Perforation of eardrum or tympanic membrane (TM) is a common clinical condition, which occurs due to infection or injury of the eardrum, and could results in varying degrees of conductive hearing loss among all ages. In this study, the authors report the combinatorial approach of designing mechanically-tunable and vascular supportive nanofibrous membranes by 3D printing-assisted electrospinning (e-spin) using polycaprolactone (PCL) and gelatin with different mass ratios suitable to repair a perforated eardrum. The physicochemical, mechanical, and biological properties of the membranes were characterized. The results show that the membrane has nanofibrous morphology with fibers are of varying size (400–600 nm in diameter) depending on processing conditions. The wettability and mechanical properties of the membrane can be tuned by regulating the gelatin content. Moreover, a biomimetic repair strategy inspired by chicken eggshell membrane, often used in wound dressings, was also presented for study and results show that the suture retention strength of the fabricated membrane can meet clinical translational requirements to promote TM healing. The vascular cell responsiveness of PCL/gelatin nanofibrous membrane was evaluated using human umbilical vein endothelial cells (HUVECs) and the results showed satisfactory biocompatibility, vascular cell responsiveness, and cell proliferation. The findings of this study demonstrate that the combinatorically engineered PCL/gelatin nanofibrous membrane has great potential for repairing perforated eardrum. © 2020 Wiley Periodicals LLC.