Department of Biological Sciences, University of North Carolina at Charlotte, Charlotte, NC.
The Center for Biomedical Engineering and Science, University of North Carolina at Charlotte, Charlotte, NC.
Transplantation. 2020 May;104(5):e125-e134. doi: 10.1097/TP.0000000000003141.
Maintaining functional vessels during preservation of vascularized composite allografts (VCAs) remains a major challenge. The University of Wisconsin (UW) solution has demonstrated significant short-term benefits (4-6 h). Here we determined whether the new hypothermic resuscitation and preservation solution HypoRP improves both structure, survival, and function of pig arteries during storage for up to 6 days.
Using porcine swine mesenteric arteries, the effects of up to 6-day incubation in a saline (PBS), UW, or HypoRP solution on the structure, cell viability, metabolism, and function were determined.
After incubation at 4°C, for up to 6 days, the structures of the arteries were significantly disrupted, especially the tunica media, following incubation in PBS, in contrast with incubation in the HypoRP solution and to a lesser extent, in UW solution. Those disruptions were associated with increased active caspase 3 indicative of apoptosis. Additionally, while incubation in PBS led to a significant decrease in the metabolic activity, UW and HypoRP solutions allowed a stable to increased metabolic activity following 6 days of cold storage. Functional responsiveness to phenylephrine (PE) and sodium nitroprusside (SNP) decreased over time for artery rings stored in PBS and UW solution but not for those stored in HypoRP solution. Moreover, artery rings cold-stored in HypoRP solution were more sensitive to ATP.
The HypoRP solution improved long-term cold storage of porcine arteries by limiting structural alterations, including the collagen matrix, reducing apoptosis, and maintaining artery contraction-relaxation functions for up to 6 days.
在保存血管化复合异体（VCAs）时保持血管功能仍然是一个主要挑战。威斯康星大学（UW）解决方案已显示出显著的短期益处（4-6 小时）。在这里，我们确定新的低温再灌注和保存液 HypoRP 是否可以改善猪动脉在长达 6 天的储存期间的结构、存活和功能。
使用猪肠系膜动脉，确定在盐水（PBS）、UW 或 HypoRP 溶液中孵育长达 6 天对结构、细胞活力、代谢和功能的影响。
在 4°C 孵育长达 6 天后，与 HypoRP 溶液孵育相比，PBS 孵育后动脉的结构明显受损，尤其是中膜，与 UW 溶液孵育相比，结构受损程度较轻。这些破坏与凋亡的活性 caspase 3 增加有关。此外，虽然在 PBS 中孵育会导致代谢活性显著下降，但 UW 和 HypoRP 溶液允许在冷储存 6 天后保持稳定或增加代谢活性。用 PBS 和 UW 溶液储存的动脉环对苯肾上腺素（PE）和硝普钠（SNP）的反应性随时间的延长而降低，但用 HypoRP 溶液储存的动脉环则没有。此外，HypoRP 溶液中冷藏的动脉对 ATP 更敏感。
HypoRP 溶液通过限制结构改变，包括胶原基质，减少细胞凋亡，并保持动脉收缩-松弛功能，在长达 6 天的时间内改善了猪动脉的长期冷藏。
