背景:连续体外灌注(ECP),或机器灌注,有望在肢体缺血再灌注损伤中延长骨骼肌保存时间。这项研究旨在使用24小时ECP方法将截肢到再植的时间窗口从目前的6小时延长到33小时。
方法:6头大白猪在全身麻醉下进行前肢截肢手术。截肢后,肢体在室温下保持9小时,然后用改良的组氨酸-色氨酸-酮戊二酸(HTK)溶液进行24小时ECP灌注.ECP之后,四肢原位移植并在体内灌注12小时。临床数据,血,收集和分析组织样本。
结果:所有6个前肢均可在室温缺血9小时后成功移植并在体内再灌注12小时,然后进行24小时ECP。通过热成像和激光多普勒成像显示,再植后观察到足够的肢体灌注。所有猪都存活下来没有出现严重的器官衰竭,并且没有发现炎性细胞因子的显著增加。宏观和组织学检查显示四肢有明显的间质性肌肉水肿,而肌纤维坏死不明显,意味着保持肌肉的完整性。
结论:使用24小时ECP已成功将肢体保存延长至33小时。修饰的组氨酸-色氨酸-酮戊二酸灌注液证明了其保护肌肉的能力。这种创新的方法不仅有利于战伤后的肢体再植,克服地理障碍,但也扩大了在各个国家和大洲匹配良好的肢体同种异体移植的前景。
BACKGROUND: Continuous extracorporeal perfusion (ECP), or machine perfusion, holds promise for prolonged skeletal muscle preservation in limb ischemia-reperfusion injury. This study aimed to extend the amputation-to-replantation time window from currently 6 hours to 33 hours using a 24-hour ECP approach.
METHODS: Six large white pigs underwent surgical
forelimb amputation under general anesthesia. After amputation, limbs were kept for 9 hours at room temperature and then perfused by 24-hour ECP with a modified histidine-tryptophan-ketoglutarate (HTK) solution. After ECP, limbs were orthotopically replanted and perfused in vivo for 12 hours. Clinical data, blood, and tissue samples were collected and analyzed.
RESULTS: All 6 forelimbs could be successfully replanted and in vivo reperfused for 12 hours after 9 hours of room temperature ischemia followed by 24 hours ECP. Adequate limb perfusion was observed after replantation as shown by thermography and laser Doppler imaging. All pigs survived without severe organ failure, and no significant increase in inflammatory cytokines was found. Macroscopy and histology showed marked interstitial muscular edema of the limbs, whereas myofiber necrosis was not evident, implying the preservation of muscular integrity.
CONCLUSIONS: The use of a 24-hour ECP has successfully extended limb preservation to 33 hours. The modified histidine-tryptophan-ketoglutarate perfusate demonstrated its ability for muscle protection. This innovative approach not only facilitates limb replantation after combat injuries, surmounting geographical barriers, but also broadens the prospects for well-matched limb allotransplants across countries and continents.