作者是感谢N. Banik博士和博士D.米切尔在这种模式发展的指导。

1。脊髓损伤前的准备工作在此过程中所需要的手术器械手术刀，皮卡和没有牙齿，止血，自护拉钩，精尖咬骨钳，针驱动器，可吸收缝线，皮肤剪辑喷头的。其他所需的手术用品手术单，手术野的无菌床单，纱布海绵，棉花尖端涂药，金属箔。高压灭菌器在手术之前，手术器械和用品。使用单独的一套工具和用品的一个动物。清洁手术区和设备（冲击，光源，立体框架，玻璃珠灭菌器，电热垫）用酒精湿巾。 打开手术布单，并使用无菌手套，悬垂外科领域小心避免污染。打开各个工具无菌，并小心地将那些在外科领域。盖个旋钮和手柄ê设备可能需要在手术过程中无菌的金属箔。开关上的玻璃珠灭菌是在操作过程中可以使用。 2。动物的制备把老鼠的实验室面积几个小时前的实际过程。管理止痛药，至少提前一个小时预期的过程（通常buprinorphine 0.006毫克/毫升皮下注射1.5毫升）。辖术前抗生素（通常为4毫克/千克拜有利皮下）。使用90毫克/公斤的氯胺酮麻醉大鼠，等到有没有脚趾捏响应。触诊最突出的胸椎棘突。这个水平通常与T10棘突。标记的位置有关T10的预期水平。在我们的实验室中，我们执行T10受伤。下列叙述中描述的技术，T10 SCI。剃须3厘米×6厘米的矩形纵向集中在T10级别。清洁皮肤三次无线次优碘溶液。应用眼科每只眼睛上的润滑剂。小心避免污染大鼠在一个舒适的位置转移到外科领域。插入直肠温度探头监控核心温度，并相应地调节加热温度接近正常（〜37.5°C）尽可能保持动物。封面用外科手术部位上面的窗口，悬垂与大鼠。 3。外科手术开始，大约4厘米的切口，使用＃10刀片集中在T10标志。继续耐心远离T9-T11棘突和椎板解剖筋膜和肌肉层。放置牵开器缩回肌肉和筋膜相差的骨。虽然稳定T9棘突大幅划分T9和T10之间的棘间韧带，用细剪刀。同样稳定的T10棘突和T10-T11之间划分棘间韧带。使用循环倍率COMPLE的德分工韧带黄韧带（ 图1），一路过关斩将。硬膜囊是显而易见的，一旦黄韧带断开。使用精尖咬骨钳，认真履行零碎在T10椎板切除双侧。采取极为谨慎，以避免咬骨钳提示硬膜囊和意外伤害的下行压力。 T10椎板和棘突完全去除。 将动物的稳定平台上的位置。使用稳定夹具夹紧然后由横向方面的T9椎体T11椎体的横向方面，固定脊柱。要小心，不要压缩大鼠平台稳定夹，因为这会限制呼吸运动的空间和增加不必要的压力动物。固定后，在脊柱上的计算机控制的冲击器的设置检查。我们通常使用3.0毫米的冲击尖在SP度的影响的深度为2毫米的4厘米/秒和停留时间为0.3秒。扩展撞击尖和降低它，直到它刚好接触线表面。缩回的尖端和降低对脊髓表面的装置2毫米。推出活塞在4厘米/秒，导致严重挫伤脊髓损伤。小心，不要制造任何不必要的压力线只用足够的压力，保持棉花尖端涂药，达到止血效果。缝合肌肉和筋膜层一个数字8针小心不要拉得太紧，使用可吸收缝合线。合的皮肤用最少2小钉书钉;多达4或5可以使用，如果切口部分保持开放后的第2个或3个订书​​钉的订书钉。 3。程序后护理将大鼠手术后24小时约33-35℃，在温暖的环境。这需要一个孵化器（而他们是无意识的）和一个加热笼的空间，一旦他们开始移动。一旦老鼠是完全AW阿克，管理5ml的生理盐水，1.5毫升（0.006毫克/毫升）buprinorphine，和0.1毫升的拜有利所有皮下。第24-48小时，拜有利皮下继续buprinorphine，一天两次，一次，连用7天。膀胱应手动表示1日3次，直到膀胱功能返回（&lt;2毫升尿液，连续3天清晨表达）。在这段时间内感染（尿中有血，白浊色泽，或难闻的气味），动物也应检查，体力活动减少或伤口愈合的问题。存在感染，应咨询当地兽医抗生素的用量增加（或重新启动）反驳。称量大鼠每日开始手术后的第二天，以评估其恢复。

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财务信息的披露无 资金披露没有。

最近，一些新的治疗方法已经显示出早期的承诺在该领域的SCI研究。，3仔细评估这些治疗脊髓损伤的临床相关模型与最大平移潜力的选择策略是必不可少的。分级计划是最近开发的临床前研究评估实力。该方案强调利用挫伤严重的脊髓损伤模型的重要性。在这里，我们描述了这样一个严重的脊髓损伤，挫伤模型一致的病变体积和功能评分，类似横断模型。这种模式可能被用来作为一个"概念证明&#39;建立疗效的神经保护和神经再生战略。 代统一挫伤SCI仍然具有挑战性。重现性，很重要的是尽可能均匀地进行损伤。必须确定一贯FR针对性的脊髓水平OM动物动物。去除的骨椎板切除应做仔细，以确保无骨碎片留在椎管，这些可能会导致压迫性损伤，并引入不必要的损伤机制的变化和恢复潜力。我们已经采取了几个步骤，以确保包括大小均匀的冲击，刚性稳定脊柱的帧装稳定武器，使用一个统一的撞击速度和撞击的深度。重要的是，大鼠舒适的位置时的稳定钳子将被锁定到椎体。脊柱排列或过剩的压力在夹紧任何变化可能会改变的生物力学影响。我们使用了一个2.5毫米的冲击，因为在本地区的利益大鼠脊髓的平均规模为2.5-3毫米。在我们的实验在0.3秒的接触时间或停留均匀设置。然而，在我们的观察和其他实验室的报告，对于严重的脊髓损伤模型的深度的影响是最关键的的参数。在本文中所描述的实验中被送到冲击在一个恒定的深度为2毫米。影响均匀性的其他因素，包括人员培训，清晰的可视化组织，以确保撞击尖直接影响脊髓组织（而不是骨性结构）。 老鼠应该不断地观察，必要的生命体征，在手术过程中最重要的核心体温和呼吸。低温麻醉管理期间和之后的主要死亡原因。监测直肠探头的核心温度和适当的加热垫，将避免这种并发症。如果呼吸不规则或的动物停止呼吸，应立即停止该过程，直到呼吸回报基线。过度疼痛或疼痛和麻醉用药不慎过量服用还可能导致呼吸困难。大鼠应仔细监测程序后。他们的瓦特八分应当准确计量，并从基线体重&gt; 20％的损失，应及时调查，食物和水的摄入量，尿路感染，皮肤破裂，后SCI肠梗阻，等兽医人员进行评估和治疗早期咨询是至关重要的在这种情况下。如前所述，大鼠14天没有恢复膀胱控制应开始回到抗生素，以避免潜在的致命性尿路感染。 脊髓损伤可以由一个位移法或恒定的力法诱导5,12本文中所描述的技术，位移法，力传递到固定的深度内的脊髓组织。允许使用计算机控制的撞击实验SCI其他常见的方法，是不是达到一定程度的控制。剪辑压缩技术不允许的钝挫伤力立即释放，而NYU影响或者依赖于重力挫伤确定力。这些技术虽然重现性好，不容许条件的模拟过程中遇到的机动车事故人类急性脊髓损伤，最常见的原因。以往的研究表明，损伤的严重程度有直接关系的影响深度比速度的影响。7因此，这种模式允许一个严重的SCI经常进行修改的严重性有潜力通过更大范围的组合力和位移。

选择适当的损伤模型是至关重要的新型治疗脊髓损伤（SCI）的临床前评价。1,2,13神经损伤挫伤模型在现场的医生和科学家在最近的一项调查，而不是对半切或完全横断模型，被普遍接受的是临床相关性。，8此意见的基础上观察，大多数在人类脊髓损伤的挫伤性质。生物学挫伤似乎也从半切或横断模型是不同的。11伊势田， 等 。 人比较软骨素酶ABC椎管内注射对半切和挫伤模型分别在神经再生的效果。观察在神经桥半切，但没有挫伤SCI组4轴突再生。半切或完全横断模型创造条件已知存在的只是一个很小的子集CL情况inical。例如，一些研究者已经采用植入支架为基础的干预病变腔半切后或完全横断，促进肝细胞再生。这种方法成为临床无关，因为创作的一腔损伤脊髓内是不切实际的，可能是不道德的。 变异功能恢复仍是一个重大的挑战挫伤模型。5,12影响力均匀交付前横贯性脊髓体积，尤其是位于腹侧运动通路，这种变化可以最小化通过使用计算机控制的冲击和稳定脊柱。必须指出的是存活轴突可塑性和抵押品的贡献是主要的脊髓损伤后的恢复机制。1因此，即使轻微挫伤技术变化可能会产生明显不同的结果。为此，我们已经开发出严重脊髓损伤产生一致挫伤体积和功能的恢复与横断模型的典范。此模型可用于神经保护和神经再生策略进行调查作为一个概念证明疗效。

<table border="1">
<tbody>
 <tr>
 <td>Instrument/Drugs</td>
 <td>Company</td>
 <td>Cat #</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>Computer controlled impactor</td>
 <td>Leica or the Infinite Horizons (formerly OSU) impactor</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Surgical instruments </td>
 </tr>
 <tr>
 <td>Scissors</td>
 <td>Fine Science Tools Inc
 </td>
 <td>14094-11 or 14060-09</td>
 <td></td>
 </tr>
 <tr>
 <td>Forceps</td>
 <td>Fine Science Tools Inc </td>
 <td>11006-12 and 11027-12 or 			11506-12</td>
 <td></td>
 </tr>
 <tr>
 <td>Hemostats</td>
 <td>Fine Science Tools Inc </td>
 <td>13009-12</td>
 <td></td>
 </tr>
 <tr>
 <td>Retractors</td>
 <td>Fine Science Tools Inc </td>
 <td>17011-10</td>
 <td></td>
 </tr>
 <tr>
 <td>Rongeurs</td>
 <td>Fine Science Tools Inc </td>
 <td>16020-14</td>
 <td></td>
 </tr>
 <tr>
 <td>Needle driver</td>
 <td>Fine Science Tools Inc </td>
 <td>12001-13</td>
 <td></td>
 </tr>
 <tr>
 <td>Stereotactic frame</td>
 <td> Leica or RWD Life Science Co. or
 TSE systems</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Buprinorphine</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Baytril</td>
 <td>Bayer</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Ketamine</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 </tbody>
</table>

a contusion model of severe spinal cord injury in rats

平移潜在的新的治疗方法，应追究严重的脊髓损伤（SCI）挫伤模型。描述了详细的方法，获得了一致的模型严重的脊髓损伤。使用一个立体框架和计算机控制的冲击，允许创建重复性损伤。低温及尿路感染在手术后期间构成重大挑战。仔细监测动物的日增重记录和膀胱的表达可以早期发现手术后的并发症。这个挫伤模型的功能结果相当于横断模型。挫伤模型可以被用来评价疗效的神经保护和神经再生的方法。

病灶体积按照上面描述的技术，我们所获得的大的和一致的病变体积。使用一个快速Luxol蓝色染色病灶体积平均为2.04毫米3（95％CI为1.9-2.18）组（n = 5只）。 图2所示平均病灶体积具有代表性的染色使用Luxol蓝快病变震中。 功能评分巴索，比蒂，布雷斯纳汉（BBB）规模作为衡量行为得分， 如图3所示。在12周的对照组大鼠BBB评分取得了平均2.2±1.1组（n = 10只）。 其他参数根据我们最近的32个动物手术经验，这种技术的成活率为93.4％。所有的死亡率相关，其次是动物的牺牲，以避免持久性尿路感染（UTI）<html>过度的痛苦和苦难。结束后7天疗程的抗生素，16.7％的动物开发尿路感染。用于检测尿路感染手术后的平均天数为14.6±7.6天。有趣的是，动物达到控制膀胱的平均伤后第13天，平均（13.33±3.6天）。大鼠UTI需要较长的时间来实现相比，无尿路感染（UTI大鼠膀胱控制 - 16±1天和12.8±3.7天的大鼠膀胱控制无尿路感染，P = 0.03）大鼠膀胱控制。在队列达到膀胱控制在14天内没有随后UTI检测。 如图4所示，有一个体重的初始增益，在脊髓损伤后有可能从液体潴留。其后的重量下降，并稳定在一个较低的水平（约10-12％，低于基线水平），10天左右。 <html> / files/ftp_upload/50111/50111fig1highres.jpg的"/&gt; 图1。椎解剖鼠从展示的棘突和椎板后路（左）中。黄韧带的位置（LF）的薄片之间的空间中描绘。中矢状部分（右），黄韧带和棘间韧带有关脊髓的相对位置。 <img src="/files/ftp_upload/50111/50111fig2.jpg" alt="图2" fo:content-width="4in" fo:src="/files/ftp_upload/50111/50111fig2highres.jpg"/> 图2。条形图显示病灶体积（单位:mm 3，Y轴）在接收严重脊髓损伤（左）的动物。具有代表性的纵向剖面震中受伤后Luxol快蓝染色（右）。 g3highres.jpg"/&gt; 图3。平均BBB（Y轴）得分超过3个月随访的过程中（X轴）的大鼠展示典型严重的脊髓损伤最小的恢复。 <img src="/files/ftp_upload/50111/50111fig4.jpg" alt="图4" fo:content-width="3.5in" fo:src="/files/ftp_upload/50111/50111fig4highres.jpg"/> 图4。线图变化超过2个星期的时间（X轴）诱导脊髓损伤后的大鼠的日增重（G，Y轴）。

Watch this Scientific Journal Video about A Contusion Model of Severe Spinal Cord Injury in Rats at JoVE.com

A Contusion Model of Severe Spinal Cord Injury in Rats

脊髓损伤严重挫伤模型描述。详细的手术前，手术和手术后的步骤进行了说明，以取得一致的模型。

挫伤模型脊髓严重损伤的保护作用

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

a-contusion-model-of-severe-spinal-cord-injury-in-rats

Universidad San Sebastian

Research

JoVE Journal

Medicine

30.3K 次观看.  Medical University of South Carolina. 脊髓损伤严重挫伤模型描述。详细的手术前，手术和手术后的步骤进行了说明，以取得一致的模型。

视频: A Contusion Model of Severe Spinal Cord Injury in Rats

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.

在大鼠脊髓损伤后的急性和慢性触觉测试

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

科研

医学

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.

评价呼吸肌慢性脊髓损伤个人使用呼吸电机控制评估（RMCA）

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.

控制皮层影响模型创伤性脑损伤

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.

无线电遥测系统，以监测大鼠脊髓横断和胚胎神经干细胞移植在心血管功能

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.

在脊髓损伤的实验挫伤型号神经干细胞移植

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.

脊髓压迫损伤的校准钳型

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

Contrast Enhanced Ultrasound Imaging for Assessment of Spinal Cord Blood Flow in Experimental Spinal Cord Injury

Reduced spinal cord blood flow (SCBF) (i.e., ischemia) plays a key role in traumatic spinal cord injury (SCI) pathophysiology and is accordingly an important target for neuroprotective therapies. Although several techniques have been described to assess SCBF, they all have significant limitations. To overcome the latter, we propose the use of real-time contrast enhanced ultrasound imaging (CEU). Here we describe the application of this technique in a rat contusion model of SCI. A jugular catheter is first implanted for the repeated injection of contrast agent, a sodium chloride solution of sulphur hexafluoride encapsulated microbubbles. The spine is then stabilized with a custom-made 3D-frame and the spinal cord dura mater is exposed by a laminectomy at ThIX-ThXII. The ultrasound probe is then positioned at the posterior aspect of the dura mater (coated with ultrasound gel). To assess baseline SCBF, a single intravenous injection (400 &#181;l) of contrast agent is applied to record its passage through the intact spinal cord microvasculature. A weight-drop device is subsequently used to generate a reproducible experimental contusion model of SCI. Contrast agent is re-injected 15 min following the injury to assess post-SCI SCBF changes. CEU allows for real time and in-vivo assessment of SCBF changes following SCI. In the uninjured animal, ultrasound imaging showed uneven blood flow along the intact spinal cord. Furthermore, 15 min post-SCI, there was critical ischemia at the level of the epicenter while SCBF remained preserved in the more remote intact areas. In the regions adjacent to the epicenter (both rostral and caudal), SCBF was significantly reduced. This corresponds to the previously described &#8220;ischemic penumbra zone&#8221;. This tool is of major interest for assessing the effects of therapies aimed at limiting ischemia and the resulting tissue necrosis subsequent to SCI.

Contrast Enhanced Ultrasound imaging is a reliable in-vivo tool for quantifying spinal cord blood flow in an experimental rat spinal cord injury model. This paper contains a comprehensive protocol for application of this technique in association with a contusion model of thoracic spinal cord injury.

超声造影成像脊髓血流量的评估实验脊髓损伤

Reduced spinal cord blood flow (SCBF) (i.e., ischemia) plays a key role in traumatic spinal cord injury (SCI) pathophysiology and is accordingly an ...

Photothrombosis-induced Focal Ischemia as a Model of Spinal Cord Injury in Mice

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) below the site of injury. The secondary ischemia that develops following the initial mechanical insult is a serious complication of the SCI and severely impairs the function and viability of surviving neuronal and non-neuronal cells in the SC. In addition, ischemia is also responsible for the growth of lesion during chronic phase of injury and interferes with the cellular repair and healing processes. Thus there is a need to develop a spinal cord ischemia model for studying the mechanisms of ischemia-induced pathology. Focal ischemia induced by photothrombosis (PT) is a minimally invasive and very well established procedure used to investigate the pathology of ischemia-induced cell death in the brain. Here, we describe the use of PT to induce an ischemic lesion in the spinal cord of mice. Following retro-orbital sinus injection of Rose Bengal, the posterior spinal vein and other capillaries on the dorsal surface of SC were irradiated with a green light resulting in the formation of a thrombus and thus ischemia in the affected region. Results from histology and immunochemistry studies show that PT-induced ischemia caused spinal cord infarction, loss of neurons and reactive gliosis. Using this technique a highly reproducible and relatively easy model of SCI in mice can be achieved that would serve the purpose of scientific investigations into the mechanisms of ischemia induced cell death as well as the efficacy of neuroprotective drugs. This model will also allow exploration of the pathological changes that occur following SCI in live mice like axonal degeneration and regeneration, neuronal and astrocytic Ca2+ signaling using two-photon microscopy.

Photothrombosis is a minimally invasive and highly reproducible procedure to induce focal ischemia in the spinal cord and serves as a model of spinal cord injury in mice.

Photothrombosis引起的局部脑缺血脊髓损伤的小鼠模型

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) ...

A Neonatal Mouse Spinal Cord Compression Injury Model

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal cord. A major current area of research is focused on the mechanisms of adaptive plasticity that underlie spontaneous or induced functional recovery following SCI. Spontaneous functional recovery is reported to be greater early in life, raising interesting questions about how adaptive plasticity changes as the spinal cord develops. To facilitate investigation of this dynamic, we have developed a SCI model in the neonatal mouse. The model has relevance for pediatric SCI, which is too little studied. Because neural plasticity in the adult involves some of the same mechanisms as neural plasticity in early life1, this model may potentially have some relevance also for adult SCI. Here we describe the entire procedure for generating a reproducible spinal cord compression (SCC) injury in the neonatal mouse as early as postnatal (P) day 1. SCC is achieved by performing a laminectomy at a given spinal level (here described at thoracic levels 9-11) and then using a modified Yasargil aneurysm mini-clip to rapidly compress and decompress the spinal cord. As previously described, the injured neonatal mice can be tested for behavioral deficits or sacrificed for ex vivo physiological analysis of synaptic connectivity using electrophysiological and high-throughput optical recording techniques1. Earlier and ongoing studies using behavioral and physiological assessment have demonstrated a dramatic, acute impairment of hindlimb motility followed by a complete functional recovery within 2 weeks, and the first evidence of changes in functional circuitry at the level of identified descending synaptic connections1.

This article describes a method for generating a reproducible spinal cord compression injury (SCI) in the neonatal mouse. The model provides an advantageous platform for studying mechanisms of adaptive plasticity that underlie spontaneous functional recovery.

新生儿小鼠脊髓压迫损伤模型

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal ...

A Tissue Displacement-based Contusive Spinal Cord Injury Model in Mice

Producing a consistent and reproducible contusive spinal cord injury (SCI) is critical to minimizing behavioral and histological variabilities between experimental animals. Several contusive SCI models have been developed to produce injuries using different mechanisms. The severity of the SCI is based on the height that a given weight is dropped, the injury force, or the spinal cord displacement. In the current study, we introduce a novel mouse contusive SCI device, the Louisville Injury System Apparatus (LISA) impactor, which can create a displacement-based SCI with high injury velocity and accuracy. This system utilizes laser distance sensors combined with advanced software to produce graded and highly-reproducible injuries. We performed a contusive SCI at the 10th thoracic vertebral (T10) level in mice to demonstrate the step-by-step procedure. The model can also be applied to the cervical and lumbar spinal levels.

We introduce a tissue displacement-based contusive spinal cord injury model that can produce a consistent contusive spinal cord injury in adult mice.

基于组织位移的小鼠脊髓损伤模型

Producing a consistent and reproducible contusive spinal cord injury (SCI) is critical to minimizing behavioral and histological variabilities between ...