Combination therapy profoundly improved skin flap survival by modulating KATP channels and nitric oxide.

A potential therapeutic approach on skin flap necrosis is to target parallel pathways involved in necrosis. Azelaic Acid, Minoxidil and Caffeine combination was tried on skin flap survival by their possible interaction with ATP sensitive potassium (KATP) channels and nitric oxide pathway.Sprauge-Dawley rats were divided into 8 groups for skin flap surgery. Azelaic acid, minoxidil, caffeine, or their combination were applied topically in different groups. Two additional groups were treated with L-NAME or glibenclamide in addition to the combination therapy. Percentage of flap necrosis was calculated and flap samples were removed to measure tissue malondialdehyde (MDA) and nitric oxide (NO) and expression of inducible nitric oxide synthase (iNOS), Bcl-2 and Bax proteins.Combination therapy profoundly decreased skin flap necrosis, tissue MDA contents, and expression of the pro-apoptotic protein Bax (p < 0.05 vs. single treatments). These effects were reversed by L-NAME and glibenclamide pre-treatments. Further evaluations showed combination therapy increases flap tissue NO content and iNOS expression (p < 0.05 vs. single treatments).Beneficial effect of the combination therapy with azelaic acid, minoxidil and caffeine therapy on rescuing the flap from necrosis by targeting parallel signaling pathways suggested potential applications in clinical practice.