Wound dressings provide therapeutic and protective features and promotes natural healing process when applied to a wound area. Being non-toxic and immunologically inert, natural biopolymers have potential in fabrication of wound dressings. Growth factors and antibiotics can also be used in functionalization of wound dressings as well as plant extracts. Olive leaf extract has gained attraction due to its dual antimicrobial and antioxidant effect. By clearing pathogenic microorganisms and scavenging against increased amount of reactive oxygen species in the wound area, it has high potential in wound healing. In this study, olive leaf extract incorporated zein fibers were prepared as a model of wound dressing contact layer. In this regard, crude olive leaf extract was fractionated and characterized in terms of antioxidant capacity, total phenol content and antimicrobial activity. Crude extract and its fractions were also subjected to wound scratch assay in the presence of hydrogen peroxide. Oleuropein, as the most abundant component in crude extract, was found to promote cell migration better and close the wound area at a higher rate than other components. On the other hand, crude olive leaf extract exhibited higher percentage of wound closure than its fractions within the same time period, which may be attributed to synergistic effect of unidentified phenolics. Crude olive leaf extract also provided crosslinking effect when incorporated into zein fibers, as well as promoting cell spreading behaviour.