感谢Ivan Laermans，Rosita Kinart，Ann Lissens（外科技术中心，比利时鲁汶KU Leuven）。 Jo Verbinnen和Kristof Reyniers（比利时鲁汶KU Leuven医学院Vesalius解剖学研究所）在实验过程中提供了技术支持。感谢Leen Mortier对数据和手稿管理的帮助。我们感谢FEG Textiltechniken制造原型网格，对其进行消毒，并无条件捐赠给研究机构。

该实验的伦理批准是从鲁汶大学动物实验伦理委员会（P065 / 2013）获得的。根据现行国家动物福利准则处理动物。 材料与实验动物<ol><li> 手术准备 <ol><li>在外科手术室中覆盖一张无菌悬垂的桌子，并准备一个无菌弯曲的套管针（ 图1 ， 图 A），无菌手术器械，缝合线和无菌纱布。如果实验包括随访，则在无菌条件下进行整个外科手术。将所有仪器放在桌子上，准备在手术期间使用。 </li><li>从无菌包装中取出一个无菌的矩形植入物和/或具有锚固臂的植入物，并将它们放在用无菌悬垂覆盖的桌子上（ 图1B ， C和D ）。 </li></ol></li></ol> 图1:套管针和植入物。 （ 一 ）套管针示意图。 （ B ）H型聚偏二氟乙烯（PVDF）植入物，具有中心部分的细节（图C ）。其形状受到目前可用于经阴道脱垂修复的四臂网格的启发。矩形体（30×40mm 2 ）通过四个伸出的臂（150×10mm 2 ）横向延伸。武器的尺寸设计得足够长，刺破相关悬架结构的基础上，早期的解剖学研究17。 （ D ）矩形植入物（30×40mm 2 ）。两种植入物均为聚偏氟乙烯;纺织品特性和性能见表1 。 <ol start="2"><li> 实验动物（Ewe，45-60公斤） <ol><li>在外科手术前30分钟，管理1毫升15毫克/毫升硫酸阿托品和1毫升/ 50公斤肌内（im）的甲苯噻嗪。 </li><li> 30分钟后，确保预防措施使羊昏昏欲睡。 </li><li>将静脉内导管插入颈静脉，给予0.075 mL / kg氯胺酮100mg / mL HCl。通过观察对疼痛刺激的反应缺乏确认深度麻醉。 </li><li>将动物移动到手术台上，并通过插管固定其气道。用5％异氟烷在5 L / min氧气中维持麻醉。 </li><li>将静脉管线插入颈静脉，并以150 mL / h的流速供应500 mL盐水溶液。 </li><li>根据当地方案，管理预防性抗生素（阿莫西林克拉维酸，7 mg / kg）和术后镇痛药（丁丙诺啡和氯甲酚，1 mL）或等效抗生素。 </li><li>把动物放在灯光下髋关节位置在手术台的末端，并使用绳索（ 图2 ， 图 A）固定其四肢，臀部处于超屈曲状态。 </li><li>通过阴道推动膀胱和直肠手术。 </li><li>剃视会阴部，大腿的中间部分，尾部折叠并用聚维酮7.5％消毒（ 图2 ， 图 B和C）。 </li></ol></li></ol><img alt="图2" src="/files/ftp_upload/55706/55706fig2.jpg" /> 图2:动物手术。 （ A ）放置在仰卧位的羊，臀部通过固定下肢而弯曲。 （ B ）套管针插入的外部入口点位于腹侧（空箭头），侧背侧折叠（全箭头）。 （ C ）腹侧插入点的位置;中间的虚线代表动物的正中飞机。 （ D ）解剖直肠阴道隔膜。 （ E ）腹侧套管穿过大腿内侧，闭孔和阴道空间的肌肉插入。穿刺套管针的轨迹由手指控制。 （ F和G ）一旦套针针就位，电线吊带（打开箭头）前进，并加载阴道网的臂。 （ H ）腹侧（全箭头）和背部（空箭头）臂的最终位置。 （ 一 ）中央部位在阴道壁与直肠外膜之间无张力放置。 <ol start="10" style="margin-left: 40px;"><li>准备在无菌条件下进行手术的人员。戴上手术帽和口罩，手洗手术，并穿上手术衣和无菌手套。 </li><li>用无菌悬垂覆盖动物，并在生殖器上方开一个开口裂孔。 </li></ol>外科手术<ol><li> 直肠阴道隔膜的制备 <ol><li>使用Allis镊子将背侧阴道壁3厘米颅骨到hymeneal环。 </li><li>取一个装有10mL盐水的注射器，并装上22 G针头。将其穿过阴道上皮（约3-4mm深）并插入直肠阴道隔膜的中线，1.5厘米颅骨至阴沟。 </li><li>通过在阴道隔膜11注射盐水进行"水解"。 </li><li>在阴道上皮做一个3厘米长的中线切口，开始尾部到Allis镊子（步骤2.1.1），并使用手术刀在hymeneal戒指结束。通过这个切口进入正阴阴道。 </li><li>将自固位牵开器（见材料表 ）放置在会阴上，并将四个尖锐的钩钩放在阴道切口中以保持开口。 </LI&gt; <li>用你的手指，将阴道筋膜从阴道壁向侧面朝向盆腔侧壁直肠解剖，并直到头部的尾部。为网格的30 x 40 mm 2中心部分创建合适的空间（ 图2 ， 图 D）。 </li><li>必要时用止血钳或十字形止血结扎器进行止血。 注意:小排气管可以用止血钳钳住。这破坏了血管并启动了自然凝血级联。为了更强的出血，用镊子抓住出血血管并放置十字交叉扎结，用方形结固定。在这一点上，可以插入矩形植入物（步骤2.2）或继续进行解剖，以用固定臂插入植入物（步骤2.3）。 </li></ol></li><li> 平面网格插入 <ol><li>将阴道牵开器插入阴道切口，以便更好地观察解剖区域的头部。 </li><li>在解剖的阴道阴道空间的最颅侧面的左侧和右侧，用简单的中断的3/0聚丙烯缝合线缝合植入物的左右颅角。切割残留的缝合材料。保持缝合线远离阴道腔（ 即不渗透阴道壁）。 注意:植入物总是缝合到包括直肠阴道隔膜的结缔组织。如果缝线材料不能在阴道中看到，阴道壁就不会渗透。 </li><li>沿着植入物的颅骨部位中途添加一个额外的简单中断缝合线。 </li><li>用简单的中断3/0聚丙烯将植入物的侧边缘缝合到周围的结缔组织上。保持植入物尽可能平坦，无张力。 </li><li>在左侧和右侧用简单的3/0聚丙烯缝合线缝合左右尾角直肠阴道空间最尾端的两侧。 </li><li>沿着植入物的尾部方向，在中途添加一个简单的中断缝合线。 </li><li>用运行中的3/0聚乳酸酶缝线闭合阴道切口。 </li></ol></li><li> 植入物插入和锚定（套管针引导技术）。 <ol><li>继续解剖步骤2.1中创建的正阴阴部空间，以达到闭孔孔的内侧，这可以很容易触发。 </li><li>解剖空间caudo横向到达骶骨韧带和尾部定位的尾骨肌肉的尾部。 </li><li>没有。 24叶片，在外阴四侧切开1厘米宽的切口，穿过皮肤和浅层肌筋膜（ 图2 ， 图 B和C）。 </li><li>在大腿的内侧形成两个"腹侧"切口，距离尾部边界近4厘米颅骨f坐标系（ 即联合下边界）和距离中线3厘米（ 图2 ， 图 C）。 </li><li>在插入尾部褶皱的水平上进行两个"背侧"切口，其中可以容易触摸到块茎的2厘米内侧（ 图2 ， 图 B）。 </li><li>将一个弯曲的套管穿过其中一个腹侧切口（ 图2 ， 图 E）。 </li><li>通过套管针穿过内收肌肉肌肉，外部闭孔和封闭孔的内侧。 </li><li>用插入阴道切口的手指控制套管针的进展。将其尖端指向提肌肛肌的腱弧（ 图2 ， 图 E）。 </li><li>将引导线暴露在阴道壁切口中，并用相应的同侧颅网臂（ 图2 ， 图 F）装载。 </li><li>拉起套管针​​，装上m通过上述结构。保持手臂无张力。 </li><li>通过第二个颅臂通过动物另一侧的腹侧切口重复该过程。 </li><li>通过背部切口之一，将套管针穿过尾骨肌肉，仅在骶骨韧带的远端（ 图2 ， 图 G）。 </li><li>将导丝穿过阴道切口，抓住网状物的背部手臂，将其拉出。保持手臂无张力和重复在另一边。 </li><li>通过使其平坦化并对臂施加张力来调整网格的位置，但保持网格无张力（ 图2， 图 I; 图3 ）。 </li><li>用一个简单的中断的3/0聚丙烯缝合线固定在网状物的尾部中部，将其固定在周围的结缔组织上。 </li><li>将手臂切开皮肤水平，并用简单中断的3/0 polygl关闭所有皮肤切口ecaprone缝线（ 图2 ， 图 H）。 </li><li>用跑步的3/0聚吡咯胶缝合阴道切口。 </li></ol></li></ol><img alt="图3" src="/files/ftp_upload/55706/55706fig3.jpg" /> 图3:桡骨穿过闭孔器和穿过尾部折叠的尾部的桡骨的示意图。广泛的骶骨韧带呈蓝色。更小的面板在缩短过量的材料之前说明了臂在放置位置的动物上的位置。主面板显示相同但皮肤和肌肉被去除。

<ol>
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Pelvic Floor Dysfunct. 18 (10), 1201-1206 (2007).</li><li>Schaller, O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Illustrated Veterinary Anatomical Nomenclature. , (2007).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Serious+Complications+Associated+with+Transvaginal+Placement+of+Surgical+Mesh+for+Pelvic+Organ+Prolapse.">Serious Complications Associated with Transvaginal Placement of Surgical Mesh for Pelvic Organ Prolapse.</a> FDA Safety Communication. , (2016).</li><li>Reinier, M., Groep, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Final+Opinion+on+the+use+of+meshes+in+urogynecological+surgery.">Final Opinion on the use of meshes in urogynecological surgery.</a> SCENIHR- European Commission. , (2016).</li></ol>

这项关于羊模型的研究计划得到了Medri和Blasingame，Burch，Garrard和Ashley（美国亚特兰大GA）的无条件补助。协议由鲁汶研发中心办公室处理。赞助商没有干扰本实验的规划，执行或报告，也没有拥有结果。 NS和LH是FP7框架（Bip-Upy项目; NMP3-LA-2012-310389）中EC的拨款的接收者。 AF获得了欧盟委员会在行业 - 学术合作伙伴计划（251356）的资助。

在这里，我们描述了羊的实验过程，旨在模拟具有或不具有锚固臂的植入物的阴道解剖和经阴道网插入。后续的步骤和仪器通过POP和压力性尿失禁15，16，19，20进行的外科手术的启发。在初步的解剖学解剖之后，在实验中插入网孔时仍然存在一些问题。在第一只动物中，发现了一个穿孔在腹腔内的脱水囊水平。这已经在临床上在妇女中描述了21 。在随后的过程中，通过更多地侧向（ 即，更靠近骨盆侧壁）引导套管针的尖端来避免这种情况。在后来的羊中，没有观察到其他并发症。 最可怕的解剖结构我们再次闭塞神经和动脉。在人类身上，套管针/植入物与闭孔器之间的距离为1.9 - 3.0厘米22 。以前通过腿部或针头轨迹的确切位置来解释套管针/植入物位置在女性中的高度变异性。因此，羊的后肢在臀部的高屈曲中被固定以允许更多地进入阴道。在这个位置，腓肠肌被颅骨移动。因此，套管针穿过内收肌肌肉，这是绵羊23中不分开而发育良好的肌肉。套管针的更多颅骨通道是可能的，但可能损害闭孔器的结构。 类似于临床上描述的，臂将植入物固定到给定的解剖位置，对应于绵羊23中的直肠阴道隔膜的自然附着物。植入物平坦，显然支撑p口腔阴道壁不会更侧向延伸。因此，它没有折叠的倾向。可能使用较大的植入物。然而，如在羊之前证实，较大的植入物具有高达50％的收缩和多个本地接枝相关并发症9相关联。 以前对各种动物模型中的新种植体的研究包括将羊作为阴道手术的模型。这项工作的一个目标是以类似于临床实践的方式来固定植入物（ 即，通过将网状物的手臂转移到骨盆中的解剖结构）。此过程在羊模型中之前没有描述。这种手术遵循类似于在临床实践中引入新的针刺和针灸辅助手术的策略15 。然而，这些发现似乎与几年前相关，因为使用阴道植入物，因此使用套管针导向程序由于FDA和SCENIHR 24，25连续的健康警快速下探。 虽然我们在这里描述了这项技术，但是本实验中的动物数量非常有限，因此解剖学变异性的全部水平可能不在此表示。另一个限制是该技术描述了后隔室程序，其可能较少实践。有关羊13前房手术的报道很少，但通常使用较小的植入物，并发症较为频繁。虽然这些发现可能足以规划进一步的实验，但是前阴道网布置的可行性也可能是有益的。 总之，这是对阴道手术的羊动物模型中安全可行的手术技术的描述，其允许经阴道和针灸引导的张力 -自由阴道植入物插入。相关的可比结构可能被盲目刺穿，对血管，神经或器官损伤没有明显的危险。该模型当然也可以用于使用天然组织的模拟阴道手术。

盆腔器官脱垂在一半以上的妇女临床诊断，至少有一名阴道分娩，但从主观上讲，其中一半的女性整体感染1例 。治疗的主要是使用任一天然组织或植入材料手术重建，但每一种方法都有其局限性，包括复发或局部并发症2，3，4。理想的植入物尚未确定;因此，在向市场推出新产品和技术之前，对产品创新的持续需求以及开发临床前实验的适当管道。一个在此轨道中的步骤是在合适的动物模型5实验评估，6。理想情况下，它们应该模仿解剖，生物力学和生物环境。当谈到th对新型植入物的实验评估，通常在较小的模型中首先测试，用于生物相容性或重建腹壁缺陷。这种类型的实验受到批评，因为植入物没有插入感兴趣的区域（ 即阴道） 7 。阴道手术模型更为稀缺，当实验的目标是记录外植体的生物力学特征时，肯定是这样。因此，有一只兔从兔子移到了羊8 。成年母羊是大型动物模型，具有相当大小和可及的阴道。它们可被用于新的植入物的中期评估，并且能够再现阴道暴露与某些材料9，10，11，12，13。不仅尺寸和解剖学的阴道和盆底与人类相当，但也发生在发生在15％的母羊中的脱垂的自发性发生。脱出的危险因素是重叠的（ 即，multiparity，POP的既往史，增加更高的体重引起的腹内压或丘陵，且（植物）的雌激素效果可比放牧时）6，14。在欧洲，羊是唯一合理的选择，因为对非人灵长类动物的研究几乎完全被禁止。在这里，通过模拟使用套管针和导向器的植入物的经阴道插入，将网格进一步放置到正阴 - 阴道隔膜中，进一步模型。这之后，通过固定用带通过肌肉的韧带，其可以被认为等同于临床实践15,16臂锚定植入物。到目前为止，这种技术虽然许多人认为由于使用这些较长的条和/或解剖结构的刺穿可能会发生特定的并发症。 在早期详细的解剖学研究中，将卵巢盆底与女性骨盆17进行了比较。当涉及到植入物的锚固时，羊不具有骶棘状韧带，但它们确实具有非常发达和广泛的骶骨韧带。阴部神经在其上方运行，使得这个地标作为悬浮点不安全。相反，尾骨肌及其筋膜以及闭孔膜可通过直肠阴道空间进入。这里提出了用于固定锚固臂的解剖结构的进入和位置。讨论了可用于定位网格的仪器。最后，武器或套管针对相邻解剖结构（如血管和神经）的关系，a也有潜在的术中并发症。

<table><tbody><tr><td>Animals:&amp;nbsp;</td><td></td><td></td><td></td></tr><tr><td>parous female sheep (45 - 65 kg)</td><td>Zo&amp;ouml;technical Institute of the KU Leuven</td><td>NA</td><td>experimetnal animal</td></tr><tr><td>Sterile clothing:&amp;nbsp;</td><td></td><td></td><td></td></tr><tr><td>sterile drape 45 x 75 cm</td><td>Lohmann &amp;amp; Rauscher, Regensdorf, Germany</td><td>33002</td><td>other material</td></tr><tr><td>sterile OR drape 150 x 180 cm</td><td>Lohmann &amp;amp; Rauscher, Regensdorf, Germany</td><td>33009</td><td>other material</td></tr><tr><td>sterile glowes 2x</td><td>Lohmann &amp;amp; Rauscher, Regensdorf, Germany</td><td>16652</td><td>other material</td></tr><tr><td>sterile surgical gown 2x</td><td>Lohmann &amp;amp; Rauscher, Regensdorf, Germany</td><td>19342</td><td>other material</td></tr><tr><td>surgical head cap 2x</td><td>Lohmann &amp;amp; Rauscher, Regensdorf, Germany</td><td>17427</td><td>other material</td></tr><tr><td>surgical face mask 2x</td><td>Lohmann &amp;amp; Rauscher, Regensdorf, Germany</td><td>11983</td><td>other material</td></tr><tr><td>Other surgical material</td><td></td><td></td><td></td></tr><tr><td>implant</td><td>FEG Textiltechnik GmbH, Aachen, Germany</td><td>NA</td><td>purposely designed implant</td></tr><tr><td>3/0 polypropylene suture</td><td>Prolene, Ethicon, Diegem, Belgium</td><td>8762H</td><td>suture material</td></tr><tr><td>3/0 polygecaprone suture</td><td>Vicryl, Ethicon</td><td>J311H</td><td>suture material</td></tr><tr><td>gauze swabs 10 x 10 cm 10x, 12-ply</td><td>Lohmann &amp;amp; Rauscher, Regensdorf, Germany</td><td>11574</td><td>other material</td></tr><tr><td>syringe 10 mL</td><td>Becton Dickinsosn S.A., Madrid, Spain</td><td>300613</td><td>aqua-dissection</td></tr><tr><td>needle 16 gauge</td><td>Terumo, Leuven, Belgium</td><td>NN-2238R</td><td>aqua-dissection</td></tr><tr><td>Surgical equipment:</td><td></td><td></td><td></td></tr><tr><td>blade no.22</td><td>Fine science isntruments, Heidelberg, Germany</td><td>10022-00</td><td>surgical instruments</td></tr><tr><td>Allis tissue forceps 1x</td><td>Fine science isntruments, Heidelberg, Germany</td><td>11091-15</td><td>surgical instruments</td></tr><tr><td>Standart pattern forceps 1 x 2 theeth 1x</td><td>Fine science isntruments, Heidelberg, Germany</td><td>11023-14</td><td>surgical instruments</td></tr><tr><td>Standart pattern forceps straight serrated 1x</td><td>Fine science isntruments, Heidelberg, Germany</td><td>11000-14</td><td>surgical instruments</td></tr><tr><td>Scalpel handle 1x</td><td>Fine science isntruments, Heidelberg, Germany</td><td>10004-13</td><td>surgical instruments</td></tr><tr><td>Halstead-Mosquito forceps 2x</td><td>Fine science isntruments, Heidelberg, Germany</td><td>13008-12</td><td>surgical instruments</td></tr><tr><td>Standart pattern scissors 1x</td><td>Fine science isntruments, Heidelberg, Germany</td><td>14001-14</td><td>surgical instruments</td></tr><tr><td>Metzenbaum scissors 1x</td><td>Fine science isntruments, Heidelberg, Germany</td><td>14016-18</td><td>surgical instruments</td></tr><tr><td>Crile Wood needle holder 1x</td><td>Fine science isntruments, Heidelberg, Germany</td><td>12003-15</td><td>surgical instruments</td></tr><tr><td>Kell forceps 1x</td><td>Fine science isntruments, Heidelberg, Germany</td><td>13018-14</td><td>surgical instruments</td></tr><tr><td>Long Starr Self-Retaining Retractor with eight 5 mm sharp stay hooks</td><td>Cooper Surgical, Tumbull, USA</td><td>3704</td><td>surgical instruments</td></tr><tr><td>Heaney Simon Vaginal Retractor</td><td>Medical supplies &amp;amp; equipments co., Katy, Texas, USA</td><td>403-129FSI</td><td>surgical instruments</td></tr><tr><td>Trocar (Insnare)</td><td>Bard, West Sussex, United Kingdom</td><td>NA</td><td>any trocar on market for transvaginal mesh implantation</td></tr><tr><td>Medication:</td><td></td><td></td><td></td></tr><tr><td>amoxilicilline clavulanate 1000 mg / 300 mL (Ampiciline)</td><td>GSK, Wavre, Belgium</td><td>NA</td><td>antibiotics</td></tr><tr><td>buprenorfin 0.3 mg/mL + chlorocresol 1.35 mg/mL (Vetregesic)</td><td>Ecuphar, Oostkamp, Belgium</td><td>NA</td><td>analgesia</td></tr><tr><td>ketamin HCL 100 mg/mL (Ketamine 1000)</td><td>Ceva Sante Animale, Brussels, Belgium</td><td>NA</td><td>anesthesia</td></tr><tr><td>isoflurane (IsoFlo)</td><td>Abbott Laboratories Ltd, Maidenhead, Berkshire, UK</td><td>NA</td><td>anesthesia</td></tr><tr><td>polyvidone iodium 7.5% (Braunol)</td><td>B. Braun Medical, Machelen, Belgium</td><td>NA</td><td>local desinfection</td></tr><tr><td>saline solution 500 mL</td><td>B. Braun Medical, Machelen, Belgium</td><td>NA</td><td>aqua-dissection</td></tr><tr><td>Xxylazine HCl , 1 mL/50 kg&amp;nbsp;</td><td>Vexylan, Ceva Sante Animale, Belgium</td><td>NA</td><td>premedication</td></tr><tr><td>atropine Sulfate 15 mg/mL (),</td><td>Viatris, Belgium</td><td>NA</td><td>premedication</td></tr></tbody></table>

transvaginal mesh insertion in the ovine model

该方案描述了使用单一的阴道切口技术将丝网插入到绵羊的直肠阴道隔膜中，在有和没有针刺引导下插入锚固臂。可汗绵羊进行直肠阴道隔膜的解剖，然后插入有或没有四个锚定臂的植入物，两者设计成适合羊的解剖结构。使用套管针和"外在"技术将锚固臂放置在适当位置。颅臂通过闭孔器，小腿肌和内收肌肌肉。尾部臂固定在骶骨韧带附近，通过尾骨肌肉。这种技术允许模仿在患有盆腔器官脱垂的妇女中进行的外科手术。解剖空间和元素容易识别。手术最关键的部分是插入颅内套管针，可以很容易地穿透腹膜腔或周围的盆腔器官。氏可以通过更广泛的腹膜后解剖和更多侧向引导套管针来避免。这种方法仅用于大型动物模型中的新型植入物的实验测试，因为临床上目前没有使用针灸引导插入。

管理在更长的观察设置 在外科手术之后，可以插入阴道包装（在手术后立即插入阴道中的盐水溶液浸泡的纱布包装）24小时以固定植入物位置。羊应该放在恢复笼中，其呼吸功能直到完全恢复。后来，它可以放置在稳定的羊，并允许其自由移动和喝酒， 自由进食 。阴道包装，如果存在，必须在手术后24小时取出。绵羊应在术后至少三天内接受镇痛药（丁丙诺芬和氯甲酚，1 mL，im）。在术后第一周，每天应检查动物，然后每周进行一次实验。 外科杂志可行性在手术过程中，任何动物的网状插入都没有问题。直肠阴道隔膜和阴道间隙解剖几乎没有出血。通过解剖可以确定闭孔孔的内侧。此外，套针针插入是直接的，其中较柔顺的肌肉和单个肌肉的更强的筋膜之间的抵抗力有所不同。虽然通过肌肉的初始轨迹较少受到控制，但一旦与闭孔肌接触，套管针的尖端便可触及。没有并发症或障碍物放置背部手臂。 后续解剖中的网格定位和结果 调查适当定位使用锚定臂的植入物被认为更难实现，通过静脉内注射1.0mL的盐酸布替米星 - 盐酸丁卡因混合物将三只动物安乐死。在安乐死之后，仔细解剖手术区域以探查网状位置和通过相关解剖结构插入网状物的手臂的效果。用标尺测量腹壁从闭孔动脉和神经到内部阴部血管的最短距离，并研究其与肛提肌腱弧的关系。确定了阴部神经和阴内动脉，并用尺子测量距离。 任何一只母羊都没有出现相关的出血，也没有手术中的神经，肠道或膀胱损伤。在第一只羊中，颅臂通过尾部asp不过，肠道保持不变。在下一只羊中，通过将套管针的尖端更远地离开死胡同，避免了这种情况。在以前描述的解剖结构中识别其他手臂通道，远离骨盆神经和血管。颅臂通过封闭孔的尾部（ 图4 ， 图 A）。套管针的入口点位于肛提肌腱弧的水平位置，2至2.5厘米尾到封闭管和闭孔器和神经（ 图4 ， 图 B）。一旦在阴道狭窄处，手臂位于阴部动脉和静脉的腹侧1至1.5厘米，阴道动脉侧面1厘米。尾部手臂通过1厘米的尾部到达广泛的骶骨韧带的尾部，直接通过尾骨肌。在那个地方，任何附近都没有主要的船只或神经。阴部神经位于o在骶骨韧带的尾部的内表面。 将网格的中央部分放置平坦，其颅部分沿腹部隔膜向下逐渐伸展到尾部尾部和尾部的尾部。没有发生直肠穿孔（ 图4 ， 图 D）。 <img alt="图4" src="/files/ftp_upload/55706/55706fig4.jpg" /> 图4. 解剖解剖对盆腔血液透析 。 A:去除顶叶腹膜后腹膜后脂肪组织的外侧骨盆侧壁。耻骨（P）位于图的顶部，阴道（V）被中间移动以显露颅臂（全箭头）的过程。手臂通过肛提肌的腱弧。绿色针脚对应于内部p的位置udendal血管，蓝色的针标记阴道动脉，黄色的针被放置在提肛ani。 B:取下手臂后的侧方骨盆侧壁（课程标有黄线）。入口处位于封闭孔的尾部（标有四个蓝色针脚）。闭孔神经和血管（红线）通过其颅骨。内部阴部和阴道血管（分别为绿色和蓝色线）为背侧背部。也指示闭塞肌（OM）和提肌肛肌（LAM）。 C:通过大腿内侧肌肉的手臂（开放箭头）的过程。小腿肌被中间移动以通过半腱肌和内收肌大肌显示手臂的过程。 D:植入物的中心部分（开放箭头）位于阴道（V），顶叶腹膜和直肠（R）之间。 <table fo:keep-together.within-page="1" fo:keep-with-next.within页= "总是"&gt; <tbody><tr><td></td><td rowspan="2"> 矩形网格 </td><td colspan="2"> 胳膊网 </td></tr><tr><td></td><td> 中央身体 </td><td> 武器 </td></tr><tr><td> 尺寸（mm x mm） </td><td> 30 x 40 </td><td> 30 x 40 </td><td> 10 x 150 </td></tr><tr><td> 厚度（mm） </td><td> 0.54 </td><td> 0.54 </td><td> 0.7 </td></tr><tr><td> 重量（g / m 2 ） </td><td> 83 </td><td> 83 </td><td> 73 </td></tr><tr><td> 孔径（mm 2 ） </td><td> 2.5 x 2.5 </td><td> 2.5 x 2.5 </td><td> 1.0 x 1.4 </td></tr><tr><td> 刚度（N / mm） </td><td> 0.3 </td><td> 0.3 </td><td> 14.7 </td></tr><tr><td> 各向异性指数60; </td><td> 1.3 </td><td> 1.3 </td><td> 7.5 </td></tr></tbody></table> 表1:干料特性。 该表显示了矩形网格和具有锚固臂的网格的材料特性。刚度和各向异性指数由Maurer 等[ 18]得到 。

Watch this Scientific Journal Video about Transvaginal Mesh Insertion in the Ovine Model at JoVE.com

Transvaginal Mesh Insertion in the Ovine Model

该方案描述了使用单一阴道切口技术在羊直肠阴道隔膜中进行网状植入，有无套管针引导插入锚固臂。

经阴道网格插入在牛模型中

This protocol describes mesh insertion into the rectovaginal septum in sheep using a single vaginal incision technique, with and without the trocar-guided ...

transvaginal-mesh-insertion-in-the-ovine-model

Universidad San Sebastian

Research

JoVE Journal

Medicine

15.9K 次观看.  Clinical Specialties Research Groups, Faculty of Medicine, KU Leuven. 该方案描述了使用单一阴道切口技术在羊直肠阴道隔膜中进行网状植入，有无套管针引导插入锚固臂。 

视频: Transvaginal Mesh Insertion in the Ovine Model

An Orthotopic Endometrial Cancer Model with Retroperitoneal Lymphadenopathy Made From In Vivo Propagated and Cultured VX2 Cells

Endometrial cancer is the most common gynecologic malignancy in North America and the incidence is rising worldwide. Treatment consists of surgery with or without adjuvant therapy depending on lymph node involvement as determined by lymphadenectomy. Lymphadenectomy is a morbid procedure, which has not been shown to have a therapeutic benefit in many patients, and thus a new method to diagnose lymph node metastases is required. To test novel imaging agents, a reliable model of endometrial cancer with retroperitoneal lymph node metastases is needed. The VX2 endometrial cancer model has been described frequently in the literature; however, significant variation exists with respect to the method of model establishment. Furthermore, no studies have reported on the use of cultured VX2 cells to create this model as only cells propagated in vivo have been previously used. Herein, we present a standardized surgical method and post-operative monitoring method for the establishment of the VX2 endometrial cancer model and report on the first use of cultured VX2 cells to create this model.

This protocol presents a standardized method to grow VX2 cells in culture and to create an orthotopic VX2 model of endometrial cancer with retroperitoneal lymph node metastases in rabbits. Orthotopic endometrial cancer models are important for the pre-clinical study of novel imaging modalities for the diagnosis of lymph node metastases.

矫形子宫内膜癌模型，具有从 Vivo 传播和培养的 VX2 细胞中制成的再生淋巴病

Endometrial cancer is the most common gynecologic malignancy in North America and the incidence is rising worldwide. Treatment consists of surgery with or ...

科研

癌症研究

Anterior Cervical Discectomy and Fusion in the Ovine Model

Anterior cervical discectomy and fusion (ACDF) is the most common surgical operation for cervical radiculopathy and/or myelopathy in patients who have failed conservative treatment1,5. Since the operation was first described by Cloward2 and Smith and Robinson6 in 1958, a variety refinements in technique, graft material and implants have been made3. In particular, there is a need for safe osteoinductive agents that could benefit selected patients. The ovine model has been shown to have anatomical, biomechanical, bone density and radiological properties that are similar to the human counterpart, the most similar level being C3/44. It is therefore an ideal model in which preclinical studies can be performed. In particular this methodology may be useful to researchers interested in evaluating different devices and biologics, including stem cells, for potential application in human spinal surgery.

This video demonstrates the technique of anterior cervical discectomy and fusion in the ovine model.

颈椎前路椎间盘切除术和融合在绵羊模型

Anterior cervical discectomy and fusion (ACDF) is the most common surgical operation for cervical radiculopathy and/or myelopathy in patients who have ...

生物学

Methods for Studying Uterine Contributions to Pregnancy Establishment in an Ovariectomized Mouse Model

For pregnancy to be established, a viable blastocyst must successfully interact with a receptive uterine lining (endometrium) to facilitate implantation and placenta formation and enable ongoing pregnancy. The limitations to pregnancy success caused by embryonic defects are well known and have been largely overcome in recent decades with the rise of in vitro fertilization (IVF) and assisted reproductive technologies. As yet, however, the field has not overcome the limitations caused by an inadequately receptive endometrium, thus resulting in stagnating IVF success rates. Ovarian and endometrial functions are closely intertwined, as hormones produced by the ovary are responsible for the endometrium's menstrual cyclicity. As such, when using rodent models of pregnancy, it can be difficult to ascertain whether an observed result is due to an ovarian or uterine deficit. To overcome this, an ovariectomized mouse model was developed with embryo transfer or artificial decidualization to allow the study of uterine-specific contributions to pregnancy. This article will provide instructions on how to perform ovariectomy and offer insights into various techniques for supplying exogenous hormones to support successful artificial decidualization or pregnancy following embryo transfer from healthy donors. These techniques include subcutaneous injection, slow-release pellets, and osmotic mini pumps. The key advantages and disadvantages of each method will be discussed, enabling researchers to choose the best study design for their specific research question.

Pregnancy establishment is a dynamic process involving complex embryo and uterine crosstalk. The precise contributions of the maternal uterine environment to these processes remain an active area of investigation. Here, detailed protocols are provided to aid in designing in vivo animal models to address these research questions.

研究卵巢切除小鼠模型中子宫对妊娠建立的贡献的方法

For pregnancy to be established, a viable blastocyst must successfully interact with a receptive uterine lining (endometrium) to facilitate implantation ...

发育生物学

In Vivo Real-Time Study of Drug Effects on Carotid Blood Flow in the Ovine Fetus

The ability of an organism to maintain a constant blood flow to the brain in response to sudden surges in systemic blood pressure (BP) is known as cerebral autoregulation (CAR), which occurs in the carotid artery. In contrast to full-term neonates, preterm neonates are unable to reduce the cerebral blood flow (CBF) in response to increased systemic BP. In preterm neonates, this exposes the fragile cerebral vessels to high perfusion pressures, leading to their rupture and brain damage. Ex vivo studies using wire myography have demonstrated that carotid arteries from near-term fetuses constrict in response to the activation of adrenergic alpha1 receptors. This response is blunted in the preterm fetus. Thus, to examine the role of alpha1-AR in vivo, presented here is an innovative approach to determine the effects of drugs on a carotid arterial segment in vivo in an ovine fetus during the developmental progression of gestation. The presented data demonstrate the simultaneous measurement of fetal blood flow and blood pressure. The perivascular delivery system can be used to conduct a long-term study over several days. Additional applications for this method could include viral delivery systems to alter the expression of genes in a segment of the carotid artery. These methods could be applied to other blood vessels in the growing organism in utero as well as in adult organisms.

In Vivo Real-Time Study of Drug Effects on Carotid Blood Flow in the Ovine Fetus

The present protocol describes a method to deliver drugs and gene expression-modifying agents perivascularly in an in utero developing fetus. Importantly, the effect of drugs/agents on blood flow can be measured with the progression of pregnancy.

体内 药物对产卵胎颈动脉血流影响的实时研究

The ability of an organism to maintain a constant blood flow to the brain in response to sudden surges in systemic blood pressure (BP) is known as ...

Ovine Lumbar Intervertebral Disc Degeneration Model Utilizing a Lateral Retroperitoneal Drill Bit Injury

Intervertebral disc degeneration is a significant contributor to the development of back pain and the leading cause of disability worldwide. Numerous animal models of intervertebral disc degeneration have been developed. The ideal animal model should closely mimic the human intervertebral disc with regard to morphology, biomechanical properties and the absence of notochordal cells. The sheep lumbar intervertebral disc model fulfils these criteria. We present an ovine model of intervertebral disc degeneration utilizing a drill bit injury through a lateral retroperitoneal approach. The lateral approach significantly reduces the incision and potential morbidity associated with the traditional anterior approach to the ovine spine. Utilization of a drill-bit method of injury affords the ability to produce a consistent and reproducible injury, of precise dimensions, that initiates a consistent degree of intervertebral disc degeneration. The focal nature of the annular and nucleus pulposus defect more closely mimics the clinical condition of focal intervertebral disc herniation. Sheep recover rapidly following this procedure and are typically mobile and eating within the hour. Intervertebral disc degeneration ensues and sheep undergo necropsy and subsequent analysis at periods from eight weeks. We believe that the drill bit injury model of intervertebral disc degeneration offers advantages over more conventional annular injury models.

Intervertebral disc degeneration is a significant contributor to back pain and a leading cause of disability worldwide. Numerous animal models of intervertebral disc degeneration exist. We demonstrate an ovine model of intervertebral disc degeneration, utilizing a drill bit, which achieves a consistent disc injury and reproducible level of disc degeneration.

Ovine Lumbar椎间盘退变模型利用横向腹膜后位钻伤

Intervertebral disc degeneration is a significant contributor to the development of back pain and the leading cause of disability worldwide. Numerous ...

医学

Intravitreal Injections in the Ovine Eye

There are several methods for the delivery of therapeutic agents to the retina, including intravitreal (IVT), subretinal, suprachoroidal, periocular, or topical administration. IVT drug delivery involves an injection into the vitreous humor of the eye, a gelatinous substance that fills the posterior chamber of the eye and maintains the shape of the eye globe. Although the IVT route is less specifically targeted than subretinal delivery, it is much less invasive and is widely used in clinical settings for a range of ocular diseases. 
We previously demonstrated the efficacy of intravitreal delivery of an adeno-associated virus (AAV)-mediated gene therapy product (AAV9.CLN5) in sheep with a naturally occurring CLN5 form of neuronal ceroid lipofuscinosis (NCL). Affected sheep received IVT gene therapy in one eye, with the other untreated eye serving as an internal control. Retinal structure and function were maintained in the treated eye up to 15 months after treatment, while the untreated eye displayed progressively declining function and severe atrophy during postmortem examination. Based on the sheep studies, the CLN5 gene therapy product was cleared as a candidate investigational new drug (IND) by the United States Food and Drug Administration in September 2021. This paper details the surgical protocol for IVT delivery of a therapeutic viral vector to the ovine eye.

Intravitreal injections were performed in the sheep eye with the aim of delivering viral-mediated gene therapy to the retina.

绵羊眼玻璃体内注射

There are several methods for the delivery of therapeutic agents to the retina, including intravitreal (IVT), subretinal, suprachoroidal, periocular, or ...

Development of a Uterosacral Ligament Suspension Rat Model

Pelvic organ prolapse (POP) is a common pelvic floor disorder (PFD) with the potential to significantly impact a woman's quality of life. Approximately 10%-20% of women undergo pelvic floor repair surgery to treat prolapse in the United States. PFD cases result in an overall $26.3 billion annual cost in the United States alone. This multifactorial condition has a negative impact on the quality of life and yet the treatment options have only dwindled in the recent past. One common surgical option is uterosacral ligament suspension (USLS), which is typically performed by affixing the vaginal vault to the uterosacral ligament in the pelvis. This repair has a lower incidence of complications compared to those with mesh augmentation, but is notable for a relatively high failure rate of up to 40%. Considering the lack of standard animal models to study pelvic floor dysfunction, there is an urgent clinical need for innovation in this field with a focus on developing cost-effective and accessible animal models. In this manuscript, we describe a rat model of USLS involving a complete hysterectomy followed by fixation of the remaining vaginal vault to the uterosacral ligament. The goal of this model is to mimic the procedure performed on women to be able to use the model to then investigate reparative strategies that improve the mechanical integrity of the ligament attachment. Importantly, we also describe the development of an in situ tensile testing procedure to characterize interface integrity at chosen time points following surgical intervention. Overall, this model will be a useful tool for future studies that investigate treatment options for POP repair via USLS.

Pelvic organ prolapse affects millions of women worldwide and yet some common surgical interventions have failure rates as high as 40%. The lack of standard animal models to investigate this condition impedes progress. We propose the following protocol as a model for uterosacral ligament suspension and in vivo tensile testing.

子宫骶韧带悬浮大鼠模型的开发

Pelvic organ prolapse (POP) is a common pelvic floor disorder (PFD) with the potential to significantly impact a woman's quality of life. Approximately ...

Chronic Ovine Model of Right Ventricular Failure and Functional Tricuspid Regurgitation

The pathophysiology of severe functional tricuspid regurgitation (FTR) associated with right ventricular dysfunction is poorly understood, leading to suboptimal clinical results. We set out to establish a chronic ovine model of FTR and right heart failure to investigate the mechanisms of FTR. Twenty adult male sheep (6-12 months old, 62 &#177; 7 kg) underwent a left thoracotomy and baseline echocardiography. A pulmonary artery band (PAB) was placed and cinched around the main pulmonary artery (PA) to at least double the systolic pulmonary artery pressure (SPAP), inducing right ventricular (RV) pressure overload and signs of RV dilatation. PAB acutely increased the SPAP from 21 &#177; 2 mmHg to 62 &#177; 2 mmHg. The animals were followed for 8 weeks, symptoms of heart failure were treated with diuretics, and surveillance echocardiography was used to assess for pleural and abdominal fluid collection. Three animals died during the follow-up period due to stroke, hemorrhage, and acute heart failure. After 2 months, a median sternotomy and epicardial echocardiography were performed. Of the surviving 17 animals, 3 developed mild tricuspid regurgitation, 3 developed moderate tricuspid regurgitation, and 11 developed severe tricuspid regurgitation. Eight weeks of pulmonary artery banding resulted in a stable chronic ovine model of right ventricular dysfunction and significant FTR. This large animal platform can be used to further investigate the structural and molecular basis of RV failure and functional tricuspid regurgitation.

Right ventricular failure and functional tricuspid regurgitation are associated with left-sided heart disease and pulmonary hypertension, which contribute significantly to morbidity and mortality in patients. Establishing a chronic ovine model to study right ventricular failure and functional tricuspid regurgitation will help in understanding their mechanisms, progression, and possible treatments.

右心室衰竭和功能性三尖瓣反流的慢性绵羊模型

The pathophysiology of severe functional tricuspid regurgitation (FTR) associated with right ventricular dysfunction is poorly understood, leading to ...

<meta charset="utf8"></head><body>优化克隆猪的输卵管胚胎移植

Embryo Transfer Surgery via Laparotomy in Gilts

This protocol aims to demonstrate the surgical technique for transferring cloned pig embryos to the oviduct, a method widely used in the production of genetically modified pigs for biomedical research. Nine gilts underwent hormonal synchronization and laparotomy for the transfer of cloned embryos produced by somatic cell nuclear transfer (SCNT) at stages of up to 4 cells on day 2 to the oviduct. Gestational diagnosis was conducted via ultrasound examination 30 days post-transfer surgery. Six out of the nine operated gilts exhibited signs of pregnancy on ultrasound examination. However, as there was no progression in fetal development as assessed by echography, the gilts underwent necropsy at 60 days for the collection of biological material and assessment of the reproductive system. Adhesions were observed in the uterine horns, ovaries, and oviducts. From the uterine lumen of two of the euthanized gilts, one and four embryonic structures with gestational ages ranging between 12 to 20 days were obtained. Despite the absence of live piglets, likely attributed to the low-efficiency rate of transferring cloned pig embryos, which is influenced by various factors, including the number and quality of transferred embryos, the presented surgical technique proved to be rapid and safe.

<meta charset="utf8"></head><body>作者聚焦:输卵管克隆猪胚胎移植的手术方法和结果

This protocol describes the application of the surgical technique used to transfer cloned pig embryos via laparotomy in gilts.

通过剖腹手术对后备母猪进行胚胎移植手术

This protocol aims to demonstrate the surgical technique for transferring cloned pig embryos to the oviduct, a method widely used in the production of ...

经阴道网格插入在牛模型中

摘要

探索更多视频

此视频中的章节

矫形子宫内膜癌模型，具有从 Vivo 传播和培养的 VX2 细胞中制成的再生淋巴病

颈椎前路椎间盘切除术和融合在绵羊模型

研究卵巢切除小鼠模型中子宫对妊娠建立的贡献的方法

体内药物对产卵胎颈动脉血流影响的实时研究

Ovine Lumbar椎间盘退变模型利用横向腹膜后位钻伤

绵羊眼玻璃体内注射

子宫骶韧带悬浮大鼠模型的开发

右心室衰竭和功能性三尖瓣反流的慢性绵羊模型

通过剖腹手术对后备母猪进行胚胎移植手术

通过剖腹手术对后备母猪进行胚胎移植手术

经阴道网格插入在牛模型中

摘要

探索更多视频

此视频中的章节

矫形子宫内膜癌模型，具有从 Vivo 传播和培养的 VX2 细胞中制成的再生淋巴病

颈椎前路椎间盘切除术和融合在绵羊模型

研究卵巢切除小鼠模型中子宫对妊娠建立的贡献的方法

体内 药物对产卵胎颈动脉血流影响的实时研究

Ovine Lumbar椎间盘退变模型利用横向腹膜后位钻伤

绵羊眼玻璃体内注射

子宫骶韧带悬浮大鼠模型的开发

右心室衰竭和功能性三尖瓣反流的慢性绵羊模型

通过剖腹手术对后备母猪进行胚胎移植手术

通过剖腹手术对后备母猪进行胚胎移植手术

体内药物对产卵胎颈动脉血流影响的实时研究