Silk sericin (SS) can be used as biomaterials in the development of scaffolds for skin regeneration as it has important properties such as antioxidant capacity and biocompatibility. However, SS-based materials have weak structural properties, thereby making its application difficult. The effect of freezing temperature (−35 °C and −80 °C) on the properties of freeze-dried SS scaffolds is evaluated in this study. The secondary structure, morphology, and thermal stability of the scaffolds were determined x-ray diffraction and Fourier-transform infrared spectroscopy; scanning electron microscopy; and thermogravimetric analysis and differential scanning calorimetry, respectively. Its water absorption capacity was also assessed. The results indicated that scaffolds manufactured at −35 °C have a higher β-sheet content (37.08%), lower pore density, and larger porosity than those frozen at −80 °C. However, the water absorption and thermal properties showed no statistically significant differences between both samples.