Comparison of Acellular Solutions for Ex-situ Perfusion of Amputated Limbs
Comparison of Acellular Solutions for Ex-situ Perfusion of Amputated Limbs
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DOI：
10.1093/milmed/usaa160
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发表时间：
2020-11-01
影响因子：
1.2
通讯作者：
Pomahac, Bohdan
中科院分区：
文献类型：
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作者：
Haug, Valentin;Kollar, Branislav;Pomahac, Bohdan
低温离体机械灌注（MP）是一种替代静态冷藏（SCS）的有前途的器官保存方法，可用于移植和带血管复合同种异体移植。在严峻的环境中用基于血液的灌注溶液灌注是有问题的，因为它们需要适当的储存和短的保质期，使得其对于军事和紧急使用是不切实际的。脱细胞灌注已被证明是有效的，但截肢MP的理想灌注液尚未确定。本研究的目的是评估替代灌注液溶液的有效性，例如富含葡聚糖的Phoxilium，Steen，材料和方法将来自约克郡猪（n = 8）的截肢前肢在SCS（n = 2）中4 ℃保存12小时或在机器中保存12小时。在10 ℃下用含氧灌注溶液（n = 6）以恒定流速灌注12小时。使用的灌注液包括改良的Steen溶液、Phoxilium（PHOX）或富含葡聚糖-40的Phoxilium（PHODEX）。灌注1小时和6小时后更换灌注液。连续记录机器数据。在特定时间点采集用于临床化学、血气分析和肌肉活检的灌注液样本，随后进行分析。在这个半体内研究中，断肢再植尚未performed.ResultsAfter截肢，每肢被成功转移，并连接到我们的灌注设备。平均总缺血时间为77.5 ± 5.24分钟。灌注溶液的温度保持在10.18 +/- 2.01 ℃，灌注压力保持在24.48 +/- 10.72 mmHg。12小时后，SCS组的肢体重量增加了3%，PHODEX组增加了36%，Steen组增加了25%，PHOX组增加了58%。与SCS组相比，PHOX组的这种增加是显著的。由于水肿，所有灌注组的压力随时间增加10.99 mmHg。除Steen组和PHODEX组外，所有组的HIF-1a水平均随时间推移而下降。灌注液样本中肌肉损伤的生物标志物，如肌酸激酶和乳酸脱氢酶，显示出组间的显著差异，PHODEX组的值最高。血气分析结果无显著差异。ConclusionWith显着较高水平的肌酸激酶和乳酸脱氢酶，MP与葡聚糖富集Phoxilium提供了类似的结果，市售灌注液，如Steen，而不需要冷藏，并在大约5%的成本Steen解决方案。有必要进行进一步的大规模再植研究，以评估富含葡聚糖的Phoxilium作为替代灌注液的有效性。
IntroductionHypothermic ex-situ machine perfusion (MP) has been shown to be a promising alternative to static cold storage (SCS) for preservation of solid organs for transplantation and vascularized composite allotransplantation. Perfusion with blood-based perfusion solutions in austere environments is problematic due to their need for appropriate storage and short shelf life, making it impractical for military and emergency use. Acellular perfusion has been shown to be effective, but the ideal perfusate solution for MP of amputated limbs is yet to be determined. The purpose of this study is to evaluate the efficacy of alternative perfusate solutions, such as dextran-enriched Phoxilium, Steen, and Phoxilium in ex-vivo hypothermic MP of amputated limbs in a porcine model.Materials and methodsAmputated forelimbs from Yorkshire pigs (n = 8) were preserved either in SCS (n = 2) at 4 degrees C for 12 hours or machine-perfused at 10 degrees C for 12 hours with oxygenated perfusion solutions (n = 6) at a constant flow rate. The perfusates used include modified Steen-solution, Phoxilium (PHOX), or Phoxilium enriched with dextran-40 (PHODEX). The perfusate was exchanged after 1 and 6 hours of perfusion. Machine data were recorded continuously. Perfusate samples for clinical chemistry, blood gas analysis, and muscle biopsies were procured at specific timepoints and subsequently analyzed. In this semi in-vivo study, limb replantation has not been performed.ResultsAfter amputation, every limb was successfully transferred and connected to our perfusion device. The mean total ischemia time was 77.5 +/- 5.24 minutes. The temperature of the perfusion solution was maintained at 10.18 +/- 2.01 degrees C, and perfusion pressure at 24.48 +/- 10.72 mmHg. Limb weight increased by 3% in the SCS group, 36% in the PHODEX group, 25% in the Steen group, and 58% in the PHOX group after 12 hours. This increase was significant in the PHOX group compared with the SCS group. All perfusion groups showed a pressure increase of 10.99 mmHg over time due to edema. The levels of HIF-1a decreased over time in all groups except the Steen and the PHODEX group. The biomarkers of muscle injury in the perfusate samples, such as creatine kinase and lactate-dehydrogenase, showed a significant difference between groups, with highest values in the PHODEX group. No significant differences were found in the results of the blood gas analysis.ConclusionWith the exception of significantly higher levels of creatine kinase and lactate dehydrogenase, MP with dextran-enriched Phoxilium provides similar results as that of the commercially available perfusates such as Steen, without the need for cold storage, and at circa 5% of the cost of the Steen solution. Further large-scale replantation studies are necessary to evaluate the efficacy of dextran-enriched Phoxilium as an alternate perfusate solution.