Name: creation of two saccular elastase-digested aneurysms with different hemodynamics in one rabbit
Uploaded: 2021-04-15T15:00:00
Description: Watch this Scientific Journal Video about Creation of Two Saccular Elastase-Digested Aneurysms with Different Hemodynamics in One Rabbit at JoVE.com

作者感谢Hans Rudolf Widmer教授，Luca Remonda博士和Javier Fandino教授对这项工作的科学支持和技术贡献。特别感谢Olgica Beslac在手术过程中的建议和Kay Nettelbeck的帮助。此外，我们感谢Daniela Casoni DVM，博士和医学兽医。Luisana Garcia，PD Alessandra Bergadano博士和Carlotta Detotto博士，感谢他们敬业的兽医支持。

注意：该实验已获得瑞士伯尔尼州地方动物护理委员会的批准（申请号BE108/16），所有动物护理和程序均按照机构指南和3R原则15，16进行。数据根据 REACH 指南报告。围手术期管理由董事会认证的兽医麻醉师进行。在这项研究中，雌性新西兰白兔，平均体重为4.0（±0.3）公斤，平均年龄为25（±5）周，被饲养在22-24°C的室温下，进行12小时的光照/黑暗循环?...

<ol>
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最常见的动脉瘤形成技术包括通过开放或血管内方法在右侧CCA的起源处创建残端动脉瘤。该模型已被验证为一种稳定的非生长动脉瘤，随时间20，21 保持开放状态。第二种可能的技术涉及通过手术在左侧吻合右侧CCA并在分叉处缝合动脉瘤袋14，22，23来建立动脉分叉动脉瘤。尽管这两?...

颅内动脉瘤是一种严重的疾病，破裂后死亡率达到50%，10-20%的患者长期残疾1。在过去的十年中，血管内治疗方案发展迅速，但与此同时，复发率也在增加，卷曲后动脉瘤再通率高达33% 2，3。为了更好地了解动脉瘤闭塞和再通的潜在病理生理学，以及开发和测试新的血管内装置，目前需要可靠的临床前模型，其血流动力学、形态学和组织学特征与人颅内动脉瘤相似4，5，6.截至今天，还没有明确的模型作为临床前试验的标准，研究人员可以使用大量的物种和技术7，8。
然而，兔子是一个特别感兴趣的物种，因为它的颈部动脉和人类脑血管之间的大小和血流动力学相似性，以及其相似的凝血和溶栓特征。颈总动脉 （CCA） 上具有弹性蛋白酶消化囊状动脉瘤的几种模型在流动条件、几何特征和壁特征方面显示出与人颅内动脉瘤的定性和定量相似性9，10，11，12。本研究旨在描述一种在同一动物中同时使用残肢和分叉弹性蛋白酶消化动脉瘤创建新的兔动脉瘤模型的技术。手术技术的灵感来自Hoh等人13和Wanderer等人14的技术，略有修改，以提供良好的标准化和可重复性，并确保低死亡率和发病率。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>4-0 resorbable suture</td>
			<td>Ethicon Inc., USA</td>
			<td>VCP292ZH</td>
			<td></td>
		</tr>
		<tr>
			<td>4-0 resorbable suture</td>
			<td>Ethicon Inc., USA</td>
			<td>VCP304H</td>
			<td></td>
		</tr>
		<tr>
			<td>6-0 non absorbable suture</td>
			<td>B. Braun, Germany</td>
			<td>C0766070</td>
			<td></td>
		</tr>
		<tr>
			<td>9-0 non absorbable suture</td>
			<td>B. Braun, Germany</td>
			<td>G1111140</td>
			<td></td>
		</tr>
		<tr>
			<td>Adrenaline</td>
			<td>Amino AG</td>
			<td>1445419</td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Amiodarone</td>
			<td>Helvepharm AG</td>
			<td>5078567</td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Anesthesia machine</td>
			<td>Dr&#228;ger</td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Aspirin</td>
			<td>Sanofi-Aventis (Suisse) SA</td>
			<td>622693</td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Atropine</td>
			<td>Labatec Pharma SA</td>
			<td>6577083</td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Bandpass filter blue</td>
			<td>Thorlabs</td>
			<td>FD1B</td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Bandpass filter green</td>
			<td>Thorlabs</td>
			<td>FGV9</td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Biemer vessel clip (2x)</td>
			<td>B. Braun Medical AG, Aesculap, Switzerland</td>
			<td>FD560R</td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Bipolar forceps</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Bispectral index (neonatal)</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Blood pressure cuff (neonatal)</td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Bycilces spotlight</td>
			<td></td>
			<td></td>
			<td>any other</td>
		</tr>
		<tr>
			<td>Clamoxyl</td>
			<td>GlaxoSmithKline AG</td>
			<td>758808</td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Dexmedetomidine</td>
			<td>Ever Pharma</td>
			<td>136740-1</td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Elastase</td>
			<td>Sigma Aldrich</td>
			<td>E7885</td>
			<td></td>
		</tr>
		<tr>
			<td>Electrocardiogram electrodes</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ephedrine</td>
			<td>Amino AG</td>
			<td>1435734</td>
			<td></td>
		</tr>
		<tr>
			<td>Esmolol</td>
			<td>OrPha Swiss GmbH</td>
			<td>3284044</td>
			<td></td>
		</tr>
		<tr>
			<td>Fentanyl (intravenous use)</td>
			<td>Janssen-Cilag AG</td>
			<td>98683</td>
			<td></td>
		</tr>
		<tr>
			<td>Fentanyl (transdermal)</td>
			<td>Mepha Pharma AG</td>
			<td>4008286</td>
			<td></td>
		</tr>
		<tr>
			<td>Fluoresceine</td>
			<td>Curatis AG</td>
			<td>5030376</td>
			<td></td>
		</tr>
		<tr>
			<td>Fragmin</td>
			<td>Pfizer PFE Switzerland GmbH</td>
			<td>1906725</td>
			<td></td>
		</tr>
		<tr>
			<td>Heating pad or heating forced-air warming system</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Isotonic sodium chloride solution (0.9%)</td>
			<td>Fresenius KABI</td>
			<td>336769</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Pfizer PFE Switzerland GmbH</td>
			<td>342261</td>
			<td></td>
		</tr>
		<tr>
			<td>lid retractor</td>
			<td></td>
			<td></td>
			<td>Approach</td>
		</tr>
		<tr>
			<td>Lidocaine</td>
			<td>Streuli Pharma AG</td>
			<td>747466</td>
			<td></td>
		</tr>
		<tr>
			<td>Longuettes</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Metacam</td>
			<td>Boehringer Ingelheim</td>
			<td>P7626406</td>
			<td>Medication</td>
		</tr>
		<tr>
			<td>Methadone</td>
			<td>Streuli Pharma AG</td>
			<td>1084546</td>
			<td>Sedaton</td>
		</tr>
		<tr>
			<td>Micro-forceps&#160; curved</td>
			<td>Ulrich Swiss, Switzerland</td>
			<td>U52-015-15</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro-forceps&#160; straight 2x</td>
			<td>Ulrich Swiss, Switzerland</td>
			<td>U52-010-15</td>
			<td></td>
		</tr>
		<tr>
			<td>Microscissors</td>
			<td>Ulrich Swiss , Switzerland</td>
			<td>U52-327-15</td>
			<td></td>
		</tr>
		<tr>
			<td>Midazolam</td>
			<td>Accord Healthcare AG</td>
			<td>7752484</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle 23 G</td>
			<td></td>
			<td></td>
			<td>arteriotomy</td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>O2-Face mask</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Operation microscope</td>
			<td>Wild Heerbrugg</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Papaverin</td>
			<td>Bichsel</td>
			<td></td>
			<td>topical application</td>
		</tr>
		<tr>
			<td>Povidone iodine</td>
			<td>Mundipharma Medical Company</td>
			<td></td>
			<td>any generic</td>
		</tr>
		<tr>
			<td>Prilocaine-lidocaine creme</td>
			<td>Emla</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Propofol</td>
			<td>B. Braun Medical AG, Switzerland</td>
			<td></td>
			<td>General anesthesia</td>
		</tr>
		<tr>
			<td>Pulse oxymeter</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Rectal temperature probe (neonatal)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ringer Lactate</td>
			<td>Bioren Sintetica SA</td>
			<td></td>
			<td>Infusion</td>
		</tr>
		<tr>
			<td>Ropivacain</td>
			<td>Aspen Pharma Schweiz GmbH</td>
			<td>1882249</td>
			<td>Local anesthesia</td>
		</tr>
		<tr>
			<td>Scalpell</td>
			<td>Swann-Morton</td>
			<td>210</td>
			<td></td>
		</tr>
		<tr>
			<td>Small animal shaver</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Soft tissue forceps</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Soft tissue spreader</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Stainless steel sponge bowls</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile micro swabs</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Stethoscope</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Surgery drape</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical scissors</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Syringes 1 mL, 2 mL, and 5 mL</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Tris-Buffer</td>
			<td>Sigma Aldrich</td>
			<td>93302</td>
			<td>Elastase solution</td>
		</tr>
		<tr>
			<td>Vascular clip applicator</td>
			<td>B. Braun, Germany</td>
			<td>FT495T</td>
			<td></td>
		</tr>
		<tr>
			<td>Vein and arterial catheter 22 G</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>vessel loop</td>
			<td></td>
			<td></td>
			<td>Approach</td>
		</tr>
		<tr>
			<td>video camera or smartphone</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Vitarubin</td>
			<td>Streuli Pharma AG</td>
			<td>6847559</td>
			<td></td>
		</tr>
		<tr>
			<td>Yasargil titan standard clip (2x)</td>
			<td>B. Braun Medical AG, Aesculap, Switzerland</td>
			<td>FT242T</td>
			<td></td>
		</tr>
	</tbody>
</table>

creation of two saccular elastase-digested aneurysms with different hemodynamics in one rabbit

具有接近人颅内动脉瘤的血流动力学、形态学和组织学特征的临床前动物模型在理解病理生理过程以及开发和测试新的治疗策略方面起着关键作用。这项研究旨在描述一种新的兔动脉瘤模型，该模型允许在同一动物内创建两个具有不同血流动力学条件的弹性蛋白酶消化的囊状动脉瘤。
五只平均体重为4.0（±0.3）公斤，平均年龄为25（±5）周的雌性新西兰白兔接受了显微外科残端和分叉动脉瘤的形成。一个动脉瘤（残端）是由右颈总动脉（CCA）暴露在其起源头躯干造成的。在CCA原点应用了一个临时夹子，另一个在2厘米以上。用局部注射100U弹性蛋白酶处理该段20分钟。通过将弹性蛋白酶处理的动脉袋缝合到右侧CCA与左侧CCA的端对侧吻合口中，产生了第二个动脉瘤（分叉）。创建后立即通过荧光血管造影控制通畅性。
平均手术时间为221分钟。在同一只动物中产生两个动脉瘤在所有兔子中都是成功的，没有并发症。除一个分叉动脉瘤外，所有动脉瘤在手术后立即未愈合，由于弹性蛋白酶孵育和立即腔内血栓形成，该动脉瘤显示出极端的组织反应。在手术和长达一个月的随访期间未观察到死亡。发病率仅限于一过性前庭综合征（一只兔子），该综合征在一天内自发恢复。
这里首次证明了创建具有残端和分叉血流动力学特征以及高度退化壁条件的双动脉瘤兔模型的可行性。该模型允许在不同流动条件下基于动脉瘤生物学研究自然病程和潜在的治疗策略。

残端和分叉动脉瘤的产生在所有5只新西兰大白兔中均成功，没有术中并发症。在手术期间或24±2天的随访期间未观察到死亡。一只兔子经历了术后并发症，伴有前庭综合征和右侧失明。动物在24小时后完全自发恢复。这种并发症不会干扰其正常活动（自由运动，水和食物摄入，与其他动物的相互作用），并且不需要任何特殊治疗。没有自发性动脉瘤破裂。
平均手术时间为221分...

Watch this Scientific Journal Video about Creation of Two Saccular Elastase-Digested Aneurysms with Different Hemodynamics in One Rabbit at JoVE.com

Creation of Two Saccular Elastase-Digested Aneurysms with Different Hemodynamics in One Rabbit

该协议描述了创建具有两个具有不同血流动力学（残端和分叉星座）的弹性蛋白酶消化动脉瘤的兔子模型的步骤。这允许在单个动物内具有不同血管结构和血流动力学状况的动脉瘤中测试新型血管内装置。

在一只兔中产生两个具有不同血流动力学的囊状弹性蛋白酶消化动脉瘤

Preclinical animal models with hemodynamic, morphologic, and histologic characteristics close to human intracranial aneurysms play a key role in the ...

我们的协议描述了一种可重复的技术，以创建兔动脉瘤模型，该模型在单个动物中以及在单个手术方法中具有残端和分叉血流动力学以及退化壁状况，具有低发病率和死亡率以及高通畅率。该模型允许直接比较囊状动脉瘤的自然病程与血流动力学的作用，以及在两种不同的动脉瘤配置和流动条件下测试血管内装置。这种方法的主要缺点仍然与创建经典分叉和残端动脉瘤模型相同，需要复杂的实验室设备和特定的显微外科技能。使用手术刀从舌骨到胸骨尾部 1.5 厘米处做一个正中皮肤切口。在进行细致止血的同时，从内侧切口准备皮下和脂肪组织。将胸头肌从粘附结缔组织中释放出来，局部应用利多卡因以避免肌阵挛。暴露胸头肌内侧的右颈总动脉或CCA，并用湿拭子保持湿润。准备胸头肌的外侧和近端，并用血管环将其内侧缩回以暴露 CCA。识别颈外静脉，并用湿的微型拭子保护它。沿着CCA仔细解剖结缔组织，直到头臂干分叉以暴露动脉。如果小分支从动脉中冒出，请用烧灼器凝固它们。应用两个临时夹子，第一个夹子在CCA的原点，第二个在CCA的远端两厘米处。将橡胶垫放在容器下方，并用盐酸罂粟碱冲洗以舒张血管。使用显微剪刀小心地取出外膜。使用 22 号四导管在远端夹下方进行动脉切开术，并将导管尾部插入近端夹。用肝素氯化钠腔内冲洗节段，直到没有可见的血液。然后用 4-0 结扎固定导管。通过导管，将0.1至0.2毫升弹性蛋白酶注入动脉段，并孵育20分钟。用弹性蛋白酶孵育20分钟后，清除弹性蛋白酶溶液，并更换注射器用0.9%盐水冲洗动脉段约10次。应用两个结扎，第一个在近端夹子远端 5 毫米处，第二个在动脉切开术下方的近端。在第一个结扎上方约三毫米处切开血管，在第二个结扎和远端夹之间再切一次。将这种自体移植物保持在肝素化溶液中，直到产生分叉动脉瘤。小心地打开第一个近端夹，并测量动脉瘤。要形成分叉动脉瘤，请小心地切除外膜，并在右侧CCA的残端横向做一个约两毫米的纵向切口。在左侧CCA上应用两个临时夹子以划定约一厘米的段，并去除中间的外膜。用 23 号针头进行动脉切开术。然后，用肝素化氯化钠冲洗该段。使用显微剪刀将动脉切开术扩大到约四到五毫米，以便缝合正确的CCA和动脉瘤袋。要进行吻合术，从左侧CCA动脉切开术的近端边缘开始，用五针缝合右颈动脉钝器的近端后壁。然后，从左侧CCA动脉切开术的远端边缘开始，用四到五针缝合动脉瘤袋的背面。继续在鱼口切口水平处的远端背部与动脉瘤移植物的垂直背面缝合三针。用三针缝合鱼嘴切口的前侧，从向上开始向下移动。完成左CCA和动脉瘤移植物正面之间的前缝合线，右CCA缝合约六针。在完成吻合术之前，用肝素化的0.9%盐水溶液腔内冲洗血管。取下右侧CCA上的夹子，同时用微型拭子对吻合口施加一些压力以进行止血。然后，继续从左侧 CCA 中移除远端夹。如果没有大出血，继续取出左侧CCA上的近端夹子以允许血液流动。随访时，通过磁共振血管造影证实动脉瘤通畅。显示了通过聚焦颈部动脉的三特斯拉 MRI 获得的三维 TOF 序列。残端动脉瘤在右锁骨下动脉上观察到，分叉动脉瘤在左锁骨下动脉吻合右CCA产生的分叉处观察到。组织提取后的肉眼检查也证实了动脉瘤通畅性。夹子上的大凹槽表示 1 毫米，小凹槽表示 0.5 毫米。用苏木精-伊红染色样品后，对残端和分叉动脉瘤进行显微镜分析。手术过程中有两个步骤至关重要，首先是右颈动脉的暴露和解剖，直到头臂干，其次是保护气管、颈静脉和喉神经等重要结构。

Research

JoVE Journal

Medicine

Creation of Reversible Cholestatic Rat Model

可逆的胆汁淤积大鼠模型的建立

Cholestasis is a clinical condition commonly encountered by both surgeons and gastroenterologists. Cholestasis can cause various physiological changes and ...

科研

医学

Microsurgical Venous Pouch Arterial-Bifurcation Aneurysms in the Rabbit Model: Technical Aspects

显微静脉袋分岔动脉瘤在兔模型：技术问题

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials ...

Isolation, Characterization and Comparative Differentiation of Human Dental Pulp Stem Cells Derived from Permanent Teeth by Using Two Different Methods

人牙髓干细胞的分离，鉴定和比较性恒牙使用两种不同的方法，得出

Developing wisdom teeth are easy-accessible source of stem cells during the adulthood which could be obtained by routine orthodontic treatments. Human ...

Non-invasive Optical Measurement of Cerebral Metabolism and Hemodynamics in Infants

婴幼儿的脑代谢和血流动力学的非侵入式光学测量

Perinatal brain injury remains a significant cause of infant mortality  and morbidity, but there is not yet an effective bedside tool that can  accurately ...

Ascending Aortic Constriction in Rats for Creation of Pressure Overload Cardiac Hypertrophy Model

大鼠建立压力超负荷心肌肥厚模型在升主动脉缩窄

Ascending aortic constriction is the most common and successful surgical model for creating pressure overload induced cardiac hypertrophy and heart ...

Creation of Abdominal Adhesions in Mice

小鼠腹腔粘连的创建

Abdominal adhesions consist of fibrotic tissue that forms in the peritoneal space in response to an inflammatory insult, typically surgery or ...

Treatment with Vancomycin Loaded Calcium Sulphate and Autogenous Bone in an Improved Rabbit Model of Bone Infection

万古霉素滴入硫酸钙和自体骨治疗兔骨感染的改进模型

Bone infection results from bacterial invasion, which is extremely difficult to treat in clinical, orthopedic, and traumatic surgery. The bone infection ...

Invasive Hemodynamic Monitoring of Aortic and Pulmonary Artery Hemodynamics in a Large Animal Model of ARDS

ards 大型动物模型主动脉和肺动脉血流动力学的侵袭血流动力学监测

One of the leading causes of morbidity and mortality in patients with heart failure is right ventricular (RV) dysfunction, especially if it is due to ...

Creation of Colonic Anastomosis in Mice

小鼠结肠吻合术的建立

Intestinal anastomoses are commonly performed in both elective and emergent operations. Even so, anastomotic leaks are a highly feared complications of ...

One-anastomosis Gastric Bypass (OAGB) in Rats

One-anastomosis Gastric Bypass OAGB in Rats

大鼠胃旁路 (oagb) 的一次吻合

The goal of this protocol is to set up a preclinical model of bariatric surgery and, more specifically, OAGB in obese rats. Based on this preclinical ...