In order to develop biodegradable silk fibroin (SF) –polyurethane (PU) composite materials, the biodegradable PU based on polycaprolactone (PCL) was synthesized. The structure and dynamics of the crystalline and non-crystalline regions of biodegradable PU and SF – PU composite fiber were studied in the dry and hydrated states using three kinds of 13C solid-state NMR, i.e.,13C CP/MAS, 13C DD/MAS and 13C r-INEPT NMR methods. In 13C r-INEPT NMR spectra which can observe only highly mobile hydrated components, only small peaks from non-crystalline region of PCL components in PU could be observed, but higher intensity peaks from the non-crystalline region could be observed for PU in the regenerated SF-PU composite fiber. On the other hand, the structure and dynamics of SF in SF-PU fiber was investigated by observing three kinds of 13C solid-state NMR spectra of [3-13C] Ser, [3-13C] Tyr and [3-13C] Ala-labeled SF. The fraction of random coil of SF, especially the fraction of hydrated random coil, increased for hydrated SF-PU composite fiber compared with random coil fraction of hydrated pure SF fiber. Thus, the interaction between SF and PU caused increase fraction of non-crystalline region of PCL in the PU and slightly amorphous SF region in the hydrated state.