本研究得到了国家自然科学基金第82102314号（至ZSJ）和32170977（至HSL）和广东省自然科学基金（编号：2022A1515010438（至ZSJ）和2022A1515012306（至HSL）的支持。本研究由暨南大学附属第一医院临床前沿技术项目支持，编号：JNU1AF-CFTP-2022-a01206（至HSL）。本研究得到了广州市科技计划项目 第202201020018号（至HSL）、2023A04J1284（至ZSJ）和2023A03J1024（至HSL）的支持。

涉及动物的研究由暨南大学伦理委员会审查和批准。
1.动物麻醉和T10椎板切除术
<ol><li>使用 8 周龄的雌性年轻成年 C57/6J 小鼠进行本研究。通过腹膜内注射氯胺酮（100mg / kg）和地西泮（5mg / kg）麻醉小鼠。检查麻醉是否成功，表现为疼痛反射丧失。在眼睛上涂抹兽医软膏，以防止麻醉下干燥。</li>
	<li>用剃须刀剃掉老鼠背部的毛发，露出皮肤。用三轮碘磷和酒精交替消毒皮肤。</li>
	<li>用手术刀在背侧皮肤上做 2.5 厘米的内侧纵向切口，并使用镊子在 T9-T11 水平暴露脊柱。</li>
	<li>使用脊柱固定器双侧固定 T10 小关节。确保脊柱稳定固定。确保剥离椎旁肌肉，并使用微磨钻去除棘突和椎板，以暴露 T10 节段的脊髓。</li>
</ol>2. 使用 G smart SCI 系统对 T10 脊髓进行挫伤
<ol><li>打开开关，等待设备自动恢复到原始状态。将脊柱固定器放入 G smart SCI 系统中，并用螺钉固定。</li>
	<li>使用操作触摸屏（图2A），设置损伤参数，包括冲击速度（1 m / s），冲击深度（三组不同的小鼠为0.5 mm，0.8 mm和1.1 mm）和停留时间（500 ms）15。</li>
	<li>通过移动平台将激光测距仪对准暴露的脊髓中心。（图2B）</li>
	<li>单击触摸屏上的 Ready 按钮（图 2C）。冲击头将根据设置参数自动调整到特定高度。托架台自动将脊髓撞击部位移动到撞击头下方。</li>
	<li>手动按压冲击头以进一步确定冲击部位。单击 “开始 ”按钮，冲击头将根据设置的参数撞击脊髓。</li>
	<li>将小鼠从设备中取出并在体视显微镜（20x）下观察以确定脊髓损伤（图3）。为了确定模型开发是否成功，请观察局部充血、塌陷和脊髓膜破裂。</li>
	<li>使用 3-0 缝合线逐层缝合肌肉、筋膜和皮肤。将小鼠放入温暖的盒子中，等待它们恢复。</li>
</ol>3. 术后护理
<ol><li>手术后每天皮下注射美洛昔康（5mg/kg），持续 7 天。每8小时手动排空膀胱，直到膀胱功能恢复。</li>
	<li>手术后 14 天，移除缝合线。</li>
</ol>4. 脊髓损伤的检测效果
<ol><li>从术后第一天开始计算小鼠的BMS评分16,17。</li>
	<li>术后第30天，进行动物行为实验，包括走秀、足部断层和旋转16,17。走秀：创纪录的45厘米距离;最大运行持续时间 8 秒;相机增益：28.02;强度阈值 0.01。脚故障：为每只鼠标记录 60 步。旋转杆：速度 20 rpm。记录鼠标下落的时间，并记录为 120 秒以上 120 秒。</li>
	<li>术后第31天，腹膜内注射氯胺酮（100mg/kg）和地西泮（5mg/kg）麻醉小鼠，然后用4%PFA灌注对小鼠实施安乐死。小心地取出脊髓，并在受伤部位上方和下方截取 5 毫米以进行石蜡包埋。在小鼠脊髓损伤中心做一个5μm的切片，并进行苏木精和伊红染色17。</li>
	<li>对于统计分析，请使用商业软件。将数据表示为均值±均值标准误差 （SEM），并使用单因素方差分析进行比较;p < 0.05 被认为是显著的。</li>
</ol>

用于精确小鼠模型的脊髓损伤挫伤装置

<ol><li>Venkatesh, K., Ghosh, S. K., Mullick, M., Manivasagam, G., Sen, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Spinal+cord+injury%3A+pathophysiology%2C+treatment+strategies%2C+associated+challenges%2C+and+future+implications%5BTitle%5D)+AND+%22Cell+and+Tissue+Research%22%5BJournal%5D)">Spinal cord injury: pathophysiology, treatment strategies, associated challenges, and future implications.</a> Cell and Tissue Research. 377 (2), 125-151 (2019).</li>
<li>Chiu, C. W., Cheng, H., Hsieh, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Contusion+Spinal+Cord+Injury+Rat+Model%5BTitle%5D)+AND+%22Bio+Protocol%22%5BJournal%5D)">Contusion Spinal Cord Injury Rat Model.</a> Bio Protocol. 7 (12), e2337(2017).</li>
<li>Thygesen, M. M., Guldbæk-Svensson, F., Rasmussen, M. M., Lauridsen, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Contusion+Spinal+Cord+Injury+via+a+Microsurgical+Laminectomy+in+the+Regenerative+Axolotl%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments%22%5BJournal%5D)">Contusion Spinal Cord Injury via a Microsurgical Laminectomy in the Regenerative Axolotl.</a> Journal of Visualized Experiments. (152), 60337(2019).</li>
<li>Anderson, T. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+controlled+pneumatic+technique+for+experimental+spinal+cord+contusion%5BTitle%5D)+AND+%22Journal+of+Neuroscience+Methods%22%5BJournal%5D)">A controlled pneumatic technique for experimental spinal cord contusion.</a> Journal of Neuroscience Methods. 6 (4), 327-333 (1982).</li>
<li>Allen, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((SURGERY+OF+EXPERIMENTAL+LESION+OF+SPINAL+CORD+EQUIVALENT+TO+CRUSH+INJURY+OF+FRACTURE+DISLOCATION+OF+SPINAL+COLUMN%3A+A+PRELIMINARY+REPORT%5BTitle%5D)+AND+%22Journal+of+the+American+Medical+Association%22%5BJournal%5D)">SURGERY OF EXPERIMENTAL LESION OF SPINAL CORD EQUIVALENT TO CRUSH INJURY OF FRACTURE DISLOCATION OF SPINAL COLUMN: A PRELIMINARY REPORT.</a> Journal of the American Medical Association. LVII (11), 878-880 (1911).</li>
<li>Cheriyan, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Spinal+cord+injury+models%3A+a+review%5BTitle%5D)+AND+%22Spinal+Cord%22%5BJournal%5D)">Spinal cord injury models: a review.</a> Spinal Cord. 52 (8), 588-595 (2014).</li>
<li>Yan, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+modified+impactor+for+establishing+a+graded+contusion+spinal+cord+injury+model+in+rats%5BTitle%5D)+AND+%22Annals+of+Translational+Medicine%22%5BJournal%5D)">A modified impactor for establishing a graded contusion spinal cord injury model in rats.</a> Annals of Translational Medicine. 10 (8), 436(2022).</li>
<li>Gruner, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+monitored+contusion+model+of+spinal+cord+injury+in+the+rat%5BTitle%5D)+AND+%22Journal+of+Neurotrauma%22%5BJournal%5D)">A monitored contusion model of spinal cord injury in the rat.</a> Journal of Neurotrauma. 9 (2), 123-126 (1992).</li>
<li>Ghnenis, A. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Evaluation+of+the+Cardiometabolic+Disorders+after+Spinal+Cord+Injury+in+Mice%5BTitle%5D)+AND+%22Biology+%28Basel%29%22%5BJournal%5D)">Evaluation of the Cardiometabolic Disorders after Spinal Cord Injury in Mice.</a> Biology (Basel). 11 (4), 495(2022).</li>
<li>Scheff, S. W., Rabchevsky, A. G., Fugaccia, I., Main, J. A., Lumpp, J. E. Jr <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Experimental+modeling+of+spinal+cord+injury%3A+characterization+of+a+force-defined+injury+device%5BTitle%5D)+AND+%22Journal+of+Neurotrauma%22%5BJournal%5D)">Experimental modeling of spinal cord injury: characterization of a force-defined injury device.</a> Journal of Neurotrauma. 20 (2), 179-193 (2003).</li>
<li>Hong, Y. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Ultrasound+stimulation+improves+inflammatory+resolution%2C+neuroprotection%2C+and+functional+recovery+after+spinal+cord+injury%5BTitle%5D)+AND+%22Scientific+Reports%22%5BJournal%5D)">Ultrasound stimulation improves inflammatory resolution, neuroprotection, and functional recovery after spinal cord injury.</a> Scientific Reports. 12 (1), 3636(2022).</li>
<li>Noyes, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Electromechanical+impactor+for+producing+experimental+spinal+cord+injury+in+animals%5BTitle%5D)+AND+%22Medical+%26+Biological+Engineering+%26+Computing%22%5BJournal%5D)">Electromechanical impactor for producing experimental spinal cord injury in animals.</a> Medical & Biological Engineering & Computing. 25 (3), 335-340 (1987).</li>
<li>Stokes, B. T., Noyes, D. H., Behrmann, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+electromechanical+spinal+injury+technique+with+dynamic+sensitivity%5BTitle%5D)+AND+%22Journal+of+Neurotrauma%22%5BJournal%5D)">An electromechanical spinal injury technique with dynamic sensitivity.</a> Journal of Neurotrauma. 9 (3), 187-195 (1992).</li>
<li>Pearse, D. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Histopathological+and+behavioral+characterization+of+a+novel+cervical+spinal+cord+displacement+contusion+injury+in+the+rat%5BTitle%5D)+AND+%22Journal+of+Neurotrauma%22%5BJournal%5D)">Histopathological and behavioral characterization of a novel cervical spinal cord displacement contusion injury in the rat.</a> Journal of Neurotrauma. 22 (6), 680-702 (2005).</li>
<li>Wu, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+Tissue+Displacement-based+Contusive+Spinal+Cord+Injury+Model+in+Mice%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments%22%5BJournal%5D)">A Tissue Displacement-based Contusive Spinal Cord Injury Model in Mice.</a> Journal of Visualized Experiments. (124), 54988(2017).</li>
<li>Forgione, N., Chamankhah, M., Fehlings, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+Mouse+Model+of+Bilateral+Cervical+Contusion-Compression+Spinal+Cord+Injury%5BTitle%5D)+AND+%22Journal+of+Neurotrauma%22%5BJournal%5D)">A Mouse Model of Bilateral Cervical Contusion-Compression Spinal Cord Injury.</a> Journal of Neurotrauma. 34 (6), 1227-1239 (2017).</li>
<li>Ji, Z. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Highly+bioactive+iridium+metal-complex+alleviates+spinal+cord+injury+via+ROS+scavenging+and+inflammation+reduction%5BTitle%5D)+AND+%22Biomaterials%22%5BJournal%5D)">Highly bioactive iridium metal-complex alleviates spinal cord injury via ROS scavenging and inflammation reduction.</a> Biomaterials. 284, 121481(2022).</li>
<li>Chen, C., Qiao, X., Liu, W., Fekete, C., Reinhardt, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Epidemiology+of+spinal+cord+injury+in+China%3A+A+systematic+review+of+the+chinese+and+english+literature%5BTitle%5D)+AND+%22Spinal+Cord%22%5BJournal%5D)">Epidemiology of spinal cord injury in China: A systematic review of the chinese and english literature.</a> Spinal Cord. 60 (12), 1050-1061 (2022).</li>
<li>Flack, J. A., Sharma, K. D., Xie, J. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Delving+into+the+recent+advancements+of+spinal+cord+injury+treatment%3A+a+review+of+recent+progress%5BTitle%5D)+AND+%22Neural+Regeneration+Research%22%5BJournal%5D)">Delving into the recent advancements of spinal cord injury treatment: a review of recent progress.</a> Neural Regeneration Research. 17 (2), 283-291 (2022).</li>
<li>Khuyagbaatar, B., Kim, K., Kim, Y. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Conversion+Equation+between+the+Drop+Height+in+the+New+York+University+Impactor+and+the+Impact+Force+in+the+Infinite+Horizon+Impactor+in+the+Contusion+Spinal+Cord+Injury+Model%5BTitle%5D)+AND+%22Journal+of+Neurotrauma%22%5BJournal%5D)">Conversion Equation between the Drop Height in the New York University Impactor and the Impact Force in the Infinite Horizon Impactor in the Contusion Spinal Cord Injury Model.</a> Journal of Neurotrauma. 32 (24), 1987-1993 (2015).</li>
<li>Alizadeh, A., Dyck, S. M., Karimi-Abdolrezaee, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Traumatic+Spinal+Cord+Injury%3A+An+Overview+of+Pathophysiology%2C+Models+and+Acute+Injury+Mechanisms%5BTitle%5D)+AND+%22Frontiers+in+Neurology%22%5BJournal%5D)">Traumatic Spinal Cord Injury: An Overview of Pathophysiology, Models and Acute Injury Mechanisms.</a> Frontiers in Neurology. 10, 282(2019).</li>
<li>Bilgen, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+new+device+for+experimental+modeling+of+central+nervous+system+injuries%5BTitle%5D)+AND+%22Neurorehabilitation+and+Neural+Repair%22%5BJournal%5D)">A new device for experimental modeling of central nervous system injuries.</a> Neurorehabilitation and Neural Repair. 19 (3), 219-226 (2005).</li>
<li>Khan, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((GSNOR+and+ALDH2+alleviate+traumatic+spinal+cord+injury%5BTitle%5D)+AND+%22Brain+Research%22%5BJournal%5D)">GSNOR and ALDH2 alleviate traumatic spinal cord injury.</a> Brain Research. 1758, 147335(2021).</li>
</ol>

作者声明没有相互竞争的经济利益。

脊髓损伤可导致感觉和运动缺陷，从而导致严重的身体和精神损伤。在中国，不同省份的脊髓损伤发病率从14.6/100万到60.6不等18。脊髓损伤患病率的增加将给医疗保健系统带来更大的压力。目前，脊髓损伤的有效治疗选择有限，因为其病理机制和修复过程尚未完全了解19。有必要创建准确且可重复的脊髓损伤动物模型，以研究脊髓损伤的病理机制和修复过程。为此，我们开发了一种准确、可重复和简单的脊髓损伤仪器。
许多设备，包括 NYU-MASCIS 撞击器、无限地平线撞击器和 OSU 撞击器，都用于创建脊髓损伤的挫伤模型 8,9,12,13,20。这些设备操作复杂，并且由于人员的熟练程度不同，可能会产生较大的错误。此外，这些设备的设计存在各种缺陷，使它们的准确性和可重复性降低6.脊髓损伤的具体位置决定了其严重程度，因此在建立脊髓损伤模型时，定位方法决定了建模的准确性21。G smart SCI系统使用激光测距仪定位脊髓损伤部位，并根据测距仪数据调整冲击头高度，设定打击深度。另一种使用激光定位的敲击设备是印第安纳大学医学院 15 的路易斯维尔损伤系统装置 （LISA） 撞击器。虽然LISA使用激光定位，但它需要手动确定和调整冲击头的高度，以实现可控的打击深度，这增加了人为干预。同时，LISA采用气动打击和笔记本电脑控制，需要更多的实验空间，增加运营成本10。G smart SCI系统通过半自动化操作过程减少人为错误，并且由于其轻巧的设计而易于移动。
基于设定的参数，该设备可以准确地影响小鼠的脊髓，从而创建具有不同程度脊髓损伤的模型。在这项研究中，不同程度脊髓损伤的小鼠后肢功能表现出显着差异。重要的是，这些检测具有可重复性，并且可以始终如一地生成 SCI 模型。
在协议中，最关键的步骤包括准确进行椎板切除术，稳定小鼠的脊柱，以及使用激光进行精确定位，以确保实验的准确性和可重复性。在设计过程中，我们对设备进行了一些改进。我们发现在罢工之后，舞台无法轻松返回测距点，因此，我们添加了一个返回测距点的按钮。此外，所需的参数无法快速调整，因此，我们添加了一个数字键盘进行输入。第一版中的慢速充电电磁驱动也得到了改进。该装置目前仅限于建立胸椎脊髓损伤挫伤小鼠模型。应进行研究以支持使用该仪器建立大鼠脊髓损伤模型或颈椎脊髓损伤模型。
此外，堪萨斯大学医学院的 Bilgen 博士描述了一种计算机控制的撞击器装置，它可以诱发中枢神经系统 （CNS） 损伤，包括创伤性脑损伤 （TBI） 和脊髓损伤 （SCI）22。与我们的设备类似，该设备也使用了各种商业化的设备和系统，因此它成功商业化并使用了23。我们描述的设备具有自动化、准确和方便的特点，有望在未来实现商业化，让更多的脊髓损伤研究人员受益。 
总之，我们设计了一种自动小鼠脊髓撞击器来创建 SCI 挫伤模型。该设备通过激光测距仪提高了精度，并通过自动化操作过程减少了人为错误。此外，G smart SCI系统比其他设备更易于操作和携带，为脊髓损伤研究带来了便利。重要的是，该设备可以根据实验的需要准确且可重复地创建不同类别的SCI小鼠模型。

脊髓损伤通常会导致损伤节段以下的永久性脊髓功能障碍。它多是由物体撞击脊柱和脊柱过度伸展引起的，例如交通事故和跌倒1.由于脊髓损伤的有效治疗方案有限，使用动物模型阐明脊髓损伤的发病机制将有助于制定适当的治疗方法。由脊髓撞击引起的脊髓损伤挫伤模型导致开发了与大多数临床脊髓损伤病例具有相似病理的动物模型 2,3。因此，为脊髓损伤挫伤生成准确、可重复和方便的动物模型非常重要。
自 Allen 于 1911 年发明第一个脊髓损伤动物模型以来，用于建立脊髓损伤动物模型的仪器的发展取得了重大进展 4,5。根据损伤机制，脊髓损伤模型分为挫伤、压迫、牵引、脱位、横断或化学6。其中，利用外力移位和损伤脊髓的挫伤模型最接近大多数脊髓损伤患者的临床病因。因此，挫伤模型已被许多研究人员用于脊髓损伤研究 3,7。各种仪器用于开发脊髓损伤挫伤模型。纽约大学 （NYU） 多中心动物脊髓损伤研究 （MASCIS） 撞击器通过减重装置产生脊髓损伤挫伤8.经过几个更新版本后，MASCIS撞击器被广泛用于开发脊髓损伤挫伤动物模型9。然而，当MASCIS的冲击杆掉落并撞击脊髓时，可能会发生多处损伤，这会影响脊髓损伤模型的损伤程度。此外，实现机械精度以确保仪器的精度和制造模型的可重复性也具有挑战性。无限地平线撞击器通过控制施加在脊髓上的力而不是重跌落来引起挫伤10.它使用连接到传感器的计算机直接测量撞击器和脊髓之间的冲击力。当达到阈值时，冲击器立即缩回，从而避免重量反弹并提高精度10,11。然而，使用这种精细运动方式造成损伤会导致不一致的损伤和功能缺陷6.俄亥俄州立大学（OSU）设备通过电磁驱动器12,13以瞬态速率压缩脊髓的背面。该装置类似于无限地平线撞击器，因为它使用短距离按压来引起脊髓损伤。然而，它有各种局限性，因为零点的初始确定会由于脑脊液的存在而引起误差6,14。综上所述，可用于开发脊髓损伤挫伤动物模型的仪器很多，但它们都存在一些局限性，导致动物模型的准确性和可重复性不足。因此，为了更准确、更方便、更可重复地创建脊髓损伤小鼠挫伤模型，需要一种自动化、智能化的脊髓损伤撞击器。
我们提出了一种新型的脊髓损伤冲击器，广州暨南大学智能脊髓损伤系统（G smart SCI system; 图1），用于生成脊髓损伤挫伤模型。该设备使用激光测距仪作为定位装置，结合自动化移动平台，根据设定的打击参数（包括打击速度、打击深度和停留时间）自动进行打击。自动化操作减少了人为因素，提高了动物模型的准确性和可重复性。

<table><tbody><tr><td>0.01M PBS (powder, pH7.2-7.4)</td><td>Solarbio Life Sciences</td><td>P1010</td><td></td></tr><tr><td>2,2,2-Tribromoethanol</td><td>Macklin</td><td>75-80-9</td><td></td></tr><tr><td>4% paraformaldehyde tissue fixative</td><td>Biosharp life science</td><td>BL539A</td><td></td></tr><tr><td>Biomicroscope</td><td>Leica</td><td>LCC50 HD</td><td></td></tr><tr><td>CatWalk&amp;nbsp;</td><td>Noldus Information Technology</td><td>CatWalk XT 9.1</td><td></td></tr><tr><td>Cover glass</td><td>CITOTEST Scientific</td><td>10212432C</td><td></td></tr><tr><td>Embedding machine</td><td>Changzhou Zhongwei Electronic Instrument</td><td>BMJ-A</td><td></td></tr><tr><td>Ethanol absolute</td><td>DAMAO</td><td>64-17-5</td><td></td></tr><tr><td>FootFaultScan</td><td>Clever Sys Inc.</td><td>-</td><td></td></tr><tr><td>Glass slide</td><td>CITOTEST Scientific</td><td>80302-2104</td><td></td></tr><tr><td>Hematoxylin and Eosin Staining Kit</td><td>Beyotime Biotechnology</td><td>C0105S</td><td></td></tr><tr><td>micro-grinding drill&amp;nbsp;</td><td>FEIYUBIO</td><td>19-7010</td><td></td></tr><tr><td>Mouse spinal fixator</td><td>RWD Life Science</td><td>68094</td><td></td></tr><tr><td>Paraffin microtome</td><td>Thermo</td><td>shandon finesse 325</td><td></td></tr><tr><td>RotaRod for Mice</td><td>Ugo Basile</td><td>47600</td><td></td></tr><tr><td>Stereomicroscope</td><td>KUY NICE</td><td>SZM-7045</td><td></td></tr><tr><td>Tert-Amyl alcohol</td><td>Macklin</td><td>75-85-4</td><td></td></tr><tr><td>Xylene</td><td>China National Pharmaceutical</td><td>#10023418</td><td></td></tr></tbody></table>

automated impactor for contusive spinal cord injury model in mice

由车祸和跌倒等创伤引起的脊髓损伤 （SCI） 与永久性脊髓功能障碍有关。通过撞击脊髓来创建脊髓损伤的挫伤模型会导致与临床实践中大多数脊髓损伤相似的病理。准确、可重复和方便的脊髓损伤动物模型对于研究脊髓损伤至关重要。我们提出了一种新型的小鼠自动脊髓损伤挫伤装置，即广州暨南大学智能脊髓损伤系统，该系统可以生成具有准确性、可重复性和便利性的脊髓损伤挫伤模型。该系统 通过 激光距离传感器与自动化移动平台和高级软件相结合，精确地生成不同程度的脊髓损伤模型。我们使用该系统创建了三个级别的脊髓损伤小鼠模型，确定了它们的 Basso 小鼠量表 （BMS） 评分，并进行了行为和染色测定以证明其准确性和可重复性。我们展示了使用该设备开发损伤模型的每个步骤，形成了一个标准化的程序。该方法可产生可重复的脊髓损伤挫伤小鼠模型，并通过方便的处理程序减少人为操作因素。开发的动物模型对于研究脊髓损伤机制和相关治疗方法是可靠的。

如上所述，对 24 只雌性小鼠（8 周龄）进行椎板切除术。假手术组（n=6）小鼠未发生脊髓损伤，其余小鼠，包括0.5 mm组（n=6）、0.8 mm组（n=6）和1.1 mm组（n=6）均受到不同深度的脊髓撞击。定期记录 BMS 评分，直至术后 1 个月（图 4）。不同组小鼠术后BMS评分差异有统计学意义。1个月后，0.5mm组小鼠术后评分为4-6分，恢复到与假手术组相似的水平。0.8 mm 和 1.1 mm 组的小鼠术后评分为 1 至 2 分。1个月后，0.8毫米组恢复到4至6分，而1.1毫米组的小鼠几乎没有恢复。
1个月后，进行动物行为试验，包括足部故障、旋转和走秀。在足部断层测定（图5A）中，0.5 mm组和假肢组之间的后肢足部断层没有显着差异。然而，0.8 mm组的后肢足断层与所有其他组有显著差异。1.1 mm组小鼠足部故障率为100%，因为后肢无法支撑地面动物，与其他组有显著差异。在rotarod测试（图5B）中，我们记录了不同小鼠组的潜伏期。0.8 mm组和1.1 mm组均与其他组有显著差异，但假手术组与0.5 mm组的结果相似。在走秀测试（图6）中，我们记录并分析了不同组小鼠的规律指数和后方最大接触面积。不同组小鼠间规律指数和后方最大接触面积存在显著差异，表明不同脊髓损伤深度的小鼠在行走功能方面存在显著差异。因此，可以使用开发的设备创建具有不同脊髓损伤深度且后肢功能显着差异的小鼠模型。
最后，我们切除了小鼠的脊髓（图7A），并制作了苏木精和伊红（HE）染色的切片（图7C）。脊髓图像和HE染色切片均存在不同程度的损伤。综上所述，我们使用所提出的仪器准确地开发了不同等级的脊髓损伤小鼠模型。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/65656/65656fig01.jpg"> 图1：广州暨南大学智能脊髓损伤系统（G smart SCI系统）。 将脊柱固定器固定在托架上，用于固定鼠标。 通过 触摸屏设置冲击参数。使用横向微驱动器调整托架的横向位置，使用触摸屏调整正面位置。激光测距仪确认撞击位置并测量撞击高度，使撞击头能够根据设定的撞击位置和深度准确撞击。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/65656/65656fig01large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/65656/65656fig02.jpg"> 图 2：操作触摸屏。 （A） 在参数页面，可以设置损伤参数，包括冲击速度（0.5-2.0 m/s）、冲击深度（0 -3 mm）和停留时间（500 -2 ms）。（B）在移动页面中，可以设置冲击头的高度和脊柱固定器的位置。（C） 在准备页面中，单击 “就绪 ”按钮将导致根据设置的参数自动将冲击头和托架工作台调整到特定位置。通过单击 “开始 ”按钮，冲击头将根据设置的参数撞击脊髓。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/65656/65656fig02large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/65656/65656fig03.jpg"> 图3：受伤后的脊髓。 （A） 假组。脊柱固定器固定T10椎骨，椎板切除术后脊髓暴露无损伤。（B） 0.5 mm组。脊髓在0.5毫米深处被击中后受轻伤，并有少量充血。（C） 0.8 mm组。脊髓中度损伤，在0.8 mm深处被击中后有明显的充血。 （D） 1.1 mm组。脊髓在1.1毫米深处被击中后严重受伤，并且充血严重。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/65656/65656fig03large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/65656/65656fig04.jpg"> 图 4：BMS 分数。 记录小鼠从术后第一天到术后1个月的BMS评分（n =6 /组）。*p< 0.05， ** p < 0.01， *** p < 0.001 与单因素方差分析进行比较。日期表示为平均值±均值的标准误差 （SEM）。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/65656/65656fig04large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/65656/65656fig05.jpg"> 图 5：脚步测试和旋转测试。 （A）不同程度脊髓损伤小鼠的后肢足部缺陷（n =6 /组）。p <0.001 比较使用单因素方差分析。（B）比较不同程度脊髓损伤小鼠（n =6 /组）在加速旋转期间跌倒的潜伏期。* p < 0.05，*** p < 0.001 与单因素方差分析进行比较。日期表示为平均值±均值的标准误差 （SEM）。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/65656/65656fig05large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/65656/65656fig06.jpg"> 图6：走秀测试。 （A，C）不同程度脊髓损伤小鼠的规律性指数（n = 6 /组）。* p < 0.05，*** p < 0.001 与单因素方差分析进行比较。（乙、丁）使用软件自动分析后方最大接触面积（n = 6 /组）。** p < 0.01，*** p < 0.001 与单因素方差分析进行比较。日期表示为平均值±均值的标准误差 （SEM）。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/65656/65656fig06large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/65656/65656fig07.jpg"> 图7：脊髓和H&E染色。 （A）手术后1个月不同程度损伤小鼠的脊髓。（B） 放大数字。（C）不同损伤程度小鼠脊髓损伤部位H&E染色。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/65656/65656fig07large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>

Watch this Scientific Journal Video about Automated Impactor for Contusive Spinal Cord Injury Model in Mice at JoVE.com

作者聚焦：深入了解脊髓损伤研究的创新

这里介绍的是一种新型的小鼠自动脊髓损伤挫伤装置，可以准确生成不同程度的脊髓损伤挫伤模型。

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 ...

我的研究范围包括脊髓损伤的发病机制和治疗方法。我们最近的研究发现，各种生物材料可以通过抗氧化应激促进小鼠脊髓损伤的修复。生物材料传感器、技术和基因修饰技术的发展，以及脊髓损伤模型的优化正在推进我们的研究领域。由于脊髓损伤的压迫和机制不完全了解，目前很难使用单一干预措施来治愈脊髓损伤。未来，研究可能需要多种干预方法。我们发现精神即刻敷可以通过抗氧化应激参与动物脊髓损伤的治疗和修复。

automated-impactor-for-contusive-spinal-cord-injury-model-in-mice

Nanyang Technological University

Research

JoVE Journal

Neuroscience

Automated Impactor for Contusive Spinal Cord Injury Model in Mice

1.8K 次观看.  Jinan University. 我的研究范围包括脊髓损伤的发病机制和治疗方法。我们最近的研究发现，各种生物材料可以通过抗氧化应激促进小鼠脊髓损伤的修复。生物材料传感器、技术和基因修饰技术的发展，以及脊髓损伤模型的优化正在推进我们的研究领域。由于脊髓损伤的压迫和机制不完全了解，目前很难使用单一干预措施来治愈脊髓损伤。未来，研究可能需要多种干预方法。我们发现精?...

视频: 作者聚焦：深入了解脊髓损伤研究的创新

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 ...

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 ...

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 Repetitive Concussive Head Injury Model in Mice

Despite the concussion/ mild traumatic brain injury (mTBI) being the most frequent occurrence of traumatic brain injury, there is still a lack of knowledge on the injury and its effects. To develop a better understanding of concussions, animals are often used because they provide a controlled, rigorous, and efficient model. Studies have adapted traditional animal models to perform mTBI to stimulate mild injury severity by changing the injury parameters. These models have been used because they can produce morphologically similar brain injuries to the clinical condition and provide a spectrum of injury severities. However, they are limited in their ability to present the identical features of injuries in patients. Using a traditional impact system, a repetitive concussive injury (rCHI) model can induce mild to moderate human-like concussion. The injury degree can be determined by measuring the period of loss of consciousness (LOC) with a sign of a transient termination of breathing. The rCHI model is beneficial to use for its accuracy and simplicity in determining mTBI effects and potential treatments.

Concussion presents the most common type of traumatic brain injury. Therefore, a repetitive concussive animal model, which replicates the important features of an injury in patients, may provide a means to study concussion in a rigorous, controlled, and efficient manner.

重复震荡颅脑损伤模型小鼠

Despite the concussion/ mild traumatic brain injury (mTBI) being the most frequent occurrence of traumatic brain injury, there is still a lack of ...

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 ...

Murine Model of Controlled Cortical Impact for the Induction of Traumatic Brain Injury

The Centers for Disease Control and Injury Prevention estimate that almost 2 million people sustain a traumatic brain injury (TBI) every year in the United States. In fact, TBI is a contributing factor to over a third of all injury-related mortality. Nonetheless, the cellular and molecular mechanisms underlying the pathophysiology of TBI are poorly understood. Thus, preclinical models of TBI capable of replicating the injury mechanisms pertinent to TBI in human patients are a critical research need. The controlled cortical impact (CCI) model of TBI utilizes a mechanical device to directly impact the exposed cortex. While no model can full recapitulate the disparate injury patterns and heterogeneous nature of TBI in human patients, CCI is capable of inducing a wide range of clinically applicable TBI. Furthermore, CCI is easily standardized allowing investigators to compare results across experiments as well as across investigative groups. The following protocol is a detailed description of applying a severe CCI with a commercially available impacting device in a murine model of TBI.

Here we describe a protocol for the induction of murine traumatic brain injury via an open-head controlled cortical impact.

创伤性脑损伤诱导受控皮质冲击的母模型

The Centers for Disease Control and Injury Prevention estimate that almost 2 million people sustain a traumatic brain injury (TBI) every year in the ...

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, ...

Electromagnetic Controlled Closed-Head Model of Mild Traumatic Brain Injury in Mice

Highly reproducible animal models of traumatic brain injury (TBI), with well-defined pathologies, are needed for testing therapeutic interventions and understanding the mechanisms of how a TBI alters brain function. The availability of multiple animal models of TBI is necessary to model the different aspects and severities of TBI seen in people. This manuscript describes the use of a midline closed head injury (CHI) to develop a mouse model of mild TBI. The model is considered mild because it does not produce structural brain lesions based on neuroimaging or gross neuronal loss. However, a single impact creates enough pathology that cognitive impairment is measurable at least 1 month after injury. A step-by-step protocol to induce a CHI in mice using a stereotaxically guided electromagnetic impactor is defined in the paper. The benefits of the mild midline CHI model include the reproducibility of the injury-induced changes with low mortality. The model has been temporally characterized up to 1 year after the injury for neuroimaging, neurochemical, neuropathological, and behavioral changes. The model is complementary to open skull models of controlled cortical impact using the same impactor device. Thus, labs can model both mild diffuse TBI and focal moderate-to-severe TBI with the same impactor.

The protocol describes mild traumatic brain injury in a mouse model. In particular, a step-by-step protocol to induce a mild midline closed head injury and the characterization of the animal model is fully explained.

小鼠轻度创伤性脑损伤的电磁控制闭头模型

Highly reproducible animal models of traumatic brain injury (TBI), with well-defined pathologies, are needed for testing therapeutic interventions and ...