Abstract <p>Urethral stenosis is a common urinary system medical condition that significantly impacts the quality of life. Tissue engineering is one of the proposed methods for treating this condition, with the primary aim of designing and creating a tissue structure to repair urethral damage. In this work, we designed a tubular tissue engineering scaffold featuring a composite gradient of polymers, taking inspiration from native tissue. To achieve this, collagen and silk fibroin were extracted, and their chemical structures were confirmed using FTIR. Subsequently, a two-nozzle electrospinning device was utilized to fabricate scaffolds composed of PCL, collagen, and silk fibroin (PCSFG). A composite gradient scaffold was created by changing the flow rate of silk fibroin. The average fiber diameter of PCSFG measured 217 ± 8 nm. The scaffold exhibited an approximately 8.85% degradation after 30 days. The ultimate tensile strength of the gradient scaffold was 5.56 MPa, within the elastic zone. Cell viability on days 7 was higher in the gradient scaffold than the other scaffolds. Furthermore, PI staining of the cultured cells on both the inner and outer layers of the PCSFG tubular scaffold demonstrated cell growth, proliferation, and infiltration from the inner to the outer layer and vice versa. Consequently, this gradient scaffold holds promise as an option for urethral tissue engineering.</p> Graphical abstract <p></p>