Name: laminotomy for lumbar dorsal root ganglion access and injection in swine
Uploaded: 2017-10-10T15:00:00.000+00:00
Duration: 9 min
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一种方式。例如, 严格的无菌技术是遵循和过期的材料永远不应该使用. </li>
	<li> 在实验环境中支持此方法的重现性, 制定内部标准操作程序, 并控制猪品种、体重、性别和不同人群之间的年龄变化. 
	注: 本协议的设计基于对 38-53 公斤重的猪的使用. </li>
</ol> 2。手术前的动物护理 <ol> <li> 管理预防性肌肉 (IM) 呋, 在手术前1天5毫克/千克. </li>
	<li> 固体食物的快速动物和限制动物从化妆品治疗, 即 油浴, 12 小时在做法之前. </li>
	<li> 诱导全麻1小时内的程序, 使用 im tiletamine 和唑, 给 Telazol 在5毫克/千克, 和 IM 嗪, 给2毫克/千克. </li>
	<li> 一旦诱导, 管理皮下 (SC) 丁丙诺啡持续释放 (SR), 给0.18 毫克/千克. </li>
	<li> 放置耳静脉导管并执行快速顺序插管以放置气管导管. </li>
	<li> 将心跳血与心率监测功能连接到舌头, 以监测氧和心率. </li>
	<li> 将动物置于俯卧位置, 用电动剪刀将皮肤夹在背部。在一个大的, 双边的区域, 从中线矢状面延伸到中线冠状面, 纵向从骶骨顶端到肩胛骨棘。使用胶带去除头发和自由漂浮的皮肤. </li>
	<li> 用温水和肥皂擦洗修剪过的区域3次, 用无绒毛巾擦干皮肤. </li>
	<li> 使用外科标记笔标记双边解剖标志。标记最后的肋骨、髂嵴、棘突和横突. 
	注: 通过标记最后的肋骨, 髂嵴, 和横向的过程, 腰椎是沿其上级, 下位, 和横向界限, 分别。本议定书的目的是指导访问和注射任何腰椎背节的利益。为参考, 优越的髂嵴与 L3 或 L4 椎水平一致. </li>
	<li> 用温暖的毯子盖住动物以运送到手术套件. </li>
</ol> 3。在操作套件中定位 <ol> <li> 轻轻地将动物抬起, 并将其置于带有填充腹部光圈的修改过的大型人性化动物吊索中. 
	注: 腹孔可减少腹部压力, 类似于人体脊柱手术期间使用的威尔逊框架。反过来, 这减少了术中出血从脊髓血管。吊带是有利的, 因为腿可以通过软垫开口自由悬挂, 保护动物免受周围神经撞击。然而, 由于吊索架是由金属制成的, 所以应该用绝缘材料来填充, 以避免电气短路和无意的动物灼伤。毯子辊可以放置在一个舒适的位置, 根据动物的大小头和颈部. </li>
	<li> 使用 1-3% 吸入 (滴定) 异氟醚维持全身麻醉, 效果良好。用眼部药膏滋润眼睛, 然后轻轻地把它们封起来. </li>
	<li> 将重要标志监视的线放置到记录温度、血压、心率和氧合状态。通过二氧化碳监视通风. </li>
	<li> 将粘合剂、一次性电分散电极置于左侧或右侧肩胛骨上. </li>
	<li> 管理温暖的林格铃声和 #39; 通过耳静脉导管维持液体。给液体以 5-10 毫升/千克/小时的速度. </li>
	<li> 在胸部和颈部放置一个强制空气预热装置, 避免覆盖最后一根肋骨. </li>
</ol> 4。左侧注射操作场无菌制剂的研究 注意: 从这一点出发, 以严格的无菌方式进行. 
<ol> <li> 根据制造商和 #39 的说明, 从0.7% 碘 povacrylex 和74% 异丙醇的广泛应用开始, 准备在腰椎上覆的皮肤。为了确保导针以后可以放在无菌的方式, lateralize 应用到侧面的计划注射通过延长防腐对中线冠状面越过明显的横向过程. </li>
	<li> 将一次性的手术巾置于长方形的方式下, 勾勒出计划中的切口部位, 这是沿着明显的腰椎棘突的中线. </li>
	<li> 在操作毛巾和裸露的皮肤上应用粘合剂抗菌切割褶皱。夹紧窗帘, 将窗帘的边缘延伸到操作现场. </li>
	<li> 在操作现场和监控技术员之间的吊索头上固定一个垂直的悬垂. </li>
	<li> 在操作场内通过夹紧不育的窗帘来确保吸入和电的安全线。通过无菌场的管子和电线的自由端. </li>
</ol> 5。皮肤切口和骨膜解剖 <ol> <li> 触沿中线进行腰椎棘突, 并确定3连续的椎体水平. </li>
	<li> 使用 #15 手术刀打开 8-12 厘米中线矢状切口通过切割褶皱直接后的棘过程。使用纱布填塞和单电保持止血. 
	注意: 应采取不偏离中线, 因为切口是先进的前向, 因为这限制了从旁肌肉出血。对棘突的定期触诊有助于推进。可根据需要放置并重新定位自护 Weitlaner、Meyerding 或 Gelpi 器, 以利于解剖。吸力用于保持能见度. </li>
	<li> 使用单电解剖皮下组织和脂肪, 直到胸腰椎筋膜到达。触的棘突深至胸腰椎筋膜, 并沿中线切开筋膜, 露出棘突之间的冈韧带. 
	注意: 胸腰椎筋膜被确定为一个有组织, 睑鞘与结缔组织颗粒, 交织在一个斜, 侧向内侧方向。在这一点上, 切口可以延长, 无论是上级或下级的方向, 以确保3棘过程是完全可见的中心棘过程在中心的解剖场. </li>
	<li> 使用 #15 刀片将一个2毫米深的旁切口通过冈韧带后, 每个棘过程。将每个切口沿棘突后表面的左侧三分之一处放置. </li>
	<li> 在每个级别轻轻地释放冈韧带视力使用5毫米更自由的电梯. </li>
	<li> 识别骨膜平面并在该平面内沿每个棘突的侧面进行解剖. </li>
	<li> 在每个棘突过程中以平行的方式执行骨膜解剖, 以确保一个温和的、甚至是解剖的实现. </li>
	<li> 使用单电与骨膜解剖一起在棘空间切割旁肌附着物. </li>
	<li> 识别每层的椎板并继续骨膜解剖, 以达到2间关节的外侧边界, 连接3裸露的椎骨, 并到达关节之间的椎板侧边, 称为峡. 
	注意: 峡是椎间孔的后缘, 其中根节位于。偶尔, 一个小静脉从位于椎板后表面的孔中产生。这些静脉在骨膜解剖时有破裂的倾向。用双极电和骨蜡联合应用于孔内, 可以很容易地实现止血. </li>
</ol> 6。单级 Laminotomy <ol> <li> 将 Laminotomy 的目标标识为位于2间关节之间和内侧的中心椎板. </li>
	<li> 将叶片追踪到它的下边缘, 使其与尾间关节的下侧关节过程中的一个点相连. </li>
	<li> 使用5毫米更自由的电梯或刮触 caudalmost 边缘与中央运河之间的过渡. 
	注意: 小心不要强迫触摸的仪器前, 因为这将接触硬脑膜囊和脊髓。请注意, 猪的脊髓延伸过腰椎 18 。椎间盘钳可用于清除覆盖此区域的额外软组织, 以方便触诊. </li>
	<li> 使用2毫米高咬, 45 度 Kerrison 钳以分段的方式提取骨骼。沿着棘突的底部, 将骨移到椎弓根的尾部, 并在其全部范围内向外侧. </li>
	<li> 使用倾斜的骨骼 rongeurs 来帮助清除骨骼。离开与椎板连接的下关节过程, 直到 laminotomy 基本完成. </li>
	<li> 确认下关节过程是自由移动的, 仅由间关节囊附着。使用 #15 或 #11 刀片切割胶囊. </li>
	<li> 以分段的方式去除下关节的过程, 但将相邻的高级关节过程保持不变. 
	注: 当 laminotomy 完成时, 止血用双极电。单电由于神经结构的接近而不被使用。骨蜡可以放置在出血部位, 从裸露的骨和可吸收的明胶海绵可以用来获得止血近软组织。Cottonoid 是一个有用的工具, 以灯芯浆液和血液远离解剖. </li>
</ol> 7。解剖的背根背节 <ol> <li> 从表面到深的开始内侧从分段的方式疏散硬膜外脂肪, 并进行侧向。用双极钳和吸入 6-10 法国弗雷泽吸入提示去除脂肪的温和解剖. 
	注: 放大镜放大或使用解剖显微镜有助于提供安全疏散硬膜外脂肪所需的详细程度, 并使用双极性电实现硬膜外静脉丛的细致止血. </li>
	<li> 识别沿中线运行的 superoinferior 方向的硬脑膜囊, 与皮肤切口的轴线平行。在硬脑膜囊中取出硬膜外脂肪, 直到可以看到硬脑膜囊产生硬脑膜神经根袖. </li>
	<li> 通过硬膜外脂肪的排空, 将硬脑膜套筒侧向和下, 直到其周围有扩大的根节. 
	注: 标识其椭圆形形状的背根, 黄色至橙色。在 mid-lumbar 脊柱, 背根通常是 4-6 毫米的大小, 最长的内侧向侧向方向, 并位于直接下位或 2-3 毫米内侧到其各自的蒂。一个钝的, 直角的神经钩可以用来轻轻触的椎弓根. </li>
	<li> 从侧面转移硬膜外脂肪, 越过根节, 直到发现毗邻的脊髓神经. 
	注: 如果 durotomy 发生, 用6-0 尼龙缝合和光滑的微针驱动, 在一个简单的运行施蒂希修复它. </li>
</ol> 8。注射背根节 <ol> <li> 使用22口径的脊椎针引导32口径对流增强型针的轨迹。刺穿22口径引导针通过皮肤和旁的肌肉. 
	注: 阴极电泳针的设计是为了实现组织中的流体对流, 也称为散流, 由于压力梯度 19 , 20 。
	<li> 将导针沿着接近根背根纵轴的轨迹方向对准, 并在解剖场的侧旁壁上产生针尖. </li>
	<li> 微调针路径, 直到针腔与根节的中心对齐. 
	注: 不应允许导针与根节接触. </li>
	<li> 将无菌 injectate 放入无菌注射器中, 并将注射器连接到无菌输液管. </li>
	<li> 将油管固定在阴极电泳针上, 并将注射器从无菌场中取出。将注射器连接到可编程注射器泵. 
	注: 油管的长度为5英尺, 以确保保持不育和流动性。此外, 最重要的是, 没有空气泡沫被引入到解决方案中. </li>
	<li> 前进 injectate 直到从阴极电泳针尖上看到表达式. </li>
	<li> 将阴极电泳针放在导针腔内, 慢慢地推进阴极电泳针, 直到它从导针上露出。确保在针对齐时不刺穿背根. </li>
	<li> 微调引导针位置沿其轨迹的长轴, 以确定阴极电泳尖端的最终位置. </li>
	<li> 使用联锁针集线器将导针和阴极电泳针固定在一起, 可以实现导轨和阴极电泳针的深度和对准. </li>
	<li> 确认所有的注塑设备连接都已完全安全, 包括导针、阴极电泳针和装有 injectate 的连接油管. </li>
	<li> 将导针沿其长轴前进, 以近似于电泳针尖和背根. </li>
	<li> 用电泳针尖刺穿背根. </li>
	<li> Submerse 将阴极电泳针尖插入根节的三维中心。注意: 由于背根是一个三维结构的可变大小和形状, 背节的曝光必须是完整的, 以便准确地把阴极电泳针尖在真正的中心的背节。真正的背根中心位于它的三解剖轴的交叉点, 即前到后, 侧向内侧, 并优于下位轴. </li>
	<li> 提供100和 #956; injectate 由土木工程署使用的毕业率和3步骤的数量. </li>
	<li> 交付4和 #956; l 在2和 #956; 第一步的 l/分钟。提供8和 #956; l 在4和 #956; 第二步的 l/分钟。交付88和 #956; l 在8和 #956; l/分钟的第三和最后一步. 
	注意: 允许在步骤和最后一步之间进行3分钟的停顿, 以实现压力平衡. </li>
	<li> 在最后的注射步骤后取出注射装置, 并在平滑、柔和的运动中沿其长轴暂停3分钟. 
	注: 对于无色的注入溶液, 在溶液中含有0.1% 重量/体积的染料, 以协助目视评估 injectate 分布的 12 。此外, 重要染料4和 #39;, 6-diamidino-2-吲 (DAPI) 是包括在溶液中的浓度为0.25 和 #956; g/和 #956; 当研究设计需要 injectate 分布的组织学评估 5 . </li>
</ol> 9。关闭 <ol> <li> 在关闭前向外科站点应用3轮温热盐水冲洗, 以调动和冲洗碎片的位置, 即 骨碎片。用吸力来回收盐水和碎屑. 
	注: 冲洗时要保证细致的止血。止血剂 (明胶海绵) 和 cottonoid 在这个时候被删除。确保所有材料和仪器在关闭前从切口部位清除. </li>
	<li> 使用3层技术关闭. </li>
	<li> 缝合胸腰椎筋膜使用0缝合简单, 中断, noninverted 的时尚。每 5-8 毫米放置一个施蒂希, 以实现水密封闭. </li>
	<li> 缝合皮下组织使用2-0 缝合在一个简单的, 中断的, 倒置的时尚, 每 5-到8毫米针放置, 以获得足够的力量. </li>
	<li> 使用0缝合线以简单、连续或中断的方式关闭皮肤. </li>
	<li> 使用针计数器可确保没有任何锐尖. </li>
	<li> 用盐水冲洗皮肤, 干燥皮肤, 并将绷带条与切口垂直放置. </li>
	<li> 在绷带条上放置纱布, 并附上最后的粘合剂抗菌切割褶皱. </li>
</ol> 10。术后动物护理 <ol> <li> Extubate, 用暖和的毯子盖住, 并将动物运送到恢复期. </li>
	<li> 按照标准的机构操作程序进行术后监测和从生存手术中恢复。至少, 观察动物每15分钟, 直到意识返回, 每小时, 直到完全麻醉恢复达到, 并每天两次. </li>
	<li> 通过管理 IM 或口服卡洛芬提供术后疼痛管理, 4 毫克/千克, 每日5天, 从术后0天开始。管理 SC 丁丙诺啡 SR, 在0.18 毫克/千克, 一次在术后天 2. </li>
	<li> 以5毫克/千克为呋, 在术后4天一次进行术后防腐管理. </li>
	<li> 清除术后 5-7 天的绷带。当伤口愈合完成时, 通常在术后 10-14 天切除缝合线. </li>
	在达到研究终点后, <li> 人道地按照标准的机构操作程序安乐动物. </li>
</ol>

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W., Huang, L. Y. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficiencies+of+transgene+expression+in+nociceptive+neurons+through+different+routes+of+delivery+of+adeno-associated+viral+vectors.">Efficiencies of transgene expression in nociceptive neurons through different routes of delivery of adeno-associated viral vectors.</a> Hum Gene Ther. 14 (9), 897-906 (2003).</li><li>Puljak, L., Kojundzic, S. L., Hogan, Q. H., Sapunar, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+delivery+of+pharmacological+agents+into+rat+dorsal+root+ganglion.">Targeted delivery of pharmacological agents into rat dorsal root ganglion.</a> J Neurosci Methods. 177 (2), 397-402 (2009).</li><li>Mason, M. R. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+AAV+serotypes+for+gene+delivery+to+dorsal+root+ganglion+neurons.">Comparison of AAV serotypes for gene delivery to dorsal root ganglion neurons.</a> Mol Ther. 18 (4), 715-724 (2010).</li><li>Jelicic Kadic, A., Boric, M., Kostic, S., Sapunar, D., Puljak, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+intraganglionic+injection+of+calcium/calmodulin-dependent+protein+kinase+II+inhibitors+on+pain-related+behavior+in+diabetic+neuropathy.">The effects of intraganglionic injection of calcium/calmodulin-dependent protein kinase II inhibitors on pain-related behavior in diabetic neuropathy.</a> Neurosci. 256, 302-308 (2014).</li><li>Greenberg, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Handbook of neurosurgery. , (2010).</li><li>Yu, H., Fischer, G., Hogan, Q. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=AAV-mediated+gene+transfer+to+dorsal+root+ganglion.">AAV-mediated gene transfer to dorsal root ganglion.</a> Methods Mol Biol. 1382, 251 (2016).</li><li>Yaksh, T. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacology+and+toxicology+of+chronically+infused+epidural+clonidine+HCL+in+dogs.">Pharmacology and toxicology of chronically infused epidural clonidine HCL in dogs.</a> Toxicol Sci. 23 (3), 319-335 (1994).</li><li>Federici, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+technique+for+spinal+cord+delivery+of+therapies:+demonstration+of+procedure+in+gottingen+minipigs.">Surgical technique for spinal cord delivery of therapies: demonstration of procedure in gottingen minipigs.</a> J Vis Exp. (70), e4371 (2012).</li><li>Lee, J. H. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+porcine+model+of+traumatic+thoracic+spinal+cord+injury.">A novel porcine model of traumatic thoracic spinal cord injury.</a> J Neurotrauma. 30 (3), 142-159 (2013).</li><li>Raore, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cervical+multilevel+intraspinal+stem+cell+therapy:+assessment+of+surgical+risks+in+Gottingen+minipigs.">Cervical multilevel intraspinal stem cell therapy: assessment of surgical risks in Gottingen minipigs.</a> Spine. 36 (3), e164 (2011).</li><li>Riley, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraspinal+stem+cell+transplantation+in+amyotrophic+lateral+sclerosis:+A+phase+I+safety+trial,+technical+note,+and+lumbar+safety+outcomes.">Intraspinal stem cell transplantation in amyotrophic lateral sclerosis: A phase I safety trial, technical note, and lumbar safety outcomes.</a> Neurosurg. 71 (2), 405-416 (2012).</li><li>Olmarker, K., Holm, S., Rosenqvist, A., Rydevik, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+nerve+root+compression.++A+model+of+acute,+graded+compression+of+the+porcine+cauda+equina+and+an+analysis+of+neural+and+vascular+anatomy.">Experimental nerve root compression. A model of acute, graded compression of the porcine cauda equina and an analysis of neural and vascular anatomy.</a> Spine. 16 (1), 61-69 (1991).</li><li>Bobo, R. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Convection-enhanced+delivery+of+macromolecules+in+the+brain.">Convection-enhanced delivery of macromolecules in the brain.</a> Proc Natl Acad Sci U.S.A. 91 (6), 2076-2080 (1994).</li><li>Lonser, R. R., Sarntinoranont, M., Morrison, P. F., Oldfield, E. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Convection-enhanced+delivery+to+the+central+nervous+system.">Convection-enhanced delivery to the central nervous system.</a> J Neurosurg. 122 (3), 697-706 (2015).</li><li>Shen, J., Wang, H. Y., Chen, J. Y., Liang, B. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Morphologic+analysis+of+normal+human+lumbar+dorsal+root+ganglion+by+3D+MR+imaging.">Morphologic analysis of normal human lumbar dorsal root ganglion by 3D MR imaging.</a> AJNR Am J Neuroradiol. 27 (0195–6108 (Print)), 2098-2103 (2006).</li><li>Pleticha, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Minimally+invasive+convection-enhanced+delivery+of+biologics+into+dorsal+root+ganglia:+validation+in+the+pig+model+and+prospective+modeling+in+humans.">Minimally invasive convection-enhanced delivery of biologics into dorsal root ganglia: validation in the pig model and prospective modeling in humans.</a> J Neurosurg. 121 (4), 851-858 (2014).</li></ol>

没有;作者没有与本研究相关的利益冲突。

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><tbody><tr><td>Large humane animal sling</td><td>Britz &amp;amp; Company</td><td>002539</td><td>Modified to include abdominal aperture</td></tr><tr><td>Adhesive patient return electrode - 9 inch</td><td>Medtronic</td><td>E7506</td><td>-</td></tr><tr><td>Ranger blood &amp;amp; fluid warming system</td><td>3M</td><td>24500</td><td>-</td></tr><tr><td>Lactated Ringer&#039;s fluid</td><td>Hospira</td><td>0409-7953-09</td><td>-</td></tr><tr><td>Force air warming device</td><td>3M</td><td>77500</td><td>-</td></tr><tr><td>Duraprep solution with applicator, 26 mL (0.7% iodine povacrylex, 74% isopropyl alcohol)</td><td>3M</td><td>8630</td><td>-</td></tr><tr><td>Sterile disposable surgical towels</td><td>Medline</td><td>MDT2168286</td><td>-</td></tr><tr><td>Ioban 2 incise drape</td><td>3M</td><td>6651EZSB</td><td>-</td></tr><tr><td>Disposable suction canister and tubing</td><td>Medline</td><td>DYND44703H</td><td>-</td></tr><tr><td>Button switch electrosurgical monopolar pencil</td><td>Medtronic</td><td>E2450H</td><td>-</td></tr><tr><td>Fine smooth straight bipolar electrosurgical forceps, 4 1/2 inch</td><td>Bovie</td><td>A826</td><td>-</td></tr><tr><td>#15 blade</td><td>Miltex</td><td>4-315</td><td>-</td></tr><tr><td>#11 blade</td><td>Miltex</td><td>4-311</td><td>-</td></tr><tr><td>4 x 4 surgical gauze</td><td>Dynarex</td><td>3262</td><td>-</td></tr><tr><td>Weitlaner self-retaining retractor, 8 inch</td><td>Miltex</td><td>11-618</td><td>-</td></tr><tr><td>Meyerding self-retaining retractor, 1 x 2 3/8 inch</td><td>Sklar</td><td>42-2078</td><td>-</td></tr><tr><td>Gelpi self-retaining retractor, 7 inch</td><td>Sklar</td><td>60-6570</td><td>-</td></tr><tr><td>Freer elevator, 5 mm</td><td>Medline</td><td>MDS4641518F</td><td>-</td></tr><tr><td>Bone wax</td><td>Ethicon</td><td>W31G</td><td>-</td></tr><tr><td>Spurling intervertebral disc rongeur, 3 mm</td><td>Sklar</td><td>42-2852</td><td>-</td></tr><tr><td>Spurling 45-degree, up-biting Kerrison rongeur, 2 mm</td><td>Medline</td><td>MDS4052802</td><td>-</td></tr><tr><td>Leksell angled rongeur, 2 mm</td><td>Sklar</td><td>40-4097</td><td>-</td></tr><tr><td>Gelfoam, size 50</td><td>Pfizer</td><td>AZL32301</td><td>-</td></tr><tr><td>Cottonoid patty</td><td>Medtronic</td><td>8004007</td><td>-</td></tr><tr><td>Frazier suction tip, 6 Fr</td><td>Sklar</td><td>50-2006</td><td>-</td></tr><tr><td>Frazier suction tip, 10 Fr</td><td>Sklar</td><td>50-2010</td><td>-</td></tr><tr><td>Dandy blunt right angle nerve hook</td><td>Medline</td><td>MDS4005220</td><td>-</td></tr><tr><td>Nylon suture, 6-0</td><td>Ethicon</td><td>697G</td><td>-</td></tr><tr><td>Castroviejo smooth micro needle holder</td><td>Medline</td><td>MDG2428614</td><td>-</td></tr><tr><td>22 gauge Quinke point spinal needle</td><td>Halyard Health</td><td>18397</td><td>-</td></tr><tr><td>32 gauge CED needle with locking Luer hub</td><td>See comments</td><td>n/a</td><td>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><td>Polyethylene tubing, 5 feet</td><td>Scientific Commodities</td><td>BB31695-PE/05</td><td>-</td></tr><tr><td>Monoject syringe, 3 mL</td><td>Kendall</td><td>SY15352</td><td>-</td></tr><tr><td>NanoJet syringe pump</td><td>Chemyx</td><td>10050</td><td>-</td></tr><tr><td>DAPI</td><td>Sigma-Aldrich</td><td>D9542</td><td>-</td></tr><tr><td>Fast Green FCF</td><td>Sigma-Aldrich</td><td>F7252</td><td>-</td></tr><tr><td>Bulb irrigation syringe</td><td>Medline</td><td>DYND20125</td><td>-</td></tr><tr><td>Fine-toothed Adson forceps</td><td>Medline</td><td>MDS1000212</td><td>-</td></tr><tr><td>Vicryl suture, 0</td><td>Ethicon</td><td>J603H</td><td>-</td></tr><tr><td>Vicryl suture, 2-0</td><td>Ethicon</td><td>J317H</td><td>-</td></tr><tr><td>Needle counter</td><td>Medline</td><td>NC20FBRGS</td><td>-</td></tr><tr><td>Steri-strip skin closure, 1/2x4 inch</td><td>3M</td><td>R1547</td><td>-</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 ...

laminotomy-for-lumbar-dorsal-root-ganglion-access-injection

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

12.7K 次观看.  Mayo Clinic, Translational Science Track, Mayo Graduate School. 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。

视频: Laminotomy for Lumbar Dorsal Root Ganglion Access and Injection in Swine

Intraspinal Cell Transplantation for Targeting Cervical Ventral Horn in Amyotrophic Lateral Sclerosis and Traumatic Spinal Cord Injury

Respiratory compromise due to phrenic motor neuron loss is a debilitating consequence of a large proportion of human traumatic spinal cord injury (SCI) cases 1 and is the ultimate cause of death in patients with the motor neuron disorder, amyotrophic laterals sclerosis (ALS) 2.
ALS is a devastating neurological disorder that is characterized by relatively rapid degeneration of upper and lower motor neurons. Patients ultimately succumb to the disease on average 2-5 years following diagnosis because of respiratory paralysis due to loss of phrenic motor neuron innnervation of the diaphragm 3. The vast majority of cases are sporadic, while 10% are of the familial form. Approximately twenty percent of familial cases are linked to various point mutations in the Cu/Zn superoxide dismutase 1 (SOD1) gene on chromosome 21 4. Transgenic mice 4,5 and rats 6 carrying mutant human SOD1 genes (G93A, G37R, G86R, G85R) have been generated, and, despite the existence of other animal models of motor neuron loss, are currently the most highly used models of the disease.
Spinal cord injury (SCI) is a heterogeneous set of conditions resulting from physical trauma to the spinal cord, with functional outcome varying according to the type, location and severity of the injury 7. Nevertheless, approximately half of human SCI cases affect cervical regions, resulting in debilitating respiratory dysfunction due to phrenic motor neuron loss and injury to descending bulbospinal respiratory axons 1. A number of animal models of SCI have been developed, with the most commonly used and clinically-relevant being the contusion 8.
Transplantation of various classes of neural precursor cells (NPCs) is a promising therapeutic strategy for treatment of traumatic CNS injuries and neurodegeneration, including ALS and SCI, because of the ability to replace lost or dysfunctional CNS cell types, provide neuroprotection, and deliver gene factors of interest 9.
Animal models of both ALS and SCI can model many clinically-relevant aspects of these diseases, including phrenic motor neuron loss and consequent respiratory compromise 10,11. In order to evaluate the efficacy of NPC-based strategies on respiratory function in these animal models of ALS and SCI, cellular interventions must be specifically directed to regions containing therapeutically relevant targets such as phrenic motor neurons. We provide a detailed protocol for multi-segmental, intraspinal transplantation of NPCs into the cervical spinal cord ventral gray matter of neurodegenerative models such as SOD1G93A mice and rats, as well as spinal cord injured rats and mice 11.

Neural precursor transplantation is a promising strategy for protecting and/or replacing lost/dysfunctional cervical phrenic motor neurons in spinal cord injury (SCI) and the motor neuron disorder, amyotrophic laterals sclerosis (ALS). We provide a protocol for cell delivery to cervical spinal cord ventral horn in rodent models of ALS and SCI.

椎管内针对肌萎缩性脊髓侧索硬化症和外伤性脊髓损伤颈腹角细胞移植

Respiratory compromise due to phrenic motor neuron loss is a debilitating consequence of a large proportion of human traumatic spinal cord injury (SCI) ...

科研

医学

Live Imaging of Dorsal Root Axons after Rhizotomy

The primary sensory axons injured by spinal root injuries fail to regenerate into the spinal cord, leading to chronic pain and permanent sensory loss. Regeneration of dorsal root (DR) axons into spinal cord is prevented at the dorsal root entry zone (DREZ), the interface between the CNS and PNS. Our understanding of the molecular and cellular events that prevent regeneration at DREZ is incomplete, in part because complex changes associated with nerve injury have been deduced from postmortem analyses. Dynamic cellular processes, such as axon regeneration, are best studied with techniques that capture real-time events with multiple observations of each living animal. Our ability to monitor neurons serially in vivo has increased dramatically owing to revolutionary innovations in optics and mouse transgenics. Several lines of thy1-GFP transgenic mice, in which subsets of neurons are genetically labeled in distinct fluorescent colors, permit individual neurons to be imaged in vivo1. These mice have been used extensively for in vivo imaging of muscle2-4 and brain5-7, and have provided novel insights into physiological mechanisms that static analyses could not have resolved. Imaging studies of neurons in living spinal cord have only recently begun. Lichtman and his colleagues first demonstrated their feasibility by tracking injured dorsal column (DC) axons with wide-field microscopy8,9. Multi-photon in vivo imaging of deeply positioned DC axons, microglia and blood vessels has also been accomplished10. Over the last few years, we have pioneered in applying in vivo imaging to monitor regeneration of DR axons using wide-field microscopy and H line of thy1-YFP mice. These studies have led us to a novel hypothesis about why DR axons are prevented from regenerating within the spinal cord11.
In H line of thy1-YFP mice, distinct YFP+ axons are superficially positioned, which allows several axons to be monitored simultaneously. We have learned that DR axons arriving at DREZ are better imaged in lumbar than in cervical spinal cord. In the present report we describe several strategies that we have found useful to assure successful long-term and repeated imaging of regenerating DR axons. These include methods that eliminate repeated intubation and respiratory interruption, minimize surgery-associated stress and scar formation, and acquire stable images at high resolution without phototoxicity.

An in vivo imaging protocol to monitor primary sensory axons following dorsal root crush is described. The procedures utilize wide-field fluorescence microscopy and thy1-YFP transgenic mice, and permit repeated imaging of axon regeneration over 4 cm in the PNS and axon interactions with the interface of the CNS.

实时成像背根轴突后根切断术

The primary sensory axons injured by spinal root injuries fail to regenerate into the spinal cord, leading to chronic pain and permanent sensory loss. ...

神经科学

Surgical Technique for Spinal Cord Delivery of Therapies: Demonstration of Procedure in Gottingen Minipigs

This is a compact visual description of a combination of surgical technique and device for the delivery of (gene and cell) therapies into the spinal cord. While the technique is demonstrated in the animal, the procedure is FDA-approved and currently being used for stem cell transplantation into the spinal cords of patients with ALS. While the FDA has recognized proof-of-principle data on therapeutic efficacy in highly characterized rodent models, the use of large animals is considered critical for validating the combination of a surgical procedure, a device, and the safety of a final therapy for human use. The size, anatomy, and general vulnerability of the spine and spinal cord of the swine are recognized to better model the human. Moreover, the surgical process of exposing and manipulating the spinal cord as well as closing the wound in the pig is virtually indistinguishable from the human. We believe that the healthy pig model represents a critical first step in the study of procedural safety.

Short visual description of the surgical technique and device used for the delivery of (gene and cell) therapies into the spinal cord. The technique is demonstrated in the animal but is entirely translatable and currently being used for human application.

外科技术治疗脊髓交付：在哥廷根小型猪演示程序

This is a  compact visual description of a combination of surgical technique and device  for the  delivery of (gene and cell) therapies into the spinal ...

A Method of Nodose Ganglia Injection in Sprague-Dawley Rat

Afferent signaling via the vagus nerve transmits important general visceral information to the central nervous system from many diverse receptors located in the organs of the abdomen and thorax. The vagus nerve communicates information from stimuli such as heart rate, blood pressure, bronchopulmonary irritation, and gastrointestinal distension to the nucleus of solitary tract of the medulla. The cell bodies of the vagus nerve are located in the nodose and petrosal ganglia, of which the majority are located in the former. The nodose ganglia contain a wealth of receptors for amino acids, monoamines, neuropeptides, and other neurochemicals that can modify afferent vagus nerve activity. Modifying vagal afferents through systemic peripheral drug treatments targeted at the receptors on nodose ganglia has the potential of treating diseases such as sleep apnea, gastroesophageal reflux disease, or chronic cough. The protocol here describes a method of injection neurochemicals directly into the nodose ganglion. Injecting neurochemicals directly into the nodose ganglia allows study of effects solely on cell bodies that modulate afferent nerve activity, and prevents the complication of involving the central nervous system as seen in systemic neurochemical treatment. Using readily available and inexpensive equipment, intranodose ganglia injections are easily done in anesthetized Sprague-Dawley rats.

Afferent vagal signaling transmits important information to central nervous system from receptors located in organs of the abdomen and thorax. The nodose ganglia of vagus nerves contain many types of receptors that modulate vagal activity. This protocol describes a method of local injections of neurochemicals into the nodose ganglia.

结状神经节注射液只SD大鼠的方法

Afferent signaling via the vagus nerve transmits important general visceral information to the central nervous system from many diverse receptors located ...

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

Achieving axon regeneration after nervous system injury is a challenging task. As different parts of the central nervous system (CNS) differ from each other anatomically, it is important to identify an appropriate model to use for the study of axon regeneration. By using a suitable model, we can formulate a specific treatment based on the severity of injury, the neuronal cell type of interest, and the desired spinal tract for assessing regeneration. Within the sensory pathway, DRG neurons are responsible for relaying sensory information from the periphery to the CNS. We present here a protocol that uses a DRG injection with a viral vector and a concurrent dorsal root crush injury in the lower cervical spinal cord of an adult rat as a model to study sensory axon regeneration. As demonstrated using a control virus, AAV5-GFP, we show the effectiveness of a direct DRG injection in transducing DRG neurons and tracing sensory axons into the spinal cord. We also show the effectiveness of the dorsal root crush injury in denervating the forepaw as an injury model for evaluating axon regeneration. Despite the requirement for specialized training to perform this invasive surgical procedure, the protocol is flexible, and potential users can modify many parts to accommodate their experimental requirements. Importantly, it can serve as a foundation for those in search of a suitable animal model for their studies. We believe that this article will help new users to learn the procedure in a very efficient and effective manner.

This protocol presents the use of a dorsal root ganglion (DRG) injection with a viral vector and a concurrent dorsal root crush injury in an adult rat as a model to study sensory axon regeneration. This model is suitable for investigating the use of gene therapy to promote 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 ...

In Vivo SiRNA Transfection and Gene Knockdown in Spinal Cord via Rapid Noninvasive Lumbar Intrathecal Injections in Mice

This report describes a step-by-step guide to the technique of acute intrathecal needle injections in a noninvasive manner, i.e. independent of catheter implantation. The technical limitation of this surgical technique lies in the finesse of the hands. The injection is rapid, especially for a trained experimenter, and since tissue disruption with this technique is minimal, repeated injections are possible; moreover immune reaction to foreign tools (e.g. catheter) does not occur, thereby giving a better and more specific read out of spinal cord modulation. Since the application of the substance is largely limited to the target region of the spinal cord, drugs do not need to be applied in large dosages, and more importantly unwanted effects on other tissue, as observed with a systemic delivery, could be circumvented1,2. Moreover, we combine this technique with in vivo transfection of nucleic acid with the help of polyethylenimine (PEI) reagent3, which provides tremendous versatility for studying spinal functions via delivery of pharmacological agents as well as gene, RNA, and protein modulators.

In Vivo SiRNA Transfection and Gene Knockdown in Spinal Cord via Rapid Noninvasive Lumbar Intrathecal Injections in Mice

This report describes a simple and rapid technique of intrathecal needle puncture for a localized transfection of siRNA in the lumbar spinal cord in mouse under short lasting light anesthesia.

体内转染和基因敲除在脊髓通过快速无创腰椎鞘内注射小鼠

This report describes a step-by-step guide to the technique of acute intrathecal needle injections in a noninvasive manner, i.e. independent of catheter ...

生物学

Real-Time Assessment of Spinal Cord Microperfusion in a Porcine Model of Ischemia/Reperfusion

Spinal cord injury is a devastating complication of aortic repair. Despite developments for the prevention and treatment of spinal cord injury, its incidence is still considerably high and therefore, influences patient outcome. Microcirculation plays a key role in tissue perfusion and oxygen supply and is often dissociated from macrohemodynamics. Thus, direct evaluation of spinal cord microcirculation is essential for the development of microcirculation-targeted therapies and the evaluation of existing approaches in regard to spinal cord microcirculation. However, most of the methods do not provide real-time assessment of spinal cord microcirculation. The aim of this study is to describe a standardized protocol for real-time spinal cord microcirculatory evaluation using laser-Doppler needle probes directly inserted in the spinal cord. We used a porcine model of ischemia/reperfusion to induce deterioration of the spinal cord microcirculation. In addition, a fluorescent microsphere injection technique was used. Initially, animals were anesthetized and mechanically ventilated. Thereafter, laser-Doppler needle probe insertion was performed, followed by the placement of cerebrospinal fluid drainage. A median sternotomy was performed for exposure of the descending aorta to perform aortic cross-clamping. Ischemia/reperfusion was induced by supra-celiac aortic cross-clamping for a total of 48 min, followed by reperfusion and hemodynamic stabilization. Laser-Doppler Flux was performed in parallel with macrohemodynamic evaluation. In addition, automated cerebrospinal fluid drainage was used to maintain a stable cerebrospinal pressure. After completion of the protocol, animals were sacrificed, and the spinal cord was harvested for histopathological and microsphere analysis. The protocol reveals the feasibility of spinal cord microperfusion measurements using laser-Doppler probes and shows a marked decrease during ischemia as well as recovery after reperfusion. Results showed comparable behavior to fluorescent microsphere evaluation. In conclusion, this new protocol might provide a useful large animal model for future studies using real-time spinal cord microperfusion assessment in ischemia/reperfusion conditions.

Spinal cord microcirculation plays a pivotal role in spinal cord injury. Most methods do not allow real-time assessment of spinal cord microcirculation, which is essential for the development of microcirculation-targeted therapies. Here, we propose a protocol using Laser-Doppler-Flow Needle probes in a large animal model of ischemia/reperfusion.

猪缺血/再灌注模型中脊髓微灌注的实时评估

Spinal cord injury is a devastating complication of aortic repair. Despite developments for the prevention and treatment of spinal cord injury, its ...

Advanced Imaging of Dorsal Root Ganglion Neurons for Sensory Modality Research

In Vivo Calcium Imaging of Dorsal Root Ganglia Neurons' Response to Somatic and Visceral Stimuli

A technique is described for surgically exposing the dorsal root ganglion (DRG) of the lumbar-6 in a live, anesthetized laboratory mouse, along with the protocol for in vivo calcium imaging of the exposed DRG in response to various visceral and somatic stimuli. Pirt-GCaMP6s mice or C57BL6 mice intrathecally injected with AAV viruses packaged with GCaMP6s were utilized to capture Ca2+ transients. The amplitude of these transients indicates sensitivity to specific sensory modalities. Afferent fibers originate from internal organs, with primary neuronal cell bodies in spinal or vagal ganglia. Studies on visceral nociception and acupuncture analgesia can potentially be conducted on primary sensory neurons using advanced imaging technologies like in vivo calcium imaging, allowing for the recording of neuronal activity ensembles in the intact animal during stimulation or intervention. The responses of DRG neuron ensembles to somatic and visceral stimuli applied to their corresponding receptive fields were recorded. This technique illustrates how neuronal populations react to various types of somatic and visceral stimuli. It is possible to comprehensively compare neuronal ensemble responses to different stimuli, which is a particularly valuable approach in research on visceral pain and segmental mechanisms of somatic stimulation, such as acupuncture.

The present protocol outlines in vivo calcium imaging for measuring the responses of ensembles of lumbar-6 DRG neurons to somatic and visceral stimuli. Thorough comparisons can be made among neurons responding to different stimuli. This protocol is valuable for investigating mechanisms of visceral pain and somatic stimulation, such as acupuncture.

A technique is described for surgically exposing the dorsal root ganglion (DRG) of the lumbar-6 in a live, anesthetized laboratory mouse, along with the ...

幼年大鼠腰椎穿刺 AAV9 递送用于 CNS 基因治疗

Intrathecal Vector Delivery in Juvenile Rats via Lumbar Cistern Injection

Gene therapy is a powerful technology to deliver new genes to a patient for the treatment of disease, be it to introduce a functional gene, inactivate a toxic gene, or provide a gene whose product can modulate the biology of the disease. The delivery method for the therapeutic vector can take many forms, ranging from intravenous infusion for systemic delivery to direct injection into the target tissue. For neurodegenerative disorders, it is often desirable to skew transduction towards the brain and/or spinal cord. The least invasive approach to target the entire central nervous system involves injection into the cerebrospinal fluid (CSF), allowing the therapeutic to reach a large fraction of the central nervous system. The safest approach to deliver a vector into the CSF is the lumbar intrathecal injection, where a needle is introduced into the lumbar cistern of the spinal cord. This technique, also known as a lumbar puncture, has been widely used in neonatal and adult rodents and in large animal models. While the technique is similar across species and developmental stages, subtle differences in size, structure, and elasticity of tissues surrounding the intrathecal space require accommodations in the approach. This article describes a method for performing lumbar puncture in juvenile rats to deliver an adeno-associated serotype 9 vector. Here, 25-35 &#181;L of vector were injected into the lumbar cistern, and a green fluorescent protein (GFP) reporter was used to evaluate the transduction profile resulting from each injection. The benefits and challenges of this approach are discussed.

作者聚焦：评估幼年大鼠的鞘内基因治疗疗效

A surgical procedure is described to perform injections into the lumbar cistern of the juvenile rat. This approach has been used for the intrathecal delivery of gene therapy vectors, but it is anticipated that this approach can be used for a variety of therapeutics, including cells and drugs.

通过腰椎池注射在幼年大鼠中鞘内载体给药

Gene therapy is a powerful technology to deliver new genes to a patient for the treatment of disease, be it to introduce a functional gene, inactivate a ...

Laminotomy 对猪腰背根神经节通路和注射的治疗

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此视频中的章节

椎管内针对肌萎缩性脊髓侧索硬化症和外伤性脊髓损伤颈腹角细胞移植

实时成像背根轴突后根切断术

外科技术治疗脊髓交付：在哥廷根小型猪演示程序

结状神经节注射液只SD大鼠的方法

背根神经节内注射和背根挤压伤为感官轴突再生模型

体内转染和基因敲除在脊髓通过快速无创腰椎鞘内注射小鼠

猪缺血/再灌注模型中脊髓微灌注的实时评估

In Vivo Calcium Imaging of Dorsal Root Ganglia Neurons' Response to Somatic and Visceral Stimuli

通过腰椎池注射在幼年大鼠中鞘内载体给药

通过腰椎池注射在幼年大鼠中鞘内载体给药