这项工作是由卫生[NS36350，NS52290和NS50243以X-MX]美国国家研究院​​的支持;霍曼马里乔治捐赠基金;印第安纳州;和露丝L.基施施泰国家研究服务奖（NRSA）1F31NS071863到CLW

1.曝光颈椎椎板的<ol><li>清洁外科表面用70％的乙醇，预热用加热垫。放置消毒纱布，棉签和蒸压手术工具在手术区域前盖上无菌手术悬垂表面。使用微珠消毒的手术工具间手术消毒。 </li><li>麻醉与氯胺酮（87.7毫克/千克）/甲苯噻嗪（12.3毫克/千克）腹腔（IP）的大鼠。麻醉的正常平面达到当动物不再到脚趾捏刺激响应。皮下注入0.01-0.05毫克/公斤丁丙诺啡和5毫克/公斤卡洛芬之前外科手术。丁丙诺啡然后应每天一次给药，每8-12小时，卡洛芬，第4-7天手术后。 </li><li>适用于保护软膏动物的眼睛，以防止手术过程中角膜干燥。 </li><li>刮胡子手术区的背从中间胸部区域的鼠的头部的后面，快船的表面。用配有HEPA过滤器真空除去剃毛皮。 </li><li>应用优碘溶液的剃区域作为外科擦洗然后清洗，用70％异丙醇擦拭的面积。 </li><li>使用手术刀刀片从所述头部的基部尾端至中期胸廓进行3-4厘米中线切口，在皮肤上。 </li><li>确定筋膜和皮下肌肉前的中线冬眠腺体在较低的颈部;切穿斜方肌和其他肌肉沿中线以减少出血。 </li><li>发现脂肪组织的肌肉底层的两个区域的中线;使用小组织牵缩器之前，胸椎T2被达到棘突的水平切脊柱旁肌肉尾端严格沿中线和单独的肌肉层。 </li><li>识别并切掉连接到T2棘突肌肉利用THIS结构作为解剖标志。 </li><li>取出T2棘突的软骨尖，改善颈椎的知名度。 </li><li>从棘突和C4-T1的椎板横向分开椎旁肌肉;然而，饶肌肉覆盖在C3椎板，以防止出血。 </li><li>切开肌肉过度从C4-T1中的薄片沿横向朝对脊柱两侧的面。 </li><li>后脊柱薄层露出，将动物在稳定器的U形通道的腹面。 </li><li>通过从T 2的地标为T1，C7，C6和C5的最后吻侧计数棘突识别C5椎骨。 </li></ol> 2.稳定的椎骨和表演的撞击伤<ol><li>定位稳定器的两个不锈钢臂通过将臂的锯齿状边缘的拉泰拉下方暂停动物在C5-6椎（ 图1C）升面。固定臂与椎骨到位（ 图2B）之后，调节稳定装置，以确保脊柱是水平和居中。最后，通过拧紧稳定的指旋螺丝锁定的怀抱。 </li><li>切断C4-5和C5-6棘突和薄片之间的韧带，以确定碳五薄层的边缘。 </li><li>用微型咬骨钳，夹远离薄片的一半处的C5右侧为用于脊髓损伤（ 图5C-E）。椎板切除后，运输的动物与伤病设备下的稳定。与稳定剂上的安装（ 图3A-C）的固定的动物一起精确地对准柱塞上使用侧向微型操纵器（图3）脊髓目标。 </li><li>在高倍率下，找到了C5和C6背根进入区（DREZs）在未与杜暴露脊髓背侧表面rotomy。瞄准柱塞在两个识别DREZs的中间，并且中途中线和脊髓（ 图5B）的横向边缘之间。 </li><li>使用NYU / MASCIS冲击装置配备2.5毫米直径的尖端（ 图3A＆B），由10克杆X12.5毫米高度下降（ 图2A）产生C5半挫伤（ 图5B＆E）。 </li><li>通过视觉上的青紫脊髓（ 图5E中 ，箭头）验证的伤害和检查由纽约大学软件12,17（ 图6）提供的伤害参数。 </li><li>缝合肌肉，用无菌4-0薇乔缝合软组织，然后关闭皮肤切口手术钉（EZ剪辑）。 </li><li>适用的抗菌药膏到外科手术部位。 </li><li>管理将5.0ml无菌0.9％盐水皮下给动物进行水化。 </li><li>将动物在热-controlled环境（再循环热水padcage上加热垫）），与设置在寝具（每天更换湿食品），和一个水瓶具有长喷口为方便放置在笼子的地板上。提供护理，以保证有足够的恢复动物返回家笼前。 </li><li>因为这是一个单方面的颈椎挫伤性损伤，动物很可能会失去功能同侧前肢，瞬时，开始在最初的几个星期伤后恢复。然而，对侧的功能应保持不变，因此动物应该可以吃喝无损伤，并在运动和疏导只有轻微受损。 </li></ol>

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我们什么都没有透露。

在这里，我们已经表现出了颈椎的稳定制造方法单侧挫伤SCI在C5。这种稳定化方法增加了创伤解剖学的精度，并产生一致的功能缺损13,18。在依赖于棘突背夹紧其他车型，棘突损害或脊椎夹子脱落的风险是相当高的。这些模型也可以允许相当脊柱移位和从挫伤力和脊柱和椎骨柱（ 图1A和B）的柔性性质产生。组织得到涂改柱塞组织接触时间，并导致不可预测的损伤力（ 图1A-B 6B）。我们的描述椎体稳定还提供了其它好处的外科准备：1）该方法充分稳定下的中心在C 5椎骨手术显微镜这增加椎板切除术（ 图1C）的精度; 2）安装在所述U形的稳定剂可以直接从手术位置来定制安装附件，避免了重新安装上的SCI设备动物的过程和节省时间采取内的动物; 3）稳定椎骨的伤害水平，直接背和尾部的损伤可以大大减少所造成的呼吸身体运动的目的网站，另一项措施，以减少可变性。 利用该稳定化方法的主要优点是，得到的减少量，或受冲击时的脊髓和列的腹侧运动。基于对一个挫伤​​简单的物理，冲击力和能量将从杆传送到脊髓，最好用软线在撞击的部位吸收能量。但是，如果线下方的脊柱的产量，因为有可能在背棘突夹紧方法（ 图1A＆B）所示 ，实际的力施加到被减小的线和可变的，这取决于产量的程度。 虽然这段视频显示的颈椎挫伤性脊髓损伤模型的整个过程中，这篇文章的精髓是引进我们的各种应用在我们的实验室使用的脊椎稳定的方法，专门针对SCI研究。这种稳定装置和方法的修改后的版本已被用于对小鼠SCI 23。这个简单的脊柱稳定的方法是一种用于脊髓损伤的研究非常有用的，我们先前已经用这种方法和设备，以执行胸椎挫伤以及裂伤SCI模型。另一个实验室在这22杂志最近描述了这种形式的稳定的宫颈损伤的变化。综上所述，我们介绍的这款新颖的椎体稳定的方法来几个surgic人的程序产生重复性实验SCI从椎板切除损伤的生产。这种稳定装置的优点不限于颈脊髓挫伤，因为此稳定化方法已适应于各种各样的实验，如帧内脊髓注射，细胞移植，脑脊液采集从延髓池，半切和横断损伤，胸椎挫伤损伤， 体内成像采用双光子显微镜，和脊髓电生理记录。增加的脊柱外科和外伤程序的质量，降低了实验变异性将有助于提供洞察损伤和恢复的真实机制，以及筛选对脊髓损伤的破坏性疾病不同疗法的效果。

目标脊髓组织一致，且可复制的机械力是最小化的功能和病理变化，并为建立成功的挫伤性脊髓损伤（SCI）模型1-7的关键。施加到脊髓的目标区域的力的大小取决于用于脊柱稳定的方法。目标脊柱的冲击柱塞和脊髓之间接触​​的过程中的位置偏移改变所得伤力。颈椎挫伤性SCI模型是一个更临床相关的模型比其它形式的SCI的，因为约50％的人的SCI的情况下，发生在这个级别8，以及使用动物颈椎损伤模型9-14几个SCI研究已经完成。虽然挫伤性SCI的模型通常通过夹紧棘突前部和后部的损伤部位利用某种形式的稳定化，这种制剂是困难的制造颈脊髓损伤。 ＆＃160;如在该演示中，我们开发了稳定的方法是在提高质量和挫伤的再现能力有利。具体地，建立了在试图修改的缺点和其他型号的挑战椎体稳定的这种方法：1）变异椎通过夹紧相邻背侧棘突喙和尾部的椎板切除可能发生的冲击力的作用下屈服。椎移位的程度取决于冲击和被稳定椎骨（ 图1）之间的椎关节的数量。因此，多个关节所涉及的不太稳定的脊柱变; 2）背侧棘突是脆弱的，导致钳衰竭为棘突骨折或夹具脱落的过程的结果;和3）对这些椎骨棘突极短的C3至T1椎骨相比，这些胸韦尔特的布雷，用传统的夹具，这使得难以把握棘突稳定颈椎。 在这里，我们描述了一种新的稳定的脊柱对于成年雌性Sprague-Dawley大鼠制备的C5挫伤性SCI的方法。这种方法可以用于其他级别的脊柱和脊髓，和缀合物的稳定性以及与其他挫伤性SCI的设备，包括纽约大学/多中心动物脊髓损伤研究（NYU / MASCIS）冲击15（ 图2）精密系统和仪器仪表，LLC无限地平线（IH）装置16，美国俄亥俄州立大学/电磁脊髓损伤设备1，和路易斯维尔损伤系统仪器（LISA）17，允许在SCI研究广泛使用。

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</table>

a novel vertebral stabilization method for producing  contusive spinal cord injury

临床相关的动物颈脊髓损伤（SCI）模型是用于开发和测试潜在的治疗是必不可少的;然而，生产可靠宫颈SCI是困难的，因为缺乏椎体稳定的令人满意的方法。传统的方法来稳定脊柱是通过连接到颈椎棘突夹暂停吻端和尾鳍颈椎。然而，稳定化的这种方法不能防止组织挫伤期间产生的颈椎过程太短被有效地由夹具（ 图1）固定。在这里，我们引入一个新的方法，在撞击伤的同一水平，完全稳定的颈椎。这种方法有效地在撞击的部位，从而大大提高了生产一致的SCI的最小化的脊柱运动。我们提供的设备（ 图2-4）的直观描述，方法s和一步一步的协议，用于颈椎5椎骨成年大鼠（C5）的稳定化，以执行椎板切除术（ 图5），并产生一个挫伤性SCI后。虽然我们只说明使用NYU / MASCIS冲击装置的颈部半挫伤，这种脊椎稳定技术可以应用到脊髓的其他区域，或者适用于其他的SCI的设备。改善通过椎体稳定脊髓曝光和固定可以是用于生产一致和可靠的受伤脊髓有价值。这种脊椎稳定方法也可用于细胞和示踪剂的立体定向注射，以及使用双光子显微镜在各种神经生物学研究的成像。

在下面这个协议，一致的和可重复的颈椎半挫伤SCI产生（ 图5,6）。使用脊椎稳定剂，以稳定相同椎骨在用于脊髓损伤的水平的横向过程允许这种令人满意的结果。使用这种方法，不仅在目标椎骨的C5，而且相邻的C4和C6均刚性地固定。 纽约大学/ MASCIS软件提供的读出与在x和y轴设置的曲线图，以及支持使用我们的脊椎稳定化方法，以及设备（ 图6）。稳定化的这种方法减少损伤的变异可导致从目标组织和脊柱（ 图1）的向下移动。损伤后，明确单方面偏蓝血肿的C5和C6 DREZs之间居中是可见的（ 图5E）。这些受伤的参数是一致的，从动物到ANI发作根据由纽约大学/ MASCIS软件中提供的读出（ 图6）。 由于颈椎半挫伤产生明显的前肢赤字，这种模式非常适合评估前肢功能的能力，如深远，疏导13，和对象操纵18-19。由于后肢运动障碍不太突出，巴索，比蒂和布雷斯纳汉（BBB）运动得分规模4是不适合在这个模型中使用。的功能的结果如下损伤是最显着的在同侧前爪伸肌缺陷，其中，所述鼠表现出“杵”握拳所有数字弯曲18。暴露在相同的损伤程度和脊髓级别的所有动物应该表现出类似的赤字在这个协议中说明的同侧前肢，在正确的伤害。动物不当受伤非常不同的表现形式赤字13,18和持续时间可能会出现。组织学上，这种模式会产生大量的灰质和白质损伤，在损伤震中和延髓和尾鳍受伤的部位，导致相当多病灶和空洞形成包含几乎完全脊髓伤侧内。大，主要是星形胶质细胞为基础的胶质瘢痕的形成与大量的神经元死亡18病灶边界。 <img alt="图1" fo:content-width="5in" src="/files/ftp_upload/50149/50149fig1highres.jpg" width="500" /> 图1：脊柱的灵活性期间挫伤SCI与不同的夹紧方式插图。图A和 B显示了灵活性，或者当棘突的背部夹紧，使不当影响和不一致的数据脊柱“收益”。于A所示的图显示在冲击（红色虚线更大的灵活性和大弯箭头）相比，在B中所示（更小的弯曲箭头），因为夹具是从椎板和损伤部位更远。 图C示出了侧向稳定性与我们描述的装置与稳定臂下的横突拧紧其中将执行影响的部位的椎骨。有脊椎的在此过程中没有灵活性，因为所关注的椎骨被完全稳定。 <img alt="图2" fo:content-width="5in" src="/files/ftp_upload/50149/50149fig2highres.jpg" width="500" /> 图2：NYU / MASCIS冲击和定制的稳定容器。图A显示了零件和纽约大学/ MASCIS脊髓损伤装置的功能，具有对损伤严重多个杆高度设置（插图）; 图B和C示出了U形的容器用于保存大鼠和锯齿稳定手臂手术和伤病期间安全稳定脊柱（设计和YP张生产）。 <img alt="图3" fo:content-width="5in" src="/files/ftp_upload/50149/50149fig3highres.jpg" width="500" /> 图3：自定义安装系统和横向microadjuster在NYU / MASCIS冲击图A详细介绍了U形稳定大鼠脊髓损伤的定制安装系统的不同组件。注意在图A的横向microadjuster，对于大鼠脊髓损伤为精确对准的关键。 图B和C提供的稳定剂的进一步描述无（B）和具有U形的大鼠的容器（C）的相对于其它重要伤病设备的组件（安装系统设计和YPž生产挂起）。 <img alt="图4" fo:content-width="5in" src="/files/ftp_upload/50149/50149fig4highres.jpg" width="500" /> 图4：手术稳定装置和附件的各个部件的测量尺寸定制稳定系统的每个部件被加亮显示的尺寸和尺度（A，C和D）。胸椎稳定臂（B）中示出，以显示在不同的脊柱外科模型使用这种装置的潜在应用。 <img alt="图5" fo:content-width="5in" src="/files/ftp_upload/50149/50149fig5highres.jpg" width="500" /> 图5：手术的地标和准备颈椎半挫伤脊髓损伤。图A和 B描绘正确IMPA正确地标对暴露的大鼠脊髓克拉对齐。合适的冲击点是直接的C5和C6背神经根（延髓-尾）和中线和脊髓（B）的横向边缘之间。 图CE显示，在更高的放大倍率，露出所需的一半的过程中颈髓受伤，经过周密的单侧椎板切除术。此外， 数字D和 E之前和脊髓挫伤损伤后表现出的线。请注意可见出血（E）所造成的影响（黑色箭头）。 <img alt="图6" fo:content-width="5in" src="/files/ftp_upload/50149/50149fig6highres.jpg" width="500" /> 图6：。可接受与不可接受的数据读出在与NYU / MASCIS冲击影响（C）的例子。上图（A）和顶部的数据集</s仲&gt;示出了一个非常良好的冲击的读出，以“％的误差”的冲击杆速度，初始高度，并在开始时数据的测量，如用红色箭头指示和下划线。所有值落在以及可接受误差的窗口中。相反，底部面板在“零”的冲击杆和尖端到脊髓表面之前，设置所述冲击杆的高度显示出由不当造成的稳定的脊柱（B）和误差的不正确的冲击所产生的数据（C）。注意初始高度表示的相当大的误差，并开始撞击一滴的时间，由红色箭头所示和下划线。该软件还提供了已经检测这些参数的（图C的底部），该错误的警告。

Watch this Scientific Journal Video about A Novel Vertebral Stabilization Method for Producing  Contusive Spinal Cord Injury at JoVE.com

A Novel Vertebral Stabilization Method for Producing  Contusive Spinal Cord Injury

脊椎稳定是必要的最小化可变性，以及生产一致的实验性脊髓损伤。使用与NYU / MASCIS冲击设备一起定制的稳定装置，我们在这里展示了合适的设备和程序，产生重复的半挫伤颈椎（C5）脊髓成年大鼠受伤。

一种新的椎稳定制备方法挫伤脊髓损伤

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

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17.5K 次观看.  Indiana University School of Medicine. 脊椎稳定是必要的最小化可变性，以及生产一致的实验性脊髓损伤。使用与NYU / MASCIS冲击设备一起定制的稳定装置，我们在这里展示了合适的设备和程序，产生重复的半挫伤颈椎（C5）脊髓成年大鼠受伤。 

视频: A Novel Vertebral Stabilization Method for Producing  Contusive Spinal Cord Injury

Longitudinal Evaluation of Mouse Hind Limb Bone Loss After Spinal Cord Injury using Novel, in vivo, Methodology

Spinal cord injury (SCI) is often accompanied by osteoporosis in the sublesional regions of the pelvis and lower extremities, leading to a higher frequency of fractures 1. As these fractures often occur in regions that have lost normal sensory function, the patient is at a greater risk of fracture-dependent pathologies, including death. SCI-dependent loss in both bone mineral density (BMD, grams/cm2) and bone mineral content (BMC, grams) has been attributed to mechanical disuse 2, aberrant neuronal signaling 3 and hormonal changes 4. The use of rodent models of SCI-induced osteoporosis can provide invaluable information regarding the mechanisms underlying the development of osteoporosis following SCI as well as a test environment for the generation of new therapies 5-7 (and reviewed in 8). Mouse models of SCI are of great interest as they permit a reductionist approach to mechanism-based assessment through the use of null and transgenic mice. While such models have provided important data, there is still a need for minimally-invasive, reliable, reproducible, and quantifiable methods in determining the extent of bone loss following SCI, particularly over time and within the same cohort of experimental animals, to improve diagnosis, treatment methods, and/or prevention of SCI-induced osteoporosis.
An ideal method for measuring bone density in rodents would allow multiple, sequential (over time) exposures to low-levels of X-ray radiation. This study describes the use of a new whole-animal scanner, the IVIS Lumina XR (Caliper Instruments) that can be used to provide low-energy (1-3 milligray (mGy)) high-resolution, high-magnification X-ray images of mouse hind limb bones over time following SCI. Significant bone density loss was seen in the tibiae of mice by 10 days post-spinal transection when compared to uninjured, age-matched control (na&iuml;ve) mice (13% decrease, p&lt;0.0005). Loss of bone density in the distal femur was also detectable by day 10 post-SCI, while a loss of density in the proximal femur was not detectable until 40 days post injury (7% decrease, p&lt;0.05). SCI-dependent loss of mouse femur density was confirmed post-mortem through the use of Dual-energy X-ray Absorptiometry (DXA), the current "gold standard" for bone density measurements. We detect a 12% loss of BMC in the femurs of mice at 40 days post-SCI using the IVIS Lumina XR. This compares favorably with a previously reported BMC loss of 13.5% by Picard and colleagues who used DXA analysis on mouse femurs post-mortem 30 days post-SCI 9. Our results suggest that the IVIS Lumina XR provides a novel, high-resolution/high-magnification method for performing long-term, longitudinal measurements of hind limb bone density in the mouse following SCI.

Longitudinal Evaluation of Mouse Hind Limb Bone Loss After Spinal Cord Injury using Novel, in vivo, Methodology

A longitudinal examination of bone loss in the femurs and tibiae of adult mice was performed following spinal cord injury using sequential low-dose X-ray scans. Tibia bone loss was detected throughout the study, while bone loss in the femur was not detected until 40 days post injury.

鼠标使用新型脊髓损伤的后肢骨质流失的纵向评价，在体内，方法论

Spinal cord injury (SCI) is often accompanied by osteoporosis in the sublesional regions of the pelvis and lower extremities, leading to a higher ...

科研

医学

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 ...

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.

单侧颈脊髓损伤的挫伤模型，利用无限的地平线冲击

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 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.

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 ...

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 ...