Name: ethanol-induced cervical sympathetic ganglion block applications for promoting canine inferior alveolar nerve regeneration using an artificial nerve
Uploaded: 2018-11-30T13:00:00
Description: Watch this Scientific Journal Video about Ethanol-Induced Cervical Sympathetic Ganglion Block Applications for Promoting Canine Inferior Alveolar Nerve Regeneration Using an Artificial Nerve at JoVE.c

这项工作得到了京都大学前沿医学研究所生物人工器官系的支持。我们要感谢前沿医学研究所的兽医工作人员。

这项研究是根据《动物护理和使用指导原则》进行的, 并得到京都大学动物研究委员会的批准 (日本京都; 授权号: r-16-16)。为尽量减少动物痛苦, 我们尽了一切努力, 本报告的所有章节都遵守《动物研究:活体实验报告》的准则。
1. pga-c 管的制造
<ol><li>要通过可吸收的聚乙酸 (pga) 管制造人工神经导管, 请使用装有48个主轴和 5个 pga 纤维的管状编织机, 该编织机由26个细丝?...

<ol>
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href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+superior+cervical+ganglion+resection+on+peripheral+facial+palsy+in+rats.">The effect of superior cervical ganglion resection on peripheral facial palsy in rats.</a> Journal of Anesthesia. 30 (4), 677-683 (2016).</li><li>Hanamatsu, N., Yamashiro, M., Sumitomo, M., Furuya, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effectiveness+of+cervical+sympathetic+ganglia+block+on+regeneration+of+the+trigeminal+nerve+following+transection+in+rats.">Effectiveness of cervical sympathetic ganglia block on regeneration of the trigeminal nerve following transection in rats.</a> Regional Anesthesia and Pain Medicine. 27 (3), 268-276 (2002).</li><li>Shionoya, Y., Sunada, K., Shigeno, K., Nakada, A., Honda, M., Nakamuram, T. <a target="_blank" 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Kuttenberger, J., Filippi, A., Sader, R., Zeilhofer, H. 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我们提出了一种有效的方法, 以共同作用, 生物可吸收的神经管结合乙醇诱导的 csgb, 进行 ian 再生。在这项研究中, 我们使用了狗, 因为其他动物模型, 如老鼠、老鼠和兔子, 预期寿命短, 身体体积小, 因此不能用来进行精确的外科手术。由于 ian 位于下颌骨周围的下颌骨管内, 因此在进行神经重建时, 需要一种手术技术来避免神经和血管损伤。这个程序的一个重要的技术提示是小心地取出下颌骨骨板, ?...

在许多情况下, 下牙槽神经 (ian) 的创伤是医源性的, 经常是由拔除第三磨牙或放置牙种植体 1,2,3引起的。ian 的损伤会导致热和触觉的缺陷, 以及感觉异常、麻醉、麻醉和异位。神经损伤不仅通过保守治疗治疗, 还通过其他方法治疗, 包括缝合和自体移植。然而, 这些方法有缺点, 其中往往包括缺乏症状改善和神经缺陷的捐献者部位 4,5,6。
人工神经-多糖酸胶原蛋白 (pga-c) 管最初是在日本发展起来的。它是一个生物吸收管, 其内腔充满海绵状胶原蛋白7。在动物实验中, 该管用于增强有腹膜神经缺损的猎狗的神经再生, 并被证明比自体神经移植促进了更高水平的恢复 8.pga-c 管在周围神经损伤患者中的临床应用始于2002年。此外, 在治疗三叉神经病变 (ian 和舌神经)9,10,11方面也取得了良好的临床效果。使用 pga-c 管成功神经再生的一个关键因素是周围组织的血液供应 8。颈椎交感神经节阻滞 (csgb) 在头部和颈部区域造成交感神经阻滞, 并增加血液流向各自神经支配区域12;因此, 它已被用于治疗复杂的区域疼痛综合征和循环系统功能不全 13,14,15。然而, 只有少数实验研究 csgb 在增加血液流动方面的有效性为16,17。为了确保足够的 csgb 效率, 封锁必须与局部麻醉剂一起应用, 每天一次或两次, 为期数周, 从而在生成动物模型以研究这一技术时构成挑战。为了解决这一限制, 在之前的一项研究中, 我们开发了一个犬类模型, 长期增加了口腔面部区域的血液流动。该模型是通过注入99.5% 乙醇来执行 csgb 生成的。我们用激光多普勒血流测量和红外热像仪每周评估口腔黏膜血流和鼻皮肤温度, 为期12周。我们发现, 在这个模型中, 口腔面部区域的血液流动增加了7-10周。
在本研究中, 我们评估了乙醇诱导的 csgb 对神经再生的影响。 pga-c 管被植入左边的 ian 有10毫米缝隙的猎狗体内。一周后, csgb 通过注射乙醇进行。手术三个月后, 我们进行了各种电生理、组织学和形态学研究, 以评估 csgb 对神经再生的影响。我们提供了一个详细的协议, 用于利用 pga-c 管和乙醇诱导的 csgb 重建。

<table border="0" cellpadding="0" cellspacing="0" width="1208">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:396px;">NMP Collagen PS</td>
			<td style="width:228px;">Nippon Meatpackers</td>
			<td style="width:224px;">301-84621</td>
			<td style="width:360px;">Atelocollagen extracted from young porcine skin by enzyme treatment</td>
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			<td height="42" style="height:42px;width:396px;">Surgical clippers</td>
			<td style="width:228px;">Roboz Surgical Instrument Company</td>
			<td style="width:224px;">RC-5903</td>
			<td></td>
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			<td height="21" style="height:21px;width:396px;">Disposable scalpel (No.15)</td>
			<td style="width:228px;">Kai medical&#160;</td>
			<td style="width:224px;">219ABBZX00073000</td>
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			<td style="width:228px;">Nakanishi</td>
			<td style="width:224px;">VS3-LED-HPSC, E1133</td>
			<td>Piezoelectric surgery for bone processing</td>
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			<td height="21" style="height:21px;width:396px;">4-0 nylon sutures</td>
			<td style="width:228px;">Ethicon</td>
			<td style="width:224px;">8881H</td>
			<td></td>
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			<td height="21" style="height:21px;width:396px;">8-0 nylon sutures</td>
			<td style="width:228px;">Ethicon</td>
			<td style="width:224px;">2775G</td>
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			<td height="21" style="height:21px;width:396px;">Isepamicin sulfate</td>
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			<td style="width:224px;">620005641</td>
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			<td height="21" style="height:21px;width:396px;">Disposable scalpel (No.10)</td>
			<td style="width:228px;">Kai medical&#160;</td>
			<td style="width:224px;">219ABBZX00073000</td>
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			<td height="21" style="height:21px;width:396px;">30-gauge needle&#160;</td>
			<td style="width:228px;">Nipro</td>
			<td>1134</td>
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		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:396px;">1-0 absorbable stitches</td>
			<td style="width:228px;">Ethicon</td>
			<td style="width:224px;">J347H</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:396px;">3-0 Nylon stitches</td>
			<td style="width:228px;">Ethicon</td>
			<td style="width:224px;">8872H</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:396px;">Neo Thermo</td>
			<td style="width:228px;">NEC Avio</td>
			<td style="width:224px;">TVS-700</td>
			<td>Infrared thermography&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:396px;">Neuropack &#931;&#160;</td>
			<td style="width:228px;">NIHON KOHDEN</td>
			<td style="width:224px;">MEB-5504</td>
			<td>Orthodromic recorder for electrophysiological recording</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:396px;">Toluidine Blue&#160;</td>
			<td style="width:228px;">Sigma-Aldrich&#160;</td>
			<td style="width:224px;">T3260-5G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:396px;">Light microscope</td>
			<td style="width:228px;">Keyence</td>
			<td style="width:224px;">BZ-9000</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:396px;">Mouse anti-human neurofilament protein monoclonal antibody</td>
			<td style="width:228px;">DAKO</td>
			<td style="width:224px;">N1591</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:396px;">Polyclonal rabbit anti-S100 antibody</td>
			<td style="width:228px;">DAKO</td>
			<td style="width:224px;">Z0311</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:396px;">Transmission electron microscopy</td>
			<td style="width:228px;">Hitachi High Technologies</td>
			<td style="width:224px;">Hitachi H-7000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:396px;">Dynamic cell count</td>
			<td style="width:228px;">Keyence</td>
			<td style="width:224px;">BZ-H1C</td>
			<td>Software for morphological evaluation</td>
		</tr>
	</tbody>
</table>

ethanol-induced cervical sympathetic ganglion block applications for promoting canine inferior alveolar nerve regeneration using an artificial nerve

聚甘酸胶原蛋白 (pga-c) 管是一种生物吸收神经管, 充满了多室结构的胶原蛋白, 由薄胶原蛋白膜组成。使用这些试管治疗受损的下牙槽神经 (ian) 时, 取得了良好的临床效果。使用 pga-c 管成功神经再生的一个关键因素是周围组织的血液供应。颈椎交感神经节阻滞 (csgb) 在头部和颈部造成交感神经阻滞, 从而增加该地区的血液流动。为确保充分发挥作用, 必须连续几周使用局部麻醉剂每天1至 2次;这在创建用于研究这一技术的动物模型时带来了挑战。为了解决这一限制, 我们在口腔面部区域长期增加血液流动的犬类模型中开发了乙醇诱导的 csgb。我们研究了通过 pga-c 管植入物进行的 ian 再生是否可以通过该模型得到提高。14个比格尔斯分别在左 ian 的10毫米缝隙上植入了一个 pga-c 管。ian 位于下颌骨管内, 周围是骨头, 因此我们选择了压电手术, 包括超声波, 进行骨处理, 以尽量减少神经和血管损伤的风险。这种方法获得了良好的手术效果。手术一周后, 其中7只狗通过注射乙醇离开了 csgb。乙醇诱导的 csgb 改善了神经再生, 提示血流的增加有效地促进了 ian 缺陷中的神经再生。这种犬类模型可以为进一步研究 csgb 的长期效果做出贡献。

我们观察到, 在左 csgb 后 1周, 被阻塞一侧的面部皮肤温度增加 (图 8)。
重建后 3个月, 重建区的 pga-c 管被吸收, 下肺泡神经再生在重建和重建 + csgb 群 (图 9a, b)18中观察到。
snap 在重建 + csgb 和仅重建组的重?...

Watch this Scientific Journal Video about Ethanol-Induced Cervical Sympathetic Ganglion Block Applications for Promoting Canine Inferior Alveolar Nerve Regeneration Using an Artificial Nerve at JoVE.c

Ethanol-Induced Cervical Sympathetic Ganglion Block Applications for Promoting Canine Inferior Alveolar Nerve Regeneration Using an Artificial Nerve

乙醇诱导的颈椎交交型神经节阻滞在促进犬下肺泡神经再生中的应用

我们评价了颈椎交感神经节阻滞对人工神经导管修复神经的影响。雄性猎狗分别在左下牙槽神经的10毫米缝隙上植入人工神经;左颈椎交感神经节通过侧开手术注射99.5% 乙醇, 阻断了颈椎交感神经节。

Polyglycolic acid collagen (PGA-C) tubes are bio-absorbable nerve tubes filled with collagen of multi-chamber structure, which consist of thin collagen ...

该方法有助于利用人工神经导管回答人工器官场神经再生的关键问题。该技术的主要优点是，压电手术可用于低劣的道骨神经重建，以避免神经和血管损伤。这种技术的影响延伸到治疗性神经病，因为它有促进神经再生的潜力。在确认麻醉后，对踏板反射缺乏反应，在眼睛前表面涂抹凝胶，以避免角膜磨损。将狗放在正确的横向位置。使用 27 量针将 3 毫升 1% 利多卡因注射到左曼迪贝左银黄中。接下来，使用数字15手术刀刀片，使一个五厘米的横向切口在左曼迪贝左丁吉瓦暴露的毛毛。使用28至32千赫频率的压电超声波振动，通过后心前身将压电超声波振动研磨成三厘米八毫米矩形的近向。由于低劣的道神经位于骨管内，周围有骨骼，我们使用由超声波组成的压电手术进行骨骼处理，以最大限度地降低神经和血管损伤的风险。取出毛骨板的前部，露出左下形的阿尔维外神经。使用手术刀转除下一个道长神经，以允许去除10毫米段。将被切断神经的近端和近端树桩插入神经管，深度为两毫米。使用 8-0尼龙缝合线和手术显微镜在8倍的放大率，以缝合管到近端和近端神经端。然后，将骨板返回到其原始位置，然后用4-0尼龙缝合线关闭伤口。手术后一天，对麻醉下的面部骨进行计算机断层成像，以确认骨质板处于适当位置。重建程序一周后，剃光手术场。然后，在手术区域左侧绘制一条切口线，并使用 21 量针将 5 毫升 1% 利多卡因注射到剃光的胸腔中，作为局部麻醉和镇痛剂。使用数字10手术刀刀片，在标记的线条上切口，并使用电动手术刀切化脂肪层，以暴露肌肉筋膜。暴露塞拉图斯腹和规模肌肉。将肌肉从腹体提升至后侧，露出第二根和第三根肋骨。在第二和第三个间空间进行左横向胸腔切除术，以暴露左颈同情结节。使用30量针将2毫升99.5%乙醇注射到宫颈交感结节中。宫颈同情结节被乙醇退化改变。然后，用中断的 1- 0 可吸收的缝线关闭成本间空间。用中断的3-0尼龙缝关闭皮肤，并在宫颈交感结块或CSGB块一周后用红外热成像测量面部皮肤温度。重建后三个月，在重建区吸收聚聚曲酸-胶原管，在重建组和重建加CSGB组中观察到低劣的道骨神经的再生。在重建加CSGB和神经重建组的重建方面，感觉神经作用潜力也是可测量的，在重建加CSGB动物中观察到的恢复指数和感觉神经传导速度明显高于重建组。骨髓神经纤维在重建组和重建加CSGB组再生下位低头骨神经的中枢和近段观察，尽管两组都显示出与正常对照组相比，再生骨髓神经直径较小。再生轴子和Schwann细胞的存在通过分别用抗神经丝和抗S100抗体染色，在重建加CSGB组的中心和远点部分得到进一步确认。形态学分析表明，与重建动物相比，CSGB组再生左下性肺活体神经中枢和近段的骨髓神经纤维密度和神经组织百分比明显较高，使用CSGB程序观察到的G比明显小。宫颈交感结节块导致神经再生改善，表明血流增加能有效促进神经再生。该技术有助于在原位组织工程领域开展研究，探索利用人工神经治疗原位神经病变的外科治疗。

Research

JoVE Journal

Neuroscience

An Optic Nerve Crush Injury Murine Model to Study Retinal Ganglion Cell Survival

一个视神经挤压伤的小鼠模型，研究视网膜神经节细胞生存

Injury to the optic nerve can lead to axonal degeneration, followed by a gradual death of retinal ganglion cells (RGCs), which results in irreversible ...

科研

神经科学

Reproducible Mouse Sciatic Nerve Crush and Subsequent Assessment of Regeneration by Whole Mount Muscle Analysis

重复性小鼠坐骨神经挤压和再生的后续评估整装肌肉分析

Regeneration in the peripheral nervous system (PNS) is widely studied both for its relevance to human disease and to understand the robust regenerative ...

Optogenetic Stimulation of the Auditory Nerve

听觉神经的光遗传学刺激

Direct electrical stimulation of spiral ganglion neurons (SGNs) by cochlear implants (CIs) enables open speech comprehension in the majority of implanted ...

Dorsal Root Ganglia Neurons and Differentiated Adipose-derived Stem Cells: An In Vitro Co-culture Model to Study Peripheral Nerve Regeneration

Dorsal Root Ganglia Neurons and Differentiated Adipose-derived Stem Cells: An In Vitro Co-culture Model to Study Peripheral Nerve Regeneration

背根神经节神经元和区分脂肪干细胞：一种<em&gt;在体外</em&gt;共培养模型来研究周围神经再生

Dorsal root ganglia (DRG) neurons, located in the intervertebral foramina of the spinal column, can be used to create an in vitro system facilitating the ...

Assessment of Neuromuscular Function Using Percutaneous Electrical Nerve Stimulation

神经肌肉功能的应用经皮神经电刺激评估

Percutaneous electrical nerve stimulation is a non-invasive method commonly used to evaluate neuromuscular function from brain to muscle (supra-spinal, ...

Fabrication of High Contact-Density, Flat-Interface Nerve Electrodes for Recording and Stimulation Applications

高接触密度的制作，平面接口神经电极用于记录和刺激的应用

Many attempts have been made to manufacture multi-contact nerve cuff electrodes that are safe, robust and reliable for long term neuroprosthetic ...

Quantifying Acute Changes in Renal Sympathetic Nerve Activity in Response to Central Nervous System Manipulations in Anesthetized Rats

麻醉大鼠中枢神经系统操作对肾交感神经活动的急性变化量化

Renal sympathetic nerve activity (RSNA) and mean arterial pressure are important parameters in cardiovascular and autonomic research; however, there are ...

Facial Nerve Surgery in the Rat Model to Study Axonal Inhibition and Regeneration

大鼠模型中的面部神经外科研究同氧抑制与再生

This protocol describes consistent and reproducible methods to study axonal regeneration and inhibition in a rat facial nerve injury model. The facial ...

Assessing Early Stage Open-Angle Glaucoma in Patients by Isolated-Check Visual Evoked Potential

通过隔离检查视觉诱发潜力评估患者早期开放角青光眼

Recently, the isolated-check visual evoked potential (icVEP) technique was designed and has been reported to detect glaucomatous damage earlier and ...

Purification of Fibroblasts and Schwann Cells from Sensory and Motor Nerves in Vitro

从体外感觉和运动神经中纯化成纤维细胞和施万细胞

The principal cells in the peripheral nervous system are the Schwann cells (SCs) and the fibroblasts. Both these cells distinctly express the sensory and ...