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Silk fibroin (SF), a unique and promising natural material extracted from silkworm, has gained much attention for its use as biomedical material, because of its biocompatibility, biodegradability, and nontoxicity. Despite its strength, SF alone is not appropriate because of its poor flexibility. Thermoplastic polyurethane (TPU) is a very desirable material for medical applications because of its flexibility. A solution to the problem of SF’s poor flexibility can be achieved by mixing flexible TPU with SF. The aim of this work was to examine the influence of FS content on the physical and thermal properties of TPU/FS blends. Blends of SF and TPU were prepared with melt mixing of TPU with 5, 10, 15, and 20 wt% of SF in a laboratory Brabender kneading chamber. The specimens of the neat components and their blends were moulded in a hydraulic press. The structural characteristics, thermal, and viscoelastic properties of neat TPU, SF and their blends (TPU/SF blends) were examined by Fourier transform infrared (FTIR) spectroscopy, differential scanning calorimetry (DSC), dynamic mechanical analysis (DMA), and thermogravimetric analysis (TGA). The FTIR results showed that the SF was successfully introduced to TPU matrix. Results obtained with DSC and DMA measurements showed that, when the SF content increased in the TPU/SF blends, the Tg, expressed as Teig, Tmg, and Tefg of the TPU, slightly shifted to lower temperature due to the incorporation of the amorphous phase of FS (aspartic acid) in the amorphous phase of TPU, and mobility of polymer chains increased. The melting temperature (Tm), expressed as Teim, Tpm, and Tefm, decreased because of the less organised crystal structure with the addition of SF. Thermal stability was determined by TGA, which showed that, with the addition of SF, the thermal stability decreased because of the lower thermal stability of SF. [ABSTRACT FROM AUTHOR]