The development of biocompatible patches that can provide adequate mechanical properties and mimicking the physiological relevance still has enormous challenges in the field of tissue engineering. In this study, the aim is to engineer and develop tri-layered nanofiber patches to synchronous the mechanical properties and mimic the complex structure of living cardiovascular tissues. The developed patches consisting of blended silk fibrin (SF) and polyvinyl alcohol (PVA) composite as a hydrophilic middle layer. The upper and lower layers were composed of polycaprolactone polymer (PCL) and Poly (Lactic Acid) (PLA), respectively. Layer by layer of the developed patches were fabricated by electrospinning technique. The patches characteristics such as morphology, surface topography, and physiochemical, biodegradable and mechanical stability were investigated. Furthermore, a human endothelial cell line (EA. hy926) was exploited to assess the biological properties of the fabricated patches. The results show that the patches had high endothelial cells biocompatibility, mechanical elasticity, and unique nanofibers structure.