In this study, we aimed to investigate the potential use of keratin biofiber (KBF)-reinforced biodegradable thermoplastic polyurethane (TPU) composites as tissue engineering scaffolds. The composites were prepared by a solution casting method. Combined salt leaching and thermally induced phase-separation methods were used to manufacture porous scaffold structures. The mechanical properties were investigated with tensile tests and dynamic mechanical analysis (DMA). An increase in the fiber content increased the modulus and strength of the composites. The DMA results revealed that the storage modulus of the composites compared to that of the neat TPU was improved by the addition of feathers. Scanning electron microscopy analysis indicated that the dispersion of KBF in the TPU matrix was uniform. According to the water contact angle measurements, the hydrophilicity increased with increasing KBF content up to 80%. The thermal properties of the KBF-TPU films were measured by a differential scanning calorimetry method. The increase in the amount of KBF did not significantly affect the glass-transition temperature values. We examined the cytotoxicity and biocompatibility by performing cell culturing with the L929 mouse fibroblast cell line. The cell viability was found to be greater than 80% for all of the scaffold compositions. This showed that the KBF-TPU composite scaffolds may serve as a highly potent new material for skin tissue engineering applications. (C) 2017 Wiley Periodicals, Inc.