兔动脉袋显微外科分叉动脉瘤模型
1Department of Neurosurgery,Kantonsspital Aarau, 2Cerebrovascular Research Group, Department for BioMedical Research,University of Bern, 3Division of Neuroradiology, Department of Radiology,Kantonsspital Aarau, 4Department of Neurosurgery, Neurocenter and Regenerative Neuroscience Cluster,Inselspital, Bern University Hospital, University of Bern, 5Department for Biomedical Research, Faculty of Medicine,University of Bern
Summary
开发和测试用于颅内动脉瘤治疗的血管内装置仍然非常重要。今天使用的大多数动脉瘤模型都忽略了动脉退行性壁的重要特征或真正分叉的血液动力学。因此，我们设计了一种新型的兔动脉袋分岔模型。
Abstract
颅内动脉瘤的血管内治疗在过去几十年中变得越来越重要，因此对测试血管内装置的需求增加。高度有必要使用符合流变学、血流动力学和动脉瘤壁条件的动物模型。因此，本研究的目的是设计一种新的标准化和可重复的手术技术，以在兔子中产生具有未修饰和修饰壁条件的自体动脉袋分叉动脉瘤。
分叉动脉瘤是通过左颈总动脉右侧吻合术产生的，两者都是显微外科缝合的动脉袋的母动脉。从右颈总动脉近端取出移植物，用于对照组（n = 7，立即自体再植入）或修饰组（n = 7，与100个国际单位弹性蛋白酶一起孵育20分钟，自体再植入）组。创建后立即通过荧光血管造影控制袋和母动脉通畅性。在随访（28天）时，所有兔子都接受了造影剂增强磁共振血管造影和荧光血管造影，然后进行动脉瘤收获，宏观和组织学评估。
共有16只雌性新西兰白兔作。两只动物过早死亡。在随访时，85.72%的动脉瘤仍然是专利。两组都显示动脉瘤大小随着时间的推移而增加;对照组（创建时为6.48±1.81 mm3 ，随访时为19.85±6.40 mm3 ，p = 0.037），这比修饰组（创建时为8.03±1.08 mm3 ，随访时为20.29±6.16 mm3 ）更为明显，p = 0.054）。
我们的研究结果表明，这种新的兔子模型是充分的，它允许在显微外科方法中创建具有不同壁条件的分叉动脉瘤。鉴于良好的长期通畅性和动脉瘤随时间推移生长的特性，该模型可作为新型血管内疗法临床前评估的重要工具。
Introduction
颅内动脉瘤 （IA） 破裂引起的蛛网膜下腔出血可通过血管内或显微外科闭塞技术有效控制1，2，3，4。不同的血管内疗法，克服盘绕后IA复发的主要局限性，在过去几十年中变得越来越重要，导致对测试血管内装置的需求增加。为了测试这些新的治疗方法，高度需要尊重流变学特性，血流动力学和动脉瘤壁条件的适当动物模型5，6，7。在这种情况下，临床和临床前研究已经揭示了动脉瘤壁疾病对动脉瘤破裂和闭塞后复发的重要作用，特别关注壁细胞的丧失7，8，9。
到目前为止，兔子的实验性动脉瘤通常是通过弹性蛋白酶孵育的颈总动脉（CCA）残端或缝合成人工CCA分叉的静脉袋产生的。10，11，12，13，14，15，16 因此，从未描述过真正的动脉袋分叉模型。
本研究的目的是设计一种安全，快速和标准化的技术，用于在兔子模型中创建具有不同壁条件的分叉动脉瘤的显微外科手术（图1）。这是通过将未修饰和修饰的动脉袋缝合到两个CCA的人工分叉中来实现的。
Protocol
所有兽医护理均按照机构指南进行（所有实验均由瑞士伯尔尼州动物护理地方委员会批准（BE 108/16）），并在董事会认证的兽医麻醉师的监督下进行。17，18严格遵守了ARR指南和3R原则。 注意：将所有动物饲养在 22\u201224 摄氏度 （°C） 的室温下，并保持 12 小时 （h） 的亮/暗循环。每次都提供免费的水，颗粒和随意的干草饮食。使?…
Representative Results
在由七只动物组成的试点系列之后，共有16只动物被纳入实验方案。两只动物过早死亡，因此被排除在最终分析之外（死亡率为12.5%）。在14只动物身上计算，对照组和修饰组在荧光血管造影过程中的即时动脉瘤通畅率为71.43%。四个动脉瘤必须通过连续的血栓清宫重新打开，并且在重复荧光血管造影后，所有病例都有记录的通畅性（100%）。MR和荧光血管造影组的动脉瘤通畅率为85.72%，28天后随访时?…
Discussion
我们的研究证明了在兔子中建立具有不同壁条件的真正分叉动脉瘤模型的可行性。总体而言，该研究纳入了14只平均体重为3.7±0.09 kg，平均年龄为112±3天的新西兰大白兔。85.72%的动脉瘤在28天的随访期间保持专利。两只动物过早死亡（死亡率为12.5%）。
先前的研究建议使用多种颅外动脉瘤模型来分析血管内动脉瘤治疗的管理25，26…
Declarações
The authors have nothing to disclose.
Acknowledgements
作者感谢Olgica Beslac和Kay Nettelbeck在围手术期的出色支持和技术援助，以及Alessandra Bergadano，DVM，PhD，PhD对长期动物健康的专门监督。
Materials
| 3-0 resorbable suture | Ethicon Inc., USA | VCP428G | |
| 4-0 non-absorbable suture | B. Braun, Germany | G0762563 | |
| 6-0 non-absorbable suture | B. Braun, Germany | C0766070 | |
| 9-0 non-absorbable suture | B. Braun, Germany | G1111140 | |
| Adrenaline | Amino AG | 1445419 | any generic |
| Amiodarone | Helvepharm AG | 5078567 | any generic |
| Anesthesia machine | Dräger | any other | |
| Aspirin | Sanofi-Aventis (Suisse) SA | 622693 | any generic |
| Atropine | Labatec Pharma SA | 6577083 | any generic |
| Bandpass filter blue | Thorlabs | FD1B | any other |
| Bandpass filter green | Thorlabs | FGV9 | any other |
| Bipolar forceps | any other | ||
| Bicycle spotlight | any other | ||
| Biemer vessel clip (2 x) | B. Braun Medical AG, Aesculap, Switzerland | FD560R | temporary |
| Bispectral index (neonatal) | any other | ||
| Blood pressure cuff (neonatal) | any other | ||
| Clamoxyl | GlaxoSmithKline AG | 758808 | any generic |
| Dexmedetomidine | Ever Pharma | 136740-1 | any generic |
| Electrocardiogram electrodes | any other | ||
| Elastase | Sigma Aldrich | 45125 | any generic |
| Ephedrine | Amino AG | 1435734 | any generic |
| Esmolol | OrPha Swiss GmbH | 3284044 | any generic |
| Fentanyl (intravenous use) | Janssen-Cilag AG | 98683 | any generic |
| Fentanyl (transdermal) | Mepha Pharma AG | 4008286 | any generic |
| Fluoresceine | Curatis AG | 5030376 | any generic |
| Fragmin | Pfizer PFE Switzerland GmbH | 1906725 | any generic |
| Glyco | any generic | ||
| Heating pad | any other | ||
| Isotonic sodium chloride solution (0.9%) | Fresenius KABI | 336769 | any generic |
| Ketamine | Pfizer | 342261 | any generic |
| Laboratory shaker | Stuart | SRT6 | any other |
| Lidocaine | Streuli Pharma AG | 747466 | any generic |
| Longuettes | any other | ||
| Metacam | Boehringer Ingelheim | P7626406 | any generic |
| Methadone | Streuli Pharma AG | 1084546 | any generic |
| Microtubes | any other | ||
| Micro needle holder | any other | ||
| Midazolam | Accord Healthcare AG | 7752484 | any generic |
| Needle holder | any other | ||
| O2-Face mask | any other | ||
| Operation microscope | Wild Heerbrugg | any other | |
| Papaverine | Bichsel | any generic | |
| Prilocaine-lidocaine creme | Emla | any generic | |
| Propofol | B. Braun Medical AG, Switzerland | any generic | |
| Pulse oxymeter | any generic | ||
| Rectal temperature probe (neonatal) | any other | ||
| Ropivacaine | Aspen Pharma Schweiz GmbH | 1882249 | any generic |
| Scalpell | Swann-Morton | 210 | any other |
| Small animal shaver | any other | ||
| Smartphone | any other | ||
| Soft tissue forceps | any other | ||
| Soft tissue spreader | any other | ||
| Stainless steel sponge bowls | any other | ||
| Sterile micro swabs | any other | ||
| Stethoscope | any other | ||
| Straight and curved micro-forceps | any other | ||
| Straight and curved micro-scissors | any other | ||
| Straight and curved forceps | any other | ||
| Surgery drape | any other | ||
| Surgical scissors | any other | ||
| Syringes 1 ml, 2ml and 5 ml | any other | ||
| Tris-Buffer | Sigma Aldrich | 93302 | any generic |
| Vascular clip applicator | B. Braun, Germany | FT495T | |
| Vein and arterial catheter 22 G | any generic | ||
| Vitarubin | Streuli Pharma AG | 6847559 | any generic |
| Yasargil titan standard clip (2 x) | B. Braun Medical AG, Aesculap, Switzerland | FT242T | temporary |
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Wanderer, S., Waltenspuel, C., Grüter, B. E., Strange, F., Sivanrupan, S., Remonda, L., Widmer, H. R., Casoni, D., Andereggen, L., Fandino, J., Marbacher, S. Arterial Pouch Microsurgical Bifurcation Aneurysm Model in the Rabbit. J. Vis. Exp. (159), e61157, doi:10.3791/61157 (2020).