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
1Vascularized Composite Allotransplantation Laboratory,Massachusetts General Hospital, 2Harvard Medical School, 3Department of Plastic, Reconstructive and Aesthetic Surgery, Rennes University Hospital Center (CHU de Rennes),University of Rennes 1, 4Plastic Surgery Research Laboratory,Massachusetts General Hospital, 5Shriners Children’s Boston, 6Center for Engineering in Medicine and Surgery,Massachusetts General Hospital, 7Department of Plastic Surgery and Hand Surgery,University Medical Center Utrecht, 8Department of Plastic, Reconstructive, and Aesthetic Surgery, Groupe Almaviva Santé,Clinique de l’Alma
Summary
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。
Abstract
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。
Introduction
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。
Protocol
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?…
Representative Results
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。 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?…
Discussion
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?…
Divulgations
The authors have nothing to disclose.
Acknowledgements
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）。
Materials
| 1 mL Syringe Luer-Lok Tip | BD | 309628 | |
| 3-0 Ethilon 18” Black Monofilament Nylon suture | Ethicon | ETH-663H | |
| 8-0 Ethilon 12” Black Monofilament Nylon suture | Ethicon | 1716G | |
| Adson Atraumatic Forceps | Aesculap Surgical Instruments | BD51R | |
| Akorn Fluorescein Injection USP 10% Single Dose Vial 5 mL | Akorn | 17478025310 | |
| Betadine Solution 5% Povidone-Iodine Antiseptic Microbicide | PBS Animal Health | 11205 | |
| Bipolar Cords | ASSI | ASSI.ATK26426 | |
| Buprenorphine Hydrochloride Injection | PAR Pharmaceutical | 3003406C | This concentration needs to be diluted for rodents. |
| Depilatory product – Nair Hair remover lotion | Nair | NC0132811 | |
| Ear tag applier | World Precision Instruments | NC0038715 | |
| Gauze Sponges | Curity | 6939 | |
| Isoflurane Auto-Flow Anesthesia Machine | E-Z Systems | EZ-190F | |
| Isoflurane, USP | Patterson Veterinary | 1403-704-06 | |
| Jewelers Bipolar Forceps Non-Stick 11 cm, straight pointed tip, 0.25 mm tip diameter | ASSI | ASSI.BPNS11223 | |
| Lone Star elastic stays | Cooper Surgical | 3311-1G | |
| Lone star Self-retaining retractor | Cooper Surgical | 3304G | |
| Metronidazole tablets USP | Teva | 500111-333-06 | |
| Micro spring handle scissors | AROSurgical | 11.603.14 | |
| Microscope (surgical) | Leica | M525 F40 | |
| Microsurgical clamp applying forceps | Ambler Surgical | 31-906 | |
| Microsurgical clamps (x2) | Millennium Surgical | 18-B1V | |
| Microsurgical Dumont #4 forceps | Dumont Swiss made | 1708-4TM-PO | |
| Microsurgical needle holder | ASSI | B-14-8 | |
| Needle holder | World Precision Instruments | 501246 | |
| Nosecone for Anesthesia | World Precision Instruments | EZ-112 | |
| Pixel analysis software | GNU Image Manipulation Program v2.10 | GIMP | GNU Open licence |
| PrecisionGlide Needle 27 G | BD | 305109 | |
| Ragnell Scissors | Roboz Surgical | RS-6015 | |
| Rimadyl (carprofen) | Zoetis | 10000319 | This concentration needs to be diluted for rodents |
| Scientific Elizabethan collar (e-collar) for Rats | Braintree Scientific | NC9263311 | |
| Small animal ear tag | National Band & Tag Company | Style 1005-1 | |
| Small Animal Heated Operating Table (Adjustable) | Peco Services Ltd | 69023 | |
| Sterile towel drape | Dynarex Corporation | 4410 | |
| Sterile water for injection and irrigation | Hospira | 0409488724-1 | |
| Surgical scrub – BD ChloraPrep Hi-Lite Orange 3 mL applicator with Sterile Solution | BD | 930415 | |
| UV lamp | UVP | UVL-56 | |
| Webcol Alcohol prep pads | Simply Medical | 5110 |
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Citer Cet Article
Berkane, Y., Alana Shamlou, A., Reyes, J., Lancia, H. H., Filz von Reiterdank, I., Bertheuil, N., Uygun, B. E., Uygun, K., Austen Jr., W. G., Cetrulo Jr., C. L., Randolph, M. A., Lellouch, A. G. The Superficial Inferior Epigastric Artery Axial Flap to Study Ischemic Preconditioning Effects in a Rat Model. J. Vis. Exp. (191), e64980, doi:10.3791/64980 (2023).