Name: laminotomy for lumbar dorsal root ganglion access and injection in swine
Uploaded: 2017-10-10T15:00:00
Description: Watch this Scientific Journal Video about Laminotomy for Lumbar Dorsal Root Ganglion Access and Injection in Swine at JoVE.com

这项研究是在舒尔茨家族基金会 (A.S.B.) 的支持下进行的。

此处描述的所有方法均已获得梅奥诊所机构动物护理和使用委员会 (IACUC) 的批准. 
 1. 严格性和重现性的先决条件 <ol> <li> 为了确保设计严谨, 遵循良好的实验室惯例的国家标准, 在任何动物之前都必须经过 IACUC (或类似的委员会) 的内部批准参与实验. 
	注意: 该协议的目的是保持一个临床忠实的方法。因此, 所涉及的材料、仪器和技术被描述成与人类最高临床标准相同的?...

<ol>
	<li>Millan, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+induction+of+pain:+An+integrative+review.">The induction of pain: An integrative review.</a> Prog Neurobiol. 57 (1), 1-164 (1999).</li><li>Burma, N. E., Leduc-Pessah, H., Fan, C. Y., Trang, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+chronic+pain:+Advances+and+challenges+for+clinical+translation.">Animal models of chronic pain: Advances and challenges for clinical translation.</a> J Neurosci Res. , (2016).</li><li>Pleticha, J., Maus, T. P., Beutler, A. 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我们试图描述一种方法的手术暴露的背根背根通过laminotomy 和 intraganglionic 注射液在一个健康的大型动物物种, 特别是猪。在啮齿类动物中, 一个类似的方法已经详细12 , 并用于提供常规药理剂8,10和病毒载体6,7,9,12到背根。上述小动物?...

背根神经节是解剖学上的离散, 神经元收集位于脊柱。每个根背根包含主要的感觉神经元, 对中枢神经系统 (CNS) 从特定的身体区域编码和中继周边刺激。例如, 关节炎的疼痛开始时, 疼痛受体位于一个联合感知有害刺激。这个过程被称为伤害。长期的有害刺激意识导致慢性疼痛1。
慢性疼痛是临床前研究的一个经常的主题2 , 其中一个目标是开发有用的方法, 靶向的镇痛药到背根, 如 intraganglionic 注射液3。然而, 根背根是难以访问, 因为他们居住在骨范围内的椎间孔4。几个小组成功地克服了这个障碍通过使用脊椎手术在啮齿目动物 5,6,7,8,9,10。
在临床上, 椎板切除术是一种常见的脊柱手术, 是指手术切除椎板, 从而去椎管11。纳入外科技术, 以支付直接根背根访问已成功在啮齿目动物5,12但是, 考虑到相关结构的大小差异以及如何影响药代动力学或技术可行性13,14。例如, 一项研究确定了 T10 的横断脊髓直径为3.0、7.0 和8.2 毫米, 分别为大鼠、猪和人, 各为15。因此, 大型动物模型较好地逼近了神经结构的人的维度。
在猪, Raore et al.使用多层次椎板切除术获得颈椎脊髓多椎管内注射16。该手术是很好的耐受性, 并导致了一期临床试验, 其中可比较的手术结局记录了17。这些结果鼓励继续使用前临床大型动物模型作为预测技术可行性和安全性的人。
到目前为止, 没有详细的方法, 以外科手术和注射的背根在一个大型动物物种。为了缩小这一平移间隙, 我们报告了一个协议的根节暴露和注射通过 laminotomy 猪。采用标准的神经外科技术、仪器和材料, 设计了模拟现代外科手术的方法。我们演示 intraganglionic 注射使用水溶液的腰椎背根, 并确认通过组织学成功分娩后21天。

<table border="0" cellpadding="0" cellspacing="0" width="1311">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Large humane animal sling</td>
			<td style="width:167px;">Britz &#38; Company</td>
			<td style="width:119px;">002539</td>
			<td style="width:484px;">Modified to include abdominal aperture</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Adhesive patient return electrode - 9 inch</td>
			<td style="width:167px;">Medtronic</td>
			<td style="width:119px;">E7506</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Ranger blood &#38; fluid warming system</td>
			<td style="width:167px;">3M</td>
			<td style="width:119px;">24500</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Lactated Ringer&#39;s fluid</td>
			<td style="width:167px;">Hospira</td>
			<td style="width:119px;">0409-7953-09</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Force air warming device</td>
			<td style="width:167px;">3M</td>
			<td style="width:119px;">77500</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:543px;">Duraprep solution with applicator, 26 ml (0.7% iodine povacrylex, 74% isopropyl alcohol</td>
			<td style="width:167px;">3M</td>
			<td style="width:119px;">8630</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Sterile disposable surgical towels</td>
			<td style="width:167px;">Medline</td>
			<td style="width:119px;">MDT2168286</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Ioban 2 incise drape</td>
			<td style="width:167px;">3M</td>
			<td style="width:119px;">6651EZSB</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Disposable suction canister and tubing</td>
			<td style="width:167px;">Medline</td>
			<td style="width:119px;">DYND44703H</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Button switch electrosurgical monopolar pencil</td>
			<td style="width:167px;">Medtronic</td>
			<td style="width:119px;">E2450H</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Fine smooth straight bipolar electrosurgical forceps, 4 1/2 inch</td>
			<td style="width:167px;">Bovie</td>
			<td style="width:119px;">A826</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">#15 blade</td>
			<td style="width:167px;">Miltex</td>
			<td style="width:119px;">4-315</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">#11 blade</td>
			<td style="width:167px;">Miltex</td>
			<td style="width:119px;">4-311</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">4x4 surgical gauze</td>
			<td style="width:167px;">Dynarex</td>
			<td style="width:119px;">3262</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Weitlaner self-retaining retractor, 8 inch</td>
			<td style="width:167px;">Miltex</td>
			<td style="width:119px;">11-618</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Meyerding self-retaining retractor, 1x2 3/8 inch</td>
			<td style="width:167px;">Sklar</td>
			<td style="width:119px;">42-2078</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Gelpi self-retaining retractor, 7 inch</td>
			<td style="width:167px;">Sklar</td>
			<td style="width:119px;">60-6570</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Freer elevator, 5 mm</td>
			<td style="width:167px;">Medline</td>
			<td style="width:119px;">MDS4641518F</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Bone wax</td>
			<td style="width:167px;">Ethicon</td>
			<td style="width:119px;">W31G</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Spurling intervertebral disc rongeur, 3 mm</td>
			<td style="width:167px;">Sklar</td>
			<td style="width:119px;">42-2852</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Spurling 45-degree, up-biting Kerrison rongeur, 2 mm</td>
			<td style="width:167px;">Medline</td>
			<td style="width:119px;">MDS4052802</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Leksell angled rongeur, 2 mm</td>
			<td style="width:167px;">Sklar</td>
			<td style="width:119px;">40-4097</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Gelfoam, size 50</td>
			<td style="width:167px;">Pfizer</td>
			<td style="width:119px;">AZL32301</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Cottonoid patty</td>
			<td style="width:167px;">Medtronic</td>
			<td style="width:119px;">8004007</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Frazier suction tip, 6 Fr</td>
			<td style="width:167px;">Sklar</td>
			<td style="width:119px;">50-2006</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Frazier suction tip, 10 Fr</td>
			<td style="width:167px;">Sklar</td>
			<td style="width:119px;">50-2010</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Dandy blunt right angle nerve hook</td>
			<td style="width:167px;">Medline</td>
			<td style="width:119px;">MDS4005220</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Nylon suture, 6-0</td>
			<td style="width:167px;">Ethicon</td>
			<td style="width:119px;">697G</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Castroviejo smooth micro needle holder</td>
			<td style="width:167px;">Medline</td>
			<td style="width:119px;">MDG2428614</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">22 gauge Quinke point spinal needle</td>
			<td style="width:167px;">Halyard Health</td>
			<td style="width:119px;">18397</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="105">
			<td height="105" style="height:105px;width:543px;">32 gauge CED needle with locking Luer hub</td>
			<td style="width:167px;">See comments</td>
			<td style="width:119px;">n/a</td>
			<td style="width:484px;">As in: Pleticha, J., Maus, T.P., Christner, J.A., Marsh, M.P., Lee, K.H., Hooten, W.M., Beutler, A.S. Minimally invasive convection-enhanced delivery of biologics into dorsal root ganglia: validation in the pig model and prospective modeling in humans. Technical note. J Neurosurg. 121(4), 851-8 (2014).</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Polyethylene tubing, 5 feet</td>
			<td style="width:167px;">Scientific Commodities</td>
			<td style="width:119px;">BB31695-PE/05</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Monoject syringe, 3 ml</td>
			<td style="width:167px;">Kendall</td>
			<td style="width:119px;">SY15352</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">NanoJet syringe pump</td>
			<td style="width:167px;">Chemyx</td>
			<td style="width:119px;">10050</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">DAPI</td>
			<td style="width:167px;">Sigma-Aldrich</td>
			<td style="width:119px;">D9542</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Fast Green FCF</td>
			<td style="width:167px;">Sigma-Aldrich</td>
			<td style="width:119px;">F7252</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Bulb irrigation syringe</td>
			<td style="width:167px;">Medline</td>
			<td style="width:119px;">DYND20125</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Fine-toothed Adson forceps</td>
			<td style="width:167px;">Medline</td>
			<td style="width:119px;">MDS1000212</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Vicryl suture, 0</td>
			<td style="width:167px;">Ethicon</td>
			<td style="width:119px;">J603H</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Vicryl suture, 2-0</td>
			<td style="width:167px;">Ethicon</td>
			<td style="width:119px;">J317H</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Needle counter</td>
			<td style="width:167px;">Medline</td>
			<td style="width:119px;">NC20FBRGS</td>
			<td style="width:484px;">-</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:543px;">Steri-strip skin closure, 1/2x4 inch</td>
			<td style="width:167px;">3M</td>
			<td style="width:119px;">R1547</td>
			<td style="width:484px;">-</td>
		</tr>
	</tbody>
</table>

laminotomy for lumbar dorsal root ganglion access and injection in swine

背根神经节是解剖学上定义良好的结构, 包含所有初级感觉神经元下的头部。这一事实使根背根有吸引力的目标, 注射新疗法旨在治疗慢性疼痛。在小动物模型中, 椎板切除术被用来促进节段注射, 因为它涉及手术切除脊椎周围的每个根节。我们展示了一种在大型动物中 intraganglionic 注射腰椎背根的技术, 即猪。Laminotomy 是为了允许使用标准的神经外科技术、仪器和材料直接进入根背根。与通过椎板切除术更广泛的骨清除相比, 我们实施 laminotomy, 以保存脊柱解剖, 同时达到足够的根节通路。术中应用无毒性染料对根节注射进行监测。在术后21天的安乐死后, 注射的成功是由组织学决定的 intraganglionic 分布 4 ", 6-diamidino-2-吲 (DAPI)。我们注入一个生物不活跃的解决方案来演示该协议。该方法可应用于今后的临床前研究, 以靶向治疗。我们的方法应该有助于测试 intraganglionic 小动物范式在大型动物物种中的可译性。此外, 该协议可作为一个关键的资源, 为那些规划前根节注射猪的前期研究。

injectate 传播的组织学评价 成功交付 injectate 到背根是由组织学评估的 DAPI 传播。该技术涉及定位针尖端在三维中心的背根。因此, 成功的分娩是通过评估 DAPI 染色的程度, 从组织学切片近 (中央节段) 和远处 (周围的背根节) 针尖。图 1A和图 1B表示成功地插入了一个背节。DAPI 染色均匀分布于中央和周围的根背根实质。因此, 一个成功的背根...

Watch this Scientific Journal Video about Laminotomy for Lumbar Dorsal Root Ganglion Access and Injection in Swine at JoVE.com

Laminotomy for Lumbar Dorsal Root Ganglion Access and Injection in Swine

我们描述了一种猪 laminotomy 的方法, 它提供了对 intraganglionic 注射液的腰椎背根神经节的通路。注射进展监测术和组织学证实21天后手术。本议定书可用于未来的临床研究, 涉及根背根注射液。

Laminotomy 对猪腰背根神经节通路和注射的治疗

Dorsal root ganglia (DRG) are anatomically well defined structures that contain all primary sensory neurons below the head. This fact makes DRG attractive ...

Research

JoVE Journal

Neuroscience

Chromatin Immunoprecipitation from Dorsal Root Ganglia Tissue following Axonal Injury

染色质免疫沉淀从以下轴索损伤的背根神经节组织

Axons in the central nervous system (CNS) do not regenerate while those in the peripheral nervous system (PNS) do regenerate 
to a limited extent after ...

科研

神经科学

Genetic Study of Axon Regeneration with Cultured Adult Dorsal Root Ganglion Neurons

培养成年的背根神经节神经元轴突再生的遗传研究

It is well known that mature neurons in the central nervous system (CNS) cannot regenerate their axons after injuries due to diminished intrinsic ability ...

Measuring Spinal Presynaptic Inhibition in Mice By Dorsal Root Potential Recording In Vivo

Measuring Spinal Presynaptic Inhibition in Mice By Dorsal Root Potential Recording In Vivo

测量脊髓突触前抑制小鼠背根电位记录<em&gt;在体内</em

Presynaptic inhibition is one of the most powerful inhibitory mechanisms in the spinal cord. The underlying physiological mechanism is a depolarization of ...

In Vitro Modeling of Cancerous Neural Invasion: The Dorsal Root Ganglion Model

In Vitro Modeling of Cancerous Neural Invasion: The Dorsal Root Ganglion Model

在癌神经侵袭的体外模型：背根神经节型号

One way that solid tumors disseminate is through neural invasion. This route is well-known in cancers of the head and neck, prostate, and pancreas. These ...

An Approach to Enhance Alignment and Myelination of Dorsal Root Ganglion Neurons

一种方法来提高背根神经节的对齐和髓鞘

Axon regeneration is a chaotic process due largely to unorganized axon alignment. Therefore, in order for a sufficient number of regenerated axons to ...

Dorsal Root Ganglion Injection and Dorsal Root Crush Injury as a Model for Sensory Axon Regeneration

背根神经节内注射和背根挤压伤为感官轴突再生模型

Achieving axon regeneration after nervous system injury is a challenging task. As different parts of the central nervous system (CNS) differ from each ...

Analysis of Immune Cells in Single Sciatic Nerves and Dorsal Root Ganglion from a Single Mouse Using Flow Cytometry

应用流式细胞仪分析单侧坐骨神经和背根神经节的免疫细胞

Nerve-resident immune cells in the peripheral nervous system (PNS) are essential to maintaining neuronal integrity in a healthy nerve. The immune cells of ...

Investigating Mammalian Axon Regeneration: In Vivo Electroporation of Adult Mouse Dorsal Root Ganglion

Investigating Mammalian Axon Regeneration: In Vivo Electroporation of Adult Mouse Dorsal Root Ganglion

哺乳动物轴突再生: 成年小鼠背根神经节体内电穿孔的研究

Electroporation is an essential non-viral gene transfection approach to introduce DNA plasmids or small RNA molecules into cells. A sensory neuron in the ...

Use of In Vivo Single-fiber Recording and Intact Dorsal Root Ganglion with Attached Sciatic Nerve to Examine the Mechanism of Conduction Failure

使用 Vivo 单纤维记录和带附骨神经的固定背道根结条，以检查传导失败机制

Single-fiber recording has been a classical and effective electrophysiological technique over the last few decades because of its specific application for ...

Rapid Isolation of Dorsal Root Ganglion Macrophages

多尾根结鱼巨噬细胞的快速隔离

There are growing interests to study the molecular and cellular interactions among immune cells and sensory neurons in the dorsal root ganglia after ...