It has been shown that vasoactive mediators such as nitric oxide are involved in the period of I/R and are likely initiators of a cascade of events that ultimately lead to late phase I/R injury protection. Twenty-seven Sprague-Dawley rats were used in 3 arms of the study involving isolation of the neurovascular pedicle and induction of ischemia in the extensor digitorum longus muscle. Experimental animals systemically received a nitric oxide donor, spermine-NONOate, 1 hour prior to a 6 hour “flap” ischemia period involving the extensor digitorum longus (EDL) muscle. Twenty-one days after the ischemic insult, the EDL muscle was analyzed by tetanic force testing, followed by histology. Ischemia/reperfusion-only animals (I/R) and control animals underwent identical analysis. Data demonstrate functional benefits in tetanic force testing in animals treated with spermine-NONOate 30 minutes before muscle flap ischemia compared to animals undergoing ischemia/reperfusion only (p<0.0001). Histology reveals attenuation in the stigmata of organ and cellular ischemic injury in the spermine-NONOate group compared to the I/R group. In conclusion, spermine NONOate prior to 6 hours of ischemia with reperfusion in a rat muscle flap model confers physiologic and histologic advantages 21 days after injury.