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大隐静脉内皮的多光子显微镜评估及其用新溶液GALA的保存

Multi-photon microscopic evaluation of saphenous vein endothelium and its preservation with a new solution, GALA.

作者信息

Thatte Hemant S, Biswas Kunda S, Najjar Samer F, Birjiniuk Vladimir, Crittenden Michael D, Michel Thomas, Khuri Shukri F

机构信息

Department of Surgery, VA Boston Healthcare System, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts, USA.

出版信息

Ann Thorac Surg. 2003 Apr;75(4):1145-52; discussion 1152. doi: 10.1016/s0003-4975(02)04705-7.

Abstract

BACKGROUND

Injury to endothelium can compromise the patency of bypass grafts harvested during coronary artery bypass graft (CABG) surgery. Maintaining structural and functional viability of endothelium in grafts may lead to improved long-term patency. The information gained from the application of multi-photon microscopy in transmission and epifluorescence mode was used to assess the structural and functional integrity of human saphenous vein segments stored in multiple preservation solutions, and to design a superior storage solution.

METHODS

Multi-photon microscopy was used to image deep within saphenous vein tissue harvested from patients undergoing CABG for analysis of endothelial structure and function. Endothelial cell structural viability, calcium mobilization, and nitric oxide generation were determined using specific fluorescence markers.

RESULTS

Within 60 minutes of harvest and storage in standard preservation solutions, calcium mobilization and nitric oxide generation were markedly diminished with more than 90% of endothelial cells no longer viable in the vein. In contrast, veins could be stored for 24 hours without substantial loss in cell viability in a newly formulated heparinized physiologic buffered salt solution containing glutathione, ascorbic acid, and L-arginine (GALA).

CONCLUSIONS

Standard solutions in clinical use today led to a profound decline in saphenous vein endothelial cell viability, whereas the newly designed physiologic salt solution (GALA) maintained endothelial function and structural viability for up to 24 hours. The improvements seen from using GALA as a vessel storage medium may lead to greater long-term vein graft patency following CABG surgery.

摘要

背景

内皮损伤会影响冠状动脉旁路移植术(CABG)中获取的旁路移植物的通畅性。维持移植物中内皮的结构和功能活性可能会提高长期通畅率。应用多光子显微镜在透射和落射荧光模式下获得的信息用于评估储存在多种保存溶液中的人隐静脉段的结构和功能完整性,并设计一种更优的保存溶液。

方法

使用多光子显微镜对从接受CABG的患者获取的隐静脉组织进行深度成像,以分析内皮结构和功能。使用特定荧光标记物测定内皮细胞的结构活性、钙动员和一氧化氮生成。

结果

在收获并储存在标准保存溶液中60分钟内,钙动员和一氧化氮生成明显减少,静脉中超过90%的内皮细胞不再存活。相比之下,在一种新配制的含有谷胱甘肽、抗坏血酸和L-精氨酸(GALA)的肝素化生理缓冲盐溶液中,静脉可以保存24小时而细胞活力没有实质性损失。

结论

当今临床使用的标准溶液导致隐静脉内皮细胞活力大幅下降,而新设计的生理盐溶液(GALA)可维持内皮功能和结构活性长达24小时。使用GALA作为血管保存介质所观察到的改善可能会导致CABG术后静脉移植物的长期通畅率更高。

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