Estimation of wound age is of at most importance in medicolegal death cases. Wound age can be evaluated using morphological, cytological, and molecular biological techniques. The purpose of this study was to evaluate the role of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) in the determination of wound age in autopsy cases. The present study included (20) autopsy cases; (16) with chronologically dated stab wounds (wound age varied from 30 to 300 minutes) and (4) cases without any injury to provide skin specimens serving as a negative control (Control group). Injured cases were examined, full-thickness skin specimens were collected and classified chronologically according to age of wound into 4 groups each group consisted of 4 specimens: Group I: (Time of injury ≤ 30 min), Group II: (Time of injury 31-60 min), Group III: (Time of injury 61-120 min), Group IV: (Time of injury >120min). Each group of injured cases was divided into two subgroups: Subgroup A: At uninjured contralateral site, Subgroup B: At the site of injury. All specimens were stained and studied and subjected to histological, immunohistochemical followed by a histomorphometric. The times passed since injury infliction (stab wound) caused a change in the area percentage of the released cytokines (TNF-α and IL6) whereas, early detection of (TNF-α and IL6) were at 30 min and a marked reaction became obvious at 31-60 min and from 30 min to 300 min, there was a significant and progressive time-dependent increase in the area percentage of the two markers, evaluated semi-quantitatively. We concluded that pro-inflammatory cytokines (IL-6) and (TNF-α) could serve as a useful tool for the estimation of wound age, in particular in the early post-traumatic interval prior to leukocyte reaction.