Objective: This project’s primary purpose was to create engineered vascular scaffolds using polyurethane, polycaprolactone, and pullulan polymers, along with suitable mechanical-dynamic conditions. Therefore, electrospun scaffolds with optimized intrinsic physiological properties and the ability to support endothelial cells were prepared in vitro, and cell viability was studied in PCL-PU and PCL-PU scaffolds containing Pullulan. The main methods: The electrospinning method has been used to prepare PCL-PU and PCL-PU scaffolds containing Pullulan. The scaffold’s surface morphology was evaluated using SEM microscopic imaging. The scaffolds’ physicochemical properties were prepared using ATR-FTIR, strain stress, and water contact angle tests, and the biocompatibility of PCL-PU and PU-PCL-Pl nanofibers was evaluated using the MTT test. Principal findings: The test results showed that PCL-PU scaffolds containing Pullulan have more suitable mechanical properties such as stress-strain, water contact angle, swelling rate, biocompatibility, fiber diameter, and pore size compared to PU-PCL. The culture of endothelial cells under static conditions on these scaffolds did not cause cytotoxic effects under static conditions compared to the control group. SEM images confirmed the ability of endothelial cells to attach to the scaffold surface. Summary and conclusion: The results showed that PCL-PU substrate containing pullulan could stimulate endothelial cells’ proliferation under static conditions.