Viability of Endothelial Cells in Preserved Human Saphenous Vein Allografts

Chee, Hyun Keun; Kim, Yong Jin

Abstract

BACKGROUND: Autogenous vein is the preferred vascular graft for patients who require coronary artery bypass surgery or peripheral arterial bypass surgery. When an autogenous vein is not available, an allograft saphenous vein can be used as an alternative conduit. Although arterial homograft has been under investigation since the beginning of this century, the viability of endothelial cells and the optimum mode of storage for the venous and arterial allografts is controversial. In addition, with the recently gained knowledge of vascular endothelial functions, such as the production of nitric oxide or thrombomodulin, the viability and antigenicity of endothelial cells are being studied again. The purpose of this study was to evaluate the viability of endothelial cells in the preserved human saphenous veins. MATERIAL AND METHOD: The veins were stored in a 4 degrees C RPMI (Roswell Park Memorial Institute) 1640 solution including 10% fetal calf serum, for one, three, five, seven or fourteen days. After the completion of the storage period, the veins were divided into two groups: Group I: studied immediately at 4 degrees C (cold) storage (I-1, I-3, I-5, I-7, I-14), and Group II: studied after storage at -196 degrees C liquid nitrogen tank (cryopreservation) in an RPMI 1640 solution containing 10% DMSO for two weeks (II-1, II-3, II-5, II-7, II-14). Light microscopy and scanning electron microscopy (SEM), trypan blue exclusion testing, and thrombomodulin immunohistochemistry were performed. RESULT: In a morphometric study using SEM, there was statistically significant increase in Gundry Score in Groups I-7, I-14, II-5, II-7, and II-14 and showed cellular destruction (p<0.05). In the thrombomodulin immunohistochemistry study, there was reactivity in Groups I-1, I-3, and I-5, but the cryopreserved group revealed decreased reactivity (p<0.05). The trypan blue exclusion testing also showed superior viability in cold storage Group I.
CONCLUSION
Venous allografts preserved in a 4 degrees C RPMI 1640 solution showed well preserved endothelial cellular integrity and thrombomodulin expression at up to seven days of preservation. Although cryopreservation of venous allografts stored in 10% DMSO -RPMI 1640 solution maintained the endothelial cellular structure on SEM, immunohistochemistry from the thrombomodulin and trypan blue exclusion testing showed decreased viability. It remains to be seen whether the decreased thrombomodulin reactivity could be restored, and what the nature to the relationship is between thrombomodulin and long-term patency of allografts.