Summary

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

Published: January 27, 2023
doi:

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%1,23而椎弓根皮瓣重建的失败率高达20%456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8,910

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111213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POD5-7111316?…

Declarações

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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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).

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