Retained structural integrity of collagen and elastin within cryopreserved human heart valve tissue as detected by two-photon laser scanning confocal microscopy
Retained structural integrity of collagen and elastin within cryopreserved human heart valve tissue as detected by two-photon laser scanning confocal microscopy
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DOI：
10.1016/j.cryobiol.2009.06.012
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发表时间：
2009-10-01
期刊：
影响因子：
2.7
通讯作者：
Heacox, Albert E.
中科院分区：
文献类型：
--
作者：
Gerson, Cindy J.;Goldstein, Steven;Heacox, Albert E.
冷冻保存通常用于心脏瓣膜同种异体移植物的长期储存。尽管冷冻保存的同种异体移植物具有优异的血流动力学性能和耐久性，但有报道质疑冷冻保存是否会影响瓣膜结构蛋白、胶原蛋白和弹性蛋白。本研究使用双光子激光扫描共聚焦显微镜（LSCM），以评估冷冻保存对人主动脉瓣和肺动脉瓣的瓣叶和管道内胶原蛋白和弹性蛋白完整性的影响。为了允许对新鲜和冷冻保存的组织进行成对比较，将试验瓣膜纵向一分为二，其中一个片段新鲜成像，另一个片段在冷冻保存和短暂储存在液氮中后成像。胶原蛋白通过二次谐波产生（SHG）刺激检测，弹性蛋白通过自发荧光激发检测。所有所得图像的定性分析表明，冷冻保存后瓣叶和导管内的胶原蛋白和弹性蛋白结构得以维持。胶原蛋白和弹性蛋白的全动态范围成像所需的优化激光透射率（OPLT）的分析表明，OPLT观察结果在新鲜和冷冻保存的样品之间具有高度可变性。响应于冷冻保存的供体特异性平均OPLT的变化没有表现出一致的方向趋势。供体聚集结果主要显示，由于冷冻保存，胶原蛋白和弹性蛋白平均OPLT无统计学显著变化。由于OPLT与结构信号强度呈反比关系，因此这些结果表明新鲜组织和冷冻保存组织之间的胶原蛋白和弹性蛋白信号强度基本上没有统计学差异。总的来说，这项研究表明，人类心脏瓣膜同种异体移植物的常规冷冻保存不会对它们的胶原蛋白和弹性蛋白结构完整性产生不良影响。(C)2009 Elsevier Inc. All rights reserved.
Cryopreservation is commonly used for the long-term storage of heart valve allografts. Despite the excellent hemodynamic performance and durability of cryopreserved allografts, reports have questioned whether cryopreservation affects the valvular structural proteins, collagen and elastin. This study uses two-photon laser scanning confocal microscopy (LSCM) to evaluate the effect of cryopreservation on collagen and elastin integrity within the leaflet and conduit of aortic and pulmonary human heart valves. To permit pairwise comparisons of fresh and cryopreserved tissue, test valves were bisected longitudinally with one segment imaged fresh and the other imaged after cryopreservation and brief storage in liquid nitrogen. Collagen was detected by second harmonic generation (SHG) stimulation and elastin by auto-fluorescence excitation. Qualitative analysis of all resultant images indicated the maintenance of collagen and elastin structure within leaflet and conduit post-cryopreservation. Analysis of the optimized percent laser transmission (OPLT) required for full dynamic range imaging of collagen and elastin showed that OPLT observations were highly variable among both fresh and cryopreserved samples. Changes in donor-specific average OPLT in response to cryopreservation exhibited no consistent directional trend. The donor-aggregated results predominantly showed no statistically significant change in collagen and elastin average OPLT due to cryopreservation. Since OPLT has an inverse relationship with structural signal intensity, these results indicate that there was largely no statistical difference in collagen and elastin signal strength between fresh and cryopreserved tissue. Overall, this study indicates that the conventional cryopreservation of human heart valve allografts does not detrimentally affect their collagen and elastin structural integrity. (C) 2009 Elsevier Inc. All rights reserved.