In this study, a wound dressing based on polyurethane (PU) blended with copper sulphate nanofibers was developed using an electrospinning technique. The prepared PU and PU nanocomposites showed smooth fibers without any bead defects. The prepared nanocomposites showed smaller fiber (663 +/- 156.30 nm) and pore (888 +/- 70.93 nm) diameter compared to the pristine PU (fiber diameter 1159 +/- 147.48 nm and pore diameter 1087 +/- 62.51 nm). The interaction of PU with copper sulphate was evident in the infrared spectrum through hydrogen-bond formation. Thermal analysis displayed enhanced weight residue at higher temperature suggesting interaction of PU with copper sulphate. The contact angle measurements revealed the hydrophilic nature of the prepared nanocomposites (71 degrees +/- 2.309 degrees) compared with pure PU (100 degrees +/- 0.5774 degrees). The addition of copper sulphate into the PU matrix increased the surface roughness, as revealed in the atomic force microscopy (AFM) analysis. Mechanical testing demonstrated the enhanced tensile strength behavior of the fabricated nanocomposites (18.58 MPa) compared with the pristine PU (7.12 MPa). The coagulation assays indicated the enhanced blood compatibility of the developed nanocomposites [activated partial thromboplastin time (APTT)-179 +/- 3.606 s and partial thromboplastin time (PT)-105 +/- 2.646 s] by showing a prolonged blood clotting time compared with the pristine PU (APTT-147.7 +/- 3.512 s and PT-84.67 +/- 2.517 s). Furthermore, the hemolysis and cytotoxicity studies suggested a less toxicity nature of prepared nanocomposites by displaying low hemolytic index and enhanced cell viability rates compared with the PU membrane. It was observed that the fabricated novel wound dressing possesses better physicochemical and enhanced blood compatibility properties, and may be utilized for wound-healing applications.