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Limited efficacy of adipose stromal cell secretome-loaded skin-derived hydrogels to augment skin flap regeneration in rats.

Insufficient vascularization is a recurring cause of impaired pedicled skin flap healing. The administration of adipose tissue-derived stromal cells' (ASC) secretome is a novel approach to augment vascularization. Yet, the secretome comprised soluble factors that require a sustained release vehicle to increase residence time. We hypothesized that administration of a hydrogel derived from decellularized extracellular matrix (ECM) of porcine skin with bound trophic factors from ASCs, enhances skin flap viability and wound repair in a rat model. Porcine skin was decellularized and pepsin-digested to form a hydrogel at 37°C. Conditioned medium (CMe) of human ASC was collected, concentrated twentyfold and mixed with the hydrogel. Sixty Wistar rats were included. A dorsal skin flap (caudal based) of 3 x 10 cm was elevated for topical application of: DMEM medium (group I), a pre-hydrogel with or without ASC CMe (group II and III) or ASC CMe (group IV). After 7, 14 and 28 days, perfusion was measured and skin flaps were harvested for wound healing assessment and immunohistochemical analysis. Decellularized skin ECM hydrogel contained negligible amounts of DNA (11.6 ± 0.6 ng/mg), was noncytotoxic and well-tolerated by rats. Irrespective of ASC secretome, ECM hydrogel application resulted macroscopically and microscopically in similar dermal wound healing in terms of proliferation, immune response and matrix remodeling as the control group. However, ASC CMe alone increased vessel density after seven days. Concluding, porcine skin derived ECM hydrogels loaded with ASC secretome are non-cytotoxic but demand optimization to significantly augment wound healing of skin flaps.