Ovine Lumbar椎间盘退变模型利用横向腹膜后位钻伤
1Department of Surgery,Monash University, 2Department of Neurosurgery,Monash University, 3The Ritchie Centre,Hudson Institute of Medical Research, 4Proteobioactives, Pty Ltd, 5Department of Neurosurgery,St Vincent’s Hospital, 6Australian Institute for Bioengineering and Nanotechnology,University of Queensland, 7School of Chemical Engineering,University of Queensland, 8Department of Neurosurgery,Monash Health
Summary
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。
Abstract
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。
Introduction
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） 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 。
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”> 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。
Protocol
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。 快羊在麻醉前18小时。在运动前6-12小时，让动物进入水中。 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。 <br…
Representative Results
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 。 图1：手术前3T MRI（ A ）和术中横向射线?…
Discussion
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…
Disclosures
The authors have nothing to disclose.
Acknowledgements
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。
Materials
| Medetomidine Hydrochloride (10 mL Injection) | Therapon/Zoetis | PFIDOM10 | Multiple suppliers: Zoetis/Ilium |
| Thiopentone | Troy | Triothiopentone | Multiple suppliers: Neon Laboratories, Jagsonphal Pharmaceuticals |
| Isoflurane (2-3 % in oxygen) | Baxter | AHN3636 | Multiple suppliers: Baxter/VetOne |
| Amoxicillin parenteral | GlaxoSmithKline | JO1CA04 | Multiple suppliers: GlaxoSmithKline/Merck |
| Bupivacaine (0.5% Injection 20 mL) | Pfizer | 005BUP001 | Multiple suppliers: Pfizer/AstraZeneca |
| PVD Iodine Solution | Jurox | 61330 | Multiple suppliers: Jurox/Orion |
| Chlorhexidine 5%w/v | Jurox | Chlorhex C 5L (SCRUB) | Multiple suppliers: Jurox/Pfizer |
| Transdermal Fental Patch (75 μg/h) | Janssen-Cilag | S8-Dur7.5 | Multiple suppliers: Sandoz |
| Buprenorphine iv | Jurox | 504410 | Multiple suppliers: LGM Pharma |
| Atipamezole (Antisedan 0.06 mg/kg – 0.08 mg/kg) | Zoetis | PFIANT10 | Multiple suppliers: Ilium |
| Oster Golden A5 2-Speed Clippers | Oster | 078005-140-003 | Oster |
| 20 ml luer lock syringe | Terumo | 6SS+20L | Multiple suppliers: Medshop Australia/Terumo |
| 21 G IV needle | Terumo | SG3-1225 | Multiple suppliers:Medshop Australia/Terumo |
| #4 scalpel handle | Austvet | AD010/04 | Multiple suppliers: Austvet/SurgicalInstruments |
| #22 scalpel baldes | Austvet | ||
| Gillies tissue forceps | Austvet | AB430/15 | Multiple suppliers: Austvet/SurgicalInstruments |
| Metzenbaum curved dissecting scissors | Austvet | AC101/14 | Multiple suppliers: Austvet/SurgicalInstruments |
| Deaver retractor | Surgical Instruments | 23.75.03 | Multiple suppliers: Surgical Instrument/Austvet |
| Hohmann retractor | Austvet | KA173/35 | Multiple suppliers: Austvet/SurgicalInstruments |
| Mayo suture scissors | Austvet | AC911/14 | Multiple suppliers: Austvet/SurgicalInstruments |
| Needleholder 14 cm | EliteMedical | 18-1030 | Multiple suppliers: EliteMedical/Austvet |
| CMT 3.5 mm Brad-Point Drill | Carbatec | 516-035-51 | Multiple suppliers: Southeast Tool/Carbatec |
| Drill Bit Stop 4 mm | Drill Warehouse | 20121600 | Multiple suppliers: Amazon |
| Bosch 10.8 V Cordless Angle Drill | Get Tools Direct | GWB10.8V-LIBB | Multiple suppliers:Bunnings/Get Tools Direct |
| Autoclavable veterinary drill bag | AustVet | DRA043-AV | AustVet |
| 2-0 absorbable synthetic braided sutures | Ethicon | VCP335H | Ethicon |
| 3-0 absorbable synthetic braided sutures | Ethicon | VCP232H | Ethicon |
| Siemens 3 Tesla Skyra Widebore MRI | Siemens | N/A | Siemens |
| 9.4 Tesla Agilent (Varian) MRI | Agilent Technologies | N/A | Agilent Technologies |
| Atomscope HF 200 A Radiogaph | Radlink | 330003A | Multiple Suppliers: Radlink/DLC Australia |
| Veterinary Pulse Oximiter | DLC | 192500A | Multiple suppliers: DLC Australi Pty Ltd/AustVet |
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Cite This Article
Lim, K., Daly, C. D., Ghosh, P., Jenkin, G., Oehme, D., Cooper-White, J., Naidoo, T., Goldschlager, T. Ovine Lumbar Intervertebral Disc Degeneration Model Utilizing a Lateral Retroperitoneal Drill Bit Injury. J. Vis. Exp. (123), e55753, doi:10.3791/55753 (2017).