Fabrication of heparinized bi-layered vascular graft with PCL/PU/gelatin co-electrospun and chitosan/silk fibroin/gelatin freeze-dried hydrogel for improved endothelialization and enhanced mechanical properties

Fabricating a biocompatible small-diameter vascular graft (< 6 mm) with mechanical properties similar to the natural vein and adding good anti-thrombogenic, endothelialization, and hyperplasia properties remains a challenge. To this end, we fabricated a heparinized bilayer graft to address this problem. The proposed bilayer sample consisted of a heparinized polycaprolactone (PCL), polyurethane (PU), and gelatin (G) co-electrospun inner layer and chitosan, gelatin, and silk fibroin freeze-dried hydrogel crosslinked with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) outer layer. The samples exhibited great ultimate stress, Young's module, and suture retention of 4.16 +/- 0.25 MPa, 8.24 +/- 2.59 MPa and 4.83 +/- 0.31 N, respectively. The heparin release assay indicated a sustained release profile of around 70% after 4weeks, which can be attributed to the excellent control via emulsion. Furthermore, the heparinized samples demonstrated good anti-thrombogenic properties investigated in the platelet adhesion assay. For the outer layer, the hydrogel crosslinked with non-toxic materials was prepared through the freeze-drying method to achieve high porosity (64.63%), suitable for smooth muscle cell activity. Moreover, inner and outer layers showed high cell viability toward endothelial (78.96%) and smooth muscle cells (57.77%), respectively. Overall, the proposed heparinized graft exhibited excellent potential for vascular graft regeneration.