In this study, first regeneration of silk fibroin was conducted. After that with a new method, in one step and without using special apparatus and/or additional materials that need to be removed in the following steps and without using methods that have significant implications, porous water-insoluble (stable) structures, were fabricated. For this purpose prepared solutions with two concentrations of 14% and 20% (w/w) were freezed primarily in three different temperatures, including -196 , -70 to -80 and -10 . After this initial freezing, samples were kept in a freezer with temperature of about -10 , for some days (actually third group don’t have primary freezing). Over this time porous and water-insoluble structures were formed. Pore size, pore morphology, micro structure and mechanical properties of fabricated structures were studied. The average pore size was obtained in the range of 4-27 µm. Most structures based on FTIR results, have relatively low ?-sheet content, which can be an advantage for applications that require a more rapid degradation rate. Based on the results of scanning electron microscopy, samples that prepared from 14% solutions with the primary freezing temperature of -70 to -80 , have lamellar or stacked layer morphology and seems different from other samples. In the case of mechanical characteristics, compressive modules change in a relatively wide range, from 105 to 980 kPa. Behavior of samples were also assessed under cyclic loading and were observed, even after one time loading and unloading, behavior of samples were changed considerably and seems approaching elastic behavior. Key Words Biomedical, Scaffold, Fibroin, Sponge, phase separati