Chris Daly博士是"手术基金会Richard Jepson研究奖学金"的获得者。作者要感谢Anne Gibbon博士，Dong Zhang博士和Monash University Monash Animal Services的工作人员对动物手术和护理的协助。

本手稿中详细列出的方案遵循蒙纳士大学动物伦理学的动物保健指南。该议定书的动物伦理批准已经由蒙纳士大学动物伦理学授予。伦理批准号:MMCA / 2014/55 绵羊准备注意:使用二至四岁的母羊。 <ol><li>快羊在麻醉前18小时。在运动前6-12小时，让动物进入水中。 </li><li>通过静脉注射盐酸米托咪啶（0.015-0.020 mg / kg）镇静动物，以方便转移到手术套件。 注意:盐酸米托咪定用于减少与其他动物分离相关的动物压力和搅动，以转移到手术套件。 </li><li>注射硫代戊酮（10-13 mL / kg），用于在到达手术套件时诱导麻醉。 </li><li>管理预防性内静脉注射抗生素（阿莫西林1 g IV）。 </li><li>使用尺寸7.5-9毫米（内径）气管插管20的插管绵羊。 </li><li>使用吸入式异氟烷（2-3％）在100％氧气中以2L / min的流速维持麻醉。将脉搏血氧计安装在羊的耳朵上。 </li><li>密切监测绵羊的生命体征（心率，呼吸频率和氧饱和度通过脉搏血氧仪和观察）和意识水平。 注意:轻度麻醉如自发咀嚼，主动反流和自发运动的指标应提高麻醉水平。指示紧急减轻麻醉的红旗标志包括呼吸困难和严重的心动过缓。眼睛的旋转不是绵羊深度麻醉的一致指标19 。 </li></ol>椎间盘水平和切口<ol><li>收集s该程序所需的紧急工具:兽医剪刀，20 mL鲁尔锁注射器，21G IV针，＃4手术刀手柄，＃22手写刀刀片，Gillies组织钳，Metzenbaum弯曲解剖剪刀，Deaver牵开器，Hohmann牵开器刀片，3.5 mm Brad点钻头，钻头停止，钻头，可高压灭菌的兽用钻袋，针架，2-0可吸收合成编织缝线，3-0可吸收合成编织缝线和Mayo缝合剪刀。 </li><li>准备操作套件。用70％乙醇清洁手术台和仪器架。在手术前高压灭菌所有手术器械。进行手术前麻醉检查。 </li><li>将羊放在手术台上的右侧位置。 </li><li>使用电子剪刀，剃刮由下肋骨下部由髂骨上部定义的区域，中间由对侧腰椎横突进行，并且同侧腰椎横突方向约10厘米。 </li><li>掌握髂嵴，腰椎横突（L1-6）和成像脊椎角度为手术切口部位的标志。用无菌笔标记这些地标。 </li><li>用氯己定和碘化碘消毒洗涤剂进行消毒，准备外侧腹部。 </li><li> 在整个手术过程中观察标准手术无菌技术。手术组在手术前擦洗。在手术部位放置无菌的开窗方形悬垂物，并在头顶上放置一个大的无菌方巾。 <ol><li>在手术前对所有在手术部位使用的物品进行灭菌。在整个手术过程中监测和维持手术部位的无菌性。确保引入无菌领域的所有物品都是无菌的，并以无菌方式转移。 </li></ol></li><li>使用手术放大镜放大镜和前照灯，以便于手术过程中的可视化。 </li><li>做一个纵切口使用＃2手术刀刀片连接到＃4手术刀手柄上，平行于前方1厘米至1厘米的腰椎横突，高于和低于感兴趣的椎间盘水平。 注意:关于切口规划的更多信息可以在讨论中找到。 </li><li>使用单极透热疗法分割下面的皮下组织和腹壁肌肉组织的外侧;将切开的方向指向感兴趣的椎间盘上方和下方的腰椎横突的尖端。 </li><li>将胸廓筋膜纵向分割成横向处理。 </li><li>可视化并保存下面的正方形体 ，腰肌和横穿神经血管束。 </li><li>通过使用透热疗法的手术保持止血。 </li><li>在暴露的椎间盘后，将手指扫到腹膜平面和腹后壁肌肉组织之间sc水平进行数字钝性解剖。 </li><li>使用Deaver牵开器将后方肌肉和腰肌肌间退缩，以进一步暴露椎间盘。 </li><li>用于椎间盘突出的凹椎椎体和凸椎间盘。 </li><li>将收缩器立即放在光盘上，并注意确保腰部血管未损坏。 </li><li>使用头灯的手术放大镜放大，识别位于下端板约1厘米尾的腰部血管。 </li><li>执行术中横向X线以确认椎间盘水平。 21 注意:射线照相设置:47kV; 4 mAs 21 </li><li> 根据所需的椎间盘水平，通过如下分离周围的结构和附件来暴露椎间盘。 <ol><li>对于L3 / 4级及以上级别，请将肌肉附件放在一边sc使用数字钝器解剖。 </li><li>对于L4 / 5及以下级别，使用双极透热和剪刀将较厚的肌腱肌肉附着物锐利地划分在椎间盘上。 注意:由于髂嵴阻塞，L6 / S1椎间盘可能难以进入。如果不能通过侧向方法实现进入，则可以使用前方法。 </li></ol></li></ol> 3.钻头伤害注意:术前规划包括伤害/治疗水平和控制水平的分配。有关级别分配的更多信息可以在讨论中找到。 <ol><li>通过观察椎间盘的左侧和前端来定义钻头入口点。 注意:入口点位于左前外侧象限的中点（由椎间盘前后两侧限定）。在该入口点插入钻头，其轨迹瞄准朝向椎间盘的中心并且稍微颅导向垂直。 </li><li> 在钻机上安装一个Brad-point钻头。确保钻头的直径略小于椎间盘高度， 即 60-70 kg绵羊腰椎间盘的约3.5 mm。 <ol><li>应用钻头止动器，为60-70公斤羊的腰椎间盘提供大约一半腰椎间盘直径的无保护钻头长度， 即 〜12 mm。 </li></ol></li><li>将钻头应用到入口点，并将其引导到稍微颅骨到椎间盘中心的轨迹。轻微的颅骨角度是尽量减少终板损伤的风险。 </li><li>将钻头缓慢进入椎间盘，钻头在低功率下1秒钟。如果遇到过多的阻力，请以轻微的颅骨或尾部方式调整轨迹 注意:这样过大的电阻可能表示与端板接触。 </li></ol>关闭<ol><li>一旦达到止血效果，用林格斯溶液冲洗伤口。 </li><li>执行分层闭合，优选使用2-0可吸收的合成编织缝线到侧腹壁组织和连续3-0未染色的可吸收合成编织的皮下缝合到皮肤。 </li></ol> 5.术后管理<ol><li>在腹股沟区域放置芬太尼透皮贴片（75μg/ h）进行手术后镇痛3天。 </li><li>另外，如果需要，使用静脉注射丁丙诺啡（0.005-0.01 mg / kg）补充镇痛。 </li><li>停止吸入麻醉。当发生自主呼吸时，取出气管内管。 </li><li>让动物在恒定观察下在保持笼中恢复。 注意:在重新获得足够的食物之前，动物不应无人值守保持胸骨躺卧的能力。 </li><li>一旦动物完全警觉和站立，重新引进食物和水。一旦完全恢复，将动物返回到其他动物持有笔的手术设备。 </li><li>密切观察24小时，继续观察一周。监测手术后疼痛或痛苦的证据。 注意:术后透皮芬太尼补片应用三天应提供足够的止痛。额外的止痛要求应提示动物检查。 </li><li>通常喂羊，允许绵羊不受限制地进行正常的活动。观察羊的任何神经缺陷的证据，如跛足。 注意:由钻头损伤方法产生的椎间盘缺损是椎间盘的前外侧，损伤深度受到钻头中止的钻头限制。由于神经元素位于椎间盘的后部/后外侧，继发于症状性髓核的神经妥协的风险偏远。该模型的这种解剖特征阻止了神经学检查的使用，以区分椎间盘退变与无髓核突出。 </li><li>在实验期结束时，将羊送回大学农场等待安乐死和尸检。 </li></ol> 6.安乐死<ol><li>在钻头椎间盘损伤后的适当时间间隔内进行绵羊安乐死。 </li><li>注射戊巴比妥钠（&gt; 100 mg / kg）用于安乐死。 </li></ol>

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</ol>

作者宣称他们没有竞争的经济利益来披露。

这种微创侧向通道方法是有效和安全的，没有手术后疝气，腹部伤口开裂或感染在本系列中观察到。使用具有深度停止的钻头椎间盘损伤模型提供了一种可重现的方法，引起已知尺寸（ 即本研究中的3.5mm直径×12mm深度损伤）的一致的椎间盘损伤。根据我们的经验，该方法比传统描述的羊手术刀片腰椎间盘切开术模型6,22（未发表的数据）中观察到的椎间盘退变程度低得多。这将在即将出版的出版物中描述。 在进行初始纵向皮肤切口（步骤2.9）时，切口的确切长度和​​位置应根据所需的椎间盘水平进行修改。更优异的光盘级别（T12 / L1）可以是rea通过将切口延伸到椎体外角，而延伸到髂嵴的切口将允许进入下腰椎（至L5 / L6）。 10厘米切割将有助于获得三到四个光盘级别，而对于访问单个光盘则需要5厘米的较小的聚焦切口。我们更喜欢在两级进行伤害，通常是L2 / L3和L3 / 4。这使得相邻的L1 / 2和L4 / 5椎间盘水平能够用作非受伤的内部控制。一旦技术上有信心，一只羊的外科手术可以在不到一个小时内完成，最少失血和不适18 。该技术的关键步骤和主要技术挑战是避免钻头椎间盘损伤时的终板损伤。在钻头入口处明确界定椎间盘的上下缘是非常重要的。缓慢地将低位演练进入到interverte中胸部圆盘，起始大致垂直与轻微的颅角也最大限度地减少终板损伤的风险。可能需要延长皮肤切口以获得足够的钻孔角度。 这种技术的简单修改包括钻头尺寸和深度的变化，因为这些将由动物和腰椎间盘的大小决定。这种方法可用于可靠地诱导椎间盘从T12 / L1到L5 / 6的退化。腹膜后方法可用于进入椎间盘以通过其它机制诱导变性16或施用实验治疗剂。 这种方法的局限性涉及椎间盘损伤的程度和随后由该方法引起的变性。如果一名调查人员试图诱发严重椎间盘退变，其他更积极的椎间盘损伤方法如解剖刀刀片瓣膜切除术6应考虑。通过钻头的损伤方法在椎间盘中产生的急性缺陷相对较小，并且可能不适合于在损伤时施用治疗剂。 选择脊椎椎间盘椎间盘损伤模型有几个原因。非人类灵长类动物尽管与临床状况（ 即直立和半直立姿势下的大量时间）具有解剖学和生物力学相似性，但存在足够的道德和实践考虑，以防止其在许多机构中的使用。虽然是四足的，羊腰椎间盘在解剖学上是可比的，并且暴露于对人腰椎间盘16,18的类似生物力学应力。绵羊在成年早期证明髓核细胞从髓核中的损失，人类也是这样。脊髓细胞可能具有祖细胞功能，并且已被证明通过椎间盘基质的再生来影响椎间盘退变的过程。最后，从务实的角度来看，绵羊是强硬的动物，能容忍手术，随时可用，并呈现经济上可行的选择16,18 。 山羊18是另一种腰椎间盘退变的动物模型，其呈现绵羊模型的许多优点 - 类似的尺寸，经济可行性，弹性以及在成人24中没有脊索细胞。其他大型动物模型提出了额外的挑战 - 猪模型中脊索细胞的存在以及可能与犬模型相关的伦理问题。对于interve的动物模型进行全面的回顾椎间盘退变，读者指导Daly 等人最近的一篇综述。 al 3 。 由于羊椎间盘表现出脊髓细胞的自发性损失，并且在23岁时发生进行性退化，所以必须确保实验中羊龄的一致性。我们更喜欢使用二至四岁的母羊，因为在这个年龄，脊索细胞现在不存在23 。从我们自己的经验来看，尽管失去了脊索细胞，但两岁到四岁的羊发生了最小的自发性变性。此外，绵羊椎体生长板在大约24个月时关闭，椎体生长已经停止数月前25 ，最大限度地减少了相邻生长板细胞对椎间盘再生的任何影响的风险。母羊是首选，因为他们的侵略性比他们的男性同行促进easi呃动物处理。如果使用男性羊，我们建议使用湿羊。 在张18的研究中，使用类似的钻头伤害方法，其中直径为4.5mm的钻头插入15mm深，手动旋转360°，以产生山羊椎间盘变性，射线照相术中无统计学差异损伤椎间盘的Pfirrmann退行性评分与术前图像相比。然而，有明显的组织学证据表明轻度至中度椎间盘退变26 。相比之下，在这项研究中，总体形态学和9.4T MRI分析显示腰椎间盘退变性显着变化的证据表明这种方法的显着优点。 该方法的应用和结果将在即将出版的出版物中描述，比较椎间盘损伤的钻头方法o在羊模型中建立的瓣膜切开术。这种方法也可以用于将来进行再生疗法的研究。

下背痛是全世界残疾的主要原因1 。腰椎间盘退变相关性椎间盘突出症被认为是下腰痛的重要原因之一2 。对可靠的椎间盘疾病动物模型的需求不断增加，以扩大对退行性过程的理解和潜在疗法的调查。 存在许多椎间盘退变的动物模型3 。用于调查退行性椎间盘疾病的动物模型的大小范围从小鼠4到较大的哺乳动物如狗5 ，绵羊6和非人灵长类动物7 。用于诱导椎间盘退变的方法可以大致分为机械类（ 例如椎间盘缩小术）<html> n 8或手术损伤6 ），化学（ 例如化学性核解解5 ）或较不常见的自发性变性（ 例如沙鼠9 ）。 鉴于人椎间盘退变的复杂性，不存在完美的动物模型。然而，在选择合适的动物模型以模拟这种情况的重要考虑已经被确定3 。这样的考虑包括:没有脊索细胞（原始细胞具有可能的祖细胞功能10，不存在于人，绵羊，山羊和软骨营养不良犬中的成年髓核，但存在于大多数哺乳动物中），动物相对于人和椎间盘尺寸相似，可比的生物力学临床条件，机制和伦理考虑3 。 <html>非人类灵长类动物符合上述许多标准，自发性椎间盘退变的狒狒和猕猴模型已被描述为11,12,13 ，两种物种在直立或半直立的姿势中花费大量的时间 - 一个明显的优势然而，道德和实际的考虑（ 例如费用，住房，自发性退化的延迟发作）限制在许多机构中的使用。 羊脊柱是椎间盘退变的确定模型，其优点包括细胞，生物力学和解剖学相似于人类脊柱10,14,15。尽管绵羊的四足身材，羊腰椎间盘暴露于人类椎间盘的相似应力s ="xref"&gt; 14。羊胎模型从伦理角度也比非人类灵长类动物模型更受广泛接受。已经描述了各种方法来启动退行性过程，其中许多方法需要直接进入椎间盘。由于脊髓在骶骨区域的终止以及后腰纵韧带在卵巢腰椎骨化，椎间盘后路接近技术上具有挑战性，较不常见于羊16 。通过前路或前外侧入路的羊腰椎的传统通路需要大的腹部切口，充满疝风险，损害内脏和神经血管结构16 。使用相对较小的外侧切口远离依赖性腹部区域可能会降低这种风险17 。 我们提出一个羊退行性腰椎间盘疾病使用微创外侧方法进行的钻头损伤，并受到张等的工作的启发。 al 18 。该协议的目的是使可靠的腰椎间盘损伤模型易于重复，产生一致的伤害，并且是安全和良好耐受的。这种方法非常适合于寻求诱导较轻度的腰椎间盘退变的研究者，而不是用传统手术瓣膜切开术（未发表的数据）观察椎间盘退变或再生疗法。这些发现将在即将出版的出版物中进行描述。

<table><tbody><tr><td>Medetomidine Hydrochloride (10 mL Injection)</td><td>Therapon/Zoetis</td><td>PFIDOM10</td><td>Multiple suppliers: Zoetis/Ilium</td></tr><tr><td>Thiopentone</td><td>Troy</td><td>Triothiopentone</td><td>Multiple suppliers: Neon Laboratories, Jagsonphal Pharmaceuticals</td></tr><tr><td>Isoflurane (2 - 3 % in oxygen)</td><td>Baxter</td><td>AHN3636</td><td>Multiple suppliers: Baxter/VetOne</td></tr><tr><td>Amoxicillin parenteral</td><td>GlaxoSmithKline</td><td>JO1CA04</td><td>Multiple suppliers: GlaxoSmithKline/Merck</td></tr><tr><td>Bupivacaine (0.5% Injection 20 mL)</td><td>Pfizer</td><td>005BUP001</td><td>Multiple suppliers: Pfizer/AstraZeneca</td></tr><tr><td>PVD Iodine Solution</td><td>Jurox</td><td>61330</td><td>Multiple suppliers: Jurox/Orion</td></tr><tr><td>Chlorhexidine 5%w/v</td><td>Jurox</td><td>Chlorhex C 5L (SCRUB)</td><td>Multiple suppliers: Jurox/Pfizer</td></tr><tr><td>Transdermal Fental Patch (75 &amp;mu;g/h)</td><td>Janssen-Cilag</td><td>S8-Dur7.5</td><td>Multiple suppliers: Sandoz</td></tr><tr><td>Buprenorphine iv</td><td>Jurox</td><td>504410</td><td>Multiple suppliers: LGM Pharma</td></tr><tr><td>Atipamezole (Antisedan 0.06 mg/kg - 0.08 mg/kg)</td><td>Zoetis</td><td>PFIANT10</td><td>Multiple suppliers: Ilium</td></tr><tr><td>Oster Golden A5 2-Speed Clippers</td><td>Oster</td><td>078005-140-003</td><td>Oster</td></tr><tr><td>20 mL luer lock syringe</td><td>Terumo</td><td>6SS+20L</td><td>Multiple suppliers: Medshop Australia/Terumo</td></tr><tr><td>21 G IV needle</td><td>Terumo</td><td>SG3-1225</td><td>Multiple suppliers:Medshop Australia/Terumo</td></tr><tr><td>#4 scalpel handle</td><td>Austvet</td><td>AD010/04</td><td>Multiple suppliers: Austvet/SurgicalInstruments</td></tr><tr><td>#22 scalpel baldes</td><td>Austvet</td><td></td><td></td></tr><tr><td>Gillies tissue forceps</td><td>Austvet</td><td>AB430/15</td><td>Multiple suppliers: Austvet/SurgicalInstruments</td></tr><tr><td>Metzenbaum curved dissecting scissors</td><td>Austvet</td><td>AC101/14</td><td>Multiple suppliers: Austvet/SurgicalInstruments</td></tr><tr><td>Deaver retractor</td><td>Surgical Instruments</td><td>23.75.03</td><td>Multiple suppliers: Surgical Instrument/Austvet</td></tr><tr><td>Hohmann retractor</td><td>Austvet</td><td>KA173/35</td><td>Multiple suppliers: Austvet/SurgicalInstruments</td></tr><tr><td>Mayo suture scissors</td><td>Austvet</td><td>AC911/14</td><td>Multiple suppliers: Austvet/SurgicalInstruments</td></tr><tr><td>Needleholder 14 cm&amp;nbsp;</td><td>EliteMedical</td><td>18-1030</td><td>Multiple suppliers: EliteMedical/Austvet</td></tr><tr><td>CMT 3.5 mm Brad-Point Drill</td><td>Carbatec</td><td>516-035-51</td><td>Multiple suppliers: Southeast Tool/Carbatec</td></tr><tr><td>Drill Bit Stop 4 mm</td><td>Drill Warehouse</td><td>20121600</td><td>Multiple suppliers: Amazon</td></tr><tr><td>Bosch 10.8 V Cordless Angle Drill</td><td>Get Tools Direct</td><td>GWB10.8V-LIBB</td><td>Multiple suppliers:Bunnings/Get Tools Direct</td></tr><tr><td>Autoclavable veterinary drill bag</td><td>AustVet</td><td>DRA043-AV</td><td>AustVet</td></tr><tr><td>2-0 absorbable synthetic braided sutures</td><td>Ethicon</td><td>VCP335H</td><td>Ethicon</td></tr><tr><td>3-0 absorbable synthetic braided sutures</td><td>Ethicon</td><td>VCP232H</td><td>Ethicon</td></tr><tr><td>Siemens 3 Tesla Skyra Widebore MRI</td><td>Siemens</td><td>N/A</td><td>Siemens</td></tr><tr><td>9.4 Tesla Agilent (Varian) MRI</td><td>Agilent Technologies</td><td>N/A</td><td>Agilent Technologies</td></tr><tr><td>Atomscope HF 200 A Radiogaph</td><td>Radlink</td><td>330003A</td><td>Multiple Suppliers: Radlink/DLC Australia</td></tr><tr><td>Veterinary Pulse Oximiter</td><td>DLC&amp;nbsp;</td><td>192500A</td><td>Multiple suppliers: DLC Australi Pty Ltd/AustVet</td></tr></tbody></table>

ovine lumbar intervertebral disc degeneration model utilizing a lateral retroperitoneal drill bit injury

椎间盘退变是背部疼痛发展和全球残疾的主要原因的重要因素。已经开发了许多椎间盘退变的动物模型。关于形态学，生物力学性质和无脊髓细胞的缺失，理想的动物模型应该密切地模拟人椎间盘。绵羊腰椎间盘模型符合这些标准。我们通过侧腹膜后方法，利用钻头损伤呈现椎间盘退变的羊模型。横向方法显着减少了与传统的前路方法相关的切口和潜在发病率。使用钻头方法的伤害提供了产生一致和可重复的损伤，精确尺寸的能力，从而引发一致程度的椎间盘退变。环形的焦点性质髓核缺损更密切地模拟了局限性椎间盘突出症的临床状况。羊按照这个程序迅速恢复，通常是在一小时内移动和进食。随后发生椎间盘退变，羊进行尸检和随后的分析。我们认为椎间盘退变的钻头损伤模型比较传统的环形损伤模型具有优势。

术前，羊进行基线3T磁共振成像（MRI）评估潜在的椎间盘形态和变性。羊进行额外的术中横向X线检查以确认椎间盘水平和计算椎间盘高度指数。来自3T MRI和术中X光片的术前矢状面切片如图1所示 。 <img alt="图1" src="/files/ftp_upload/55753/55753fig1.jpg" /> 图1:手术前3T MRI（ A ）和术中横向射线照相（ B ）。 （ A ）羊腰椎3T MRI（3T T2加权自旋回波序列）的矢状切片显示腰椎1/2（L1 / 2）至腰骶（L6 / S1）椎间盘。椎间盘具有均匀的高信号外观表明没有明显的手术前椎间盘退变的证据。请注意，羊腰椎通常有六个腰椎，羊脊髓终止于骶骨区域。 （ B ）外科手术器械标记L3 / L4椎间盘的术中横向X光片（设置:47 kV; 4 mAs），显示L1 / L2和L6 / S1椎间盘。刻度棒= 25mm。 <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55753/55753fig1large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> 手术后，羊通常恢复，并在1小时内独立移动。仔细观察绵羊一周，然后在椎间盘损伤后8周再次返回农田，直到尸检。没有发生不良事件。在椎间盘损伤后8周，羊进行尸检，X光检查和MRI检查以及用于组织学和生化分析的盘的加工。 损伤后8周（56天）损伤的羊腰椎间盘的总体形态学外观和放射学9.4T MRI图像的代表性手术后图像如图2所示 。总体形态学图像显示穿透纤维环并延伸到髓核中的钻头损伤道。这在9.4T MRI中也是显而易见的。将在即将出版的详细描述模型验证研究的出版物中描述对该方法的结果的综合描述和分析。 <img alt="图2" src="/files/ftp_upload/55753/55753fig2.jpg" /> 图2:损伤盘的形态和MRI图像。 （ A ）。椎间盘突出症状的形态学形态穿透纤维环（AF）并延伸到髓核（NP）中。 （ B ）。 9.4T MRI（T2加权快速自旋回波序列）也显示穿透AF进入NP的损伤部位。比例尺= 10mm。 <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55753/55753fig2large.jpg" target="_blank">请点击此处查看此图的较大版本。</a>

Watch this Scientific Journal Video about Ovine Lumbar Intervertebral Disc Degeneration Model Utilizing a Lateral Retroperitoneal Drill Bit Injury at JoVE.com

Ovine Lumbar Intervertebral Disc Degeneration Model Utilizing a Lateral Retroperitoneal Drill Bit Injury

椎间盘退变是背部疼痛的重要原因，也是全球残疾的主要原因。存在许多椎间盘退变的动物模型。我们展示了椎间盘退变的羊模型，利用钻头，实现了一致的椎间盘损伤和椎间盘退变的可重复水平。

Ovine Lumbar椎间盘退变模型利用横向腹膜后位钻伤

Intervertebral disc degeneration is a significant contributor to the development of back pain and the leading cause of disability worldwide. Numerous ...

ovine-lumbar-intervertebral-disc-degeneration-model-utilizing-lateral

Universidad San Sebastian

Research

JoVE Journal

Medicine

11.6K 次观看.  Monash University. 椎间盘退变是背部疼痛的重要原因，也是全球残疾的主要原因。存在许多椎间盘退变的动物模型。我们展示了椎间盘退变的羊模型，利用钻头，实现了一致的椎间盘损伤和椎间盘退变的可重复水平。 

视频: Ovine Lumbar Intervertebral Disc Degeneration Model Utilizing a Lateral Retroperitoneal Drill Bit Injury

Renal Ischaemia Reperfusion Injury: A Mouse Model of Injury and Regeneration

Renal ischaemia reperfusion injury (IRI) is a common cause of acute kidney injury (AKI) in patients and occlusion of renal blood flow is unavoidable during renal transplantation. Experimental models that accurately and reproducibly recapitulate renal IRI are crucial in dissecting the pathophysiology of AKI and the development of novel therapeutic agents. Presented here is a mouse model of renal IRI that results in reproducible AKI. This is achieved by a midline laparotomy approach for the surgery with one incision allowing both a right nephrectomy that provides control tissue and clamping of the left renal pedicle to induce ischaemia of the left kidney. By careful monitoring of the clamp position and body temperature during the period of ischaemia this model achieves reproducible functional and structural injury. Mice sacrificed 24 hr&#160;following surgery demonstrate loss of renal function with elevation of the serum or plasma creatinine level as well as structural kidney damage with acute tubular necrosis evident. Renal function improves and the acute tissue injury resolves during the course of 7 days following renal IRI such that this model may be used to study renal regeneration. This model of renal IRI has been utilized to study the molecular and cellular pathophysiology of AKI as well as analysis of the subsequent renal regeneration.

The mouse model of renal ischaemia reperfusion injury described here comprises of a right nephrectomy that provides control tissue and clamping of the left renal pedicle to induce ischaemia that results in acute kidney injury. This model uses a midline laparotomy approach with all steps performed via one incision.

肾缺血再灌注损伤:损伤的小鼠模型和再生

Renal ischaemia reperfusion injury (IRI) is a common cause of acute kidney injury (AKI) in patients and occlusion of renal blood flow is unavoidable ...

科研

医学

An In Vitro Organ Culture Model of the Murine Intervertebral Disc

Intervertebral disc (IVD) degeneration is a significant contributor to low back pain. The IVD is a fibrocartilaginous joint that serves to transmit and dampen loads in the spine. The IVD consists of a proteoglycan-rich nucleus pulposus (NP) and a collagen-rich annulus fibrosis (AF) sandwiched by cartilaginous end-plates. Together with the adjacent vertebrae, the vertebrae-IVD structure forms a functional spine unit (FSU). These microstructures contain unique cell types as well as unique extracellular matrices. Whole organ culture of the FSU preserves the native extracellular matrix, cell differentiation phenotypes, and cellular-matrix interactions. Thus, organ culture techniques are particularly useful for investigating the complex biological mechanisms of the IVD. Here, we describe a high-throughput approach for culturing whole lumbar mouse FSUs that provides an ideal platform for studying disease mechanisms and therapies for the IVD. Furthermore, we describe several applications that utilize this organ culture method to conduct further studies including contrast-enhanced microCT imaging and three-dimensional high-resolution finite element modeling of the IVD.

An In Vitro Organ Culture Model of the Murine Intervertebral Disc

Whole organ culture of the intervertebral disc (IVD) preserves the native extracellular matrix, cell phenotypes, and cellular-matrix interactions. Here we describe an IVD culture system using mouse lumbar and caudal IVDs in their functional spinal units and several applications utilizing this system.

一个<em&gt;体外</em鼠类的椎间盘&gt;器官培养模型

Intervertebral disc (IVD) degeneration is a significant contributor to low back pain. The IVD is a fibrocartilaginous joint that serves to transmit and ...

生物工程

Utilizing Murine Inducible Telomerase Alleles in the Studies of Tissue Degeneration/Regeneration and Cancer

Telomere dysfunction-induced loss of genome integrity and its associated DNA damage signaling and checkpoint responses are well-established drivers that cause tissue degeneration during ageing. Cancer, with incidence rates greatly increasing with age, is characterized by short telomere lengths and high telomerase activity. To study the roles of telomere dysfunction and telomerase reactivation in ageing and cancer, the protocol shows how to generate two murine inducible telomerase knock-in alleles 4-Hydroxytamoxifen (4-OHT)-inducible TERT-Estrogen Receptor (mTERT-ER) and Lox-Stopper-LoxTERT (LSL-mTERT). The protocol describes the procedures to induce telomere dysfunction and reactivate telomerase activity in mTERT-ER and LSL-mTERT mice in vivo. The representative data show that reactivation of telomerase activity can ameliorate the tissue degenerative phenotypes induced by telomere dysfunction. In order to determine the impact of telomerase reactivation on tumorigenesis, we generated prostate tumor model G4 PB-Cre4 PtenL/L p53L/L LSL-mTERTL/L and thymic T-cell lymphoma model G4 Atm-/- mTERTER/ER. The representative data show that telomerase reactivation in the backdrop of genomic instability induced by telomere dysfunction can greatly enhance tumorigenesis. The protocol also describes the procedures used to isolate neural stem cells (NSCs) from mTERT-ER and LSL-mTERT mice and reactivate telomerase activity in NSCs in vitro. The representative data show that reactivation of telomerase can enhance the self-renewal capability and neurogenesis in vitro. Finally, the protocol describes the procedures for performing telomere FISH (Fluorescence In Situ Hybridization) on both mouse FFPE (Formalin Fixed and Paraffin Embedded) brain tissues and metaphase chromosomes of cultured cells.

Telomere and telomerase play essential roles in ageing and tumorigenesis. The goal of this protocol is to show how to generate two murine inducible telomerase knock-in alleles and how to utilize them in the studies of tissue degeneration/regeneration and cancer.

在组织变性/再生和癌症的研究利用小鼠诱导端粒酶等位基因

Telomere dysfunction-induced loss of genome integrity and its associated DNA damage signaling and checkpoint responses are well-established drivers that ...

A Proinflammatory, Degenerative Organ Culture Model to Simulate Early-Stage Intervertebral Disc Disease.

Symptomatic intervertebral disc (IVD) degeneration (IDD) is a major socioeconomic burden and is characterized by inflammation and tissue degradation. Due to the lack of causative therapies, there is an urgent need for innovative experimental organ culture models to study the mechanisms involved in the progression of the disease, find therapeutic targets, and reduce the need for animal models. We here present a novel, three-dimensional organ culture model protocol mimicking the proinflammatory and catabolic microenvironment, which is present during IDD. 
Initially, bovine caudal IVDs were dissected, cleaned, and cultured in the tissue culture medium. Dynamic physiologic or pathologic loading was applied in a custom-made bioreactor for 2 hours per day. IVDs were assigned to a control group (high glucose medium, physiological loading, phosphate-buffered saline injection) and a pathological group (low glucose medium, pathological loading, tumor necrosis factor-alpha injection) for four days. Gene expression analysis from collected nucleus pulposus cells of the IVDs and enzyme-linked immunosorbent assay of the conditioned organ culture media was performed. 
Our data revealed a higher expression of inflammatory markers and reduced disc heights after loading in the pathological group compared to the control group. This protocol is reliable to simulate IVD inflammation and degeneration and can be further expanded to broaden its application scope.

This protocol presents a novel experimental model of proinflammatory, degenerative bovine organ culture to simulate early-stage intervertebral disc degeneration.

模拟早期椎间盘疾病的发炎性、退行性器官培养模型。

Symptomatic intervertebral disc (IVD) degeneration (IDD) is a major socioeconomic burden and is characterized by inflammation and tissue degradation. Due ...

<meta charset="utf8"></head><body>IDD 模型兔腰部靶向推拿刺激方案

Operational and Intervention Effects of Targeted Tuina in Lumbar Intervertebral Disc Degeneration Model Rabbits

Tui Na or massage therapy alleviates symptoms related to intervertebral disc degeneration (IDD). However, precise, repeatable, standardized instructions for Tuina manipulation are lacking. This study establishes IDD model rabbits induced by fibrous ring puncture, creates targeted Tuina stimulation protocols at the acupuncture points in the lumbar region, and describes in detail the operation methods and requirements of kneading, pointing, and flicking. New Zealand male white rabbits (n = 15) were selected and randomly divided into a blank group, a model group, and a Tuina group. The rabbits in the model group and the Tuina group were molded by fibrous ring puncture; the rabbits in the model group were only immobilized on the operating table without treatment. In contrast, the Tuina group used the "8N/10N, 30 cycles/min" prescription for kneading, pointing, and flicking to perform the intervention, using tactile sensory aids to monitor and regulate the intensity of the Tuina operation. Imaging diagnosis and pathological tests were used to assess the effect of Tuina in rabbits, and the results showed improved imaging features and significantly lowered pathology scores of lumbar disc degeneration in the Tuina group compared to the model group (P &lt; 0.01). Targeted Tuina in the lumbar region may be beneficial in the alleviation of lumbar disc degeneration, but further verification is needed. By regularly performing Tuina and recording the mechanical information involved enables reproducible manipulation prescriptions and helps to observe the basic features of the underlying mechanism of Tuina for IDD.

<meta charset="utf8"></head><body>靶向推拿对腰椎间盘退变模型兔的作及干预效果

The finger tactile measurement system quantifies tactile pressure on the finger using a highly sensitive electrosurgical pressure sensor, with software displaying and accurately recording the pressure data and video in real time. Two analysis modules unify the data processing for thumb tracking and conditioning during Tuina.

靶向推拿对腰椎间盘退变模型兔的作及干预效果

Tui Na or massage therapy alleviates symptoms related to intervertebral disc degeneration (IDD). However, precise, repeatable, standardized instructions ...

Induction of Lumbar Disc Degeneration in Rabbits Through a Transabdominal Approach

Lower back pain is an extremely common medical issue in populations worldwide. One of the main contributors to lower back pain is intervertebral disc (IVD) degeneration. An ideal animal model of IVD degeneration is essential to study the pathophysiology of lower back pain and investigate potential therapeutic strategies. Rabbit models are reliable, economical, and easily established animal models. The retroperitoneal approach has been widely used to induce IVD degeneration in rabbit models. However, there are reported complications associated with this technique, such as the avulsion of segmental arteries and nerve root injury. In this paper, we aim to show a surgical protocol using needle puncture to establish rabbit lumbar disc degeneration via a transabdominal approach. Consequently, radiological checks and histological analyses indicated that lumbar disc degeneration was successfully established in rabbits. This surgical protocol presents the precise location of target discs and high reproducibility of IVD degeneration models with fewer complications.

<meta charset="utf8"></head><body>通过经腹入路诱导兔腰椎间盘退变

Here, we describe a surgical protocol using needle puncture to establish rabbit lumbar disc degeneration via a transabdominal approach. Radiological checks and histological analyses were used to confirm the successful establishment of lumbar disc degeneration.

通过经腹入路诱导兔腰椎间盘退变

Lower back pain is an extremely common medical issue in populations worldwide. One of the main contributors to lower back pain is intervertebral disc ...

A Mouse Model of Lumbar Spine Instability

Intervertebral disc degeneration (IDD) is a common pathological change leading to low back pain. Appropriate animal models are desired for understanding the pathological processes and evaluating new drugs. Here, we introduced a surgically induced lumbar spine instability (LSI) mouse model that develops IDD starting from 1 week post operation. In detail, the mouse under anesthesia was operated by low back skin incision, L3&#8211;L5 spinous processes exposure, detachment of paraspinous muscles, resection of processes and ligaments, and skin closure. L4&#8211;L5 IVDs were chosen for the observation. The LSI model develops lumbar IDD by porosity and hypertrophy in endplates at an early stage, decrease in intervertebral disc volume, shrinkage in nucleus pulposus at an intermediate stage, and bone loss in lumbar vertebrae (L5) at a later stage. The LSI mouse model has the advantages of strong operability, no requirement of special equipment, reproducibility, inexpensive, and relatively short period of IDD development. However, LSI operation is still a trauma that causes inflammation within the first week post operation. Thus, this animal model is suitable for study of lumbar IDD.

We developed a lumbar intervertebral disc degeneration mouse model by resection of L3&#8211;L5 spinous processes along with supra- and inter-spinous ligaments and detachment of paraspinous muscles.

腰椎不稳定的鼠标模型

Intervertebral disc degeneration (IDD) is a common pathological change leading to low back pain. Appropriate animal models are desired for understanding ...

Ovine Lumbar椎间盘退变模型利用横向腹膜后位钻伤

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肾缺血再灌注损伤:损伤的小鼠模型和再生

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在组织变性/再生和癌症的研究利用小鼠诱导端粒酶等位基因

模拟早期椎间盘疾病的发炎性、退行性器官培养模型。

靶向推拿对腰椎间盘退变模型兔的作及干预效果

靶向推拿对腰椎间盘退变模型兔的作及干预效果

靶向推拿对腰椎间盘退变模型兔的作及干预效果

通过经腹入路诱导兔腰椎间盘退变

通过经腹入路诱导兔腰椎间盘退变

腰椎不稳定的鼠标模型