Name: the superficial inferior epigastric artery axial flap to study ischemic preconditioning effects in a rat model
Uploaded: 2023-01-27T14:45:00
Description: Watch this Scientific Journal Video about The Superficial Inferior Epigastric Artery Axial Flap to Study Ischemic Preconditioning Effects in a Rat Model at JoVE.com

这项工作由马萨诸塞州总医院（WGA）和Shriners Children's Boston（B.U，K.U，C.L.C）资助。Y.B和I.F.v.R由Shriners儿童医院资助（提案ID：#970280和#857829分别为）。

马萨诸塞州总医院机构动物护理和使用委员会批准了实验方案（IACUC-协议 #2022N000099）。作者遵循ARRIVE（动物研究： 报告体内 实验）指南清单进行这项工作。所有动物都按照国家卫生研究所实验动物护理和使用指南进行了人道护理。所有实验共使用12只雄性刘易斯大鼠（250-350g，8-10周龄）。
1. 动物制备
注意：大鼠代谢率高，脂肪储备有?...

<ol>
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O., Damsgaard, T. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ICG+angiography+in+immediate+and+delayed+autologous+breast+reconstructions:+peroperative+evaluation+and+postoperative+outcomes.">ICG angiography in immediate and delayed autologous breast reconstructions: peroperative evaluation and postoperative outcomes.</a> Journal of Plastic Surgery and Hand Surgery. 52 (5), 307-311 (2018).</li><li>K&#252;ntscher, M. V., 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=Ischemic+preconditioning+by+brief+extremity+ischemia+before+flap+ischemia+in+a+rat+model.">Ischemic preconditioning by brief extremity ischemia before flap ischemia in a rat model.</a> Plastic and Reconstructive Surgery. 109 (7), 2398-2404 (2002).</li><li>Liu, R. 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本文描述了在大鼠中收获的可重复的筋膜皮肤皮瓣模型，允许腹腔内评估。这种循序渐进的手术方案为研究小组提供了一个可靠的模型来测试不同的腹腔内膜方案。通过防止椎弓根以外的任何血管形成，该模型允许从伤口床和边缘研究皮瓣的新生血管形成。这项研究对POD5进行了连接，因为以前的研究已经观察到POD5-711，13，16?...

整形和重建手术在不断发展。肌肉、筋膜皮肤和穿支皮瓣的发展使得在降低发病率的同时提供更高质量的重建成为可能。将这种改进的解剖学知识与增强的技术技能相结合，重建外科医生可以在缺陷不接近任何局部解决方案时进行自由皮瓣移植。然而，虽然穿支皮瓣手术是目前重建手术中最先进的技术，但文献报道游离皮瓣转移的失败率为5%1，2，3，而椎弓根皮瓣重建的失败率高达20%4，5，6。当皮瓣的椎弓根受损时，会发生部分或全部皮瓣故障，因此必须不断寻求对当前技术的改进。提高皮瓣存活率的方法之一是促进其在伤口床上的新生血管形成，从而允许通过椎弓根以外的来源灌注。缺血性预处理（I.P.）最初已在心脏模型7中描述，表明暴露于受控缺血的器官在通过经历缺血诱导的新生血管而失去其主要血液供应后存活到更高程度。几位作者研究了这一基石原理，以优化临床前和临床模型中的皮瓣存活率8，9，10。
与其他提高皮瓣存活率的方法相比，该技术的优势在于其易于实施，包括血源的钳夹/脱夹测试。在大鼠模型中，以前的作者使用浅表下上腹部动脉（SIEA）皮瓣通过夹住主椎弓根11，12，13来研究腹腔膜外膜。尽管如此，该模型可能会遇到一些技术问题，并且文献缺乏描述良好的协议。
因此，这项工作旨在为研究人员提供大鼠SIEA皮瓣采购技术的详细描述，该技术对股血管进行了扩展解剖，以允许对轴向筋膜皮瓣模型进行腹腔研究。该模型保留了上腹部血管的完整性，而是操纵了更具弹性的股血管。我们分享我们的经验和工具，以改进对这种现象的研究，并提高该程序的可复制性。

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			<td>BD</td>
			<td>309628</td>
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			<td>3-0 Ethilon 18&#8221; Black Monofilament Nylon suture</td>
			<td>Ethicon</td>
			<td>ETH-663H</td>
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			<td>8-0 Ethilon 12&#8221; Black Monofilament Nylon suture</td>
			<td>Ethicon</td>
			<td>1716G</td>
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			<td>Adson Atraumatic Forceps</td>
			<td>Aesculap Surgical Instruments</td>
			<td>BD51R</td>
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			<td>Akorn Fluorescein Injection USP 10% Single Dose Vial 5 mL&#160;</td>
			<td>Akorn</td>
			<td>17478025310</td>
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			<td>Betadine Solution 5% Povidone-Iodine Antiseptic Microbicide</td>
			<td>PBS Animal Health</td>
			<td>11205</td>
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			<td>Bipolar Cords</td>
			<td>ASSI</td>
			<td>ASSI.ATK26426</td>
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			<td>Buprenorphine Hydrochloride Injection</td>
			<td>PAR Pharmaceutical</td>
			<td>3003406C</td>
			<td>This concentration needs to be diluted for rodents.</td>
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			<td>Depilatory product &#8211; Nair Hair remover lotion</td>
			<td>Nair</td>
			<td>NC0132811</td>
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			<td>Ear tag applier</td>
			<td>World Precision Instruments</td>
			<td>NC0038715</td>
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			<td>Gauze Sponges</td>
			<td>Curity</td>
			<td>6939</td>
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			<td>Isoflurane Auto-Flow Anesthesia Machine&#160;</td>
			<td>E-Z Systems</td>
			<td>EZ-190F</td>
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			<td>Isoflurane, USP</td>
			<td>Patterson Veterinary</td>
			<td>1403-704-06</td>
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			<td>Jewelers Bipolar Forceps Non-Stick 11 cm, straight pointed tip, 0.25 mm tip diameter</td>
			<td>ASSI</td>
			<td>ASSI.BPNS11223</td>
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			<td>Lone Star elastic stays</td>
			<td>Cooper Surgical</td>
			<td>3311-1G</td>
			<td></td>
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			<td>Lone star Self-retaining retractor&#160;</td>
			<td>Cooper Surgical</td>
			<td>3304G</td>
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			<td>Metronidazole tablets USP</td>
			<td>Teva</td>
			<td>500111-333-06</td>
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			<td>Micro spring handle scissors&#160;</td>
			<td>AROSurgical</td>
			<td>11.603.14</td>
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			<td>Microscope (surgical)</td>
			<td>Leica</td>
			<td>M525 F40</td>
			<td></td>
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			<td>Microsurgical clamp applying forceps</td>
			<td>Ambler Surgical</td>
			<td>31-906</td>
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			<td>Microsurgical clamps (x2)&#160;</td>
			<td>Millennium Surgical&#160;</td>
			<td>18-B1V</td>
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			<td>Microsurgical Dumont #4 forceps&#160;</td>
			<td>Dumont Swiss made&#160;</td>
			<td>1708-4TM-PO</td>
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			<td>Microsurgical needle holder</td>
			<td>ASSI</td>
			<td>B-14-8</td>
			<td></td>
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			<td>Needle holder</td>
			<td>World Precision Instruments</td>
			<td>501246</td>
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			<td>Nosecone for Anesthesia&#160;</td>
			<td>World Precision Instruments</td>
			<td>EZ-112</td>
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			<td>GNU Image Manipulation Program v2.10</td>
			<td>GIMP</td>
			<td>GNU Open licence</td>
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			<td>BD</td>
			<td>305109</td>
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			<td>Ragnell Scissors&#160;</td>
			<td>Roboz Surgical</td>
			<td>RS-6015</td>
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			<td>Rimadyl (carprofen)</td>
			<td>Zoetis</td>
			<td>10000319</td>
			<td>This concentration needs to be diluted for rodents</td>
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			<td>Braintree Scientific&#160;</td>
			<td>NC9263311</td>
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			<td>National Band &#38; Tag Company</td>
			<td>Style 1005-1</td>
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			<td>Peco Services Ltd</td>
			<td>69023</td>
			<td></td>
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			<td>Dynarex Corporation</td>
			<td>4410</td>
			<td></td>
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			<td>Sterile water for injection and irrigation&#160;</td>
			<td>Hospira</td>
			<td>0409488724-1</td>
			<td></td>
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			<td>BD</td>
			<td>930415</td>
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			<td>UV lamp</td>
			<td>UVP</td>
			<td>UVL-56</td>
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			<td>Webcol Alcohol prep pads&#160;</td>
			<td>Simply Medical</td>
			<td>5110</td>
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</table>

the superficial inferior epigastric artery axial flap to study ischemic preconditioning effects in a rat model

筋膜皮肤皮瓣（FCF）已成为整形和重建手术中复杂缺损重建的金标准。这种肌肉保留技术允许转移血管化组织以覆盖任何大的缺陷。FCF可用作带蒂皮瓣或自由皮瓣;然而，在文献中，有蒂FCF和游离FCF的失败率高于5%，为这些技术和该领域的进一步知识扩展留下了改进的空间。缺血性预处理（I.P.）已被广泛研究，但IP方案的机制和优化尚未确定。这种现象在整形和重建手术中确实没有得到很好的探索。本文介绍了一个手术模型，用于研究大鼠轴向筋膜皮瓣模型中的腹腔方案，描述了如何安全可靠地评估腹膜对皮瓣存活的影响。本文介绍了完整的外科手术，包括提高该模型可靠性的建议。目的是为研究人员提供一个可重复和可靠的模型来测试各种缺血性预处理方案，并评估它们对皮瓣生存能力的影响。

所有皮瓣在POD5上都是可行的，仅通过SIEA显示良好的血管形成。 图5 显示了静脉注射荧光素前后的皮瓣，显示出完全的血管化。
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/64980/64980fig05.jpg"> 图 5：立即静脉注射荧光素血管造影 （POD0）。 该评估显示皮瓣仅由SIEA形成血管化。绿色荧光?...

Watch this Scientific Journal Video about The Superficial Inferior Epigastric Artery Axial Flap to Study Ischemic Preconditioning Effects in a Rat Model at JoVE.com

The Superficial Inferior Epigastric Artery Axial Flap to Study Ischemic Preconditioning Effects in a Rat Model

该协议描述了收获，缝合和监测大鼠的筋膜皮肤皮瓣，通过钳夹和结扎股血管，可以很好地观察和操纵通过浅表下上腹部血管的血流。这对于涉及缺血性预处理的研究至关重要。

浅表下上腹部动脉轴向皮瓣研究大鼠模型中缺血预处理效应

Fasciocutaneous flaps (FCF) have become the gold standard for complex defect reconstruction in plastic and reconstructive surgery. This muscle-sparing ...

该手术方案为研究筋膜皮瓣缺血性预处理方案提供了一个可靠的模型，以改善这些手术的结果。该协议易于学习，可重复且可靠。与其他方案的主要区别在于使用股血管来引发缺血性预处理，避免伤害小血管。它最终可以导致优化游离和带蒂皮瓣重建的缺血预处理方案，以提高皮瓣生存能力并减少皮瓣脆弱性的关键时期。该方法有助于重建手术领域，并非常适合缺血再灌注损伤的研究。首先，将适当镇静的动物置于仰卧位。将腹部从腹股沟折痕下方剃到剑突水平以上。在剃须部位涂抹脱毛霜，等待五分钟后再清洁。使用无菌皮笔和尺子，首先标记动物腹部的中线，然后标记腹股沟折痕。在做切口之前，通过画一个椭圆形或矩形来指示动物身体上所需皮瓣的位置，垂直长度可达六厘米，水平宽度约为三厘米，从腹股沟折痕向颅骨延伸。现在画五六个垂直于襟翼极限的等距标记。然后用一把拉格内尔剪刀，在腹股沟折痕上做一个三到四厘米的纵向切口。一定要向上拉皮肤，以免损坏血管。使用 4 个珠宝商显微外科镊子暴露和识别股骨和上腹部血管，方法是打开和关闭镊子以分离筋膜并进入腹股沟脂肪垫下方的血管。使用腹股沟切口使用拉格内尔剪刀开始皮瓣切口。注意破坏腹部肌肉上的皮肤和结缔组织的整个厚度。当皮瓣的尖端最初从周围皮肤中释放出来时，通过使用 Ragnell 剪刀尖端将皮瓣与肌肉分开，同时烧灼皮瓣周围的任何穿支血管和真皮丛血管，从而从远端到近端进行皮瓣获取。一旦皮瓣完全收获，旨在通过轻轻向上提起皮瓣以观察血管，用皮瓣包裹上腹部浅下动脉或SIEA的整个分支。接下来，将皮瓣内侧和外侧下侧的脂肪垫分开。使用双极烧灼器仔细烧灼靠近切口边界的脂肪垫，而不会伤害浅表下上腹部蒂。在完成皮瓣收获后进行股血管准备之前，通过静脉向动物注射单剂量的17.5国际单位肝素钠。为了更好地观察血管，请将动物放置在孤星自留性牵开器中。现在在具有40倍放大倍率的手术显微镜下工作，使用几个4个显微手术钳在近端和远端解剖股血管，直到出现浅表上腹部血管。在股骨远端血管上，清洁筋膜并轻轻地将神经从动脉和静脉中解放出来。使用 8-0尼龙缝合线，通过绕过动脉和静脉下方的显微手术针架来结扎股骨远端血管，在上腹部血管出现后立即扣住缝合线并绑住这些血管，在椎弓根起源后留出一毫米的距离。在股骨近端血管上，重复清洁结缔组织的相同过程。但是，将动脉和静脉彼此分开，以便有效地钳夹。为了诱发间歇性缺血，将显微手术夹分别放在每个股动脉近端和静脉上。缺血性损伤完成后，将皮瓣缝合到术前绘制的标记的原始位置。使用3-0尼龙在腹股沟折痕内侧，皮瓣周围，并在腹股沟折痕侧侧处缝合皮瓣。为了验证皮瓣的血液供应，使用与先前注射相同的技术和工具将0.25毫升10%无菌荧光素钠注射到静脉中。三分钟后，用366纳米的长波紫外灯照射，以显示与灌注区域相对应的荧光区域。闭合和验证后，沿缝合线铺上粉碎的甲硝唑，以防止自动残缺，并在同一区域喷洒液体绷带。为了在缺血性预处理期结束时从上腹部血液供应中去除皮瓣，沿皮瓣的下缘烧灼低于脂肪垫。术后第 0 天或 POD0 立即静脉注射荧光素血管造影显示仅通过 SIEA 使皮瓣完全血管化。组织，包括整个皮瓣桨，灌注良好。POD5和POD10上阴性对照的血管造影显示襟翼桨的完全活力。该组中所有未在饲养血管上进行结扎的动物都保持健康。对于阳性对照组，结扎后立即没有绿色荧光，表明皮瓣没有灌注，证明没有新生血管形成。这在POD10上得到证实，观察到85%的皮肤桨坏死。有趣的是，远端尖端是可行的和新生血管化的。POD10襟翼表面活力统计分析显示，阴性对照组存活率为99.5%，阳性对照组存活率为11.25%。一个旨在研究缺血预处理的示例实验组显示出13.67%的存活率，表明特定的缺血预处理方案在该模型中效率低下。两个主要步骤是在显微镜下收获皮瓣和血管解剖。最重要的建议是拥有高质量的微型仪器。下一个目标是通过改变缺血持续时间和节律来尝试不同的缺血预处理方案，以选择在早期血流中断后实现最佳皮瓣存活能力的方案。该技术通过为缺血融合损伤提供良好的模型，可以导致缺血预处理应用于现代重建手术和重建移植的新发现。

Research

JoVE Journal

Immunology and Infection

Murine Superficial Lymph Node Surgery

小鼠浅表淋巴结手术

In the field of immunology, to understand the progression of an immune response against a vaccine, an infection or a tumour, the response is often ...

科研

免疫与感染

Cutaneous Leishmaniasis in the Dorsal Skin of Hamsters: a Useful Model for the Screening of Antileishmanial Drugs

在仓鼠的背部皮肤：皮肤利什曼病Antileishmanial药物筛选的一个有用的模型

Traditionally, hamsters are experimentally inoculated in the snout or the footpad. However in these sites an ulcer not always occurs, measurement of ...

An In vitro Model to Study Heterogeneity of Human Macrophage Differentiation and Polarization

An In vitro Model to Study Heterogeneity of Human Macrophage Differentiation and Polarization

一个<em在体外</em&gt;模型研究人类巨噬细胞分化和极化的异质性

Monocyte-derived macrophages represent an important cell type of the innate immune system. Mouse models studying macrophage biology suffer from the ...

Non-Invasive Model of Neuropathogenic Escherichia coli Infection in the Neonatal Rat

Non-Invasive Model of Neuropathogenic Escherichia coli Infection in the Neonatal Rat

Neuropathogenic的非侵入型<em&gt;大肠杆菌</em&gt;感染乳鼠

Investigation of the interactions between animal host and bacterial pathogen is only meaningful if the infection model employed replicates the principal ...

Human Placental and Decidual Organ Cultures to Study Infections at the Maternal-fetal Interface

人胎盘及蜕膜器官培养研究在母胎界面感染

The placenta shows a large degree of interspecies anatomic variability. To best understand biology and pathophysiology of the human placenta, it is ...

Implementation of a Permeable Membrane Insert-based Infection System to Study the Effects of Secreted Bacterial Toxins on Mammalian Host Cells

基于插入渗透膜感染系统的实现研究分泌细菌病毒对哺乳动物宿主细胞的作用

Many bacterial pathogens secrete potent toxins to aid in the destruction of host tissue, to initiate signaling changes in host cells or to manipulate ...

Flow Cytometric Analysis of Particle-bound Bet v 1 Allergen in PM10

粒子结合Bet v 1的变应原PM10的流式细胞分析

Flow cytometry is a method widely used to quantify suspended solids such as cells or bacteria in a size range from 0.5 to several tens of micrometers in ...

Visualizing the Effects of Sputum on Biofilm Development Using a Chambered Coverglass Model

可视化痰生物膜发展影响使用腔室盖玻片型号

Biofilms consist of groups of bacteria encased in a self-secreted matrix. They play an important role in industrial contamination as well as in the ...

Effects of Exposure of Formaldehyde to a Rat Model of Atopic Dermatitis Induced by Neonatal Capsaicin Treatment

甲醛暴露对大鼠新生辣椒素致过敏性皮炎模型的影响

Atopic dermatitis is chronically relapsing pruritic eczema and prevails around the world especially in developed countries. Complex interactions between ...

Stenosis of the Inferior Vena Cava: A Murine Model of Deep Vein Thrombosis

下腔静脉狭窄: 深静脉血栓形成的小鼠模型

Deep vein thrombosis (DVT) and its devastating complication, pulmonary embolism, are a severe health problem with high mortality. Mechanisms of thrombus ...