Electrospinning is a versatile technique for producing nanocomposite in biomedical applications. In this study, polyurethane (PU) cardiac patch loaded with nickel oxide (NiO) was fabricated using electrospinning technique. The morphology study revealed that the PU/NiO nanocomposites exhibited reduced fiber diameter (758 AE 157.10 nm) and pore diameter (868 AE 73.26 nm) compared to the pristine PU (fiber diameter 890 AE 116.91 nm and pore diameter 1064 AE 74.31 nm). The contact angle study indicated the decreased wettability behavior of the electrospun nanocomposite (106 AE 0.58 ) than the pure PU (100 AE 0.58 ). The incorporation of NiO into PU improved the mechanical strength (15.25 MPa) and surface roughness (448 nm) of the pristine PU (tensile strength 7.12 MPa and surface roughness 313 nm). The developed PU/NiO nanocomposites showed delayed blood clotting time and low hemolytic percentage as revealed in the coagulation studies insinuating the improved anticoagulant nature compared to the pristine PU. Moreover, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay demonstrated the nontoxic behavior of fibroblast cells in the developed nanocomposite (135 AE 1%) than the pristine PU (133.33 AE 8.96%). The newly developed nanocomposite patch rendered better physicochemical, improved blood compatibility, and nontoxic to the fibroblast cells. Hence, the electrospun PU/NiO nanocomposite might serve as a plausible scaffold for the cardiac tissue engineering.