínico reproducible, cuantificable y confiable de la insuficiencia respiratoria inducida por una lesión cervical parcial le ayudará a comprender la neuroplasticidad respiratoria y no respiratoria posterior y permiten probar estrategias de reparación putativos. 

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

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.

Intraespinal trasplante de células de Orientación asta ventral del cuello uterino en la esclerosis lateral amiotrófica y la lesión de la médula espinal

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

Investigación

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 y crónica Prueba sensorial táctil después de la lesión de la médula espinal en ratas

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.

Un modelo de la contundente unilateral Lesión Medular Cervical Uso del Impactador Infinite Horizon

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.

Controlado lesiones laceración cervical en ratones

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.

Un modelo Contusión de la lesión de la médula espinal severa en ratas

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

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.

Evaluación de activación muscular respiratoria Utilizar evaluaciones de control de motores respiratoria (RMCA) en personas con lesión de la médula espinal crónica

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

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 de Impacto cortical de la lesión 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.

Un sistema de radio-telemetría para monitorizar la función cardiovascular en ratas con la médula espinal y transección Embryonic Stem Neural injertos 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.

El uso sinérgico de las células precursoras neuronales y Péptidos de autoensamblaje en Experimental cervical Lesión de la Médula Espinal

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

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 Pinzas Modelo de Lesión Medular Compresión

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