This study was supported by a grant from the Dutch Kidney Foundation (KJPB 08.0003). 
Carolien Rothuizen is acknowledged for her contribution to the study. Hoang Pham is acknowledged for his assistance with the pathology work-up.

所有实验均通过在莱顿大学医学中心的动物福利委员会的批准。 1.动物的制备和麻醉<ol><li>麻醉在充满3-4％异氟醚持续麻醉诱导室鼠标（1-3个月）。 </li><li>剃颈部和使用电剃刀左大腿的内侧部分的腹侧和用一块胶带来去除毛发。 </li><li>上双眼眼应用软膏。 </li><li>在其上鼻子通气面罩固定在加热毯动物的位置。固定头部用胶带的鼻罩。 <ol><li>使用富氧空气集在下面的流在1.5-2.0％的浓度递送异氟烷:空气0.3升/分钟，100％氧气0.2升/分。捏脚趾之间的皮肤，以评估麻醉深度，并根据需要调整异氟醚的浓度。 </li></ol></li><li>固定的动物的加热毯用胶带上是在仰卧位置设定在约37℃的四肢。 </li><li>注入丁丙诺啡溶解在氯化钠（NaCl）0.9％，0.1毫克的剂量/在动物的侧翼0.5毫升无菌的0.9％生理盐水皮下注射一起公斤。 </li><li>申请洗必泰酊0.5％到准备好的区域。 </li></ol> 2.皮肤切口<ol><li>其对外科医生的头在显微镜下动物的位置。 </li><li>使在使用微剪刀腹侧颈部的中线的约1.5厘米的纵向切口。 </li><li>抓住用钳子唾液腺和颅取代正确的唾液腺，直到它部分是伤口之外。 </li></ol> 3.解剖和准备静脉<ol><li>确定并直言不讳地通过散布完全从它的血管周围组织解剖右颈外静脉的分支背内侧只是一个长边和在血管的方向。 </li><li>将一个循环（10.0缝合）周围的解剖静脉和应用一个结没有锁定它。 </li></ol> 4.删除胸锁乳突肌<ol><li>使用镊子，直言沿边境蔓延，从剖析其周围右胸锁乳突肌。将一对开放钳分离肌肉下，用6.0缝线结扎其近端和远端。随后，消费使用cauterizer肌肉。 </li></ol> 5.解剖和准备颈总动脉<ol><li>识别并使用镊子解剖右颈总动脉。放置一个6.0缝合线周围的动脉中的动脉的操作中提供帮助。 </li><li>作为远端和近端尽可能应用血管夹。 </li><li>使在约1mm使用专门微剪刀动脉的中间的纵向切口。 </li><li>冲洗动脉用肝素溶液[100 IU / ml的]，直到容器是明确的血液。 </li><li>测量并调整切口的长度，如果需要将血管夹[1.1 mm]的匹配的宽度。 </li></ol> 6.静脉结扎<ol><li>近端早已准备好的静脉应用血管钳夹，并将静脉周围缝合线。 </li><li>使用被放置在缝合线上和结扎静脉的远端尽可能使用10.0缝线被以前放置在止血剂应用温和尾部牵引到静脉。 </li><li>使用切剪刀刚好接近其结扎静脉。 </li><li>使用血管钳，轻轻打开静脉的腔，并用肝素溶液[100国际单位/毫升]冲洗。 </li></ol> 7.创建接合部1 <ol><li>静脉连接到动脉用2平方海里在十二点钟位置，随后由一个缝合在六点一个间断缝合（10.0）时钟的位置。确保将静脉动脉连接，而不让其绕其自身的轴线。为了提高手术曝光旋转动物。 </li><li>定位静脉周围缝合线，并用止血剂应用温和的横向牵引力。 </li><li>使用相同的间断缝合，通过将大约3-4额外缝线在吻合的可见腹侧完成吻合术。 </li></ol> 8.肝素管理<ol><li>旋转加热毯180°聚焦显微镜左侧大腿。 </li><li>通过寻找，在上腿的内侧部分纵向运行，并且可以通过皮肤可见血管结构识别股静脉/动脉/神经束。使用的约1cm直接在长度方式股​​静脉上方的微剪刀切开。 </li><li>小心解剖股静脉的血管周围组织和在剂量Ô注入肝素f在股静脉0.2国际单位/克体重。 </li></ol> 9.创建接合部2 <ol><li>返回到颈部区域，并删除被置于静脉周围的缝合线。 </li><li>放置缝合线（6.0），该随后通过颈动脉的下方，在静脉和再次低于颈动脉。 （ 图1L） </li><li>取下静脉血管钳。 </li><li>接着，通过同时加捻两个血管夹拧半完成的动静脉瘘沿着以顺时针方式颈动脉的轴线180°，并应用内侧牵引到缝合线的端部。 </li><li>如在步骤7.2中描述完成以同样的方式的吻合术。 </li></ol> 10.血管钳去除<ol><li>取出缝合线和旋转血管夹180°逆时针。 </li><li>移除远侧血管钳接着近端血管夹。 </li> &lt;LI&gt;轻轻阻塞与血管钳静脉流出道评估吻合的通畅。在专利吻合的情况下，预封闭的静脉部分将在一个脉动方式展开。重新定位合适的唾液腺到原来的解剖位置使用镊子。 </li></ol> 11.皮肤缝合和术后护理<ol><li>关闭上腿的皮肤与一个不间断缝合（6.0）。 </li><li>关闭颈部皮肤用不间断缝合（6.0）。 </li><li>从加热毯取出的动物，并注入0.5毫升0.9％生理盐水皮下注射。 </li><li>将动物在黑暗的笼子里是带加热灯加热，让它完全恢复。当动物没有完全恢复，确保它不从血流动力学的冲击，由于在手术区出血受苦。寻找迹象，如颈部肿胀和血液渗漏。 </li><li>后bupr的第一次注射后约6小时enorphine，注入的剂量是由制造商推荐，以便为附加的72小时提供足够的镇痛丁丙诺啡皮下的持续释放制剂的单次剂量。 </li></ol> 12.组织收获<ol><li>麻醉动物用含有咪达唑仑（5毫克/千克）腹腔内给药的麻醉剂混合物，美托咪定（0.5毫克/千克）和芬太尼（0.05毫克/千克）。 </li><li>通过插入通过其前爪和脸颊针头进入非加热的硅垫固定在仰卧位置的动物。 </li><li>使约1.5-2.0厘米的切口在与所述微剪刀颈部疤痕。 </li><li>剖析AVF，并把缝合线（6.0）围绕proximal-，远端动脉和静脉流出道，便于识别。 </li><li>作为该协议的第10.3节描述评估AVF的通畅。 </li><li>通过执行剖腹探查内侧打开ü腹腔唱微型剪刀。 <ol><li>通过使用一对在锁骨中线尾端开始剪刀的左和右腹侧胸廓切开。用微型剪刀，横切附加到腹侧肋骨的中间部分，并随后置换本条颅使用镊子隔膜。 </li><li>找到下腔静脉和用微剪横切它。 </li></ol></li><li>插入针23号针头在左心室和在约100毫米汞柱的压力下，用PBS灌注，直至血管内流体是近清楚。 </li><li>不除去针，现在用4％福尔马林在约100毫米汞柱的压力灌注10分钟。 </li><li>通过与微剪刀横切两个动脉和静脉切除AVF并在4％的福尔马林溶液O / N淹没它。 </li></ol> 13.组织包埋和剖分<ol><li>处理该组织根据制造商的protoco石蜡湖该协议包括施加如下: <ol><li>浸入乙醇70％，在室温1小时。重复2倍。 </li><li>浸入乙醇96％，在室温1小时。重复2倍。 </li><li>浸入乙醇99.5％在室温1小时。重复一次。 </li><li>浸入乙醇99.5％在室温1.5小时。 </li><li>在二甲苯浸入100％，在室温1小时。重复一次。 </li><li>在62℃浸入石蜡1小时。重复2倍。 </li></ol></li><li>嵌入AVF石蜡使得静脉流出道在垂直于包埋盒定向。 </li><li>用切片机，创建12个连续切片每一个5微米的厚度由30节。然后每个串行部分，放置在从一个位置以有序的方式开始多达十二开始最接近吻合的区域的单个安装玻璃一个部分。 </li></ol>

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作者什么都没有透露。

该AVF被认为是致命的血液透析治疗脚跟。不幸的是，AVF仍存有大量的失败8-10受到影响。尽管在底层机制的广泛研究，确切的病理生理机制仍然不明。为AVF失败众多鼠模型已在文献2,3,11,12说明。然而，这些模型的掺入静脉结束这一最在临床情况中使用动脉侧吻合配置。这是非常重要，因为将得到的血流动力学起着血管重塑中起重要作用。此外，一些模型中使用的合成的袖口的动脉和静脉2,11，这是不在临床上所使用的连接。 为提高临床相关性，因此，我们开发了在其中的枸骨的一个分支之间建立了一个单边静脉端侧动脉吻合的小鼠模型拉尔静脉和普通颈动脉间断缝合6。在这个模型中关键的组织形态学变化进行观察，包括向外重塑和进步内膜增生，最终导致失败AVF。 手术过程中的一个重要方面是麻醉协议。它建议使用异氟醚吸入麻醉下，因为这是用于获得麻醉的长时间的安全和容易的方法。后者是为训练阶段，其中，整个过程可能需要长达3小时尤其重要。 为了创建用于动静脉瘘的横移流出静脉必要室，同​​侧胸锁乳突肌需要被切除。 手术期间相对于该容器的处理，血管解剖应以钝方式使用镊子进行。使用专门的血管钳直为Recommended用于直接处理和船只的操纵，因为它们提供更精确和产生较少的机械损坏，由于圆尖。缝合线的使用作为容器循环用止血组合可以在安全的组织处理辅助，而且，在作为第三手这一点至关重要，以便该过程可以在没有任何直接的帮助下进行使用时，以最佳的方式呈现所述组织。 毫无疑问，在过程中最困难的一步是动脉和静脉之间的缝合展示位置。必须小心不要缝合吻合术的"背面" - 侧与"前" - 侧，因为这会导致吻合，可导致所述AVF的早期失效的狭窄。为了防止这种情况的发生，位置，以便将它们分开的容器的两个壁之间的血管钳的尖端。的技术难度可以被认为是该模型的一个限制。但是，我们并不认为日在此过程中比是目前最广泛使用的3,6-模型更具挑战性。 这很难使对未来研究的样本大小一般说明，作为所计算的样本大小并不仅仅依赖于动物之间也对干预的"强度"的可变性。我们估计约10只小鼠（不包括退出因为AVF的技术故障的研究的小鼠）的一组的大小应足以研究，着眼于在AVF失败的特定基因的作用。最近，我们能够获得与在弹力鼠AVF重塑13的作用的研究10只动物的平均组大小显著的成果。 我们的小鼠模型的一个方面，需要进一步讨论。已知的是在患者的终末期肾病的尿毒症环境有助于血管疾病的多种包括静脉内膜hyperplasi一个连前血液透析通路手术14-16。目前这个模型不包括尿毒症环境。因此，慢性肾功能衰竭17的一个模型的掺入将是有价值的添加剂步骤，以进一步改善此小鼠模型的有效性。

一个功能血管通路管道是对于依赖于长期血液透析维持生命肾功能衰竭患者至关重要。一个动静脉瘘（AVF）的建设是目前血管通路的首选。然而，AVF相关并发症构成发病的一个主要原因患者慢性血液透析。尽管进行了广泛的科学努力，没有新颖的方法，以减少AVF访问相关的并发症，并导致AVF耐用性显着改善。这一令人失望的进步的一部分涉及到血液透析访问失败的基本病理生理学的理解不够充分。 解开的AV访问失败，动物模型密切模仿人类病理学的病理生理学是极其重要的。在这方面，不但动物种类，但也吻合部位，所需的抗混凝疗法和的持续时间浪涌后跟进RY应考虑到1。虽然大型动物是最适合的目的是开发新的治疗策略的干预研究，小鼠模型已经获得在底层的AV接入失败的分子机制，更深入的了解，由于转基因小鼠的可用性的最大潜力。此外，可用于这一目的在相比更大的动物使用较低成本的大量小鼠。 AVF失败的第一鼠模型是由桂和等 2在该模型中在2004年描述的，使用的颈动脉，并使用血管内导管的端至端的方式颈静脉构建动静脉瘘。虽然端至端的配置和血管内导管的存在限制这种模型为人类动静脉瘘的有效性这种模式可能是研究在动静脉瘘的早期静脉适应有用。一种改进AVF模型由Castier和等人介绍了3，其中的端部颈动脉被连接到颈静脉的一侧。然而，在血液透析患者动静脉瘘通常由解剖静脉的端部到动脉的侧面构成。 AVF的确切配置是AV访问模型的一个重要特征，因为它决定了导管4内的血流动力学轮廓。后者是内皮功能障碍和内膜增生（IH）5的后续发展的重要因素。 一种新的小鼠模型最近被用相同的解剖形态发展为利用人类6。在这个模型中，AVF在C57BL / 6小鼠吻合颈外静脉的分支的末端间断缝合颈总动脉的一侧。在本文中，我们专注于这个模型的显微外科方法，以便于广泛使用这种小鼠模型，旨在解开复杂的病理生理机制血液透析访问失败。

<table><tbody><tr><td>Dissecting microsocpe</td><td>Leica</td><td>M80</td><td></td></tr><tr><td>Forceps</td><td>Medicon</td><td>07.61.25</td><td></td></tr><tr><td>Vascular forceps</td><td>S&amp;amp;T</td><td>JFL-3D.2</td><td></td></tr><tr><td>Vascular forceps</td><td>S&amp;amp;T</td><td>D-5a.2</td><td></td></tr><tr><td>Forceps</td><td>Roboz</td><td>SS/45</td><td></td></tr><tr><td>Micro scissor 5 mm blade</td><td>Fine science tools</td><td>15000-08</td><td></td></tr><tr><td>Micro scissor 2 mm blade</td><td>Fine science tools</td><td>15000-03</td><td></td></tr><tr><td>Scissor</td><td>Medicon</td><td>05.12.21</td><td></td></tr><tr><td>Clip applier 1</td><td>S&amp;amp;T</td><td>CAF-4</td><td></td></tr><tr><td>Vascular clamp 1</td><td>S&amp;amp;T</td><td>B-1V</td><td></td></tr><tr><td>Clip applier 2</td><td>BBraun</td><td>FE572K</td><td></td></tr><tr><td>Vascular clamp 2</td><td>BBraun</td><td>FE740K</td><td></td></tr><tr><td>Hemostatic forceps</td><td>BBraun</td><td>BH110</td><td></td></tr><tr><td>10.0 sutures</td><td>BBraun</td><td>G1117041</td><td></td></tr><tr><td>6.0 sutures</td><td>BBraun</td><td>768464</td><td></td></tr><tr><td>Cauterizer</td><td>Fine science tools</td><td>18010-00</td><td></td></tr><tr><td>Needle holder</td><td>Medicon</td><td>11.82.18</td><td></td></tr><tr><td>Ocular ointment</td><td>Pharmachemie</td><td>41821101</td><td></td></tr><tr><td>Chlorhexidine tincture 0,5%</td><td>Leiden University Medical Center</td><td>NA</td><td></td></tr><tr><td>Heparin</td><td>Leo Pharma</td><td>012866-08</td><td></td></tr><tr><td>Buprenorphin</td><td>RB Pharmaceuticals&amp;nbsp;</td><td>283732</td><td></td></tr><tr><td>Isoflurane</td><td>Pharmachemie</td><td>45,112,110</td><td></td></tr><tr><td>Anesthesia mask</td><td>Maastricht university</td><td>custom made</td><td></td></tr><tr><td>Midazolam</td><td>Actavis</td><td>AAAC6877</td><td></td></tr><tr><td>Dexmedetomidine</td><td>Orion</td><td>141-267</td><td></td></tr><tr><td>Fentanyl</td><td>Bipharma</td><td>15923002</td><td></td></tr><tr><td>Continuous anaesthetic induction chamber</td><td>Vet-tech solutions</td><td>AN010R</td><td></td></tr></tbody></table>

a novel murine model of arteriovenous fistula failure: the surgical procedure in detail

The arteriovenous fistula (AVF) still suffers from a high number of failures caused by insufficient remodeling and intimal hyperplasia from which the exact pathophysiology remains unknown. In order to unravel the pathophysiology a murine model of AVF-failure was developed in which the configuration of the anastomosis resembles the preferred situation in the clinical setting. A model was described in which an AVF is created by connecting the venous end of the branch of the external jugular vein to the side of the common carotid artery using interrupted sutures. At a histological level, we observed progressive stenotic intimal lesions in the venous outflow tract that is also seen in failed human AVFs. Although this procedure can be technically challenging due to the small dimensions of the animal, we were able to achieve a surgical success rate of 97% after sufficient training. The key advantage of a murine model is the availability of transgenic animals. In view of the different proposed mechanisms that are responsible for AVF failure, disabling genes that might play a role in vascular remodeling can help us to unravel the complex pathophysiology of AVF failure.

创建吻合（图1）后，通畅应由不久包藏有血管钳静脉流出道进行评估。当吻合是专利，血管道近端闭塞应明确扩大以脉动方式。此外，通畅是通过使用近红外荧光检查（NIRF），该有效地作为一个血管造影（ 图2）确认。在手术过程的故障会导致吻合中（图2）中描绘的闭塞。这种故障可通过太窄吻合区域，容器的扭转，肝素剂量不足或吻合的前侧连接到背面侧的意外缝线放置引起的。 在组织学水平，血管重塑在AVF的方法可以优雅使用这一模型中研究。在AVF血管重塑的发生是由于的结果增加血流量和压力。在小鼠中，该反应的特征在于在圆周从而增加在管腔面积在第2周手术后的增加（例如，向外重塑）。这些2周后，管腔面积逐渐减小由于在向外重构和内膜的正在进行的增厚停止。这些进步狭窄病变的形成术后导致的AVF 50％遮挡，在4个星期。因此，最佳的时间点收获动静脉瘘会在术后2周，因为在此阶段，在专利AVF血管响应的适当的分析仍然是可行的。手术后2周静脉流出道的免疫染色表明内膜的细胞区室主要由α-平滑肌肌动蛋白（α-SMA）阳性细胞（图3）的，如在失败的人的动静脉瘘，以及7观察。 鉴于的复杂性的<html>显微方法，这是现实的，在小鼠的比例程序的技术故障估计。在我们手中，该过程的成功率是在开始的67％。然而，随着进一步训练这个速度增加至97％。失败的主要原因是出血（60％），其次是急性血栓形成（27％）和anesthesia-相关死亡（13％）。足够的培训之后，在外科手术可以在大约1小时来进行。 <img alt="图1" src="/files/ftp_upload/53294/53294fig1.jpg" /> 图1.手术过程的详细方案（A - T）。成功建立一个AVF的关键步骤。 <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/53294/53294fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 重2"SRC ="/文件/ ftp_upload / 53294 / 53294fig2.jpg"/&gt; 图2.专利和闭塞动静脉瘘的宏观照片。（A）专利AVF与远端颈总动脉血管钳。 （B）闭塞AVF。 （白色箭头）表示血流的方向。 （C）的专利使用AVF近红外透视证实。 （红色箭头）指示动脉。 （蓝色箭头）表示静脉流出道。 <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/53294/53294fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/53294/53294fig3.jpg" /> 使用苏木，phloxin和孔雀皇形态概述- 第14天未手术相比对照血管AVF的静脉流出道图3.组织学染色 （B A）。罗恩表示AVF创建后14天明显增加血管周围及内膜增生的发展。 （C - D）的免疫组织化学染色证明大多数细胞呈现内膜增生是α-平滑肌肌动蛋白阳性。 （L）流明; （1H）内膜增生;比例尺:200微米<a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/53294/53294fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about A Novel Murine Model of Arteriovenous Fistula Failure: The Surgical Procedure in Detail at JoVE.com

A Novel Murine Model of Arteriovenous Fistula Failure: The Surgical Procedure in Detail

Here we present a murine model of arteriovenous fistula (AVF) failure in which a clinically relevant anastomotic configuration is incorporated. This model can be used to study the pathophysiology and to test possible therapeutic interventions.

动静脉内瘘失败的新型小鼠模型:在详细介绍手术过程

The arteriovenous fistula (AVF) still suffers from a high number of failures caused by insufficient remodeling and intimal hyperplasia from which the ...

a-novel-murine-model-arteriovenous-fistula-failure-surgical-procedure

Universidad San Sebastian

Research

JoVE Journal

Medicine

13.0K 次观看.  Leiden University Medical Center. Here we present a murine model of arteriovenous fistula (AVF) failure in which a clinically relevant anastomotic configuration is incorporated. This model can be used to study the pathophysiology and to test possible therapeutic interventions.

视频: A Novel Murine Model of Arteriovenous Fistula Failure: The Surgical Procedure in Detail

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

在的鼠标Aortocaval瘘的技术方面

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

科研

医学

Adult Mouse Venous Hypertension Model: Common Carotid Artery to External Jugular Vein Anastomosis.

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model creating an artificial fistula between the common carotid artery (CCA) and the external jugular vein (EJV) has been widely described and proved technically feasible. This construct provokes a consistent cerebral venous hypertension (CVH), and therefore has helped studying the contribution of venous hypertension to formation, clinical symptoms, and prognosis of brain AVMs and dural AVFs. Equivalent mice models have been only scarcely described and have shown trouble with stenosis of the fistula. An established murine model would allow the study of not only pathophysiology but also potential genetic therapies for these cerebrovascular diseases.
We present a model of arteriovenous fistula that produces a durable intracranial venous hypertension in the mouse. Microsurgical anastomosis of the murine CCA and EJV can be difficult due to diminutive anatomy and frequently result in a non-patent fistula. In this step-by-step protocol we address all the important challenges encountered during this procedure. Avoiding excessive retraction of the vein during the exposure, using 11-0 sutures instead of 10-0, and making a carefully planned end-to-side anastomosis are some of the critical steps. Although this method requires advanced microsurgical skills and a longer learning curve that the equivalent in the rat, it can be consistently developed.
This novel model has been designed to integrate transgenic mouse techniques with a previously well-established experimental system that has proved useful to study brain AVMs and dural AVFs. By opening the possibility of using transgenic mice, a broader spectrum of valid models can be achieved and genetic treatments can also be tested. The experimental construct could also be further adapted to the study of other cerebrovascular diseases related with venous hypertension such as migraine, transient global amnesia, transient monocular blindness, etc.

We describe a method for creating a reliable model of cerebral venous hypertension in the adult mouse. This model has been widely described and tested in the rat. This new counterpart in the mice opens the possibility of using genetic modified animals and thereby broadens the applications of the model.

成年小鼠静脉高压型号：颈总动脉，以颈外静脉吻合。

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model ...

A New Murine Model of Endovascular Aortic Aneurysm Repair

Endovascular aneurysm exclusion is a validated technique to prevent aneurysm rupture. Long-term results highlight technique limitations and new aspects of Abdominal aortic aneurysm (AAA) pathophysiology. There is no abdominal aortic aneurysm endograft exclusion model cheap and reproducible, which would allow deep investigations of AAA before and after treatment. We hereby describe how to induce, and then to exclude with a covered coronary stentgraft an abdominal aortic aneurysm in a rat. The well known elastase induced AAA model was first reported in 19901 in a rat, then described in mice2. Elastin degradation leads to dilation of the aorta with inflammatory infiltration of the abdominal wall and intra luminal thrombus, matching with human AAA. Endovascular exclusion with small covered stentgraft is then performed, excluding any interactions between circulating blood and the aneurysm thrombus. Appropriate exclusion and stentgraft patency is confirmed before euthanasia by an angiography thought the left carotid artery. Partial control of elastase diffusion makes aneurysm shape different for each animal. It is difficult to create an aneurysm, which will allow an appropriate length of aorta below the aneurysm for an easy stentgraft introduction, and with adequate proximal and distal neck to prevent endoleaks. Lots of failure can result to stentgraft introduction which sometimes lead to aorta tear with pain and troubles to stitch it, and endothelial damage with post op aorta thrombosis. Giving aspirin to rats before stentgraft implantation decreases failure rate without major hemorrhage. Clamping time activates neutrophils, endothelium and platelets, and may interfere with biological analysis.

Histological and biochemical modifications after aneurysm endograft exclusion are still unclear. We describe a new model of endograft implantation on a murine aneurysm. Through thrombus analysis with and without persistant circulating blood stream interface, and new endovascular biomaterials evaluation, this model will help to better understand endovascular aneurysm exclusion pathobiology.

一种新的小鼠模型主动脉瘤腔内修复

Endovascular aneurysm exclusion is a validated technique to prevent aneurysm rupture. Long-term results highlight technique limitations and new aspects of ...

Murine Model of Femoral Artery Wire Injury with Implantation of a Perivascular Drug Delivery Patch

Percutaneous interventions including balloon angioplasty and stenting have been used to restore blood flow in vessels with occlusive vascular disease. While these therapies lead to the rapid restoration of blood flow, these technologies remain limited by restenosis in the case of bare metal stents and angioplasty, or reduced healing and possibly enhanced risk of thrombosis in the case of drug eluting stents. A key pathophysiological mechanism in the formation of restenosis is intimal hyperplasia caused by the activation of vascular smooth muscle cells and inflammation due to arterial stretch and injury. Surgeries that induce arterial injury in genetically modified mice are useful for the mechanistic study of the vascular response to injury but are often technically challenging to perform in mouse models due to the their small size and lack of appropriate sized devices. We describe two approaches for a surgical technique that induces endothelial denudation and arterial stretch in the femoral artery of mice to produce robust neointimal hyperplasia. The first approach creates an arteriotomy in the muscular branch of the femoral artery to obtain vascular access. Following wire injury this arterial branch is ligated to close the arteriotomy. A second approach creates an arteriotomy in the main femoral artery that is later closed through localized cautery. This method allows for vascular access through a larger vessel and, consequently, provides a less technically demanding procedure that can be used in smaller mice. Following either method of arterial injury, a degradable drug delivery patch can be placed over or around the injured artery to deliver therapeutic agents.

We describe a surgical technique that produces wire injury in the femoral artery of mice to induce neointimal hyperplasia to serve as a model testing system for the perivascular delivery of therapeutic compounds for the inhibition of restenosis.

股动脉线损伤的小鼠模型的血管周围给药补丁植入

Percutaneous interventions including balloon angioplasty and stenting have been used to restore blood flow in vessels with occlusive vascular disease. ...

Intraluminal Drug Delivery to the Mouse Arteriovenous Fistula Endothelium

Delivery of therapeutic agents to enhance arteriovenous fistula (AVF) maturation can be administered either via intraluminal or external routes. The simple murine AVF model was combined with intraluminal administration of drug solution to the venous endothelium at the same time as fistula creation. Technical aspects of this model are discussed. Under general anesthesia, an abdominal incision is made and the aorta and inferior vena cava (IVC) are exposed. The infra-renal aorta and IVC are dissected for clamping. After proximal and distal clamping, the puncture site is exposed and a 25 G needle is used to puncture both walls of the aorta and into the IVC. Immediately after the puncture, a reporter gene-expressing viral vector was infused in the IVC via the same needle, followed by 15 min of incubation. The intraluminal administration method enabled more robust viral gene delivery to the venous endothelium compared to administration by the external route. This novel method of delivery will facilitate studies that explore the role of the endothelium in AVF maturation and enable intraluminal drug delivery at the time of surgical operation.

After puncturing the aorta through the inferior vena cava (IVC) to create an aorto-caval fistula in the mouse, solution containing a drug is infused into the IVC via the same needle, followed by incubation. This method enables more robust drug delivery to the venous endothelium compared to the external route.

腔内药物输送到鼠标动静脉瘘内皮

Delivery of therapeutic agents to enhance arteriovenous fistula (AVF) maturation can be administered either via intraluminal or external routes. The ...

Murine Model of Central Venous Stenosis using Aortocaval Fistula with an Outflow Stenosis

Central venous stenosis is an important entity contributing to arteriovenous fistula (AVF) failure. A murine AVF model was modified to create a partial ligation of the inferior vena cava (IVC) in the outflow of the fistula, mimicking central venous stenosis. Technical aspects of this model are introduced. The aorta and IVC are exposed, following an abdominal incision. The infra-renal aorta and IVC are dissected for proximal clamping, and the distal aorta is exposed for puncture. The IVC at the midpoint between the left renal vein and the aortic bifurcation is carefully dissected to place an 8-0 suture beneath the IVC. After clamping the aorta and IVC, an AVF is created by puncturing the infra-renal aorta through both walls into the IVC with a 25 G needle, followed by ligating a 22 G intra-venous (IV) catheter and IVC together. The catheter is then removed, creating a reproducible venous stenosis without occlusion. The aorta and IVC are unclamped after confirming primary hemostasis. This novel model of central vein stenosis is easy to perform, reproducible, and will facilitate studies on AVF failure.

An aortocaval fistula was created by puncturing the murine infra-renal aorta through both walls into the inferior vena cava and was followed by creation of a stenosis in its outflow via partial ligation of the inferior vena cava. This reproducible model can be used to study central venous stenosis.

中央静脉狭窄病的Murine模型,使用具有流出狭窄病的动脉静脉瘘管病

Central venous stenosis is an important entity contributing to arteriovenous fistula (AVF) failure. A murine AVF model was modified to create a partial ...

Creating Radio-cephalic Arteriovenous Fistula in the Forearm with a Modified No-Touch Technique

Autologous arteriovenous fistula (AVF) is the primary and best option to obtain vascular access for hemodialysis treatment; other options are arteriovenous graft (AVG) and central venous catheterization (CVC). The implementation of radio-cephalic autologous arteriovenous fistula (RC-AVF) in the forearm was preferred among patients with superior vascular conditions. However, there is a high rate of early fistula failure. The chosen surgical method is understood to have an effect on the maturation of the fistula. New surgical procedures such as radial artery deviation and reimplantation (RADAR) have been significantly improved for juxta-anastomotic stenosis. Nevertheless, new problems such as stenosis of arteries and narrowing of surgical indication were also found. In this report, we presented a modified no-touch technique (MNTT) to create an RC-AVF, in which the venous and arterial wall avoid devascularization and the radial artery does not sever.

We present a modified no-touch technique (MNTT) to create a radio-cephalic arteriovenous fistula (RC-AVF) in which the venous and arterial wall avoid devascularization and the radial artery does not sever.

使用改进的非接触技术在前臂中创建放射性头动静脉瘘

Autologous arteriovenous fistula (AVF) is the primary and best option to obtain vascular access for hemodialysis treatment; other options are ...

A Murine Model of Hemodialysis Access-Related Hand Dysfunction

Chronic kidney disease is a major public health problem, and the prevalence of end-stage renal disease (ESRD) requiring chronic renal replacement therapies such as hemodialysis continues to increase. Autogenous arteriovenous fistula (AVF) placement remains a primary vascular access option for ESRD patients. Unfortunately, approximately half of the hemodialysis patients experience dialysis access-related hand dysfunction (ARHD), ranging from subtle paresthesia to digital gangrene. Notably, the underlying biologic drivers responsible for ARHD are poorly understood, and no adequate animal model exists to elucidate the mechanisms and/or develop novel therapeutics for the prevention/treatment of ARHD. Herein, we describe a new mouse model in which an AVF is created between the left common iliac artery and vein, thereby facilitating the assessment of limb pathophysiology. The microsurgery includes vessel isolation, longitudinal venotomy, creation of arteriovenous anastomosis, and venous reconstruction. Sham surgeries include all the critical steps except for AVF creation. Iliac AVF placement results in clinically relevant alterations in central hemodynamics, peripheral ischemia, and impairments in hindlimb neuromotor performance. This novel preclinical AVF model provides a useful platform that recapitulates common neuromotor perturbations reported by hemodialysis patients, allowing researchers to investigate the mechanisms of ARHD pathophysiology and test potential therapeutics.

This protocol details the surgical steps of murine common iliac arteriovenous fistula creation. We developed this model to study hemodialysis access-related limb pathophysiology.

血液透析通路相关手功能障碍的小鼠模型

Chronic kidney disease is a major public health problem, and the prevalence of end-stage renal disease (ESRD) requiring chronic renal replacement ...

A Modified Technique for Arteriovenous Fistula Construction in Rabbits

Juxta-anastomotic stenosis is a challenging problem that often causes non-maturation and decreases the patency of an arteriovenous fistula (AVF). Injury to the veins and arteries during the operation and hemodynamic changes can lead to intimal hyperplasia, leading to juxta-anastomotic stenosis. To reduce injury to the veins and arteries during the operation, this study proposes a new modified no-touch technique (MNTT) for AVF construction that can decrease the rate of juxta-anastomotic stenosis and improve the AVF patency. To unravel the hemodynamic changes and mechanisms of the MNTT, this study presented an AVF procedure using this technique. Although this procedure is technically challenging, 94.4% procedural success was achieved after adequate training. Ultimately, 13 out of 34 rabbits had a functional AVF 4 weeks after the surgery, leading to a 38.2% AVF patency rate. However, at 4 weeks, the survival rate was 86.1%. Ultrasonography showed active blood flow through AVF anastomosis. Furthermore, the spiral laminar flow was observed in the vein and artery near the anastomosis, suggesting that this technique may improve the hemodynamics of the AVF. On histological observation, significant venous intimal hyperplasia was observed at the AVF anastomosis, whereas no significant intimal hyperplasia was observed at the proximal external jugular vein (EJV) of the anastomosis. This technique will improve the understanding of the mechanisms underlying the use of MNTT for AVF construction and provide technical support for the further optimization of the surgical approach in AVF construction.

The present protocol proposes the creation of an arteriovenous fistula in rabbits using a modified no-touch technique. The technique involves the side-to-side anastomosis of the common carotid artery and external jugular vein without the dissection of the perivenous tissues or cutting off the artery.

一种改良的兔动静脉瘘构建技术

Juxta-anastomotic stenosis is a challenging problem that often causes non-maturation and decreases the patency of an arteriovenous fistula (AVF). Injury ...

改良动静脉瘘手术建立 RV 体积超负荷小鼠模型

Establishment and Confirmation of a Postnatal Right Ventricular Volume Overload Mouse Model

Right ventricular (RV) volume overload (VO) is common in children with congenital heart disease. In view of distinct developmental stages,the RV myocardium may respond differently to VO in children compared to adults. The present study aims to establish a postnatal RV VO model in mice using a modified abdominal arteriovenous fistula. To confirm the creation of VO and the following morphological and hemodynamic changes of the RV, abdominal ultrasound, echocardiography, and histochemical staining were performed for 3 months. As a result, the procedure in postnatal mice showed an acceptable survival and fistula success rate. In VO mice, the RV cavity was enlarged with a thickened free wall, and the stroke volume was increased by about 30%-40% within 2 months after surgery. Thereafter, the RV systolic pressure increased, corresponding pulmonary valve regurgitation was observed, and small pulmonary artery remodeling appeared. In conclusion, modified arteriovenous fistula (AVF) surgery is feasible to establish the RV VO model in postnatal mice. Considering the probability of fistula closure and elevated pulmonary artery resistance, abdominal ultrasound and echocardiography must be performed to confirm the model status before application.

作者聚焦:出生后右心室容量超负荷小鼠模型的建立和确认  

This protocol presents the establishment and confirmation of a postnatal right ventricular volume overload (VO) model in mice with abdominal arteriovenous fistula (AVF), which can be applied to investigate how VO contributes to postnatal heart development.