作者感谢克里斯蒂安Pahrmann她的技术援助。这项研究是由德国妇儿基金会资助Herzforschung（F / 28/14）。 DW是由来自国际社会旅游奖心脏和肺移植的支持。 TD从否则-克朗 - 费森尤斯基金会（2012_EKES.04）收到否则克朗卓越助学金。 SS收到研究经费来自德意志研究联合会（DFG; DE2133 / 2-1，TD和SCHR992 / 3- 1，SCHR992 / 4-1，SS）。

动物遵照指南实验动物的原则，通过实验动物资源研究所编制和美国国立卫生研究院公布收到的人文关怀。所有动物方案是由主管地方当局（"献给金额UND GESUNDHEIT Verbraucherschutz，Hansestadt（办公室健康与消费者保护）汉堡"）的批准。 1.动物护理<ol><li>获取Lewis大鼠（LEW / CRL）大鼠和ROSA /荧光素酶LEW转基因大鼠实验动物研究所体重300-350克</li><li>保持常规条件下的老鼠在通风柜和养活他们的标准大鼠饲料和蒸压水随意。 </li><li>执行使用ROSA /荧光素酶LEW转基因大鼠作为捐助者和同系LEW / CRL大鼠作为收件人移植术。 </li></ol> 2.捐助者大鼠的制备<ol><li>使用吸纳离子腔麻醉异氟烷（2.5-3％）的老鼠。 </li><li>放置在它的后面的大鼠和保持与覆盖嘴和鼻子的面罩的麻醉。按捏后脚和验证的情况下反射的检查麻醉足够的深度。涂一些药膏兽医的眼睛，以防止干燥，而在麻醉下。 </li><li>传播后腿和用胶布固定自己的位置。 </li><li>剃腹股沟头发与头发剪，并使用聚维酮碘，随后通过80％乙醇消毒的整个区域。重复消毒一步的两倍。 注:手术区，纱布，及手术器械应灭菌。维持无菌区整个过程和穿一次性使用的，无菌手术手套，口罩，和帽。 </li><li>在显微镜下，进行沿LINEA inguinalis垂直切口。用两个镊子皮下组织轻轻分开，并从其ORI暴露腹壁浅静脉杜松子酒的股静脉。小心地从周围组织中分离出腹壁浅静脉。 </li><li>停止使用两个微夹子固定腹壁浅静脉血流。 </li><li>通过仔细地抬起与镊子分离静脉，并通过容器与microscissors切割收获静脉的大致0.5到1厘米的段。留在容器残端微夹具防止血液流失。放置在无菌纱布取下的一块静脉。小心地将一地下30针收获静脉的一端内侧和冲洗用肝素容器（50单位/ ml）。 注意:小心使用静脉，避免吊装，切割，和冲洗过程中的损坏。确保肝素适量的冲洗移植。 </li><li>保持在冰上在1％利多卡因的血管区段，直到移植到受体鼠，以防止一个容器痉挛。 </li><li>通过增加麻醉5％异氟醚安乐死供体鼠。后2-3分钟，运恩沿着白线腹部，划破膜片，并取出心脏停止流通。 </li></ol> 3.收件人鼠的制备<ol><li>麻醉并固定收件人大鼠以相同的方式作为供体鼠。 </li><li>剃腿的内侧与毛发修剪器，并使用聚维酮碘和80％乙醇消毒三次。 </li><li>监测麻醉深度，并确保它是按捏后脚时验证的情况下的反射的足够了。 </li><li>从膝盖到腹股沟褶执行中值股骨切口。在显微镜下，用2镊子股动脉从周围环境中分离出来。 </li><li>使用微夹钳阻止血液的流动。放置近侧夹紧第一，其次远侧夹子。 </li><li>剪出夹紧股动脉microscissors短段和丢弃。缩短剩下的树桩动脉与microscissors，创造一个差距是1-2毫米比静脉移植大。利用地下30针冲洗动脉残端与肝素。 注:如果外膜伸出略微超出血管断端，使用镊子小幅拉过来的船只将其取出一块。 </li><li>从步骤2.8动脉树桩之间放置收集静脉和调整长度，使其合适地配合到间隙。注静脉的方向。 </li><li>首先用一个10-0普理灵缝合执行近端吻合。在所示（ 图1D）的顺序进行单针。开始与腹侧增加三个缝合线之前的每个侧面缝合。之后，放置在容器的背侧三针来完成吻合术。 </li><li>与使用相同的技术，作为在步骤3.8中描述的近端吻合接枝连接远侧血管。再次，开始在每个横向侧的缝合线，然后将三道缝线上的腹侧的side和背侧。 </li><li>负载两个1毫升注射器用纤维蛋白胶成分1和2小心提起用镊子接枝拖放大约100μl的纤维蛋白胶部件1的接枝下，接着部件2。 注:确保组件1和2在1应用:1的比例。 </li><li>放置接枝背部在其位置，并在移植物的顶部下降一个额外的100μl的组分1和2的。可以肯定的胶水涵盖接枝并且为了防止吻合不全和静脉移植物的过度膨胀的吻合术。 </li><li>小心地打开远端夹紧，随后近端。 </li><li>确认通过检查在移植静脉和移植物的远端动脉可见光脉冲一个成功的手术。 </li><li>除去过多的胶水，阻碍皮肤缝合。使用镊子解除固化胶水和microscissors去除多余的。关闭皮肤层用5-0普理灵缝线。 </li><li>注射4-5米克/公斤卡洛芬皮下注射使鼠醒来了。不要离开无人看管的动物，直到它已经恢复了足够的意识，以保持胸骨斜卧。保持动物在一个单一的笼子里，直到它完全恢复。 </li><li>添加安乃近到饮用水（每100毫升50毫克安乃近）作为止痛药为以下3天，每日监测动物。 </li></ol> 4.多普勒超声注:使用超声duxplex可视化血流非侵入性大鼠14。 <ol><li>麻醉中的感应腔（异氟烷2％）大鼠。将在它的后面大鼠和与覆盖鼻面罩维持麻醉。 </li><li>使用电推剪及脱毛霜去除周围的大腿区域的头发。 </li><li>应用超声波凝胶大腿。确保有没有气泡。使用MS 400传感器获取多普勒超声图像（中心频率:30男赫兹）与230-400帧/秒的帧速率。 </li></ol> 5.组织病理学注:嘉实和染色与马松染色的容器形态分析15。 <ol><li>过夜固定在4％低聚甲醛的收获容器和在增加的乙醇浓度脱水它。嵌入石蜡的样品，并将其切成用切片机5微米厚的切片。 注:多聚甲醛是有毒的，应特别小心。 </li><li>用三色染色液染色之前Deparaffinize的幻灯片。脱水染色切片，用二甲苯清除它们，在安装中安装它们。干燥载玻片后，用显微镜观看样本。 </li></ol> 6.生物发光成像（BLI） 注:移植术后通过测定生物发光信号1跟踪随着时间的推移在体内6。 <ol><li>溶解1g D-荧光素钾盐在22ml的PBS中，并将其注入腹膜内到大鼠（375毫克/千克体重）。等待15分钟的荧光素在动物中循环。 </li><li>将老鼠变成一个实时定量生物发光系统并访问生物发光信号。 </li></ol>

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The authors have nothing to disclose.

本视频演示了大鼠静脉插入模型，探讨静脉移植的损失，并允许潜在的病理过程的探索和新的药物和治疗方案的测试。 麻醉的手术过程中的一个关键方面。建议连续吸入性麻醉系统，因为这是一种安全，简便的方法，尤其是在长时间的操作。这可以在训练阶段期间，当操作的时间超过1小时，是非常重要的。 相对于手术过程，重要的是不向收获及植入...

冠状动脉疾病及其并发症是世界范围内死亡的主要原因之一。当前的治疗策略集中在重新建立血流，或者通过扩张变窄容器或通过创建旁路。使用自体静脉冠状动脉旁路移植术（CABG）于1968年首次描述，并不断得到完善，多年来。除了左前的血运重建降支动脉，隐静脉导管是最常用的1。然而，移植通畅仍然隐静脉移植（SVG）的致命弱点。手术一年后，通畅率85％，下降幅度为61％，十年后2,3。因此揭幕SVG通畅损失的病理生理机制和原因的一项重要任务。 本视频演示了大鼠静脉插入模型，探讨静脉移植物损失。这种方法的总体目标是发掘潜在的病理生物学并在疾病进展和-physiological流程，制定药物或治疗方法测试一个合适的模型。通过移植的腹壁浅静脉进入动脉系统，这种模式密切模仿冠状动脉旁路移植术的临床设置。手术创伤，局部缺血，和壁应力是病理血管变化重要触发器和在所述的模型模仿。 不同型号和种类都可以探讨静脉移植通畅的损失。大动物模型，如猪4，羊5，狗6和猴7，类似于人类血管和解剖结构，从而使复杂的治疗策略，如旁路支架或新的外科技术，待测试8。然而，需要特殊的房屋，设备，人员和工作人员。此外，高的成本和手术过程中需要额外的麻醉师阻碍其更广泛的应用。钐所有的动物，包括老鼠，很容易处理，不需要特殊的住房，并有管理的成本。相比于小鼠模型9,10，大鼠模型具有较好的可操作性的成果优势，从而减少可变性。老鼠是生理和基因更类似于人类比老鼠11,12。此外，大多数的野生型小鼠仅开发有限myointima 13，其使小鼠模型容易发生II型误差。主小鼠静脉，如下腔静脉的组织学，只由几个细胞层的并呈现早期评估困难13。另一个缺点是可用于接枝恢复之后的后续分析的组织的量小。 在此视频描述的模型是可重复的，价格低廉，且容易执行，并且它可以快速而可靠地建立。它特别适合用于评估昂贵的实验治疗剂，如病毒载体用于基因疗法，以经济的方式。

<table><tbody><tr><td>Rat LEW/Crl</td><td>Charles River</td><td>Stock number 004</td><td></td></tr><tr><td>Rat LEW-Tg(Gt(ROSA)26Sor- 1&lt;br/&gt; luc)11Jmsk</td><td>Institute of laboratory animals, Kyoto University, Japan</td><td>NBPR rat number 0299</td><td>http://www.anim.med.kyoto-u.ac.jp/NBR/</td></tr><tr><td>PFA 4%</td><td>Electron Microscopy Sciences</td><td>#157135S</td><td>20%</td></tr><tr><td>hair clipper</td><td>WAHL</td><td>8786-451A ARCO SE</td><td></td></tr><tr><td>Forene</td><td>AbbVie</td><td>PZN 10182054 Art.Nr.: B506</td><td>Isoflurane</td></tr><tr><td>microsurgical clamp</td><td>Fine Science Tools</td><td>18055-04</td><td>Micro-Serrefine - 4 mm</td></tr><tr><td>clamp applicator</td><td>Fine Science Tools</td><td>18056-14</td><td></td></tr><tr><td>hair removal creme</td><td>Rufin cosmetic</td><td>27618</td><td></td></tr><tr><td>Povidone-Iodine</td><td>Betadine Purdue Pharma</td><td>NDC:67618-152</td><td></td></tr><tr><td>10-0 Ethilon suture</td><td>Ethicon</td><td>2814G</td><td></td></tr><tr><td>5-0 prolene suture</td><td>Ethicon</td><td>EH7229H</td><td></td></tr><tr><td>Rimadyl</td><td>Pfizer</td><td>400684.00.00</td><td>Carprofen</td></tr><tr><td>Novaminsulfon</td><td>Ratiopharm</td><td>PZN 03530402</td><td>Metamizole</td></tr><tr><td>Heparin</td><td>Rotexmedica</td><td>PZN: 3862340</td><td>25.000 I.E./ ml</td></tr><tr><td>Xylocain 1%</td><td>AstraZeneca</td><td>PZN: 1137907</td><td>Lidocain</td></tr><tr><td>EVICEL</td><td>J&amp;amp;J Med.Ethicon Biosur</td><td>PZN 7349697 Art. Nr.:EVK01DE</td><td>fibrin glue</td></tr><tr><td>NaCl 0.9%</td><td>B.Braun</td><td>PZN 06063042 Art. Nr.: 3570160</td><td></td></tr><tr><td>Vevo 770 high-resolution &lt;em&gt;in vivo&lt;/em&gt; micro-imaging system</td><td>VisualSonics</td><td></td><td>duplex sonography</td></tr><tr><td>Ecogel 100 ultrasound gel</td><td>Eco-med</td><td>30GB</td><td></td></tr><tr><td>D-Luciferin Firefly, potassium salt</td><td>Biosynth</td><td>L-8220</td><td></td></tr><tr><td>PBS pH 7.4</td><td>Gibco</td><td>10010023</td><td></td></tr><tr><td>Xenogen Ivis 200</td><td>Perkin Elmer</td><td></td><td>bioluminescence imaging</td></tr><tr><td>Weigerts iron hematoxylin Kit</td><td>Merck</td><td>1.15973.0002</td><td>Trichrome staining</td></tr><tr><td>Resorcine-Fuchsine Weigert</td><td>Waldeck</td><td>2.00E-30</td><td>Trichrome staining</td></tr><tr><td>Acid Fuchsin</td><td>Sigma-Aldrich</td><td>F8129-25G</td><td>Trichrome staining</td></tr><tr><td>Ponceau S solution</td><td>Serva Electrophoresis</td><td>33427</td><td>Trichrome staining</td></tr><tr><td>Azophloxin</td><td>Waldeck</td><td>1B-103</td><td>Trichrome staining</td></tr><tr><td>Molybdatophosphoric acid hydrate</td><td>Merck</td><td>1.00532.0100</td><td>Trichrome staining</td></tr><tr><td>Orange G</td><td>Waldeck</td><td>1B-221</td><td>Trichrome staining</td></tr><tr><td>Light Green SF</td><td>Waldeck</td><td>1B-211</td><td>Trichrome staining</td></tr><tr><td>Vitro-Clud</td><td>Langenbrinck</td><td>04-0001</td><td></td></tr><tr><td>Glacial Acetic Acid</td><td>Sigma-Aldrich</td><td>537020</td><td></td></tr><tr><td>37% HCl</td><td>Sigma-Aldrich</td><td>H1758</td><td></td></tr><tr><td>Xylene</td><td>Th. Geyer</td><td>3410</td><td></td></tr><tr><td>Paraffin</td><td>Leica biosystems</td><td>REF 39602004</td><td></td></tr><tr><td>Ethanol absolute</td><td>Th. Geyer</td><td>2246</td><td></td></tr><tr><td>Ethanol 96%</td><td>Th. Geyer</td><td>2295</td><td></td></tr><tr><td>Ethanol 70%</td><td>Th. Geyer</td><td>2270</td><td></td></tr><tr><td>Slide Rack</td><td>Ted Pella</td><td>21057</td><td></td></tr><tr><td>Staining dish</td><td>Ted Pella</td><td>21075</td><td></td></tr><tr><td>Bepanthen Eye and Nose ointment</td><td>Bayer</td><td>1578675</td><td>Eye ointment</td></tr></tbody></table>

vein interposition model: a suitable model to study bypass graft patency

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. Research models for bypass graft failure are essential for a better understanding of pathobiological and pathophysiological processes during graft patency loss. Large animal models, such as pigs or sheep, resemble human anatomical structures but require special facilities and equipment. This video describes a rat vein interposition model to investigate vein graft patency loss. Rats are inexpensive and easy to handle. Compared to mouse models, the convenient size of rats permits better operability and enables a sufficient amount of material to be obtained for further diverse analysis. In brief, the inferior epigastric vein of a donor rat is harvested and used to replace a segment of the femoral artery. Anastomosis is conducted via single stitches and sealed with fibrin glue. Graft patency can be monitored non-invasively using duplex sonography. Myointimal hyperplasia, which is the main cause for graft patency loss, develops progressively over time and can be calculated from histological cross sections.

大鼠静脉插入模型适用于研究myointima增生静脉移植失败的发展。动物从手术中恢复良好，显示出出色的身体条件后的操作。 图1示出了关键的手术步骤。沿着LINEA inguinalis切开皮肤后，上腹部浅静脉和股动脉被确定（ 图1A）。应仔细进行接枝的收获，而不会损坏接枝（ 图1B），因为这可能导致早期移植物衰竭和通畅损失。在受体动物（ 图1C）定位?...

Watch this Scientific Journal Video about Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency at JoVE.com

Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

This video demonstrates a model to study the development of myointimal hyperplasia after venous interposition surgery in rats.

静脉介入型号:研究旁路移植通畅一个合适的模型

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. ...

vein-interposition-model-suitable-model-to-study-bypass-graft

Research

JoVE Journal

Medicine

8.7K 次观看.  University Heart Center Hamburg. This video demonstrates a model to study the development of myointimal hyperplasia after venous interposition surgery in rats. 

视频: Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

Training a Sophisticated Microsurgical Technique: Interposition of External Jugular Vein Graft in the Common Carotid Artery in Rats

Neointimal hyperplasia is one the primary causes of stenosis in arterialized veins that are of great importance in arterial coronary bypass surgery, in peripheral arterial bypass surgery as well as in arteriovenous fistulas.1-5 The experimental procedure of vein graft interposition in the common carotid artery by using the cuff-technique has been applied in several research projects to examine the aetiology of neointimal hyperplasia and therapeutic options to address it. 6-8 The cuff prevents vessel anastomotic remodeling and induces turbulence within the graft and thereby the development of neointimal hyperplasia.
Using the superior caval vein graft is an established small-animal model for venous arterialization experiment.9-11 This current protocol refers to an established jugular vein graft interposition technique first described by Zou et al., 9 as well as others.12-14 Nevertheless, these cited small animal protocols are complicated.
To simplify the procedure and to minimize the number of experimental animals needed, a detailed operation protocol by video training is presented. This video should help the novice surgeon to learn both the cuff-technique and the vein graft interposition. Hereby, the right external jugular vein was grafted in cuff-technique in the common carotid artery of 21 female Sprague Dawley rats categorized in three equal groups that were sacrificed on day 21, 42 and 84, respectively. Notably, no donor animals were needed, because auto-transplantations were performed. The survival rate was 100 % at the time point of sacrifice. In addition, the graft patency rate was 60 % for the first 10 operated animals and 82 % for the remaining 11 animals. The blood flow at the time of sacrifice was 8&plusmn;3 ml/min. In conclusion, this surgical protocol considerably simplifies, optimizes and standardizes this complicated procedure. It gives novice surgeons easy, step-by-step instruction, explaining possible pitfalls, thereby helping them to gain expertise fast and avoid useless sacrifice of experimental animals.

Neointimal hyperplasia is the primary cause of stenosis in arterialized veins. We propose a new protocol whereby the right external jugular vein is grafted using the cuff technique in the common carotid artery of Sprague Dawley rats. The survival rate was 100 % at the time point of sacrifice.

培养一个精密的显微外科技术在大鼠颈总动脉，颈外静脉移植：介入

Neointimal hyperplasia is one the primary causes of stenosis in arterialized veins that are of great importance in arterial coronary bypass surgery, in ...

科研

医学

A Modified Heterotopic Swine Hind Limb Transplant Model for Translational Vascularized Composite Allotransplantation (VCA) Research

Vascularized Composite Allotransplantation (VCA) such as hand and face transplants represent a viable treatment option for complex musculoskeletal trauma and devastating tissue loss. Despite favorable and highly encouraging early and intermediate functional outcomes, rejection of the highly immunogenic skin component of a VCA and potential adverse effects of chronic multi-drug immunosuppression continue to hamper widespread clinical application of VCA. Therefore, research in this novel field needs to focus on translational studies related to unique immunologic features of VCA and to develop novel immunomodulatory strategies for immunomodulation and tolerance induction following VCA without the need for long term immunosuppression.
This article describes a reliable and reproducible translational large animal model of VCA that is comprised of an osteomyocutaneous flap in a MHC-defined swine heterotopic hind limb allotransplantation. Briefly, a well-vascularized skin paddle is identified in the anteromedial thigh region using near infrared laser angiography. The underlying muscles, knee joint, distal femur, and proximal tibia are harvested on a femoral vascular pedicle. This allograft can be considered both a VCA and a vascularized bone marrow transplant with its unique immune privileged features. The graft is transplanted to a subcutaneous abdominal pocket in the recipient animal with a skin component exteriorized to the dorsolateral region for immune monitoring.
Three surgical teams work simultaneously in a well-coordinated manner to reduce anesthesia and ischemia times, thereby improving efficiency of this model and reducing potential confounders in experimental protocols. This model serves as the groundwork for future therapeutic strategies aimed at reducing and potentially eliminating the need for chronic multi-drug immunosuppression in VCA.

A Modified Heterotopic Swine Hind Limb Transplant Model for Translational Vascularized Composite Allotransplantation VCA Research

Vascularized Composite Allotransplantations (VCA) have become a clinical reality. However, broad clinical application of VCA is limited by chronic multi-drug immunosuppression. The authors present a reliable and reproducible large animal model to translate novel immunomodulatory strategies that can minimize or potentially eliminate the need of immunosuppression in VCA.

平移血管复合同种异体移植（VCA）研究一种改进的异位猪后肢移植模型

Vascularized Composite Allotransplantation (VCA) such as hand and face transplants represent a viable treatment option for complex musculoskeletal trauma ...

An In Vitro Model of a Parallel-Plate Perfusion System to Study Bacterial Adherence to Graft Tissues

Various valved conduits and stent-mounted valves are used for right ventricular outflow tract (RVOT) valve replacement in patients with congenital heart disease. When using prosthetic materials however, these grafts are susceptible to bacterial infections and various host responses.
Identification of bacterial and host factors that play a vital role in endovascular adherence of microorganisms is of importance to better understand the pathophysiology of the onset of infections such as infective endocarditis (IE) and to develop preventive strategies. Therefore, the development of competent models to investigate bacterial adhesion under physiological shear conditions is necessary. Here, we describe the use of a newly designed in vitro perfusion chamber based on parallel plates that allows the study of bacterial adherence to different components of graft tissues such as exposed extracellular matrix, endothelial cells and inert areas. This method combined with colony-forming unit (CFU) counting is adequate to evaluate the propensity of graft materials towards bacterial adhesion under flow. Further on, the flow chamber system might be used to investigate the role of blood components in bacterial adhesion under shear conditions. We demonstrated that the source of tissue, their surface morphology and bacterial species specificity are not the major determining factors in bacterial adherence to graft tissues by using our in-house designed in vitro perfusion model.

An In Vitro Model of a Parallel-Plate Perfusion System to Study Bacterial Adherence to Graft Tissues

We describe an in-house designed in vitro flow chamber model, which allows the investigation of bacterial adherence to graft tissues.

平行板灌注系统的体外模型研究了细菌对移植组织的粘附性

Various valved conduits and stent-mounted valves are used for right ventricular outflow tract (RVOT) valve replacement in patients with congenital heart ...

发育生物学

Implantation of Inferior Vena Cava Interposition Graft in Mouse Model

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. The long-term clinical results showed excellent patency rates, however, with significant incidence of stenosis. To investigate the cellular and molecular mechanisms of vascular neotissue formation and prevent stenosis development in tissue engineered vascular grafts (TEVGs), we developed a mouse model of the graft with approximately 1 mm internal diameter. First, the TEVGs were assembled from biodegradable tubular scaffolds fabricated from a polyglycolic acid nonwoven felt mesh coated with &#949;-caprolactone and L-lactide copolymer. The scaffolds were then placed in a lyophilizer, vacuumed for 24 hr, and stored in a desiccator until cell seeding. Second, bone marrow was collected from donor mice and mononuclear cells were isolated by density gradient centrifugation. Third, approximately one million cells were seeded on a scaffold and incubated O/N. Finally, the seeded scaffolds were then implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. The implanted grafts demonstrated excellent patency (&#62;90%) without evidence of thromboembolic complications or aneurysmal formation. This murine model will aid us in understanding and quantifying the cellular and molecular mechanisms of neotissue formation in the TEVG.

To improve our knowledge of cellular and molecular neotissue formation, a murine model of the TEVG was recently developed. The grafts were implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. This model achieves similar results to those achieved in our clinical investigation, but over a far shortened time-course.

下腔静脉间置移植物在小鼠模型中植入

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. ...

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

In reconstructive surgery, there is a clinical need for an alternative to the current methods of autologous reconstruction which are complex, costly and trade one defect for another. Tissue engineering holds the promise to address this increasing demand. However, most tissue engineering strategies fail to generate stable and functional tissue substitutes because of poor vascularization. This paper focuses on an in vivo tissue engineering chamber model of intrinsic vascularization where a perfused artery and a vein either as an arteriovenous loop or a flow-through pedicle configuration is directed inside a protected hollow chamber. In this chamber-based system angiogenic sprouting occurs from the arteriovenous vessels and this system attracts ischemic and inflammatory driven endogenous cell migration which gradually fills the chamber space with fibro-vascular tissue. Exogenous cell/matrix implantation at the time of chamber construction enhances cell survival and determines specificity of the engineered tissues which develop. Our studies have shown that this chamber model can successfully generate different tissues such as fat, cardiac muscle, liver and others. However, modifications and refinements are required to ensure target tissue formation is consistent and reproducible. This article describes a standardized protocol for the fabrication of two different vascularized tissue engineering chamber models in vivo.

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

This is a guideline for constructing in vivo vascularized tissue using a microsurgical arteriovenous loop or a flow-through pedicle configuration inside a tissue engineering chamber. The vascularized tissues generated can be employed for organ regeneration and replacement of tissue defects, as well as for drug testing and disease modeling.

在组织工程由内在血管化<em&gt;在体内</em&gt;组织工程商会

In reconstructive surgery, there is a clinical need for an alternative to the current methods of autologous reconstruction which are complex, costly and ...

生物工程

Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement

The mainstay of contemporary therapies for extensive occlusive arterial disease is venous bypass graft. However, its durability is threatened by intimal hyperplasia (IH) that eventually leads to vessel occlusion and graft failure. Mechanical forces, particularly low shear stress and high wall tension, are thought to initiate and to sustain these cellular and molecular changes, but their exact contribution remains to be unraveled. To selectively evaluate the role of pressure and shear stress on the biology of IH, an ex&#160;vivo perfusion system (EVPS) was created to perfuse segments of human saphenous veins under arterial regimen (high shear stress and high pressure). Further technical innovations allowed the simultaneous perfusion of two segments from the same vein, one reinforced with an external mesh. Veins were harvested using a no-touch technique and immediately transferred to the laboratory for assembly in the EVPS. One segment of the freshly isolated vein was not perfused (control, day 0). The two others segments were perfused for up to 7 days, one being completely sheltered with a 4 mm (diameter) external mesh. The pressure, flow velocity, and pulse rate were continuously monitored and adjusted to mimic the hemodynamic conditions prevailing in the femoral artery. Upon completion of the perfusion, veins were dismounted and used for histological and molecular analysis. Under ex&#160;vivo conditions, high pressure perfusion (arterial, mean = 100 mm Hg) is sufficient to generate IH and remodeling of human veins. These alterations are reduced in the presence of an external polyester mesh.

Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement

The mechanisms leading to the development of intimal hyperplasia (IH) and vein graft failure are still poorly understood. This study describes an ex&#160;vivo system to perfuse human veins under controlled flow and pressure. Furthermore the efficiency of external mesh reinforcement to limit the development of IH was evaluated.

步骤人类隐静脉<em&gt;前体内</em&gt;灌注和外部强化

The mainstay of contemporary therapies for extensive occlusive arterial disease is venous bypass graft. However, its durability is threatened by intimal ...

A Rabbit Model of Durable Transgene Expression in Jugular Vein to Common Carotid Artery Interposition Grafts

Vein graft bypass surgery is a common treatment for occlusive arterial disease; however, long-term success is limited by graft failure due to thrombosis, intimal hyperplasia, and atherosclerosis. The goal of this article is to demonstrate a method for placing bilateral venous interposition grafts in a rabbit, then transducing the grafts with a gene transfer vector that achieves durable transgene expression. The method allows the investigation of the biological roles of genes and their protein products in normal vein graft homeostasis. It also allows the testing of transgenes for the activities that could prevent vein graft failure, e.g., whether the expression of a transgene prevents the neointimal growth, reduces the vascular inflammation, or reduces atherosclerosis in rabbits fed with a high-fat diet. During an initial survival surgery, the segments of right and left external jugular vein are excised and placed bilaterally as reversed end-to-side common carotid artery interposition grafts. During a second survival surgery, performed 28 days later, each of the grafts is isolated from the circulation with vascular clips and the lumens are filled (via an arteriotomy) with a solution containing a helper-dependent adenoviral (HDAd) vector. After a 20-min incubation, the vector solution is aspirated, the arteriotomy is repaired, and flow is restored. The veins are harvested at time points dictated by individual experimental protocols. The 28-day delay between the graft placement and the transduction is necessary to ensure the adaptation of the vein graft to the arterial circulation. This adaptation avoids rapid loss of transgene expression that occurs in vein grafts transduced before or immediately after grafting. The method is unique in its ability to achieve durable, stable transgene expression in grafted veins. Compared to other large animal vein graft models, rabbits have advantages of low cost and easy handling. Compared to rodent vein graft models, rabbits have larger and easier-to-manipulate blood vessels that provide abundant tissue for analysis.

This method describes the placement of interposition vein grafts in rabbits, the transduction of the grafts, and the achievement of durable transgene expression. This allows the investigation of physiological and pathological roles of transgenes and their protein products in grafted veins, and testing of gene therapies for vein graft disease.

颈动脉介入移植中颈静脉持久性转基因表达的家兔模型

Vein graft bypass surgery is a common treatment for occlusive arterial disease; however, long-term success is limited by graft failure due to thrombosis, ...

Synergizing Antegrade Endoscopic with Bridging Vein Harvesting for Improvement of Great Saphenous Vein Graft Quality from the Lower Leg

Antegrade endoscopic harvesting of autografts for bypass grafting may be an optimal strategy addressing excellent graft quality and reduced post-operative wound complications. This standardized protocol for antegrade endoscopic vein harvesting (EVH) from the lower leg has the potential to be introduced to routine coronary artery bypass grafting (CABG). Patients undergoing CABG surgery are positioned on a surgical table with two additional foam rollers below the extended legs, enabling antegrade EVH from the lower leg. Following minimally invasive surgical access through a bridging vein harvest technique, an endoscopic optical dissector is inserted antegrade into the wound. The main vessel and side branches are dissected under continuous optical control of vein quality status and the working channel. After, an endoscopic optical retractor is inserted with an internal bipolar electrocoagulation device for precise, safe, and tissue-protective interruption of side branches. After release of the vein, the vessel is cut off at the proximal and distal ends under optical control, retrieved from the wound, then cannulated and flushed with heparinized saline. Finally, all side branches of the vein graft are double-clipped. Vascular histology is analyzed in a randomized selection of vein samples. After applying this standardized EVH protocol, the learning curve was shown to be steep, and graft quality was sufficient for coronary artery bypass grafting in every case. There was no conversion to surgical harvesting and low risks for tissue damage and bleeding. Leg positioning and synergizing EVH with bridging vein harvesting improved procedural success and vein graft quality. In our hands, antegrade EVH from the lower leg was feasible, demonstrating straightforward graft dissection as well as adequate macroscopic and microscopic graft quality with preserved endothelial integrity. In conclusion, the introduced technique is safe, shows excellent vein autograft quality, and illustrates feasibility for elective and urgent isolated CABG and combined CABG scenarios.

Presented here is a protocol for antegrade endoscopic vein harvesting from the lower leg, which can safely be introduced in routine coronary artery bypass grafting. Vein grafts present excellent graft quality following this standardized protocol with positioning of the legs, minimally invasive access to the vein, and antegrade endoscopic vein harvesting.

协同前体内窥镜与桥接静脉收获，提高从下腿大萨普努斯静脉移植质量

Antegrade endoscopic harvesting of autografts for bypass grafting may be an optimal strategy addressing excellent graft quality and reduced post-operative ...

用于绵羊静脉依从性评估的 3D 血管造影

Quantifying Inferior Vena Cava Compliance and Distensibility in an In Vivo Ovine Model Using 3D Angiography

Synthetic vascular grafts overcome some challenges of allografts, autografts, and xenografts but are often more rigid and less compliant than the native vessel into which they are implanted. Compliance matching with the native vessel is emerging as a key property for graft success. The current gold standard for assessing vessel compliance involves the vessel's excision and ex vivo biaxial mechanical testing. We developed an in vivo method to assess venous compliance and distensibility that better reflects natural physiology and takes into consideration the impact of a pressure change caused by flowing blood and by any morphologic changes present. 
This method is designed as a survival procedure, facilitating longitudinal studies while potentially reducing the need for animal use. Our method involves injecting a 20 mL/kg saline bolus into the venous vasculature, followed by the acquisition of pre and post bolus 3D angiograms to observe alterations induced by the bolus, concurrently with intravascular pressure measurements in target regions. We are then able to measure the circumference and the cross-sectional area of the vessel pre and post bolus. 
With these data and the intravascular pressure, we are able to calculate the compliance and distensibility with specific equations. This method was used to compare the inferior vena cava's compliance and distensibility in native unoperated sheep to the conduit of sheep implanted with a long-term expanded polytetrafluorethylene (PTFE) graft. The native vessel was found to be more compliant and distensible than the PTFE graft at all measured locations. We conclude that this method safely provides in vivo measurements of vein compliance and distensibility.

作者聚焦:先天性心脏病研究中新生血管发育的改进成像

This protocol allows for the in vivo quantification of venous compliance and distensibility using catheterization and 3D angiography as a survival procedure allowing for a variety of potential applications.

使用 3D 血管造影量化体内绵羊模型中的下腔静脉顺应性和扩张性

Synthetic vascular grafts overcome some challenges of allografts, autografts, and xenografts but are often more rigid and less compliant than the native ...

静脉介入型号:研究旁路移植通畅一个合适的模型

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使用 3D 血管造影量化体内绵羊模型中的下腔静脉顺应性和扩张性

使用 3D 血管造影量化体内绵羊模型中的下腔静脉顺应性和扩张性