Mechanical and seeding properties of human umbilical vein – a potential scaffold for a tissue-engineered vessel graft

Abstract

Objectives: The mechanical properties and seeding with endothelial cells were investigated in fresh and cryopreserved human umbilical vein. Methods: Human umbilical veins (HUV) were frozen in Euro-Collins/1M DMSO at –1°C/min and stored in liquid nitrogen. Stress-strain relationships of fresh and thawed veins were determined in an uniaxial tension-testing rig. HUV endothelial cells (HUVEC) were seeded onto denuded HUV under static conditions and grown for 3d. Luminal surfaces were analyzed by scanning electron microscopy. Calcein-stained cells were seeded hyperconfluently to determine the cell retention capacity of fresh and cryopreserved veins. Results: The stress-strain relationships of HUV followed a biphasic pattern typical for natural vessels. Neither the failure stress (2.71±0.36 vs. 3.25±0.97 N, n=3) nor the displacement required to achieve failure (9.73±0.9 vs. 7.43±2.07mm, n=3) were altered by cryopreservation. The burst pressure was estimated as approx. 1000mm Hg within the limitations of the uniaxial model. HUVEC seeded onto denuded HUV formed patches (at 9E3 cells per cm2) or an almost confluent endothelium (at 3E4 cells per cm2) within three days. The capacity to retain seeded HUVEC of denuded HUV was not altered by cryopreservation (1.15±0.08E5 vs. 1.26±0.14E5 cells per cm2, n=6). Conclusions: The burst pressure of HUV seems to be sufficiently high for the human arterial circulation and is not altered by cryopreservation. HUVEC can establish a confluent endothelium on denuded HUV. Therefore HUV appears to be a suitable storable scaffold for vascular tissue engineering.