Department for BioMedical Research, DBMR, University of Bern, Bern 3008, Switzerland.
Department of Plastic- and Hand Surgery, Inselspital, Bern University Hospital, Bern 3010, Switzerland.
Mil Med. 2024 Aug 19;189(Suppl 3):83-92. doi: 10.1093/milmed/usae043.
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.
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.
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.
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.
连续的体外灌注(ECP)或机器灌注有望延长肢体缺血再灌注损伤中骨骼肌的保存时间。本研究旨在通过 24 小时 ECP 方法将截肢再植时间窗从目前的 6 小时延长至 33 小时。
6 只大白猪在全身麻醉下接受了前肢截肢手术。截肢后,肢体在室温下保存 9 小时,然后通过改良组氨酸-色氨酸-酮戊二酸(HTK)溶液进行 24 小时 ECP 灌注。ECP 后,肢体原位再植并在体内灌注 12 小时。收集和分析临床数据、血液和组织样本。
在室温缺血 9 小时后,6 只前肢均能成功再植并在体内再灌注 12 小时,随后进行 24 小时 ECP。再植后通过热成像和激光多普勒成像观察到足够的肢体灌注。所有猪均存活且无严重器官衰竭,且未发现炎症细胞因子显著增加。大体观察和组织学检查显示肢体明显的间质肌水肿,而肌纤维坏死不明显,提示肌肉完整性得到保存。
使用 24 小时 ECP 成功地将肢体保存时间延长至 33 小时。改良的组氨酸-色氨酸-酮戊二酸灌注液显示了其对肌肉的保护能力。这种创新方法不仅便于在战斗伤害后进行肢体再植,克服地理障碍,而且还拓宽了在国家和大陆之间进行匹配良好的肢体同种异体移植的前景。