大鼠部分异位欣德利姆移植模型
1Division of Plastic and Reconstructive Surgery,Massachusetts General Hospital, Harvard Medical School, 2Vascularized Composite Allotransplantation Laboratory, Center for Transplantation Sciences,Massachusetts General Hospital, Harvard Medical School, 3Shriners Hospital for Children, 4Service de Chirurgie Plastique, Hôpital Européen Georges Pompidou, Assistance Publique-Hôpitaux de Paris (APHP),Université de Paris
Summary
本文介绍了大鼠部分异位骨质皮瓣移植方案及其中期随访的潜在结果。
Abstract
血管化复合异位移植 （VCA） 是患者在复杂组织缺陷后没有自体手术可能性的最高级重建选项。脸部和手部移植改变了毁容患者的生活，赋予他们新的审美和功能社会器官。尽管结果大有希望，但由于终生免疫抑制合并症和传染性并发症，VCA仍然表现不佳。大鼠是研究免疫途径和移植排斥机制 的活体 研究的理想动物模型。大鼠还广泛应用于新型复合组织移植保存技术，包括灌注和低温保存研究。大鼠VCA的模型必须可重复、可靠和高效，术后发病率和死亡率较低。异位肢体移植程序符合这些标准，比矫肢移植更容易执行。掌握啮齿动物显微外科模型需要在显微外科和动物护理方面有扎实的经验。据报道，这是大鼠部分异位骨皮瓣移植、术后结果和预防潜在并发症的方法的可靠和可重复的模型。
Introduction
在过去的二十年里， VCA 已经发展成为一种革命性的治疗患者谁遭受严重毁容，包括脸部1 ，上肢截肢2 ，3 ，和其他复杂的组织缺陷4 ， 5 。然而，终生免疫抑制的后果仍然阻碍这些复杂的重建手术的广泛应用。基础研究对于改进反排斥策略至关重要。增加VCA保存时间对于改善移植物流和增加捐赠者库也至关重要（因为VCA捐赠者必须比实体器官捐献者满足更多的标准，包括肤色、解剖大小、性别）。在此背景下，大鼠肢体移植被广泛应用于6、7、新耐受诱导方案8、保存研究9、10、11等免疫排斥的研究中。因此，这些 VCA 模型是 VCA 转化研究需要掌握的关键元素。
骨皮瓣在文献中被描述为研究8、12、13、14大鼠VCA的可靠模型。虽然正位全肢移植允许长期评估移植功能，但它是一个耗时的程序，与更高的术后发病率和死亡率14。相比之下，异位肢体移植模型是非功能性的，但能够对VCA进行可重复的研究。在开始大鼠VCA移植研究之前，可以可靠地预测术后的结果。本研究报告了大鼠的部分异位骨质皮瓣移植模型，其中包括在三周的随访期内可能出现的频繁可能的结果和术后并发症。
Protocol
所有动物都根据《国家卫生研究院护理和使用实验室动物指南》得到人道护理。机构动物护理和使用委员会（IACUC-协议2017N000184）和动物护理和使用审查办公室（ACURO）批准了所有动物协议。近亲繁殖的雄性刘易斯大鼠（250-400克）用于所有实验。 1. 手术 使用异黄素吸入麻醉刘易斯大鼠。诱导感应室中5%异黄素的麻醉，通过呼吸锥体保持1.5-3%异黄酮吸入麻醉。 手…
Representative Results
在这项单操作者研究中，进行了30例同步异位部分肢体移植。成功定义为术后第21天没有VCA失败或需要安乐死的并发症。移植物的正常演变表现在图3中。接受者部分肢体采购和移植插入的平均持续时间分别为35分钟和105分钟：平均缺血时间是105分钟。在随访过程中，发生两种类型的并发症（表1）早或晚。有的需要安乐死，有的需要抢救，都与兽医人员讨论过（<st…
Discussion
在文献15、16、17中描述了啮齿动物的矫肢移植模型：然而，他们需要神经修复，肌肉重新连接，和完美的骨合成股骨，这可能是一个非常困难的步骤。这些模型也与14号啮齿动物的发病率和死亡率较高有关，特别是在短期随访中，因为移植后肢的正常功能的恢复可能需要几个月的时间。然而?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
这项工作得到了助理国防部长办公室的支持，通过国会指导的医学研究计划，根据第一奖。W81XWH-17-1-0680。意见、解释、结论和建议是作者的意见，不一定得到国防部的赞同。
Materials
| 24 GA angiocatheter | BD Insyte Autoguard | 381412 | |
| 4-0 suture Black monofilament non absorbable suture | Ethicon | 1667 | Used to suture the E-collar to the back of the neck |
| 4-0 suture Coated Vicryl Plus Antibacterial | Ethicon | VCP496 | |
| Adson Tissue Forceps, 11 cm, 1 x 2 Teeth with Tying Platform | ASSI | ASSI.ATK26426 | |
| Bipolar cords | ASSI | 228000C | |
| Black Polyamide Monofilament USP 10-0, 4 mm 3/8c | AROSurgical | T04A10N07-13 | Used to perform the microvascular anastomoses |
| Buprenorphine HCl | Pharmaceutical, Inc | 42023-179-01 | |
| Dilating Forceps | Fine science tools (FST) | 18131-12 | |
| Dissecting Scissors 15 cm, Round Handle 8 mm diameter, Straight Slender Tapered Blade 7 mm, Lipshultz Pattern | ASSI | ASSI.SAS15RVL | |
| Double Micro Clamps 5.5 x 1.5 mm | Fine science tools (FST) | 18040-22 | |
| Elizabethan collar | Braintree Scientific | EC-R1 | |
| Forceps 13.5 cm long, Flat Handle, 9 mm wide Straight Tips 0.1 mm diameter (x2) | ASSI | ASSI.JFL31 | |
| Halsey Micro Needle Holder | Fine science tools (FST) | 12500-12 | |
| Heparin Lock Flush Solution, USP, 100 units/mL | BD PosiFlush | 306424 | |
| Isoflurane | Patterson Veterinary | 14043-704-06 | |
| Jewelers Bipolar Forceps Non Stick 11 cm, straight pointed tip, 0.25 mm tip diameter | ASSI | ASSI.BPNS11223 | |
| Lone Star Elastic Stays | CooperSurgical | 3314-8G | Used to retract the inguinal ligament for femoral vessels dissection |
| Lone Star Self-Retaining Retractors | CooperSurgical | 3301G | |
| Micro-Mosquito Hemostats | Fine science tools (FST) | 13010-12 | Used to retract the inguinal fat pad distally |
| Needle Holder, 15 cm Round Handle, 8 mm diameter, Superfine Curved Jaw 0.2 mm tip diameter, without lock | ASSI | ASSI.B1582 | |
| Nylon Suture Black Monolfilament 8-0, 6.5 mm 3/8c | Ethilon | 2808G | Used to ligate collateral branches on the femoral vessels |
| Offset Bone Nippers | Fine science tools (FST) | 16101-10 | |
| S&T Vascular Clamps 5.5 x 1.5 mm | Fine science tools (FST) | 00398-02 | |
| Walton scissors | Fine science tools (FST) | 14077-09 |
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Cite This Article
Goutard, M., Randolph, M. A., Taveau, C. B., Lupon, E., Lantieri, L., Uygun, K., Cetrulo Jr., C. L., Lellouch, A. G. Partial Heterotopic Hindlimb Transplantation Model in Rats. J. Vis. Exp. (172), e62586, doi:10.3791/62586 (2021).