K. Ozer, N. L. Werner, Fares Alghanem, Stephanie L. Rakestraw, Dylan C. Sarver, B. Nicely, R. Pietroski, P. Lange, S. Rudich, C. Mendias, A. Rojas-Peña, J. Magee, R. Bartlett
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This process is time limited, as the tissue has to be revascularized within 4–6 hours to minimize ischemia reperfusion (IR) injury. Normothermic perfusion was proposed as an alternative method of preservation in solid organ transplantation Method helps to avoid complications associated with cold preservation and maintains tissue viability without inducing IR injury Using this method, previous investigators demonstrated its potential to prolong swine forelimb allograft survival up to 24 hours (4,5) In this study, we aimed to test this system on human forearm allografts Methods Five human forearms were procured from braindead adult donors under tourniquet control. Following elbow disarticulation, the brachial artery was cannulated. The limb was flushed with heparinized saline and connected to a temperature controlled (30–33 C) ex situ perfusion system (Figure) for 24 hours. The perfusate consisted of plasma and red blood cells with a target hemoglobin (Hb) concentration of 4–6 g/dL. Muscle biopsies (flexor carpi radialis) were obtained at 0, 12, and 24 hours Results Average warm ischemia time was 76 minutes Average arterial systolic pressure was 93§2 mmHg, perfusion flow 310 § 20 mL/min (»6–8% of the donor’s estimated cardiac output), and vascular resistance 153 § 16 mmHg/mL/min. Perfusate had an average pH of 743 § 004 , pCO2 32 § 1 mmHg, pO2 317 § 18 mmHg, and Hb 48 § 04 g/dL Electrolytes (sodium, potassium, chloride) remained within a physiologic range Lactate started to increase steadily throughout the experiment; however, neuromuscular electrical stimulation revealed ongoing contraction throughout the experiment H&E staining showed mild fatty infiltration on some myocytes at 24 hours There was minimal change in fiber size, likely due to variation in age and gender between donors Muscle architecture was preserved at the end of 24 hours perfusion Conclusions All limbs remained viable after 24 hours of nearnormothermic ex situ perfusion as evidenced by ongoing neuromuscular stimulation While no assumptions can be drawn about the long-term function of the extremity, this approach could help extend VCA transplantation to a wider geographic area It also has the potential to circumvent complications associated with cold preservation. CONTACT Kagan Ozer, MD kozer@umich.edu © 2016 Kagan Ozer, Nicole Werner, Fares Alghanem, Stephanie L. Rakestraw, Dylan Sarver, Bruce Nicely, Richard Pietroski, Paul Lange, Steve M. Rudich, Chris L. Mendias, Alvaro Rojas-Pena, John C. Magee, and Robert H. Bartlett. Published with license by Taylor & Francis. This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted. VASCULARIZED COMPOSITE ALLOTRANSPLANTATION 2016, VOL. 3, NOS. 1–2, 6 http://dx.doi.org/10.1080/23723505.2016.1232926","PeriodicalId":372758,"journal":{"name":"Vascularized Composite Allotransplantation","volume":"50 1","pages":"0"},"PeriodicalIF":0.0000,"publicationDate":"2016-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"2497: Near normothermic ex-situ perfusion extends human limb allograft survival up to 24 hours\",\"authors\":\"K. Ozer, N. L. Werner, Fares Alghanem, Stephanie L. Rakestraw, Dylan C. Sarver, B. Nicely, R. Pietroski, P. Lange, S. Rudich, C. Mendias, A. Rojas-Peña, J. Magee, R. Bartlett\",\"doi\":\"10.1080/23723505.2016.1232926\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"2497: Near normothermic ex-situ perfusion extends human limb allograft survival up to 24 hours Kagan Ozer, MD, Nicole Werner, MD, Fares Alghanem, BS, Stephanie L. Rakestraw, BS, Dylan Sarver, BS, Bruce Nicely, RN, MSN, Richard Pietroski, MS, Paul Lange, MD, Steve M. Rudich, MD, PhD, Chris L. Mendias, PhD, Alvaro Rojas-Pena, MD, John C. Magee, MD, and Robert H. Bartlett, MD University of Michigan, Ann Arbor, MI, USA; Gift of Life, Ann Arbor, MI, USA Background Currently vascularized composite allografts (VCA) are cold preserved (4 C) until transplantation. This process is time limited, as the tissue has to be revascularized within 4–6 hours to minimize ischemia reperfusion (IR) injury. Normothermic perfusion was proposed as an alternative method of preservation in solid organ transplantation Method helps to avoid complications associated with cold preservation and maintains tissue viability without inducing IR injury Using this method, previous investigators demonstrated its potential to prolong swine forelimb allograft survival up to 24 hours (4,5) In this study, we aimed to test this system on human forearm allografts Methods Five human forearms were procured from braindead adult donors under tourniquet control. Following elbow disarticulation, the brachial artery was cannulated. The limb was flushed with heparinized saline and connected to a temperature controlled (30–33 C) ex situ perfusion system (Figure) for 24 hours. The perfusate consisted of plasma and red blood cells with a target hemoglobin (Hb) concentration of 4–6 g/dL. Muscle biopsies (flexor carpi radialis) were obtained at 0, 12, and 24 hours Results Average warm ischemia time was 76 minutes Average arterial systolic pressure was 93§2 mmHg, perfusion flow 310 § 20 mL/min (»6–8% of the donor’s estimated cardiac output), and vascular resistance 153 § 16 mmHg/mL/min. Perfusate had an average pH of 743 § 004 , pCO2 32 § 1 mmHg, pO2 317 § 18 mmHg, and Hb 48 § 04 g/dL Electrolytes (sodium, potassium, chloride) remained within a physiologic range Lactate started to increase steadily throughout the experiment; however, neuromuscular electrical stimulation revealed ongoing contraction throughout the experiment H&E staining showed mild fatty infiltration on some myocytes at 24 hours There was minimal change in fiber size, likely due to variation in age and gender between donors Muscle architecture was preserved at the end of 24 hours perfusion Conclusions All limbs remained viable after 24 hours of nearnormothermic ex situ perfusion as evidenced by ongoing neuromuscular stimulation While no assumptions can be drawn about the long-term function of the extremity, this approach could help extend VCA transplantation to a wider geographic area It also has the potential to circumvent complications associated with cold preservation. 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2497年:接近常压的离体灌注延长人类肢体同种异体移植物存活24小时Kagan Ozer, MD, Nicole Werner, MD, Fares Alghanem, BS, Stephanie L. Rakestraw, BS, Dylan Sarver, BS, Bruce Nicely, RN, MSN, Richard Pietroski, MS, Paul Lange, MD, Steve M. Rudich, MD, PhD, Chris L. Mendias, PhD, Alvaro rojaspena, MD, John C. Magee, MD和Robert H. Bartlett, MD密歇根大学,Ann Arbor, MI, USA;目前血管化复合同种异体移植物(VCA)在移植前都是低温保存(4℃)。这个过程是有时间限制的,因为组织必须在4-6小时内重建血运,以尽量减少缺血再灌注(IR)损伤。常温灌注被提出作为实体器官移植的另一种保存方法,这种方法有助于避免与冷保存相关的并发症,并在不诱导IR损伤的情况下保持组织活力。使用这种方法,先前的研究人员证明了它有可能将猪前肢同种异体移植物的存活时间延长至24小时(4,5)。方法从脑死亡成人供体中获取5条前臂,采用止血带控制。肘关节脱臼后,将肱动脉插管。肢体用肝素化生理盐水冲洗,并连接到温度控制(30-33℃)的非原位灌注系统(图)24小时。灌注液由血浆和红细胞组成,靶血红蛋白(Hb)浓度为4-6 g/dL。结果平均热缺血时间为76分钟,平均动脉收缩压为93±2 mmHg,灌注流量为310±20 mL/min(占供体估计心输出量的6-8%),血管阻力为153±16 mmHg/mL/min。Perfusate的平均pH值为743§004,pCO2为32§1 mmHg, pO2为317§18 mmHg, Hb为48§04 g/dL。电解质(钠、钾、氯化物)保持在生理范围内。然而,在整个实验过程中,神经肌肉电刺激显示持续收缩,H&E染色在24小时时显示一些肌细胞有轻微的脂肪浸润。在24小时灌注结束时,肌肉结构得以保留结论经持续的神经肌肉刺激,所有肢体在24小时的近恒温非原位灌注后仍能存活,但不能对肢体的长期功能做出假设。这种方法可以帮助将VCA移植扩展到更广泛的地理区域,也有可能避免与冷保存相关的并发症。联系Kagan Ozer, MD kozer@umich.edu©2016 Kagan Ozer, Nicole Werner, Fares Alghanem, Stephanie L. Rakestraw, Dylan Sarver, Bruce Nicely, Richard Pietroski, Paul Lange, Steve M. Rudich, Chris L. Mendias, Alvaro ro哈斯- pena, John C. Magee和Robert H. Bartlett。由Taylor & Francis授权出版。这是一篇在知识共享署名-非商业许可(http://creativecommons.org/licenses/by-nc/3.0/)条款下发布的开放获取文章,该许可允许在任何媒体上不受限制的非商业使用、分发和复制,前提是正确引用原始作品。指定作者的精神权利得到了维护。血管化复合异体移植,2016,VOL. 3, no . 1-2, 6 http://dx.doi.org/10.1080/23723505.2016.1232926
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2497: Near normothermic ex-situ perfusion extends human limb allograft survival up to 24 hours
2497: Near normothermic ex-situ perfusion extends human limb allograft survival up to 24 hours Kagan Ozer, MD, Nicole Werner, MD, Fares Alghanem, BS, Stephanie L. Rakestraw, BS, Dylan Sarver, BS, Bruce Nicely, RN, MSN, Richard Pietroski, MS, Paul Lange, MD, Steve M. Rudich, MD, PhD, Chris L. Mendias, PhD, Alvaro Rojas-Pena, MD, John C. Magee, MD, and Robert H. Bartlett, MD University of Michigan, Ann Arbor, MI, USA; Gift of Life, Ann Arbor, MI, USA Background Currently vascularized composite allografts (VCA) are cold preserved (4 C) until transplantation. This process is time limited, as the tissue has to be revascularized within 4–6 hours to minimize ischemia reperfusion (IR) injury. Normothermic perfusion was proposed as an alternative method of preservation in solid organ transplantation Method helps to avoid complications associated with cold preservation and maintains tissue viability without inducing IR injury Using this method, previous investigators demonstrated its potential to prolong swine forelimb allograft survival up to 24 hours (4,5) In this study, we aimed to test this system on human forearm allografts Methods Five human forearms were procured from braindead adult donors under tourniquet control. Following elbow disarticulation, the brachial artery was cannulated. The limb was flushed with heparinized saline and connected to a temperature controlled (30–33 C) ex situ perfusion system (Figure) for 24 hours. The perfusate consisted of plasma and red blood cells with a target hemoglobin (Hb) concentration of 4–6 g/dL. Muscle biopsies (flexor carpi radialis) were obtained at 0, 12, and 24 hours Results Average warm ischemia time was 76 minutes Average arterial systolic pressure was 93§2 mmHg, perfusion flow 310 § 20 mL/min (»6–8% of the donor’s estimated cardiac output), and vascular resistance 153 § 16 mmHg/mL/min. Perfusate had an average pH of 743 § 004 , pCO2 32 § 1 mmHg, pO2 317 § 18 mmHg, and Hb 48 § 04 g/dL Electrolytes (sodium, potassium, chloride) remained within a physiologic range Lactate started to increase steadily throughout the experiment; however, neuromuscular electrical stimulation revealed ongoing contraction throughout the experiment H&E staining showed mild fatty infiltration on some myocytes at 24 hours There was minimal change in fiber size, likely due to variation in age and gender between donors Muscle architecture was preserved at the end of 24 hours perfusion Conclusions All limbs remained viable after 24 hours of nearnormothermic ex situ perfusion as evidenced by ongoing neuromuscular stimulation While no assumptions can be drawn about the long-term function of the extremity, this approach could help extend VCA transplantation to a wider geographic area It also has the potential to circumvent complications associated with cold preservation. CONTACT Kagan Ozer, MD kozer@umich.edu © 2016 Kagan Ozer, Nicole Werner, Fares Alghanem, Stephanie L. Rakestraw, Dylan Sarver, Bruce Nicely, Richard Pietroski, Paul Lange, Steve M. Rudich, Chris L. Mendias, Alvaro Rojas-Pena, John C. Magee, and Robert H. Bartlett. Published with license by Taylor & Francis. This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted. VASCULARIZED COMPOSITE ALLOTRANSPLANTATION 2016, VOL. 3, NOS. 1–2, 6 http://dx.doi.org/10.1080/23723505.2016.1232926