Activin and follistatin play an important role in the development and function of the skin. In particular they influence various cutaneous processes including inflammation, angiogenesis and wound healing. Androgens also have an important influence on wound healing. Although previous studies provide evidence for an interaction between testosterone and activin/follistatin signals during cutaneous wound healing, no data has been done to confirm this interrelationship. The aim of this study was to examine the relationship between testosterone and the actions of activin/follistatin in the skin during wound healing. In the study, intact and castrated male mice received two linear full-thickness incisions. Serum and skin levels of activin and follistatin were measured at 0, 3, 5, 7 and 14 days post-wounding. Macroscopic and histological analysis showed that intact male mice had a significant delayed in wound healing compared to castrated males. Following injury, intact males had a decrease in serum follistatin, whereas in castrated males, levels remained constant. Castration affected unwounded skin, with a decrease in follistatin (p<0.001) and an increase in activin (p<0.001) skin levels. When wounded, castrated males displayed a delay in the peak of activin with reduced IL-6 levels, compared to the intact males. This study provides the first evidence that the actions of androgens on wound healing are associated with changes in cutaneous levels of activin and follistatin. The male genotype is considered to be a risk factor for impaired healing, with chronic ulcers commonly found in the elderly population. The finding that activin and follistatin are participating in the effects of sex hormones during wound healing processes are of great interest since new targets for treatments of wound healing using follistatin as a regulator of activin could be considered as strategies to solve these kind of problems.