Aligned electrospun fibers used for the fabrication of tubular scaffolds possess the ability to regulate cellular alignment and relevant functional expression, with applications in tissue engineering. Despite significant progress in the fabrication of small-diameter vascular grafts (SDVGs) over the past decade, several challenges remain; one of the most problematic of these is the fabrication of aligned nanofibers for multilayer SDVGs. Furthermore, delamination between each layer is difficult to avoid during the fabrication of multilayer structures. This study introduces a new fabrication method for minute delamination four-layer tubular scaffolds (FLTSs) that consist of an interior layer with highly longitudinal aligned nanofibers, two middle layers composed of electrospun sloped and circumferentially aligned fibers, and an exterior layer comprising random fibers. These FLTSs are used to simulate the structures and functions of native blood vessels. Here, thermoplastic polyurethane (TPU)/polycaprolactone (PCL)/polyethylene glycol (PEG) were electrospun to fabricate FLTSs or tubular scaffolds with completely random fibers layer (RLTSs). The surface wettability of the TPU/PCL/PEG tubular scaffold was tested by water contact angle analysis. In particular, compared with RLTSs, FLTSs showed excellent mechanical properties, with higher circumferential and longitudinal tensile properties. Furthermore, the high viability of the human umbilical vein endothelial cells (HUVECs) on the FLTSs indicated the biocompatibility of the tubular scaffolds comparing to RLTSs. The aligned and random composite structure of the FLTSs are conducive to promoting the growth of HUVECs, and the cell adhesion and proliferation on these scaffolds was found to be superior to that on RLTSs. These results demonstrate that the fabricated FLTSs have the potential for application in vascular tissue regeneration and clinical arterial replacements.
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