A major roadblock in the development of tissue engineered vascular grafts (TEVGs) is achieving construct endothelialization that is stable under physiological stresses. The aim of the current study was to validate an approach for generating a mechanically stable layer of endothelial cells (ECs) in the lumen of TEVGs. To accomplish this goal, a unique method was developed to fabricate a thin EC layer using poly(ethylene glycol) diacrylate (PEGDA) as an intercellular "cementing" agent. This EC layer was subsequently bonded to the lumen of a tubular scaffold to generate a bi-layered construct. The viability of bovine aortic endothelial cells (BAECs) through the "cementing" process was assessed. "Cemented" EC layer expression of desired phenotypic markers (AcLDL uptake, VE-cadherin, eNOS, PECAM-1) as well as of injury-associated markers (E-selectin, SM22α) was also examined. These studies indicated that the "cementing" process allowed ECs to maintain high viability and expression of mature EC markers while not significantly stimulating primary injury pathways. Finally, the stability of the "cemented" EC layers under abrupt application of high shear pulsatile flow (~11 dyn/cm2, P avg ~ 95 mmHg, ΔP ~ 20 mmHg) was evaluated and compared to that of conventionally "seeded" EC layers. Whereas the "cemented" ECs remained fully intact following 48 h of pulsatile flow, the "seeded" EC layers delaminated after less than 1 h of flow. Furthermore, the ability to extend this approach to degradable PEGDA "cements" permissive of cell elongation was demonstrated. Combined, these results validate an approach for fabricating bi-layered TEVGs with stable endothelialization.
- Endothelial cells
- Mechanical conditioning
- Poly(ethylene glycol)
- Tissue engineered vascular grafts