这项工作部分由NIH NS059622，NS073636，DOD CDMRP W81XWH-12-1-0562支持;美国退伍军人事务部优异评审奖I01 BX002356; Craig H Neilsen基金会296749;印第安纳脊髓和脑损伤研究基金会和马里·胡尔曼乔治基金会基金（XMX）; Norton Healthcare，Louisville，KY（YPZ）;印第安纳州ISDH 13679（XW）;和神经科学基金会。

所有手术和动物处理程序均按照"实验动物护理和使用指南"（国家研究委员会）和印第安纳大学医学院动物保健和使用委员会准则进行。 1.准备动物并进行T10脊柱椎板切除术<ol><li>在高压灭菌器中灭菌手术器械和金属脊柱稳定器。清洁手术台。将加热垫加热至37°C。将加热垫放在手术台上，并用无菌手术包盖住。在整个手术过程中使用无菌技术。 </li><li> 在10周龄时使用女性年轻成人C57 / 6J小鼠进行本研究。用腹膜内（ip）注射氯胺酮（87.7mg / kg）和赛拉嗪（12.3mg / kg）混合物麻醉每只动物。通过引发对爪夹捏诱导的伤害感受刺激的反应来确认完全麻醉。 </stro<html> NG&gt; <ol><li>皮下给予丁丙诺啡（0.01-0.05mg / kg），镇痛剂和卡洛芬（5mg / kg），非类固醇抗炎药。 </li></ol></li><li>使用电动剪刀将头发刮在胸腰椎上。用betadine溶液和70％酒精擦拭液擦拭皮肤。 </li><li>将眼用软膏涂在角膜上，以防止手术中的眼睛干燥。 </li><li>用手术刀，在动物背部做一个1.5厘米的中线皮肤切口，露出第9 至第 11胸椎椎板。推拉皮下脂肪组织。解剖椎旁肌肉脱离棘突和椎板，朝向侧面的侧面。 </li><li>将鼠标放在稳定器的U形槽上（ 图2A 和2B ）。将不锈钢手臂双侧夹在T10椎骨的暴露面上图4A），并使用连接在手臂上的翼形螺丝（ 图2A ）拧紧。 </li><li>使用暴露在脊髓上的硬膜的微型骨骼去除T10棘突和椎板（椎板切除术）（ 图4B ）。 </li></ol> 2.使用LISA冲击器进行T10挫伤伤害<ol><li>转动氮气罐上压力调节器的旋钮，将压缩氮气设置为20 PSI或138 kPa（ 图1A ）。 注意:压力可从10-120 PSI调节。更高的压力将导致更高的速度冲击。直径为1.2mm的SCI装置尖端设计用于小鼠，并且针对大鼠设计直径为2.2mm的尖端。当从小鼠变为大鼠时，可以通过向金属尖端添加环（ID为1.2mm / od 2.2mm）来形成较大直径的尖端。我们在这只小鼠SCI中使用了1.2毫米的尖端tudy。使用前先对SCI尖端进行灭菌。 </li><li>打开电脑启动软件。按钮1（ 图1B ）将冲击器尖端激活到完全伸出的位置（ 图3A -1 ）。 注意:按钮1的功能是手动启动或停用气动气缸。 </li><li>将U形容器用鼠标放在舞台上（ 图2B ）。通过拧紧安装座的翼形螺钉将支架固定到位（ 图2B ）。 </li><li>在"SET ZERO LEVEL"区域（绿色）下，通过点击"开始读取"按钮（ 图3A ），用激光传感器测量与完全延伸的柱塞尖端的距离，设置零电平。该距离将显示在该区域的"范围"参数中（ 图3A ）。点击"设置零"按钮（ 例如， 图 3.29中的8.951 mm）。 </li><li>按钮1（ 图1B ）撤回冲击器尖端（ 图3B -1 ，由上箭头指示）并解锁紧固螺丝1（ 图2B ）。将螺丝拉到正确的位置（ 图3B -1 ，用横向箭头指示）将尖端远离激光束路径，顺时针旋转螺丝90°以锁定螺丝。 </li><li>通过调整正面和侧面微驱动器（ 图1C ）来移动舞台，将激光束瞄准暴露背脊髓的中心。伤害位置被定位后，通过点击"SET INJURY L"下的"START READING"按钮来测量组织距离EVEL"区域（蓝色）（ 图3B 和3B-1 ）。 </li><li> 通过垂直微驱动器（ 图1C ）缓慢地调节传感器和脊髓之间的距离，以达到"SET INJURY LEVEL"区域中所需的位移参数（ 例如， 0.500 mm，显示在"损伤"参数框中） （蓝色）（ 图3B ）。 <ol><li>当达到所需的伤害位移时，记录组织距离（ 例如， 8.451 mm，显示在"范围"参数框中）（ 图3B ）。定义所需的位移（损伤）=尖端距离（零） - 组织距离（范围）（ 图3B ）。当达到所需的伤害（ 例如，组织移位为0.500毫米）（ 图3B ）时，点击"SET INJURY LEVEL"区域下的"SET INJURY"按钮，设置伤害。 </li></ol></li><li>将螺丝1逆时针旋转90°以解锁螺丝，将冲击头推回激光束轨迹（ 图3C -1 ，箭头所示方向），并将螺丝1顺时针转动90°。 </li><li>单击红色"运行实验"区域（ 图3C ）下的运行按钮执行影响。该区域下的参数框将显示受伤时间，力（mV），速度（m / s）和伤害位移（mm）（ 图3C ）。 </li><li>记录并保存所有伤害数据后，用鼠标从舞台上取出U形槽。用手术显微镜观察脊髓损伤（ 图4C ）。 </li><li>使用3-0丝（Henry Schein，776-SK）连续缝合缝合椎旁肌肉，浅表筋膜和皮肤。 </li><li>注射e动物用1mL的0.9％盐水皮下进行水合，并将其置于温度控制的垫上，直到全部意识恢复。将鼠标放入可以进入食物和水的笼子里。 </li><li>对于术后护理，手动表达膀胱，直到自发性膀胱排尿返回。对于镇痛，注射丁丙诺啡（0.05-2.0mg / kg，SQ）8-12h /天，持续2天。如果发生膀胱感染，将Baytril（SQ，5-10mg / kg，0.1mL，每日1次）注射7-10天。如果发生局部/全身感染，将注射庆大霉素（SQ，5-8mg / kg，在1mL无菌盐水中每8-12小时稀释）4天。 </li><li>在SCI后14天取出缝线。 </li><li>损伤后第42天，通过灌注处死小鼠。在适当的麻醉1.2之后，它们将在0.01M PBS中用30ml（0.01M）磷酸盐缓冲盐水（PBS）和30ml 4％多聚甲醛灌注。一厘米脊髓包括病变史诗将进行收集和处理进行切片和组织学分析。 </li></ol>

<ol>
	<li>Young, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+cord+contusion+models.">Spinal cord contusion models.</a> Prog. Brain Res. 137, 231-255 (2002).</li><li>Gale, K., Kerasidis, H., Wrathall, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+cord+contusion+in+the+rat:+behavioral+analysis+of+functional+neurologic+impairment.">Spinal cord contusion in the rat: behavioral analysis of functional neurologic impairment.</a> Exp. Neurol. 88 (1), 123-134 (1985).</li><li>Gruner, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+monitored+contusion+model+of+spinal+cord+injury+in+the+rat.">A monitored contusion model of spinal cord injury in the rat.</a> J. Neurotrauma. 9 (2), 126-128 (1992).</li><li>Scheff, S. W., Rabchevsky, A. G., Fugaccia, I., Main, J. A., Lumpp, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+modeling+of+spinal+cord+injury:+Characterization+of+a+force-defined+injury+device.">Experimental modeling of spinal cord injury: Characterization of a force-defined injury device.</a> J. Neurotrauma. 20 (2), 179-193 (2003).</li><li>Stokes, B. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+spinal+cord+injury:+a+dynamic+and+verifiable+injury+device.">Experimental spinal cord injury: a dynamic and verifiable injury device.</a> J. Neurotrauma. 9 (2), 129-134 (1992).</li><li>Young, W., Chen, J., Xu, X. M., Xu, Z. C., Zhang, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MASCIS+spinal+cord+contusion+model.">MASCIS spinal cord contusion model.</a> Animal Models of Acute Neurological Injuries. , 411-422 (2009).</li><li>Jakeman, L. B., McTigue, D. M., Walters, P., Stokes, B. T., Chen, J., Xu, X. M., Xu, Z. C., Zhang, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Ohio+State+University+ESCID+spinal+cord+contusion+model.">The Ohio State University ESCID spinal cord contusion model.</a> Animal Models of Acute Neurological Injuries. , 433-448 (2009).</li><li>Scheff, S., Roberts, K. N., Chen, J., Xu, X. M., Xu, Z. C., Zhang, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Infinite+Horizon+spinal+cord+contusion+model.">Infinite Horizon spinal cord contusion model.</a> Animal Models of Acute Neurological Injuries. , 423-433 (2009).</li><li>Sances, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+biomechanics+of+spinal+injuries.">The biomechanics of spinal injuries.</a> Crit. Rev. Biomed. Eng. 11 (1), 1-76 (1984).</li><li>Zhang, Y. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+cord+contusion+based+on+precise+vertebral+stabilization+and+tissue+displacement+measured+by+combined+assessment+to+discriminate+small+functional+differences.">Spinal cord contusion based on precise vertebral stabilization and tissue displacement measured by combined assessment to discriminate small functional differences.</a> J. Neurotrauma. 25 (10), 1227-1240 (2008).</li><li>Walker, M. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+vertebral+stabilization+method+for+producing+contusive+spinal+cord+injury.">A novel vertebral stabilization method for producing contusive spinal cord injury.</a> J. Vis. Exp. (95), (2015).</li><li>Allen, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgery+of+experimental+lesion+of+spinal+cord+equivalent+to+crush+injury+of+fracture+dislocation+of+spinal+column.+A+preliminary+report.">Surgery of experimental lesion of spinal cord equivalent to crush injury of fracture dislocation of spinal column. A preliminary report.</a> J. A. M. A. 57, 878-880 (1911).</li><li>Jakeman, L. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Traumatic+spinal+cord+injury+produced+by+controlled+contusion+in+mouse.">Traumatic spinal cord injury produced by controlled contusion in mouse.</a> J. Neurotrauma. 17 (4), 299-319 (2000).</li><li>Rivlin, A. S., Tator, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+duration+of+acute+spinal+cord+compression+in+a+new+acute+cord+injury+model+in+the+rat.">Effect of duration of acute spinal cord compression in a new acute cord injury model in the rat.</a> Surg. Neurol. 10 (1), 38-43 (1978).</li><li>Zhang, Y. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled+cervical+laceration+injury+in+mice.">Controlled cervical laceration injury in mice.</a> J. Vis. Exp. (75), (2013).</li><li>Ma, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+controlled+spinal+cord+contusion+for+the+rhesus+macaque+monkey.">A controlled spinal cord contusion for the rhesus macaque monkey.</a> Exp. Neurol. 279, 261-273 (2016).</li></ol>

医学博士Christopher B. Shields拥有Louisville Impactor System，LLC生产的路易斯维尔损伤系统仪器（LISA）的所有权。

在1911年，艾伦描述了使用固定重量的第一个减重模型，以在狗的暴露脊髓上引起伤害12 。已经基于Allen模型开发了类似的减重模型，包括NYU / MASCIS冲击器3,6,13,14。除了减重模型，还创建了其他SCI设备。 OSU / ESCID 5,7模型使用组织置换机制来控制伤害严重程度，IH模型4,8使用力创建可分级的SCI。在这些系统中，椎体稳定是通过将​​棘突和尾部夹在受伤部位而获得的。这些装置利用低伤害速度，特别是0.33-0.9m / s（NYU / MASCIS），0.148m / s（OSU / ESCID）和0.13m / s（1H）。稳定脊柱和尾部棘突可能会导致脊柱的弹性和脊柱在冲击过程中移动，这可能会影响伤害的准确性。 LISA方法试图克服现有模型的缺点，特别是在脊柱不稳定性和低伤害速度方面。该方法使用双侧面稳定并避免与损伤相关的运动伪影。该装置利用可设置在0.5-2m / s 11,15之间的高冲击速度。激光传感器比ESCID模型中使用的Ling振动器更先进，并且精确地测量距离脊髓表面的距离，而不需要任何组织接触。该模型最初被开发用于产生大鼠SCI，并且现在已经适应于在小鼠和非人灵长类动物16上产生SCI，并进行修改。 脊椎在所有实验性SCI方法，特别是在组织置换模型中，消除减少变异性。激光距离传感器确定呼吸运动期间脊髓组织位移的大小。重要的是，激光聚焦的脊髓点应该是冲击器撞击的相同点。当冲击器尖端和激光束对准时，该步骤在校准步骤（ 图3 ）期间完成。该模型的潜在缺点是从硬脑膜表面测量组织位移的大小。尽管硬脑膜的厚度在动物之间构成微不足道的差异，但蛛网膜下腔充满脑脊髓液（CSF）可能存在明显的变异性。当使用小组织置换产生非常轻微的挫伤伤害时，可能发生损伤结果的变异性。总的来说，伤害的一致性主要依赖于关于组织位移的准确性以及柱塞的速度和组织接触时间。 组织位移范围宽（精度:0-10±0.005 mm）。根据以前的试验数据和在啮齿动物和非人灵长类动物中发表的信息，SC的前后径的20％位移产生轻度SCI，30-40％的置换产生中度SCI，&gt; 50％的位移以1 m / s的速度产生严重的SCI。根据动物种类，会有轻微的差异。停留时间可以使用时间继电器从0到5 s调整。在我们的研究中，停留时间设定为300 ms。这可以轻松调整，以复制其他SCI设备的驻留时间，包括NYU和IH型号。 总之，我们已经在成年小鼠中开发了一种基于位移的脓性SCI模型。该模型使用U形稳定器来稳定双侧脊柱面，避免绳索与激光引导测量绳表面相关的运动伪影。该模型可以产生0.5-2米/秒的高速线损伤。激光传感器比常规方法更精确，以确定速度和到冲击表面的距离。该模型可以在轻度到重度的各个层面产生脊髓损伤。修改后，该装置也可能在大鼠和大型动物如非人灵长类动物中产生损伤。

在人类中发生的最常见的脊髓损伤（SCI）是脓性SCI 1 。为了研究SCI后的损伤机制和各种治疗策略，需要在啮齿动物中进行精确，一致和可重现的迂回SCI模型。 在实验性SCI研究2,3,4,5,6中已经使用了许多具有各种损伤机制的脊髓挫伤损伤模型。三个肆意的SCI模型 - 具体来说，纽约大学（NYU）/多中心动物脊髓损伤研究（MASCIS）影响因子3,6 ，俄亥俄州立大学（OSU）冲击/电磁SCI装置（ESCID） 5 ， 7 ，ad无限景观（IH）影响器4,8在国际SCI研究领域被广泛接受。 NYU / MASCIS冲击器或等效物质通过将不同高度的固定重量下降到目标脊髓上造成伤害，造成多重伤害严重性3,6 。 OSU / ESCID通过诱导组织移位造成伤害5,7 。 IH冲击器通过向脊髓4,8施加不同的力而产生损伤。每个冲击器使用不同的速度，这是影响伤害结果的重要参数。 NYU / MASCIS设备产生的范围从0.33-0.9米/秒。 IH装置的最大速度为0.13m / s 4 。 OSU / ESCID冲击器的固定速度为0.148 m / s 5 。值得注意的是，se模型低于在临床速度观察到的模型，其通常超过1.0m / s 9 。 在这里，我们介绍一种新型的基于位移的迂回SCI装置，称为路易斯维尔损伤系统装置（LISA），以在高冲击速度10的小鼠中产生SCI。该系统包括一个脊椎稳定器，可以在损伤部位牢固地稳定椎骨，从而允许产生一个恒定的，可重复的SCI。该装置的激光传感器确保精确确定组织位移以及由此产生的SCI严重程度。在与脊髓接触点处的柱塞的速度可以从0.5至2m / s调节。这些损伤参数临床上严重复制创伤性SCI。

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</table>

a tissue displacement-based contusive spinal cord injury model in mice

产生一致和可重复的挫伤性脊髓损伤（SCI）对于最小化实验动物之间的行为和组织学变化至关重要。已经开发了几种迂回的SCI模型，以使用不同的机制产生伤害。 SCI的严重程度是基于给定体重下降的高度，伤害力或脊髓位移。在本研究中，我们引入了一种新型的小鼠瘟疫SCI装置，路易斯维尔伤害系统装置（LISA）冲击器，可以创造出具有高伤害速度和精度的基于位移的SCI。该系统利用激光距离传感器与先进的软件相结合，产生分级和高度重现的伤害。我们在小鼠的第十个胸椎（T10）水平进行了一个迂回的SCI，以证明一步一步的程序。该模型也可应用于颈椎和腰椎脊柱水平。

该装置由五个主要部件组成:（1）具有冲击头的主体（ 图1C ），（2）具有软件的计算机（ 图1B ），（3）电气控制箱（ 图1B ），（4）a脊椎稳定器（ 图2A ）和（5）用于气动控制系统的压缩空气（ 图1A ）。为了诱导精确的组织置换，系统依靠激光传感器来测量完全延伸的柱塞尖端和目标脊髓背侧表面之间的距离。由于激光束仅到达冲击器的反射面（ 图2B和图3A -1 ），该软件考虑到了尖端的4毫米厚度。有两个位置可以放置柱塞尖端:（1）in是激光束（ 图3A -1 ）或（2）在远离激光束的横向位置的路径（ 图3B -1 ）。当柱塞处于激光束路径（ 图3A -1 ）时，它测量距冲击器尖端的距离，并监测冲击器尖端在伸展和缩回之间运动时的速度。当柱塞处于远离激光束路径的横向位置（ 图3B -1 ）时，测量激光与脊髓之间的距离。 使用我们的脊椎稳定器的T10椎骨的稳定是程序的一个组成部分（ 图2A ） 10,11 。使用激光传感器的可靠距离测量取决于s目标的能力，如果存在运动可能会扭曲。为了确定该系统的准确性和一致性，8只小鼠遭受0.5mm位移损伤。这些动物表现出±0.001mm（±SD）的位移变异性，表明该系统是高度准确和可重复的。 图4显示了在手术显微镜下（ 图4B ）和之后（ 图4C ）挫伤之前的稳定器（ 图4A ）中的固定的靶椎骨和暴露的T10脊髓。 压缩空气的压力在受伤时控制冲击器的速度。我们的数据表明，在138 kPa的压力下，冲击速度为0.81±0.0345 m / s（平均值±SD）。电器盒上的旋钮（ 图1B ）控制损伤后尖端接触（停留时间）的持续时间，可以在0到5000 ms之间调整。大多数实验中的尖端停留时间设定为0.32±0.0147s（平均值±SD）（ 图5 ）。使用该装置，可以在成年小鼠中以0mm（假手术对照），0.2mm（轻度损伤），0.5mm（中度损伤）和0.8mm（严重损伤）的组织移位产生严重性依赖性挫伤性损伤（ 图6 ）。 <img alt="图1" class="xfigimg" src="/files/ftp_upload/54988/54988fig1.jpg" /> 图1:路易斯维尔伤害系统装置（LISA）。 （ A ）系统由撞击器，控制系统和压缩空气源组成。 （ B ）控制系统包括控制箱和膝上型计算机。控制箱的软件和控制按钮允许用户建立lish伤害参数。 （ C ）激光传感器是该装置的关键部件，并测量受伤目标的位置，从脊髓到传感器的距离以及受伤速度。冲击尖端的快速上下移动由压缩空气提供动力。损伤的位置和组织位移的严重程度通过微型驱动器进行调节，这可以控制三维运动。 <a href="//ecsource.jove.com/files/ftp_upload/54988/54988fig1large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> <img alt="图2" class="xfigimg" src="/files/ftp_upload/54988/54988fig2.jpg" /> 图2:稳定器和鼠标座。 （ A ）脊柱稳定器由U型槽和两个金属臂组成，用于固定鼠标脊椎。 （ B ）然后将稳定器安装在冲击器装置上。 Ť他的红线表示激光束路径。 <a href="//ecsource.jove.com/files/ftp_upload/54988/54988fig2large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> <img alt="图3" class="xfigimg" src="/files/ftp_upload/54988/54988fig3.jpg" /> 图3:产生侵入性SCI的方法。 （ A - C ）显示了具有三个伤害参数/区域的图形用户界面（GUI）软件。 （ A ， A-1 ）绿色区域（SET ZERO LEVEL）校准柱塞尖端的距离。红线表示激光束路径。 （ B ， B-1 ）蓝色区域用于设置伤害级别（SET INJURY LEVEL）。冲击器被提起并横向移动到右侧，以允许激光束到达脊髓的背表面以设定零水平。红线表示激光b<html>电子路径。 （ C ， C-1 ）在冲击之前，尖端被移回到激光束路径上以执行伤害（RUN）。伤害参数处于红色区域（RUN EXPERIMENT）。红线表示激光束路径。 <a href="//ecsource.jove.com/files/ftp_upload/54988/54988fig3large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> <img alt="图4" class="xfigimg" src="/files/ftp_upload/54988/54988fig4.jpg" /> 图4:伤害暴露和评估。 （ A ）脊椎稳定器的金属臂稳定T10椎骨。 （ B ）T10椎板切除术暴露脊髓，背部血管清晰可见。 （ C ）脊髓背表面的冲击引起的挫伤（箭头）证实了损伤。比例尺= 2mm。g4large.jpg"target ="_ blank"&gt;请点击此处查看此图的较大版本。 <img alt="图5" class="xfigimg" src="/files/ftp_upload/54988/54988fig5.jpg" /> 图5:损伤参数。一致伤害参数包括组织移位（mm），损伤速度（m / s）和末梢停留时间（s）。 N = 8，平均值±SD。 <a href="//ecsource.jove.com/files/ftp_upload/54988/54988fig5large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> <img alt="图6" class="xfigimg" src="/files/ftp_upload/54988/54988fig6.jpg" /> 图6:组织学评估。 （ A ）假手术（0毫米），（ B ）轻度（0.2毫米），（ C ），（ C） </strong&gt;）中等（0.5mm）和（ D ）使用LISA装置的T10严重（0.8mm）痉挛性SCI。损伤中心拍摄图像。刻度棒= 500μm。 <a href="//ecsource.jove.com/files/ftp_upload/54988/54988fig6large.jpg" target="_blank">请点击此处查看此图的较大版本。</a>

Watch this Scientific Journal Video about A Tissue Displacement-based Contusive Spinal Cord Injury Model in Mice at JoVE.com

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

我们介绍一种基于组织移位的挫伤脊髓损伤模型，可以在成年小鼠中产生一致的挫伤性脊髓损伤。

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 ...

a-tissue-displacement-based-contusive-spinal-cord-injury-model-in-mice

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

11.1K 次观看.  Indiana University School of Medicine. 我们介绍一种基于组织移位的挫伤脊髓损伤模型，可以在成年小鼠中产生一致的挫伤性脊髓损伤。 

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

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

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 ...

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.

控制宫颈裂伤损伤小鼠

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.

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 Novel Vertebral Stabilization Method for Producing  Contusive Spinal Cord Injury

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

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

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 ...

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 ...