Said Sayf A, Ordeñana Carlos X, Rezaei Majid, Figueroa Brian A, Dasarathy Srinivasan, Brunengraber Henri, Rampazzo Antonio, Gharb Bahar Bassiri
Cleveland Clinic Foundation, Department of Plastic Surgery, 9500 Euclid Ave, A60, Cleveland, OH 44195.
Cleveland Clinic Foundation, Department of Gastroenterology, 9500 Euclid Ave, Cleveland, OH 44195.
Mil Med. 2020 Jan 7;185(Suppl 1):110-120. doi: 10.1093/milmed/usz314.
Ex-vivo normothermic limb perfusion (EVNLP) has been proven to preserve limb viability better than standard cold storage. Perfusates containing packed red blood cells (pRBC) improve outcomes when compared to acellular perfusates. Limitations of pRBC-based perfusion include limited availability, need for cross match, mechanical hemolysis, and activation of pro-inflammatory proteins. Hemoglobin-based oxygen carrier (HBOC)-201 (Hemopure) is a solution of polymerized bovine hemoglobin, characterized by low immunogenicity, no risk of hemolytic reaction, and enhanced convective and diffusive oxygen delivery. This is a preliminary study on the feasibility of EVNLP using HBOC-201 as an oxygen carrier.
Three porcine forelimb perfusions were performed using an established EVNLP model and an HBOC-201-based perfusate. The perfusion circuit included a roller pump, oxygenator, heat exchanger, and reservoir. Electrolytes, limb temperature, weight, compartment pressure, nerve conduction, and perfusion indicated by indocyanine green angiography and infra-red thermography were monitored. Histological evaluation was performed with hematoxylin and eosin and electron microscopy.
Three limbs were perfused for 21.3 ± 2.1 hours. Muscle contractility was preserved for 10.6 ± 2.4 hours. Better preservation of the mitochondrial ultrastructure was evident at 12 hours in contrast to crystallization and destruction features in the cold-storage controls.
An HBOC-201-EVNLP produced outcomes similar to RBC-EVNLP with preservation of muscle contractility and mitochondrial structure.
已证实体外常温肢体灌注（EVNLP）比标准冷藏能更好地维持肢体活力。与无细胞灌注液相比，含浓缩红细胞（pRBC）的灌注液可改善治疗效果。基于pRBC的灌注存在一些局限性，包括可用性有限、需要交叉配型、机械性溶血以及促炎蛋白的激活。基于血红蛋白的氧载体（HBOC）-201（Hemopure）是一种聚合牛血红蛋白溶液，具有低免疫原性、无溶血反应风险以及增强的对流和扩散性氧输送等特点。这是一项关于使用HBOC-201作为氧载体进行EVNLP可行性的初步研究。
使用已建立的EVNLP模型和基于HBOC-201的灌注液对三只猪的前肢进行灌注。灌注回路包括一个滚压泵、氧合器、热交换器和储液器。监测电解质、肢体温度、重量、腔室压力、神经传导以及吲哚菁绿血管造影和红外热成像显示的灌注情况。采用苏木精和伊红染色及电子显微镜进行组织学评估。
三只肢体灌注了21.3±2.1小时。肌肉收缩力维持了10.6±2.4小时。与冷藏对照组的结晶和破坏特征相比，12小时时线粒体超微结构的保存情况明显更好。
HBOC-201-EVNLP产生的结果与RBC-EVNLP相似，能维持肌肉收缩力和线粒体结构。
