Messner Franka, Hautz Theresa, Blumer Michael J F, Bitsche Mario, Pechriggl Elisabeth J, Hermann Martin, Zelger Bettina, Zelger Bernhard, Öfner Dietmar, Schneeberger Stefan
1 Center for Operative Medicine, Department of Visceral, Transplant and Thoracic Surgery, Medical University Innsbruck, Innsbruck, Austria. 2 Department of Anatomy, Histology and Embryology, Division of Clinical and Functional Anatomy, Medical University Innsbruck, Innsbruck, Austria. 3 Department of Anesthesiology and Critical Care Medicine, Medical University Innsbruck, Innsbruck, Austria. 4 Department of Pathology, Medical University Innsbruck, Innsbruck, Austria. 5 Department of Dermatology, Medical University Innsbruck, Innsbruck, Austria.
Transplantation. 2017 Sep;101(9):e301-e310. doi: 10.1097/TP.0000000000001845.
We herein investigate critical ischemia times and the effect of novel preservation solutions such as new histidine-tryptophan-ketoglutarate (HTK-N) and TiProtec on the individual tissues of a rat limb isograft.
Orthotopic hind-limb transplantations were performed in male Lewis rats after 2 hours, 6 hours, or 10 hours of cold ischemia (CI). Limbs were flushed and stored in HTK-N, TiProtec, HTK, or saline solution. Muscle, nerve, vessel, skin, and bone samples were procured on day 10 for histology, immunohistochemistry, confocal and electron microscopy, and quantitative real-time polymerase chain reaction analysis.
Histomorphology of the muscle showed a mainly perivascular inflammatory infiltrate, fibrotic degeneration, and neovascularization after 6 hours and 10 hours of CI. However, centrally aligned nuclei observed in muscle fibers suggest for muscle regeneration in these samples. In addition to Wallerian degeneration, nerve injury was significantly aggravated (P = 0.032) after prolonged CI. Proinflammatory and regulatory cytokines were most significantly upregulated after 2-hour CI. Our data suggest no superiority of novel perfusates HTK-N and TiProtec in terms of tissue preservation, compared with HTK and saline.
Limiting CI time for less than 6 hours is the most significant factor to reduce tissue damage in vascularized tissue transplantation. Signs of muscle regeneration give rise that ischemic muscle damage in limb transplantation might be reversible to a certain extent.
我们在此研究关键缺血时间以及新型保存液（如新组氨酸 - 色氨酸 - 酮戊二酸 [HTK - N] 和TiProtec）对大鼠肢体移植各组织的影响。
在雄性Lewis大鼠中，分别在冷缺血（CI）2小时、6小时或10小时后进行原位后肢移植。将肢体冲洗后，保存在HTK - N、TiProtec、HTK或盐溶液中。在第10天采集肌肉、神经、血管、皮肤和骨样本，进行组织学、免疫组织化学、共聚焦和电子显微镜检查以及定量实时聚合酶链反应分析。
CI 6小时和10小时后，肌肉组织形态学显示主要为血管周围炎性浸润、纤维化变性和新生血管形成。然而，在肌纤维中观察到的中央排列的细胞核提示这些样本中有肌肉再生。除了华勒变性外，长时间CI后神经损伤明显加重（P = 0.032）。促炎和调节细胞因子在CI 2小时后上调最为显著。我们的数据表明，与HTK和盐溶液相比，新型灌注液HTK - N和TiProtec在组织保存方面并无优势。
将CI时间限制在6小时以内是减少血管化组织移植中组织损伤的最重要因素。肌肉再生迹象表明肢体移植中缺血性肌肉损伤在一定程度上可能是可逆的。
