In this study, amphiphilic poly(epsilon-caprolactone)-pluronic-poly(epsilon-caprolactone) (PCL-pluronic-PCL, PCFC) copolymers were synthesized by ring-opening copolymerization and then reacted with isophorone diisocyanate to form polyurethane (PU) copolymers. The molecular weight of the PU copolymers was measured by gel permeation chromatography, and the chemical structure was analyzed by H-1-nuclear magnetic resonance and Fourier transform infrared spectra. Then, the PU copolymers were processed into fibrous scaffolds by the electrospinning technology. The morphology, surface wettability, mechanical strength, and cytotoxicity of the obtained PU fibrous mats were investigated by scanning electron microscopy, water contact angle analysis, tensile test, and MTT analysis. The results show that the molecular weights of PCFC and PU copolymers significantly affected the physicochemical properties of electrospun PU nanofibers. Moreover, their good in vitro biocompatibility showed that the as-prepared PU nanofibers have great potential for applications in tissue engineering. (c) 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016, 133, 43643.