In this study, poly(vinyl alcohol) nanofibres were produced and evaluated for wound dressing applications. However, the solubility of the poly(vinyl alcohol) matrix in the aqueous medium generally limits its applications. To overcome this problem, 1,2,3,4 butanetetracarboxylic acid (BTCA) and citric acid (CA) were used as crosslinking agents. Successful bead-free nanofibres were produced, and they both preserved their fibrous structure after water treatment. The mean fibre diameters of polycarboxylic acid crosslinked nanofibres were lower compared to pure PVA nanofibres. Although the morphology of BTCA and CA crosslinked nanofibres was similar, the swelling degree of PVA/CA was found to be higher. Furthermore, toxicity and keratinocyte cell proliferation performance of produced PVA/BTCA and PVA/CA nanofibres indicated that these nanofibrous materials could be used in wound dressing applications.