Name: calibrated forceps model of spinal cord compression injury
Uploaded: 2015-04-24T15:00:00.000+00:00
Duration: 9 min 41 s
Description: Watch this Scientific Journal Video about Calibrated Forceps Model of Spinal Cord Compression Injury at JoVE.com

Funding for this work was provided by Shriners Hospitals and CIRM fellowships to AMC and AM. We would also like to acknowledge Plemel et al.7 for first designing and publishing the calibrated forceps model.

所有动物的程序及动物护理方法应该由该机构的机构动物护理和使用委员会（IACUC）批准。 1.手术准备<ol><li>收集所有必要的外科手术工具和试剂:镊子，剪刀瓦纳，roungeurs，牵引器，手术刀，剪刀，缝合，皮肤缝合器，棉签，无菌生理盐水，手术海绵和异氟醚。准备和高压灭菌的外科手术工具的完整包在手术前。如果执行手术在一个以上的小鼠，制备和高压灭菌的每只动物的工具之一包，或高压灭菌手术器械的一个分组和消毒用工具消毒器在手术之间。通常，无菌工具之一包可以用在最多5只小鼠，如果玻璃珠工具terilizer用于清洁在两者之间手术（5只动物后的工具必须重新清洗并在使用前灭菌）的工具。请与您当地的IACUC管理员instituti检查具体的指导方针。 </li><li>消毒手术区域用70％异丙醇擦拭。建立手术单，以确保在手术过程中的无菌场被维持。 </li><li>权衡手术前各小鼠。施用0.05毫克/公斤体重丁丙诺啡的剂量，皮下。 </li><li>通过在异氟醚机的感应腔室的剂量为4％施用异氟烷麻醉动物。 </li><li>一旦动物被麻醉，敷药膏对眼睛，以防止脱水，设置在加热垫的动物，在37℃，并确保该鼠标的头部适当地坐落在麻醉锥。 （注:使用热源，不会造成热灼伤; 也就是说 ，循环水毯，热水袋，或等效。）此时辖2％异氟醚的剂量动物。 </li><li>剃鼠标的背表面上，围绕着预定切口位置约1厘米。 </li><li>消毒切口s伊特通过用70％异丙醇擦拭洗涤，然后用碘溶液（10％聚维酮碘，1％有效碘）。重复3次。 </li></ol> 2.背椎板切除术<ol><li>在作出一个切口，确保动物正常通过检查用脚趾或尾巴捏法见长麻醉。 </li><li>使沿背棘与手术刀和刀片切口，然后检查反射了。拱起背部，帮助妥善可视化地标，如椎骨之间的界限。 </li><li>通过皮肤切开。插入牵开器保持皮肤和筋膜从脊髓背面。清除在脊髓任一侧的组织以暴露覆盖脊柱的肌肉。 注:外科医生应熟悉解剖标志。例如，肩胛骨的劣角对应用T7。鼠标的脊柱的自然曲线的顶部是T12和可以用作一个参考点。 </li><li>与对罗珀照明，确定椎骨之间的空间。发现T10的后端与切断肌肉和筋膜垂直于椎间盘空间。切刚够露出T10的棘突和后部薄层。 </li><li>使用一对细尖杜蒙＃5镊子，去除一些从椎板和棘突的组织的，以暴露小纱条脊髓。必要时，用组织钳脊柱稳定。 </li><li>通过插入一对小瓦瑙剪刀沿着椎骨的背外侧侧面和刚椎板下方的一侧执行椎板切除术。 <ol><li>使小，小心剪刀通过椎板的侧面切割。确保没有压力施加到脊髓。 </li><li>重复另一侧。 </li><li>轻轻推压，停止作为必要的止血用棉条或外科用纱布，同时注意不要施加压力的脊髓。 注:准备发泡胶在事件出血需要被控制浸泡在无菌盐水中。 </li><li>一旦切口已经作出，抬起该椎骨的背侧并轻轻清除任何组织的附件。使用适当的手段来控制，如果必要的出血。 </li></ol></li></ol> 3.脊髓压迫<ol><li>使用roungeurs或椎板钳，确保了脊髓的侧面是自由椎体骨，使得校准钳子用于压缩损伤可对脊髓的两侧位于。钳子的臂必须能够被放置在脊髓和相邻侧的钳子的前端硬膜外空间内必须能够到达椎管的楼层。 <ol><li>确保脊髓认为可视性良好。 </li></ol></li><li>位于校准杜蒙＃5镊子大约在脊髓的暴露段的中间。回想一下，部队的武器PS必须放置在硬膜外腔的相邻侧面和尖端必须接触椎管的底板，以产生重现的伤害。 </li><li>仔细压迫脊髓直到隔板连接。固定到位，持续15秒。 </li><li>轻轻释放压缩力和从脊髓取出校准的钳子。无菌盐水应该用来恢复稳态之前伤口闭合。 </li></ol> 4.创伤闭合<ol><li>仔细缝合肌肉层在脊髓，同时注意不要破坏或施加压力的脊髓。 </li><li>使用任一缝合线或U形钉来闭合皮肤在伤口上。 </li><li>如果使用气体麻醉，开始逐渐/关闭麻醉剂。 </li><li>辖0.1毫升每10g体重乳酸林格氏来帮助在手术时，动物是昏昏欲睡，从损伤恢复的手术后帐户脱水。该解决方案应该是WÄRMEd可在注射前的体温。 </li><li>鼠标放置在一个床上用品，免费笼。笼应搁在一个加热垫（如在1.1.5中描述）的方式，使一半的笼区域是在垫的顶部，而另一半被搁在一个RT计数器，为了得到鼠标的气候方案，一旦它走动。请注意，以确保"回收笼"坐落在一个安静的环境。密切监测动物，直到它恢复意识，此时鼠标可以被转移到一个普通笼子里的床上用品。 </li></ol> 5.术后护理<ol><li>手术后，按照需要来管理疼痛症状给药0.05毫克/千克体重丁丙诺啡皮下注射，每12小时一个剂量的第3天手术后，再。 </li><li>辖乳酸林格氏（每10g体重皮下注射0.1ml）中的第3的剂量 - 手术后第5天。给这个剂量，如果/当一个进制开始显示出这个初始时间段以外脱水的迹象。 </li><li>手动表达使用克雷德机动每日两次的动物的膀胱。轻轻触诊动物的腹部找到膀胱，然后应用温和的下行压力，直到膀胱是空的。 <ol><li>如果膀胱不空或尿液是血性或阴天，辖50毫克/公斤按体重拜有利的，以动物通过interperitoneal注射10天。 </li></ol></li><li>监测动物对自噬，脱水和过度减肥（体重大于20％）的迹象。如果动物遇到任何这些症状，请与兽医立即用问候治疗方案咨询，或安乐死的动物以人道的方式下IACUC准则。 <ol><li>鉴于流动性可能在手术后的权利的限制，采取必要的步骤，以确保动物获得食物和水。预包装湿的食物，还有一s的水凝胶，可以提供给动物在这些实例。 </li></ol></li></ol> 6.评估组织损伤的压缩损伤引起的<ol><li>如在步骤1.4和1.5中所述麻醉动物。检查麻醉深度脚趾捏和监控角膜眨眼反射。当动物是不敏感的刺激，继续执行步骤6.2。 </li><li>执行一个心内灌注10。 <ol><li>暴露胸腔和针头插入左心室。冲洗掉现有的流体用20 - 30毫升冰冷的磷酸盐缓冲盐水（PBS），随后加入15 - 40毫升冰冷的4％多聚甲醛（PFA）的。 </li></ol></li><li>取出脊髓。 <ol><li>切开皮肤背侧脊髓和清除周围的脊柱的长度的任何多余的组织。 </li><li>消费脊柱和切去任何剩余的组织。椎板切除术和我的实际水平njury可以通过计数肋的确认。 </li><li>使用万纳剪刀和镊子来置换脊柱的小部分在一个尾至头端的方向。继续砍，直到线暴露足以让安全移除。切割的位置和方法的可视化可以通过使用一个立体镜的来促进。 </li></ol></li><li>放置脊髓组织在4％PFA。允许组织向定位后在该溶液中24小时，在4℃下。 </li><li> Cryoprotect组织孵育在30％蔗糖24小时，在4℃。 </li><li>在OCT嵌入组织。简单地说，采取组织从30％蔗糖孵化和删除任何多余的解决方案。将组织放入盛有OCT一个cryomold孵育1小时，在4℃。 <ol><li>除去从4℃的模具中，确认组织的方向取向，将模具中的2-甲基丁烷浅盘（预冷却1小时在干冰），并允许OCT来完全固化。如果使用即时或STO继续干冰再在-80℃。 </li></ol></li><li>切割组织成使用低温恒温器20微米的矢状切面。直接安装到组织的幻灯片。储存在-20℃直到使用。 </li><li>执行苏木和曙红（H＆E）染色。 <ol><li>简要地说，再水合所述组织（5分钟，2次），染色用苏木精（2.5分），并用水（1分钟，2次）。 </li><li>孵育组织中的50％，然后70％的乙醇（每次3分钟），染色与曙红（45秒），并且温育在90％以上（5秒），95％（5秒）脱水，100％乙醇（2分钟）和异丙醇（2分钟）。 </li><li>清除二甲苯（5分钟，3次）。 注:H＆E染色将随具体的组织厚度和条件。因此，标准化是在进行试验的组织标本前需要。 </li></ol></li><li>覆盖组织与薄带Permount（约100微米）和盖玻片。按下盖玻片上的各方，确保流体的正确分布。让幻灯片干O / N。 </li><li>可见ualize使用使用附带的软件数码显微镜拍摄的图像组织切片。 </li></ol>

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The authors have nothing to disclose.

的脊髓损伤模型的选择是在设计实验，以确定对脊髓损伤的人类病例的治疗的疗效很重要。这些实验需要的动物模型，该模型是高度可再现的，以限制可变性可能导致不确定的数据。他们也应该是临床相关正确地评价人的条件，他们的造型。为此目的，在一个横切选择的压缩或挫伤性损伤是更临床相关的3。然而，撞击和体重下降的装置为挫伤受伤需要使用昂贵和复杂的机械。与此相反，脊髓损伤的校准钳子模型利用改性镊子，很容易从普通实验室材料来组装，并在手术仅需要一个标准的背侧椎板切除后额外的步骤，以暴露脊髓。然而，使用这种方法的一个缺点是，压缩力总是施加横向而非背侧，如是最常见于脊髓损伤9，和压缩损伤的方法，使用影响组织的更大的喙-尾程度比挫伤模型1,2-产生人类临床病例。该模型已被证明了该技术的发起者，并且我们，以生成可再现的SCI 7,11，并且非常适合于小鼠的大小。此外，这种损伤模型允许动物手术后及治疗方法使用多种行为测试，如巴索鼠标规模为运动和冯弗雷头发测试，以验证动物的人群共享相同的损伤程度进行评估和神经功能缺损7,11-13。这些相同的技术也可以用来评估的过程中调查研究给予动物治疗的功效，满足对用于评估疗法的SCI 2,7-动物模型的一般标准。 制备校准钳子的方法S为损伤模型简单，能够实现与各种不同的方法。我们已经使用了隔板11的方法，由Plemel 7所公布，并且还修改使用小螺钉，它不仅提供了一个更简单的方法，用于产生压缩装置钳子，也允许为多功能性在调整最终压缩宽度受益比较研究。在创造钳子选项的范围几乎是无限的，只要该间隔件（多个）提供始终关闭所述钳子以相同距离的稳定装置，并能承受高压灭菌和消毒。此视频中所描述的手术方法是在用户之间高度重现，但是有必要执行椎板切除和缝合动物后的程序已被执行，以使​​脊髓不会受到任何附加的压缩力，可能会增加当护理采取受伤的严重程度和混淆未来的实验。 </p&gt; 用适当的训练和练习，压迫损伤的校准钳子模型非常适合于在模仿在人体中观察到的2,3,7-临床病例的小鼠进行脊髓损伤。由于易于制造镊子，产生不同程度损伤严重程度的小鼠能够容易地完成。这将是大有裨益观察不同程度的转基因小鼠严重的遗传效应对SCI以及评价干细胞移植的疗效小鼠。大多数的文献中的研究已经在大鼠上进行，由于它们的大小，这通常使得手术更容易执行。然而由Plemel 等 7本视频出版，由我们描述的方法应该使SCI要对小鼠以极大的方便和可重复性进行。

脊髓损伤的动物模型可用于评估旨在减轻损害创伤到脊髓的后果的治疗范例的功效有价值的工具。出实验的必要性，这些模型必须提供运动和感觉的行为重复的缺陷，随意调节产生不同程度的伤害，并证明损害严重程度与神经功能缺损的观察到的程度。有SCI三种主要类型有伤害的鲜明特色:切断，挫伤和压缩1。简要地说，一个横断损伤是裂伤至脊髓，挫伤损伤起因于施加到背侧脊髓的简要，焦力，并且当有害的力被施加到脊髓发生压迫损伤，并且也可以是被称为挤压伤2。 完全横断伤是在人类3临床上较少见，而挫伤了ð压缩损伤是比较常见的。压缩损伤产生类似于被人的SCI发现引起，例如，肿瘤压迫或其他有害的压缩力的结果，并且可以使用的工具的一个简单的阵列来执行。挫伤和压缩损伤是在双方都是一个压缩力两者具有类似的病理特征，如cytoarchitectonic解体相似，并且唤起类似的内源性应答损伤1,4。挫伤损伤模型通常适用于类似SCI的人类病例来自脊柱2,5,6的冲击造成的方式使用特殊的设备，这股力量对脊髓背侧。与此相反，压缩损伤可通过各种施加力背侧或侧面的方法来生成的。的压缩损伤的方法包括校准钳7，动脉瘤夹2，或者直接放置一个重量到脊髓8。的优势动脉瘤夹是它们能提供不同量的力9。添加权重给脊髓直接8的背表面的方法中，需要的重量到位10分钟后，急剧增加了手术的长度，并导致不一致由于重量和运动的放置由于的呼吸动物。由于小尺寸的小鼠，情境动物设计用于大鼠，如撞击用于挫伤受伤使用专门的设备，可能是困难的或导致不一致的受伤7。然而，小鼠是可在一个宽范围的转基因品系，不象较大的动物如大鼠或兔，这对脊髓损伤的研究非常有用的。 使用校准钳压迫脊髓的Plemel方法生成一个可重复使用SCI损伤程度及神经功能缺损7之间的相关度很高。这种手术SCI模型使用一对修饰将在由任一金属的环氧树脂或一些其它障碍物，防止完全关闭一个限定的距离保持分开号5杜蒙钳生成的。这种工程化的间距确保了钳子将始终接近一定的宽度在多次手术和由不同的用户。所述Plemel方法的优点是，以产生校准的钳子的材料可以容易地购得，使用时需要专门的设备组装在实验室。这些镊子可以承受多轮高压灭菌和杀菌的，而且缺乏一个单独的，笨重的设备的简化手术。 在这个视频，我们证明在小鼠脊髓外科手术使用的一对校准镊子以产生一个压缩的损伤。我们也解决与脊髓受伤的实验动物的照顾，以提高他们的生活质量手术后，降低死亡率独特的担忧。

<table><tbody><tr><td>Isoflurane machine</td><td>Smiths Medical PM, Inc</td><td>VCT302</td><td></td></tr><tr><td>Isoflurane</td><td>Phoenix Pharmaceutical</td><td>NDC: 66794-013-25</td><td></td></tr><tr><td>Dissecting Scope</td><td>Seiler Precision Microscopes</td><td>SSI 202/402</td><td></td></tr><tr><td>Germinator-500 (tool sterilizer)</td><td>Thomas Scientific</td><td>3885A20</td><td></td></tr><tr><td>Puralube (eye ointment)</td><td>Dechra</td><td>NDC 17033-211-38</td><td></td></tr><tr><td>Scalpel handle (#3)</td><td>Fine Science Tools</td><td>10003-12</td><td></td></tr><tr><td>Scalpel blade (#11)</td><td>Fisher Scientific</td><td>&amp;nbsp;08-914B</td><td></td></tr><tr><td>Retractor (Colibri)</td><td>Fine Science Tools</td><td>17000-03</td><td></td></tr><tr><td>Friedman Pearson roungeur</td><td>Fine Science Tools</td><td>16021-14</td><td></td></tr><tr><td>Vanna (Castroviejo) scissors</td><td>Roboz</td><td>RS-5658</td><td></td></tr><tr><td>Tissue forceps</td><td>Fine Science Tools</td><td>11029-14</td><td></td></tr><tr><td>Laminectomy forceps (Dumont #2)</td><td>Fine Science Tools</td><td>11223-20</td><td></td></tr><tr><td>Dumont #5 forceps</td><td>Fine Science Tools</td><td>11252-20</td><td></td></tr><tr><td>Stapler</td><td>Fine Science Tools</td><td>12031-07</td><td></td></tr><tr><td>Staples (wound clips)</td><td>Reflex7</td><td>203-1000</td><td></td></tr><tr><td>Sutures</td><td>Henry Schein</td><td>101-2636</td><td></td></tr><tr><td>Needles (30 G x &amp;frac12;)</td><td>BD Biomedical</td><td>305106</td><td></td></tr><tr><td>Syringe (1 ml)</td><td>BD Biomedical</td><td>309659</td><td></td></tr><tr><td>Baytril (enrofloxacin)</td><td>Bayer</td><td>NADA 140-913</td><td></td></tr><tr><td>Buprenex (buprenorphine)</td><td>Cardinal Health</td><td>NDC 12496-0757-1</td><td></td></tr></tbody></table>

calibrated forceps model of spinal cord compression injury

小鼠脊髓压迫损伤是有价值的动物模型脊髓损伤（SCI）和脊髓再生疗法的研究。压迫损伤的校准镊子模型是一个方便，成本低，而且非常重复性的动物模型SCI。我们所用的一对修改的镊子根据由Plemel 等人发表的方法（2008），以横向地压迫脊髓，以0.35毫米的距离。在此视频中，我们将展示一个背椎板切除以暴露脊髓，其次是脊髓与改性钳子的压缩。在视频中，我们也将解决与截瘫实验动物的护理问题。这种损伤模型产生表现出的感觉障碍，以及受损的后肢运动功能的小鼠。此外，受伤的这个方法，在SCI的病理产生一致的畸变，通过免疫组化方法确定。看着这个VID后EO，观众应该能够确定用于各种严重程度的，在小鼠为设计损伤后，以减轻损害对脊髓损伤和/或治疗方法的研究产生的SCI的必要的用品和方法。

我们进行了椎板12小鼠 - 如上所述（25 30克），接着脊髓压缩为0.25毫米（N = 4），0.35毫米（N = 4）和0.55毫米（N = 4）。我们在三个牺牲的动物（N = 6）和7（6）日伤后心内灌注。脊髓从脊柱去除，制备并如上所述进行处理的组织。整个脊髓的图像用的Leica EZ4数字显微镜和附带的软件。脊髓切片图像使用Olympus数码显微镜及配套的软件采取2倍的放大倍率。 我们发现，脊髓受压产生的震中在压缩（ 图1）的部位受伤。伤病的影响，在延髓和尾鳍方向延伸几毫米。损伤的严重性随着所述间隔件之间的距离减小（0.25毫米&gt;0.35毫米&gt;0.55毫米， 图2）。压缩后的三天，有血在伤的震中和损伤后不在场7天周边地区。 0.25毫米和0.35毫米按压产生的空腔，但不是0.55毫米模型。经过7天，背，腹白质主要是缩小尺寸的震中，灰质组织被严重扭曲，和空化呈持续性。采用适当的测试，如巴索鼠标规模为运动功能和冯弗雷头发和乙基氯化物测试感觉功能正如我们在以前的出版物8展示了这些cytoarchitectonical改变被翻译成运动和感觉的改变动物的行为进行评估。 <img alt="图1" src="/files/ftp_upload/52318/52318fig1highres.jpg" width="500" /> 完整脊髓图1代表性图像之前和AF之三伤。 （A）完整的脊髓。（B）后0.35毫米压缩脊髓。箭头表示损伤的边界。星号indentifies损伤震中。 D =背，L =横向。比例尺:0.50毫米<a href="https://www.jove.com/files/ftp_upload/52318/52318fig1highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图2" src="/files/ftp_upload/52318/52318fig2highres.jpg" width="650" /> 图鼠标脊髓2.代表图像之前和压迫损伤在不同宽度压缩后。 （A）的控制脊髓矢状切面。（B）在一个0.35毫米SCI后第7天压迫损伤（dpi）的震中冠状切面（C，E，G）脊髓3 DPI为0.25宽度矢状切面0.35 0.55毫米（D，F，H）的H＆E矢状切片染色脊髓第7 DPI至0.25，0.35 0.55毫米的宽度。星号indentifies损伤震中。所有部分用H＆E。 D =背，L =横向。比例尺:1.25毫米<a href="https://www.jove.com/files/ftp_upload/52318/52318fig2highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about Calibrated Forceps Model of Spinal Cord Compression Injury at JoVE.com

Calibrated Forceps Model of Spinal Cord Compression 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 ...

calibrated-forceps-model-of-spinal-cord-compression-injury

Research

JoVE Journal

Medicine

18.4K 次观看.  University of California, Davis. 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.

视频: Calibrated Forceps Model of Spinal Cord Compression Injury

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 ...

科研

医学

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 ...

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 ...

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 ...

A Neuronal Apoptosis Model induced by Spinal Cord Compression in Rat

As a severe progressive degenerative disease, cervical spondylotic myelopathy (CSM) has a poor prognosis and is associated with physical pain, stiffness, motor or sensory dysfunction, and a high risk of spinal cord injury and acroparalysis. Thus, therapeutic strategies that promote efficient spinal cord regeneration in this chronic and progressive disease are urgently needed. Effective and reproducible animal spinal cord compression models are required to understand the complex biological mechanism underlying CSM. Most spinal cord injury models reflect acute and structural destructive conditions, whereas animal models of CSM present a chronic compression in the spinal cord. This paper presents a protocol to generate a rat spinal cord compression model, which was further evaluated by assessing the behavioral score and observing the compressed spinal cord region. The behavioral assessments showed decreased monitor motor disability, including joint movements, stepping ability, coordination, trunk stability, and limb muscle strength. Hematoxylin and eosin (H&#38;E) staining and immunostaining revealed considerable neuronal apoptosis in the compressed region of the spinal cord.

Here, we present a protocol to generate a rat spinal cord compression model, assess its behavioral score, and observe the compressed spinal cord region. The behavioral assessments showed decreased monitor motor disability. Hematoxylin and eosin staining and immunostaining revealed considerable neuronal apoptosis in the compressed region of the spinal cord.

As a severe progressive degenerative disease, cervical spondylotic myelopathy (CSM) has a poor prognosis and is associated with physical pain, stiffness, ...

神经科学

Establishing a Mouse Contusion Spinal Cord Injury Model Based on a Minimally Invasive Technique

Using minimally invasive methods to model spinal cord injury (SCI) can minimize behavioral and histological differences between experimental animals, thereby improving the reproducibility of the experiments.
These methods need two requirements to be fulfilled: clarity of the surgical anatomical pathway and simplicity and convenience of the laboratory device. Crucially for the operator, a clear anatomical pathway provides minimally invasive exposure, which avoids additional damage to the experimental animal during the surgical procedures and allows the animal to maintain a consistent and stable anatomical morphology during the experiment.
In this study, the use of a novel integrated platform called the SCI coaxial platform for spinal cord injury in small animals to expose the T9 level spinal cord in a minimally invasive way and stabilize and immobilize the vertebra of mice using a vertebral stabilizer is researched, and, finally, a coaxial gravity impactor is used to contuse the spinal cord of mice to approach different degrees of T9 spinal cord injury. Finally, histological results are provided as a reference for the readers.

Minimally invasive techniques and a simple laboratory device improve the reproducibility of the spinal cord injury model by reducing operative damage to the experimental animals and allowing anatomical morphology maintenance. The method is worthwhile because the reliable results and reproducible procedure facilitate investigations of the mechanisms of disease reparation.

基于微创技术的小鼠挫伤脊髓损伤模型的建立

Using minimally invasive methods to model spinal cord injury (SCI) can minimize behavioral and histological differences between experimental animals, ...

Establishment of Central Cord Syndrome Model in C57BL/6J Mouse

Animal models of central cord syndrome (CCS) could substantially benefit preclinical research. Identifiable anatomical pathways can give minimally invasive exposure approaches and reduce extra injury to experimental animals during operation, enabling the maintenance of consistent and stable anatomical morphology during experiments to minimize behavioral and histological differences between individuals to improve the reproducibility of experiments. In this study, the C6 level spinal cord was exposed using a spinal cord injury coaxial platform (SCICP) and combination with a minimally invasive technique. With the assistance of a vertebral stabilizator, we fixed the vertebrae and compressed the spinal cords of C57BL/6J mice with 5 g/mm2 and 10 g/mm2 weights with SCICP to induce different degrees of C6 spinal cord injury. In line with the previous description of CCS, the results reveal that the lesion in this model is concentrated in the gray matter around the central cord, enabling further research into CCS. Finally, histological results are provided as a reference for the readers.

The present protocol simulating central cord syndrome (CCS) in mice has improved repeatability and minimized operation damage to the experimental animals, avoiding disrupting the anatomical structure excessively. The strategy in this study is advantageous because it allows for research into injury mechanisms by producing consistent results.

C57BL/6J小鼠中索综合征模型的建立

Animal models of central cord syndrome (CCS) could substantially benefit preclinical research. Identifiable anatomical pathways can give minimally ...

用于精确小鼠模型的脊髓损伤挫伤装置

Automated Impactor for Contusive Spinal Cord Injury Model in Mice

Spinal cord injury (SCI) due to traumatic injuries such as car accidents and falls is associated with permanent spinal cord dysfunction. Creation of contusion models of spinal cord injury by impacting the spinal cord results in similar pathologies to most spinal cord injuries in clinical practice. Accurate, reproducible, and convenient animal models of spinal cord injury are essential for studying spinal cord injury. We present a novel automated spinal cord injury contusion device for mice, the Guangzhou Jinan University smart spinal cord injury system, that can produce spinal cord injury contusion models with accuracy, reproducibility, and convenience. The system accurately produces models of varying degrees of spinal cord injury via laser distance sensors combined with an automated mobile platform and advanced software. We used this system to create three levels of spinal cord injury mice models, determined their Basso mouse scale (BMS) scores, and performed behavioral as well as staining assays to demonstrate its accuracy and reproducibility. We show each step of the development of the injury models using this device, forming a standardized procedure. This method produces reproducible spinal cord injury contusion mice models and reduces human manipulation factors via convenient handling procedures. The developed animal model is reliable for studying spinal cord injury mechanisms and associated treatment approaches.

作者聚焦：深入了解脊髓损伤研究的创新

Presented here is a novel automated spinal cord injury contusion device for mice, which can accurately produce spinal cord injury contusion models with varying degrees.

用于小鼠挫伤脊髓损伤模型的自动撞击器

Spinal cord injury (SCI) due to traumatic injuries such as car accidents and falls is associated with permanent spinal cord dysfunction. Creation of ...