The determination of wound age and vitality has an important role in forensic practice. Objectives: Interleukin-I β (IL1- β), interleukin- 6 (IL-6) and fibronectin (Fn) were estimated (in extracts of skin samples), and histopathological examination of the skin samples in adult male rats for differentiation between antemortem (AM) and postmortem (PM) wounds and determination of wound age. Methods: forty two adult male albino rats were divided equally into 7groups. AM (group I) and PM (group II) controls: the samples were taken from the dorsum of intact skin rats. The wound samples were taken 0.5 hour (group III), 3 hours (group IV), 6 hours (group V) and 24 hours (group VI) after infliction of the incised wounds. Group VII: samples taken from PM inflected wounds. The skin biopsies were taken under anesthesia in all AM tested groups and all specimens were biopsied for biochemical and histopathological examination. Results: There was significant time dependent increase in IL-1β, IL6 and Fn levels at 0.5, 3, 6 and 24 hours after wound infliction in comparison with the AM control. Specimens from PM wounds showed insignificant changes in IL-1β, IL6 and Fn levels in comparison with PM control. The histopathological examination revealed signs of vitality in AM wound groups that were absent in PM inflected wound group. Conclusion: We concluded that the combined assay of IL-1β, IL-6 and Fn may be a useful tool in age estimation of AM inflicted skin wounds. We also concluded that vital reactions that appeared in AM skin wound sections are essential for release of these parameters.