Name: taking the next step: a neural coaptation orthotopic hind limb transplant model to maximize functional recovery in rat
Uploaded: 2020-08-30T13:00:00
Description: Watch this Scientific Journal Video about Taking the Next Step: a Neural Coaptation Orthotopic Hind Limb Transplant Model to Maximize Functional Recovery in Rat at JoVE.com

这项工作由弗兰克尔基金会和西北纪念医院麦考密克·格兰特（恢复行动）资助。本出版物中报告的研究得到了国家卫生研究院国家普通医学研究所的支持，其编号为T32GM008152。这项工作得到了西北大学显微外科核心和行为表型核心的支持。

所有实验都是根据国家卫生研究院实验室动物护理和使用指南进行的，并经西北大学动物护理和使用委员会批准。具体程序根据协议 IS00001663 执行。
注：使用两株老鼠，刘易斯大鼠和八月哥本哈根x爱尔兰（ACI）大鼠。动物被分成三个治疗组：无免疫抑制的异丙类移植（ACI到刘易斯），常规免疫抑制的异丙移植植物（ACI到刘易斯），和异种移植（刘易斯对刘易斯或ACI到ACI）。L...

<ol>
	<li>Elliott, R. M., Tintle, S. M., Levin, L. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Upper+extremity+transplantation:+current+concepts+and+challenges+in+an+emerging+field.">Upper extremity transplantation: current concepts and challenges in an emerging field.</a> Current Reviews in Musculoskeletal Medicine. 7 (1), 83-88 (2014).</li><li>Petruzzo, P., 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=The+International+Registry+on+Hand+and+Composite+Tissue+Transplantation.">The International Registry on Hand and Composite Tissue Transplantation.</a> Transplantation. 90 (12), 1590-1594 (2010).</li><li>Shapiro, R. I., Cerra, F. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+model+for+reimplantation+and+transplantation+of+a+complex+organ:+the+rat+hind+limb.">A model for reimplantation and transplantation of a complex organ: the rat hind limb.</a> Journal of Surgical Research. 24 (6), 501-506 (1978).</li><li>Brandacher, G., Grahammer, J., Sucher, R., Lee, W. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+for+basic+and+translational+research+in+reconstructive+transplantation.">Animal models for basic and translational research in reconstructive transplantation.</a> Birth Defects Research Part C: Embryo Today. 96 (1), 39-50 (2012).</li><li>Furtmuller, G. J., 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=Orthotopic+Hind+Limb+Transplantation+in+the+Mouse.">Orthotopic Hind Limb Transplantation in the Mouse.</a> Journal of Visualized Experiments. (108), e53483 (2016).</li><li>Sucher, R., 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=Orthotopic+hind-limb+transplantation+in+rats.">Orthotopic hind-limb transplantation in rats.</a> Journal of Visualized Experiments. (41), e2022 (2010).</li><li>Horner, B. M., 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=In+vivo+observations+of+cell+trafficking+in+allotransplanted+vascularized+skin+flaps+and+conventional+skin+grafts.">In vivo observations of cell trafficking in allotransplanted vascularized skin flaps and conventional skin grafts.</a> Journal of Plastic, Reconstructive &amp; Aesthetic Surgery. 63 (4), 711-719 (2010).</li><li>Fleissig, Y., 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=Modified+Heterotopic+Hindlimb+Osteomyocutaneous+Flap+Model+in+the+Rat+for+Translational+Vascularized+Composite+Allotransplantation+Research.">Modified Heterotopic Hindlimb Osteomyocutaneous Flap Model in the Rat for Translational Vascularized Composite Allotransplantation Research.</a> Journal of Visualized Experiments. (146), e59458 (2019).</li><li>Zhou, X., 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+series+of+rat+segmental+forelimb+ectopic+implantation+models.">A series of rat segmental forelimb ectopic implantation models.</a> Scientific Reports. 7 (1), (2017).</li><li>Adamson, L. A., 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+modified+model+of+hindlimb+osteomyocutaneous+flap+for+the+study+of+tolerance+to+composite+tissue+allografts.">A modified model of hindlimb osteomyocutaneous flap for the study of tolerance to composite tissue allografts.</a> Microsurgery. 27 (7), 630-636 (2007).</li><li>Ulusal, A. E., Ulusal, B. G., Hung, L. M., Wei, F. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterotopic+hindlimb+allotransplantation+in+rats:+an+alternative+model+for+immunological+research+in+composite-tissue+allotransplantation.">Heterotopic hindlimb allotransplantation in rats: an alternative model for immunological research in composite-tissue allotransplantation.</a> Microsurgery. 25 (5), 410-414 (2005).</li><li>Fries, C. A., 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=enzyme+responsive,+tacrolimus-eluting+hydrogel+enables+long-term+survival+of+orthotopic+porcine+limb+vascularized+composite+allografts:+A+proof+of+concept+study.">enzyme responsive, tacrolimus-eluting hydrogel enables long-term survival of orthotopic porcine limb vascularized composite allografts: A proof of concept study.</a> PLoS One. 14 (1), 0210914 (2019).</li><li>Cottrell, B. L., 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=Neuroregeneration+in+composite+tissue+allografts:+effect+of+low-dose+FK506+and+mycophenolate+mofetil+immunotherapy.">Neuroregeneration in composite tissue allografts: effect of low-dose FK506 and mycophenolate mofetil immunotherapy.</a> Plastic and Reconstructive Surgery. 118 (3), 5 (2006).</li><li>Benhaim, P., Anthony, J. P., Lin, L. Y., McCalmont, T. H., Mathes, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+long-term+study+of+allogeneic+rat+hindlimb+transplants+immunosuppressed+with+RS-61443.">A long-term study of allogeneic rat hindlimb transplants immunosuppressed with RS-61443.</a> Transplantation. 56 (4), 911-917 (1993).</li><li>Greene, E. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Anatomy of the rat. Volume N.S. American Philos. Soc. , 27 (1935).</li><li>Schmalbruch, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fiber+composition+of+the+rat+sciatic+nerve.">Fiber composition of the rat sciatic nerve.</a> Anatomical Record. 215 (1), 71-81 (1986).</li><li>Cheah, M., Fawcett, J. W., Andrews, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+Thermal+Pain+Sensation+in+Rats+and+Mice+Using+the+Hargreaves+Test.">Assessment of Thermal Pain Sensation in Rats and Mice Using the Hargreaves Test.</a> Bio-protocol. 7 (16), (2017).</li><li>Hargreaves, K., Dubner, R., Brown, F., Flores, C., Joris, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+and+sensitive+method+for+measuring+thermal+nociception+in+cutaneous+hyperalgesia.">A new and sensitive method for measuring thermal nociception in cutaneous hyperalgesia.</a> Pain. 32 (1), 77-88 (1988).</li><li>Hong, S. H., Eun, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+Forelimb+Allotransplantation+in+Canine+Model.">Experimental Forelimb Allotransplantation in Canine Model.</a> BioMed Research International. 2016, 1495710 (2016).</li><li>Mathes, D. W., 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+preclinical+canine+model+for+composite+tissue+transplantation.">A preclinical canine model for composite tissue transplantation.</a> Journal of Reconstructive Microsurgery. 26 (3), 201-207 (2010).</li><li>Ibrahim, Z., 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+modified+heterotopic+swine+hind+limb+transplant+model+for+translational+vascularized+composite+allotransplantation+(VCA)+research.">A modified heterotopic swine hind limb transplant model for translational vascularized composite allotransplantation (VCA) research.</a> Journal of Visualized Experiments. (80), e50475 (2013).</li><li>Fries, C. A., 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+Porcine+Orthotopic+Forelimb+Vascularized+Composite+Allotransplantation+Model:+Technical+Considerations+and+Translational+Implications.">A Porcine Orthotopic Forelimb Vascularized Composite Allotransplantation Model: Technical Considerations and Translational Implications.</a> Plastic and Reconstructive Surgery. 138 (3), 71 (2016).</li><li>Kim, M., Fisher, D. T., Powers, C. A., Repasky, E. A., Skitzki, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improved+Cuff+Technique+and+Intraoperative+Detection+of+Vascular+Complications+for+Hind+Limb+Transplantation+in+Mice.">Improved Cuff Technique and Intraoperative Detection of Vascular Complications for Hind Limb Transplantation in Mice.</a> Transplantation Direct. 4 (2), 345 (2018).</li><li>Chong, A. S., Alegre, M. L., Miller, M. L., Fairchild, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lessons+and+limits+of+mouse+models.">Lessons and limits of mouse models.</a> Cold Spring Harbor Perspectives in Medicine. 3 (12), 015495 (2013).</li><li>Sucher, R., 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=Mouse+hind+limb+transplantation:+a+new+composite+tissue+allotransplantation+model+using+nonsuture+supermicrosurgery.">Mouse hind limb transplantation: a new composite tissue allotransplantation model using nonsuture supermicrosurgery.</a> Transplantation. 90 (12), 1374-1380 (2010).</li><li>Tung, T. H., Mohanakumar, T., Mackinnon, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+mouse+model+for+heterotopic+limb+and+composite-tissue+transplantation.">Development of a mouse model for heterotopic limb and composite-tissue transplantation.</a> Journal of Reconstructive Microsurgery. 17 (4), 267-273 (2001).</li><li>Tang, J., 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+vascularized+elbow+allotransplantation+model+in+the+rat.">A vascularized elbow allotransplantation model in the rat.</a> Journal of Shoulder and Elbow Surgery. 24 (5), 779-786 (2015).</li><li>Yan, Y., 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=Nerve+regeneration+in+rat+limb+allografts:+evaluation+of+acute+rejection+rescue.">Nerve regeneration in rat limb allografts: evaluation of acute rejection rescue.</a> Plastic and Reconstructive Surgery. 131 (4), 511 (2013).</li><li>Georgiade, N. G., Serafin, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> A Laboratory Manual of Microsurgery. Fourth Printing. , (1986).</li><li>Tseng, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suppression+of+autotomy+by+N-methyl-D-aspartate+receptor+antagonist+(MK-801)+in+the+rat.">Suppression of autotomy by N-methyl-D-aspartate receptor antagonist (MK-801) in the rat.</a> Neuroscience Letters. 240 (1), 17-20 (1998).</li><li>Haselbach, D., 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=Regeneration+patterns+influence+hindlimb+automutilation+after+sciatic+nerve+repair+using+stem+cells+in+rats.">Regeneration patterns influence hindlimb automutilation after sciatic nerve repair using stem cells in rats.</a> Neuroscience Letters. 634, 153-159 (2016).</li><li>Kloos, A. D., Fisher, L. C., Detloff, M. R., Hassenzahl, D. L., Basso, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stepwise+motor+and+all-or-none+sensory+recovery+is+associated+with+nonlinear+sparing+after+incremental+spinal+cord+injury+in+rats.">Stepwise motor and all-or-none sensory recovery is associated with nonlinear sparing after incremental spinal cord injury in rats.</a> Experimental Neurology. 191 (2), 251-265 (2005).</li><li>Berryman, E. R., Harris, R. L., Moalli, M., Bagi, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Digigait+quantitation+of+gait+dynamics+in+rat+rheumatoid+arthritis+model.">Digigait quantitation of gait dynamics in rat rheumatoid arthritis model.</a> Journal of Musculoskeletal and Neuronal Interactions. 9 (2), 89-98 (2009).</li><li>Hamers, F. P., Lankhorst, A. J., van Laar, T. J., Veldhuis, W. B., Gispen, W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+quantitative+gait+analysis+during+overground+locomotion+in+the+rat:+its+application+to+spinal+cord+contusion+and+transection+injuries.">Automated quantitative gait analysis during overground locomotion in the rat: its application to spinal cord contusion and transection injuries.</a> Journal of Neurotrauma. 18 (2), 187-201 (2001).</li><li>Deumens, R., Marinangeli, C., Bozkurt, A., Brook, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+motor+outcome+and+functional+recovery+following+nerve+injury.">Assessing motor outcome and functional recovery following nerve injury.</a> Methods in Molecular Biology. 11622, 179-188 (2014).</li><li>Bozkurt, A., 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=Aspects+of+static+and+dynamic+motor+function+in+peripheral+nerve+regeneration:+SSI+and+CatWalk+gait+analysis.">Aspects of static and dynamic motor function in peripheral nerve regeneration: SSI and CatWalk gait analysis.</a> Behavioural Brain Research. 219 (1), 55-62 (2011).</li><li>Neckel, N. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+to+quantify+the+velocity+dependence+of+common+gait+measurements+from+automated+rodent+gait+analysis+devices.">Methods to quantify the velocity dependence of common gait measurements from automated rodent gait analysis devices.</a> Journal of Neuroscience Methods. 253, 244-253 (2015).</li><li>Yeh, L. S., Gregory, C. R., Theriault, B. R., Hou, S. M., Lecouter, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+functional+model+for+whole+limb+transplantation+in+the+rat.">A functional model for whole limb transplantation in the rat.</a> Plastic and Reconstructive Surgery. 105 (5), 1704-1711 (2000).</li></ol>

肢体移植，在更广泛的血管成分异位移植（VCA）类别下，具有广泛的应用治疗前景尚未实现。主要障碍在于VCA和神经运动恢复技术目前所特有的未解决的免疫学问题。新技术的发展将取决于灵活、健壮和可重复的动物建模。
在VCA中已经建立了许多动物模型，每种模型都有特定的优点。非人类灵长类动物模型为人类患者提供了有吸引力的可翻译性，但一直受?...

血管化复合异体移植（VCA）或复合组织异体移植（CTA）等更广泛范畴的肢体移植尚未实现其治疗承诺。自1998年、1999年在法国里昂和肯塔基州路易斯维尔成功进行人类手部移植手术以来，全球已对经过精心挑选的患者进行了100多例上肢移植手术。广泛的适用性已经阻碍了大量的免疫抑制和有限的功能神经运动恢复。目前的免疫抑制策略导致85%的急性排斥发生率，面对77%的机会性感染2。另一方面，手移植后功能恢复发生;手臂肩部和手部（DASH）的残疾分数从71分提高到43分，但功能水平可能仍有资格成为残疾2。鉴于肢体移植的非生命保存性质，目前的技术必须在动物模型中进行改进，才能在VCA中迈出下一步。
自1978年第一个大鼠模型的肢体移植3，许多创新的动物模型已经发展，以推进VCA 4领域，纳入血管袖口的阿内斯特莫西，以尽量减少手术时间5,5，6，异位 骨皮移植，以尽量减少生理侮辱接受动物77，8，9，10，11，,8,9,10,11和新颖的免疫方法7，12，13，14。,13,147,这里介绍的正畸右后肢中大腿移植的老鼠模型强调经过精密、久经考验的显微手术技术，如手工缝制血管肿瘤和神经整形，作为对一个稳健、可重复的模型平台的前期投资，同时研究当前VCA限制的两个方面：免疫抑制策略和功能神经运动恢复。

<table>
	<tbody>
		<tr>
			<td>Anesthesia machine</td>
			<td>Vet Equip</td>
			<td>911103</td>
			<td></td>
		</tr>
		<tr>
			<td>0.5cc syringe</td>
			<td>Exel</td>
			<td>26018</td>
			<td></td>
		</tr>
		<tr>
			<td>18-gauge needle</td>
			<td>BD</td>
			<td>305196</td>
			<td></td>
		</tr>
		<tr>
			<td>1cc syringe</td>
			<td>BD</td>
			<td>309659</td>
			<td></td>
		</tr>
		<tr>
			<td>22-gauge needle</td>
			<td>BD</td>
			<td>305156</td>
			<td></td>
		</tr>
		<tr>
			<td>24-gauge angiocatheter</td>
			<td>Sur-Vet</td>
			<td>SROX2419V</td>
			<td></td>
		</tr>
		<tr>
			<td>25-gauge needle</td>
			<td>Exel</td>
			<td>26403</td>
			<td></td>
		</tr>
		<tr>
			<td>3 cc syringe</td>
			<td>BD</td>
			<td>309657</td>
			<td></td>
		</tr>
		<tr>
			<td>5cc syringe</td>
			<td>Exel</td>
			<td>26230</td>
			<td></td>
		</tr>
		<tr>
			<td>Alcohol</td>
			<td>Fisher Scientific</td>
			<td>HC-600-1GAL</td>
			<td></td>
		</tr>
		<tr>
			<td>Anesthesia induction chamber</td>
			<td>Vet Equip</td>
			<td>941443</td>
			<td></td>
		</tr>
		<tr>
			<td>Anesthetic gas scavenger system</td>
			<td>Vet Equip</td>
			<td>931401</td>
			<td></td>
		</tr>
		<tr>
			<td>Bipolar electrocautery</td>
			<td>Aura</td>
			<td>26-500</td>
			<td></td>
		</tr>
		<tr>
			<td>Bitter Spray Mist</td>
			<td>Henry Schein</td>
			<td>5553</td>
			<td></td>
		</tr>
		<tr>
			<td>Bone wax</td>
			<td>CP Medical</td>
			<td>CPB31A</td>
			<td></td>
		</tr>
		<tr>
			<td>Breathing circuit</td>
			<td>Vet Equip</td>
			<td>921413</td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenophine</td>
			<td>Reckitt Benckiser</td>
			<td>12496075705</td>
			<td></td>
		</tr>
		<tr>
			<td>Castro-Viejos needle drivers</td>
			<td>Roboz</td>
			<td>RS-6416</td>
			<td></td>
		</tr>
		<tr>
			<td>Cordless rotary saw</td>
			<td>Dremel</td>
			<td>8050-N/18</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton swab stick</td>
			<td>Fisher Scientific</td>
			<td>23-400-101</td>
			<td>For hemostasis</td>
		</tr>
		<tr>
			<td>DigiGait Appparatus and Software</td>
			<td>Mouse Specifics</td>
			<td>MSI-DIG, DIG-SOFT</td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont forceps (#4)</td>
			<td>Roboz</td>
			<td>RS-4972</td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont forceps (#5)</td>
			<td>Roboz</td>
			<td>RS-5035</td>
			<td></td>
		</tr>
		<tr>
			<td>Enrofloxacin</td>
			<td>Norbrook</td>
			<td>ANADA 200-495</td>
			<td></td>
		</tr>
		<tr>
			<td>FK-506</td>
			<td>Astellas</td>
			<td>301601</td>
			<td></td>
		</tr>
		<tr>
			<td>Gauze</td>
			<td>Kendall</td>
			<td>1903</td>
			<td></td>
		</tr>
		<tr>
			<td>Gauze</td>
			<td>Covidien</td>
			<td>8044</td>
			<td></td>
		</tr>
		<tr>
			<td>Gloves</td>
			<td>Microflex</td>
			<td>DGP-350-M</td>
			<td></td>
		</tr>
		<tr>
			<td>Hair clippers</td>
			<td>Oster</td>
			<td>078005-010-003</td>
			<td></td>
		</tr>
		<tr>
			<td>Handheld monopolar electrocautery</td>
			<td>Bovie</td>
			<td>AA00</td>
			<td></td>
		</tr>
		<tr>
			<td>Hargreaves Apparatus</td>
			<td>Ugo Basile S.R.L. Gemonio, Italy</td>
			<td>37370</td>
			<td></td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Walgreens</td>
			<td>126987</td>
			<td></td>
		</tr>
		<tr>
			<td>Heparin</td>
			<td>Fresenius Kabi</td>
			<td>42592K</td>
			<td></td>
		</tr>
		<tr>
			<td>Hot plate</td>
			<td>Corning</td>
			<td>PC-351</td>
			<td>For warming resusscitation fluid</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Henry Schein</td>
			<td>29405</td>
			<td></td>
		</tr>
		<tr>
			<td>Lactated ringers</td>
			<td>Baxter</td>
			<td>2B2074</td>
			<td></td>
		</tr>
		<tr>
			<td>Large petri dish</td>
			<td>Fisher Scientific</td>
			<td>FB0875713</td>
			<td>For donor graft while in chilled saline</td>
		</tr>
		<tr>
			<td>Meloxicam</td>
			<td>Henry Schein</td>
			<td>49755</td>
			<td></td>
		</tr>
		<tr>
			<td>micro Collin Hartmann retractor</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Micro dissecting scissors</td>
			<td>Roboz</td>
			<td>RS-5841</td>
			<td></td>
		</tr>
		<tr>
			<td>Microfibrillar collagen powder</td>
			<td>BD</td>
			<td>1010590</td>
			<td>For hemostasis</td>
		</tr>
		<tr>
			<td>Microvascular clips</td>
			<td>Roboz</td>
			<td>RS-5420</td>
			<td></td>
		</tr>
		<tr>
			<td>Normal saline</td>
			<td>Baxter</td>
			<td>2F7124</td>
			<td></td>
		</tr>
		<tr>
			<td>Opthalmic lube</td>
			<td>Dechra</td>
			<td>IS4398</td>
			<td></td>
		</tr>
		<tr>
			<td>Rapmycin</td>
			<td>MedChem Express</td>
			<td>HY-10219</td>
			<td></td>
		</tr>
		<tr>
			<td>Small petri dish</td>
			<td>Fisher Scientific</td>
			<td>FB0875713A</td>
			<td>For warmed resusscitation fluid</td>
		</tr>
		<tr>
			<td>Sterile drapes</td>
			<td>ProAdvantage</td>
			<td>N207100</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical gown</td>
			<td>Cardinal Health</td>
			<td>9511</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical mask</td>
			<td>3M</td>
			<td>1805</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical microscope, optic model OPMIMD</td>
			<td>Zeiss</td>
			<td>169756</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical microscope, Universal S3</td>
			<td>Zeiss</td>
			<td>243188</td>
			<td></td>
		</tr>
		<tr>
			<td>Suture 10-0 nylon</td>
			<td>Covidien</td>
			<td>N2512</td>
			<td></td>
		</tr>
		<tr>
			<td>Suture 5-0 vicryl</td>
			<td>Ethicon</td>
			<td>J213H</td>
			<td></td>
		</tr>
		<tr>
			<td>Suture 7-0 silk tie</td>
			<td>Teleflex</td>
			<td>103-S</td>
			<td></td>
		</tr>
		<tr>
			<td>Tape</td>
			<td>3M</td>
			<td>1530-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasonic instrument cleaner</td>
			<td>Roboz</td>
			<td>RS-9911</td>
			<td></td>
		</tr>
		<tr>
			<td>Vessel dilation forceps</td>
			<td>Roboz</td>
			<td>RS-5047</td>
			<td></td>
		</tr>
	</tbody>
</table>

taking the next step: a neural coaptation orthotopic hind limb transplant model to maximize functional recovery in rat

特别是肢体移植和血管化复合异体移植（VCA），一般有广泛的治疗前景，由于目前免疫抑制和功能神经运动恢复的限制，已经阻碍了这种前景。许多动物模型已经开发用于研究VCA的独特特性，但在这里我们提出了一个强大的可重复模型的老鼠正交后肢移植，旨在同时研究当前VCA限制的两个方面：免疫抑制策略和功能神经运动恢复。该模型的核心是致力于细致，久经考验的显微外科技术，如手工缝制血管肿瘤和手工缝制神经共生股股骨神经和坐骨神经。这种方法可以进行持久的肢体重建，使活更长的动物能够恢复、恢复日常活动和功能测试。通过传统免疫抑制剂的短期治疗，异体移植动物在移植后存活了70天，而异体移植动物在术后200天后提供长寿对控制。神经功能恢复的证据在术后30天存在。该模型不仅为询问VCA和神经再生特有的免疫问题提供了一个有用的平台，还允许对专为VCA量身定做的新治疗策略进行体内测试。

生存和恢复取决于细致的手术技术。注意血管性肿瘤和神经性肿瘤，以及上述骨合一，是最大化此模型成功的关键。操作设计和具有代表性的原子结果如图 1所示。
总体死亡率取决于免疫抑制策略，大多数异移植动物达到100-200个术后研究终点，如图 2所示。一旦走出急性术后窗口，治疗的异种移植动物可以体验到生存长达58个术后?...

Watch this Scientific Journal Video about Taking the Next Step: a Neural Coaptation Orthotopic Hind Limb Transplant Model to Maximize Functional Recovery in Rat at JoVE.com

Taking the Next Step: a Neural Coaptation Orthotopic Hind Limb Transplant Model to Maximize Functional Recovery in Rat

该协议提出了一个强大的，可重复的模型血管化复合异体移植（VCA），旨在同时研究免疫学和功能恢复。时间投入到细致的技术，在右中大腿后肢正交移植与手工缝制血管肿瘤和神经共性产生研究功能恢复的能力。

采取下一步：神经整形正交后肢移植模型，以最大限度地提高大鼠的功能恢复

Limb transplant in particular and vascularized composite allotransplant (VCA) in general have wide therapeutic promise that have been stymied by current ...

本视频将提供详细的描述，如何执行正畸右后肢，中大腿移植大鼠。捐赠大鼠右后肢在中大腿水平被截肢，然后储存在冰上。接受大鼠右后肢在中大腿水平也截肢。然后，供体大鼠肢体附着在受助鼠上。我们将描述如何重新连接骨骼、动脉、静脉、神经、肌肉和皮肤系统。肢体移植，更具体地说，血管化复合异体移植，VCA，有广泛的治疗前景。但它目前受到技术、免疫抑制和功能差的神经运动恢复的有限限制。我们的模型强调经过时间考验的显微手术技术，如手工缝制血管解剖学和神经共生。一年来，我们进行了200多次手术。虽然有一个初始学习曲线到开始微外科医生，通过使用这个视频来学习我们的方法和他们对专用实践的致力于，任何微外科医生将能够掌握这个模型。所有实验都是按照国家卫生研究院实验室动物护理和使用指南进行的，并经西北大学动物护理和使用委员会批准。在协议 IS 00001663 上执行了特定过程。选择一只8至16周大的老鼠，手术时体重在250到400克之间。在修剪大鼠的后肢毛皮之前，通过蒸发器用5%等值的麻醉诱导全身麻醉。使用鼻锥保持麻醉在 2 到 2.5% 之间，将大鼠放在操作板上，下面有加热器。用70%的酒精消毒无毛皮肤，用无菌纱布保护手术场。使用适当的显微外科显微镜和此处显示的显微手术仪器，以及易于获取的双极和单极电卡，您将准备好开始解剖。使用剪刀在右后肢周围做圆周和皮肤切口。立即从内侧折痕开始，其水平与内侧韧带大致相同，并横向延伸后侧，以完成圆周切口。然后暴露切口下方的肌肉层，解剖和烧灼的肤浅的表层胃，导致从肌肉层到刚刚创建的近近皮瓣。将近位皮瓣超位反射到内侧韧带和侧皮皮瓣推断到膝盖。使用钢丝缩回器或卷纱布帮助露出磁场。观察大鼠的内向解剖与人类相似;从横向到中枢，在于神经、动脉和静脉。剖析股骨神经，在内侧韧带近侧与分叉之间急剧分割，如果可能的话。低劣地收回分裂的神经， 保持它的安全远离的方式， 覆盖在潮湿的纱布下。将注意力转向股动脉和静脉使用四厘米长的 7-0 丝状领带来创伤性地缩回血管，而不是直接处理它们。现在密切关注将改善你最后的解剖学结果。用7-0丝带将股骨容器的所有分支都拉上。在领带之间划分分支。动脉和静脉分支，需要分裂包括表面的环弹性的腹腔和肌肉血管。表面的环皮动物通常是最大的，似乎潜入深，就像人类的富伦达股骨一样，但大鼠中不存在。股骨血管的更近源分支，如最高的根和盐状分支通常不需要分割。现在用 7 - 0 丝状动脉尽可能与内关节韧带连接， 并在连接之间划分。类似于动脉，拉大和分割股骨静脉。反射动脉和静脉低劣， 安全的方式， 覆盖在潮湿的纱布下与股骨神经覆盖之前.解剖腹体肌肉群，注意烧灼任何可见的血管。现在注意血脂将最大限度地减少接受者在以后的再发性输血期间的失血。深到腹骨肌肉群，识别和尖锐地划分坐骨神经近到它的分支。所有三个坐骨神经科通常可见，三头，胸膜和耳骨。这三者都应保留在捐赠者肢体中。在此解剖中通常未看到第四个皮肤分支。完成在中大腿水平的其余腹体和背部肌肉群的分部与细致的海莫西。可能需要以医疗方式收回肢体以完成肌肉的分割。使用手持式无绳旋转锯在中轴上转换股骨。肢体现在从捐赠者那里释放。从捐赠者那里取出肢体移植物后，从移植侧股股动脉和静脉树桩上切开丝束的末端，从而重新打开血管。将一个24测量的血管导管插入移植动脉树桩，用250个国际单位的肝素冲洗，用5毫升的冰冷普通盐水稀释，小心观察它流出开放的静脉。慢慢地，轻轻冲洗移植物约三分钟。过度的强力冲洗可能会损坏内皮。将移植物放在冷藏盐水盘中，嵌套在冰桶中，直到移植。用双边胸腔切除术对捐赠鼠进行安乐死。接受者手术开始类似于供体手术，麻醉大鼠，修剪毛皮，在内毛折痕上做一个包皮切口，反射皮瓣，解剖股骨神经，动脉和静脉。并连接与前面描述的相同的分支容器。然而，在手术过程中，与捐赠者更分裂股骨神经，但如果可能的话，则与分体神经分开。分割和释放股动脉和静脉，并用微手术斗牛犬夹分别夹在内侧韧带的水平。继续手术，如前所述，分裂大腿的腹骨支持肌肉在中大腿水平与细致的海莫西，同时收回肢体医疗必要的。确定坐骨神经近向分支点并分割，并使用旋转锯在中轴处横贯股骨。移除接收者原生右后肢，并按照当地动物护理和使用委员会准则进行适当处置。在这个阶段，通过鼻锥将异氟兰降低为 1 到 1.5%非常重要。在手术中这种不太痛苦的部分，太多的麻醉会导致老鼠的死亡。是时候把骨头拼凑起来。使用手持式旋转锯，剃掉捐赠者和接受者股骨切割端的任何不规则现象。切断18表针的轮毂端，该针将成为股骨内杆。在操作骨骼之前，将少量骨蜡涂抹到股骨的接受者切端，以减少骨髓出血。使用18量针作为内杆，将捐赠者和接受者股骨进行注射。一些力是必要的，但不要把骨骼都用，以使皮层断裂。根据需要，取出针头并将其修剪到不适当的长度，使两个骨骼平滑地在针上，骨骼之间没有显示任何针头。将小支撑，如纱布垫或小岩石或建模粘土下方的捐赠者肢体，以保持其紧张。用8到10个简单的中断5-0多糖蛋白缝合线重新建立腹体肌肉群，使移植物不会围绕股骨针旋转。这为动物提供了肢体稳定性。现在开始阿纳斯特莫西。定期用冰冷的盐水灌溉移植物和手术场，以便更好地可视化，减少缺血性再灌注损伤。使用简单的中断 10-0 尼龙缝合线，以端到端的方式对齐供体和接受者股动脉和原子体，避免张力和循环。动脉平均需要六个缝合线。与动脉类似，以端到端的方式使捐赠者和接受者股骨静脉具有原子性。静脉需要六到八个缝合线。慷慨的冷盐水灌溉，创伤容器处理技术，留下长尾巴作为容器缩回的停留缝合线，是有效微外科麻醉的重要工具。将适量的止血纤维素粉放在两个麻醉剂上，然后去除静脉和动脉上的近端微外科斗牛犬夹。检查两种麻醉和流动。使用棉签棒轻轻搅拌静脉，并检查两个静脉上有无一好血栓。当两个原点被确认为满意时，修剪任何剩余的长停留缝合尾，以匹配其他。将大鼠重新定位到左侧脱花位置，使用自由电烧机获得任何再灌注肌肉出血的细致止血。一旦肌肉痉挛得到保证，注意力就转向神经神经。用简单中断的 5-0 多糖蛋白缝合线，重新建立直接位于坐骨神经下的后肌群。修剪回任何神经切割端，看起来不规则。用10-0尼龙缝合线重新建立坐骨神经，争取肾上腺素叮咬。8 到 10 个简单的中断缝合线通常就足够了。用5-0多糖蛋白缝合线重新建立剩余的后体肌肉群。然后用4-0多糖蛋白连续运行缝合关闭后层皮肤。重新定位大鼠至苏皮位置，用10-0尼龙缝合重新建立股骨神经，再次争取肾上腺素叮咬。两到三个简单的中断缝合线通常就足够了。以与后体皮肤相同的方式关闭剩余的腹体皮肤。避免多余的缝合尾巴，这一旦醒来，可能会刺激大鼠。一旦动物离开术后立即窗口，他们可以测试神经运动恢复。哈格里夫斯测试用于测试热感觉。将大鼠放在测试容器中，并允许其适应20分钟。仪器玻璃被确认清洁，热源被证实与调查员的手指工作。测试前，确认大鼠是否清醒，测试的爪子位于红外运动探测器上。在强度级别 90 时传输热能。记录动物移动爪子远离热源的时间延迟。如果在 20 秒内未发生任何移动，则中止测试以防止受伤。在计算每种动物平均戒断延迟时间之前，每个测试肢体获得五次试验，不包括最高值和最低值。下面是以卡通格式描述的操作设计。捐赠鼠和受养大鼠的右后肢在中大腿水平被截肢，作为坐骨横截面。然后，在接受者手术中必须重新连接股动脉、静脉和神经，以及坐骨神经和股骨。该模型可用于显示免疫抑制和移植策略的差别生存效应。有了这种技术，长期生存是可以实现的。该模型可用于研究感觉神经恢复，这可能在术后四周内显现出来。也可以使用步态分析平台对电机功能进行询问。在这里，我们提出了一个具有良好生存特性的老鼠模型平台，我们用它来探测感官和运动恢复，如程序图中所看到的。肢体移植在更广泛的VCA范畴下，血管化成分异体移植已得到广泛应用的治疗承诺尚未实现。主要路障线目前的免疫抑制和神经运动恢复技术。新技术的发展将取决于灵活、健壮和可重复的动物建模。利用这个平台，我们希望调查人员能够推进肢体移植的多个方面。感谢您的收看，并祝您下一步好运。

Research

JoVE Journal

Medicine

Orthotopic Aortic Transplantation: A Rat Model to Study the Development of Chronic Vasculopathy

主动脉原位移植大鼠模型，研究慢性血管病变的发展

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant ...

科研

医学

A Modified Heterotopic Swine Hind Limb Transplant Model for Translational Vascularized Composite Allotransplantation (VCA) Research

A Modified Heterotopic Swine Hind Limb Transplant Model for Translational Vascularized Composite Allotransplantation VCA Research

平移血管复合同种异体移植（VCA）研究一种改进的异位猪后肢移植模型

Vascularized Composite Allotransplantation (VCA) such as hand and face transplants represent a viable treatment option for complex musculoskeletal trauma ...

Demonstration of the Rat Ischemic Skin Wound Model

在大鼠缺血性皮肤创伤模型的演示

The propensity for chronic wounds in humans increases with ageing, disease conditions such as diabetes and impaired cardiovascular function, and ...

Functional Human Liver Preservation and Recovery by Means of Subnormothermic Machine Perfusion

功能人类肝脏保存和恢复由Subnormothermic机灌注手段

There is currently a severe shortage of liver grafts available for transplantation.  Novel organ preservation techniques are needed to expand the pool of ...

Orthotopic Hind Limb Transplantation in the Mouse

原位后肢移植中的鼠标

In vivo animal model systems, and in particular mouse models, have evolved into powerful and versatile scientific tools indispensable to basic and ...

Heterotopic Renal Autotransplantation in a Porcine Model: A Step-by-Step Protocol

异位自体肾移植的猪模型：有步骤，分步协议

Kidney transplantation is the treatment of choice for patients suffering from end-stage renal disease. It offers better life expectancy and higher quality ...

Method of Direct Segmental Intra-hepatic Delivery Using a Rat Liver Hilar Clamp Model

直接节段性肝内交付使用大鼠肝门部夹具模型方法

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ...

Preclinical Model of Hind Limb Ischemia in Diabetic Rabbits

糖尿病家兔欣德肢体缺血的临床前模型

Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular ...

A Cryoinjury Model to Study Myocardial Infarction in the Mouse

研究小鼠心肌梗死的低温损伤模型

The use of animal models is essential for developing new therapeutic strategies for acute coronary syndrome and its complications. In this article, we ...

Reduced Complications after Arterial Reconnection in a Rat Model of Orthotopic Liver Transplantation

矫形肝移植大鼠模型中动脉重联后并发症减少

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation ...