Substitution or bypass is the most effective treatment for vascular occlusive diseases.The demand for artificial blood vessels has seen an unprecedented rise due to the limited supply of autologous blood vessels. Tissue engineering is the best approach to provide artificial blood vessels. In this study, a new type of small-diameter artificial blood vessel with good mechanical and biological properties was designed by using electrospinning coaxial fibers. Four groups of coaxial fibers vascular membranes having polyurethane/gelatin core-shell structure were cross-linked by the EDC-NHS system and characterized. The core-shell structure of the coaxial vascular fibers was observed by transmission electron microscope. After the crosslinking, the stress and elastic modulus increased and the elongation decreased, burst pressure of 0.11 group reached the maximum (2844.55 ± 272.65 mmHg) after cross-linking, which acted as the experimental group. Masson staining identified blue-stained ring or elliptical gelatin ingredients in the vascular wall. The cell number in the vascular wall of the coaxial group was found in muscle embedding experiment significantly higher than that of the non-coaxial group at all time points (p < 0.001). Our results showed that the coaxial vascular graft with the ratio of 0.2:0.11 had better mechanical properties (burst pressure reached 2844.55 ± 272.65 mmHg); Meanwhile its biological properties were also outstanding, which was beneficial to cell entry and offered good vascular remodeling performance. Polyurethane (PU); Gelatin (Gel); Polycaprolactone (PCL); polylactic acid (PLA);1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC); N-Hydroxy succinimide (NHS); 4-Morpholine-ethane-sulfonic (MES); phosphate buffered saline (PBS); fetal calf serum (FCS); Minimum Essential Medium (MEM); Dimethyl sulfoxide (DMSO); hematoxylineosin (HE).

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A completely confluent endothelial cell (EC) monolayer is required to maintain proper vascular function in small diameter tissue-engineered vascular graft (TEVG). However, the most effective method for EC attachment to the luminal surface and formation of an entire endothelium layer that works in vitro remains a complicated challenge that requires urgent resolution. Although pulsatile flow has been shown to be better suited for the generation of functional endothelium, the optimal frequency setting is unknown. Several pulsatile flow frequencies were used to implant rat bone mesenchymal stem cells (MSC) into the lumen of decellularized porcine carotid arteries. The endothelium's integrity and cell activity were investigated in order to determine the best pulse frequency settings. The results showed that MSC were maximally preserved and exhibited maximal morphological changes with improved endothelialization performance in response to increased pulse stimulation frequency. Increased pulse frequency stimulation stimulates the expression of mechanoreceptor markers, cytoskeleton reorganization in the direction of blood flow, denser skeletal proteins fibronectin, and stronger intercellular connections when compared to constant pulse frequency stimulation. MSC eventually develops an intact endothelial layer with anti-thrombotic properties on the inner wall of the decellularized tubular lumen. Conclusion: The decellularized vessels retain the three-dimensional structure of the vasculature, have a surface topography suitable for MSC growth, and have good mechanical properties. By increasing the frequency of pulsed stimulation, MSC endothelialize the lumen of the decellularized vasculature. It is expected to have anti-thrombotic and anti-neointimal hyperplasia properties after implantation, ultimately improving the patency of TEVG.