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冷冻保存可降低动脉同种异体移植物在保存期间的血栓形成性。

Cryopreservation moderates the thrombogenicity of arterial allografts during storage.

机构信息

Department of Vascular and Endovascular Surgery, Heart and Vascular Center, Semmelweis University, Budapest, Hungary.

Department of Biochemistry, Semmelweis University, Budapest, Hungary.

出版信息

PLoS One. 2021 Jul 22;16(7):e0255114. doi: 10.1371/journal.pone.0255114. eCollection 2021.

Abstract

INTRODUCTION

Management of vascular infections represents a major challenge in vascular surgery. The use of cryopreserved vascular allografts could be a feasible therapeutic option, but the optimal conditions for their production and use are not precisely defined.

AIMS

To evaluate the effects of cryopreservation and the duration of storage on the thrombogenicity of femoral artery allografts.

METHODS

In our prospective study, eleven multi-organ-donation-harvested human femoral arteries were examined at five time points during storage at -80°C: before cryopreservation as a fresh native sample and immediately, one, twelve and twenty-four weeks after the cryopreservation. Cross-sections of allografts were perfused with heparin-anticoagulated blood at shear-rates relevant to medium-sized arteries. The deposited platelets and fibrin were immunostained. The thrombogenicity of the intima, media and adventitia layers of the artery grafts was assessed quantitatively from the relative area covered by fibrin- and platelet-related fluorescent signal in the confocal micrographs.

RESULTS

Regression analysis of the fibrin and platelet coverage in the course of the 24-week storage excluded the possibility for increase in the graft thrombogenicity in the course of time and supported the hypothesis for a descending trend in fibrin generation and platelet deposition on the arterial wall. The fibrin deposition in the cryopreserved samples did not exceed the level detected in any of the three layers of the native graft. However, an early (up to week 12) shift above the native sample level was observed in the platelet adhesion to the media.

CONCLUSIONS

The hemostatic potential of cryopreserved arterial allografts was retained, whereas their thrombogenic potential declined during the 6-month storage. The only transient prothrombotic change was observed in the media layer, where the platelet deposition exceeded that of the fresh native grafts in the initial twelve weeks after cryopreservation, suggesting a potential clinical benefit from antiplatelet therapy in this time-window.

摘要

简介

血管感染的处理是血管外科学面临的重大挑战。使用冷冻保存的血管同种异体移植物可能是一种可行的治疗选择,但对于其生产和使用的最佳条件尚未明确界定。

目的

评估冷冻保存和储存时间对股动脉同种异体移植物血栓形成的影响。

方法

在我们的前瞻性研究中,在 -80°C 下储存时,在五个时间点检查了 11 个人多器官捐献收获的人类股动脉:冷冻保存前作为新鲜的天然样本,以及冷冻保存后立即、一、十二和二十四周。用肝素抗凝的血液对同种异体移植物的横截面进行灌注,以达到与中等大小动脉相关的剪切速率。用免疫染色法对沉积的血小板和纤维蛋白进行染色。从共聚焦显微镜图像中纤维蛋白和血小板相关荧光信号的相对面积评估动脉移植物的内皮层、中膜和外膜层的血栓形成。

结果

24 周储存过程中纤维蛋白和血小板覆盖的回归分析排除了移植物血栓形成随时间增加的可能性,并支持了纤维蛋白生成和血小板在动脉壁上沉积呈下降趋势的假设。冷冻保存样本中的纤维蛋白沉积未超过天然移植物任何一层的检测水平。然而,在中膜层观察到早期(直至 12 周)对天然样本水平的增加,表明在冷冻保存后的最初 12 周内血小板黏附到中膜的可能性更大。

结论

冷冻保存的动脉同种异体移植物的止血潜力得以保留,而在 6 个月的储存过程中其血栓形成潜力下降。仅在中膜层观察到短暂的促血栓形成变化,在冷冻保存后最初 12 周内,血小板沉积超过新鲜天然移植物,提示在此时间窗口内抗血小板治疗可能具有潜在的临床获益。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3bd3/8297765/02953709629b/pone.0255114.g001.jpg

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