Rostami Sara, Xu Michael, Datta Shaishav, Haykal Siba
Department of Surgery, Latner Thoracic Surgery Research Laboratories, Toronto General Hospital Research Institute, University Health Network, University of Toronto, Toronto, ON, Canada.
Division of General Surgery, Department of Surgery, University of Toronto, Toronto, ON, Canada.
Transplant Direct. 2021 Dec 13;8(1):e1251. doi: 10.1097/TXD.0000000000001251. eCollection 2022 Jan.
Ischemia-reperfusion injury plays an important role in vascularized composite allotransplantation (VCA). Currently, there is no ideal preservation solution for VCA. In this study, we investigated the effects of 4 different preservation solutions on different tissues within an allogeneic hindlimb rat model.
Sprague Dawley rat hindlimbs were flushed and placed at 4°C for 6 h in heparinized saline, histidine-tryptophan-ketoglutarate, University of Wisconsin (UW), and Perfadex and heterotopically transplanted for ease of ambulation. Apoptosis, necrosis, and the extracellular matrix of the tissues within the allograft were analyzed 2 h posttransplantation using immunohistochemistry, terminal deoxynucleotidyl transferase 2'-deoxyuridine 5'-triphosphate nick-end labeling (TUNEL) assay, and enzyme-linked immunoassay.
Higher expression of cleaved caspase 3, a significant increase of high-mobility group box 1 and TUNEL-positive apoptotic cells were observed in the muscle and vessels preserved with heparinized saline compared with UW and Perfadex following reperfusion. Higher expression of TUNEL-positive apoptotic cells was observed in the skin at 12 h of ischemia and in the nerve following reperfusion with histidine-tryptophan-ketoglutarate as a preservation solution.
Our data suggest that UW and Perfadex are preferred solutions in VCA. The vessels within the allografts appear to be very susceptible, with laminins and CD31 playing a role in ischemia-reperfusion injury.
缺血再灌注损伤在血管化复合组织异体移植（VCA）中起重要作用。目前，尚无理想的VCA保存液。在本研究中，我们在同种异体大鼠后肢模型中研究了4种不同保存液对不同组织的影响。
将Sprague Dawley大鼠后肢冲洗后，置于含肝素的生理盐水、组氨酸-色氨酸-酮戊二酸、威斯康星大学（UW）保存液和Perfadex保存液中，于4°C保存6小时，然后进行异位移植以便于活动。移植后2小时，使用免疫组织化学、末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）法和酶联免疫吸附测定法分析同种异体移植物内组织的凋亡、坏死和细胞外基质。
再灌注后，与UW保存液和Perfadex保存液相比，用含肝素的生理盐水保存的肌肉和血管中，裂解的半胱天冬酶3表达更高，高迁移率族蛋白盒1显著增加，TUNEL阳性凋亡细胞显著增多。以组氨酸-色氨酸-酮戊二酸作为保存液时，缺血12小时的皮肤和再灌注后的神经中观察到TUNEL阳性凋亡细胞表达更高。
我们的数据表明，UW保存液和Perfadex保存液是VCA中的首选保存液。同种异体移植物中的血管似乎非常敏感，层粘连蛋白和CD31在缺血再灌注损伤中起作用。
