A preliminary study on polycaprolactone and gelatin-based bilayered tubular scaffolds with hierarchical pore size constructed from nano and microfibers for vascular tissue engineering

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Abstract
Due to the insufficient endothelialization and the poor colonization of smooth muscle cells (SMCs), small-diameter vascular tissue engineering is still challenging. An ideal vascular graft is expected to effectively support the formation of endothelial monolayer and the colonization of SMCs. In this study, we proposed a bilayered scaffold with hierarchical pore size constructed from nano and microfibers by electrospinning PCL-PEG-PCL (PCE) and a blend of PCE and gelatin (PCEG) sequentially. The structural features of nano and microfibers were tuned by the concentration of PCE and the proportion of PCE/gelatin in electrospun solution respectively. The results demonstrated the best nanofiber morphology and relatively high mechanical properties were achieved in 18% PCE (w/v) (PCE18) and PCE and gelatin with a weight ratio of 7:3 (P7G3) at a concentration of 18% (w/v) electrospun membranes. The in vitro co-culturing studies of cells and membranes indicated all the PCE membranes supported the proliferation and spreading of endothelial cells and the further endothelialization of the membranous surface, while PCEG membranes facilitated the migration inward of SMCs. Taking the porosity and mechanical properties into consideration, PCE18 and P7G3 were chosen to construct the inner and outer layers of the bilayered scaffold with hierarchical pore size respectively. The circumferential ring test demonstrated that the bilayered scaffold has good mechanical property both in dry and wet state. After cells were co-cultured with this bilayered scaffold for 7 days, the results manifested a continuous endothelial monolayer has formed on the luminal surface and the SMCs have started to colonized from outer layers, indicating the vast potential of this bilayered scaffold in vascular remodeling and regeneration.

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