Characterization of Ground Silk Fibroin through Comparison of Nanofibroin and Higher Order Structures

[Image: see text] Silk fibroin, a biodegradable component of silk, is increasingly used for various applications and studied intensively. Recently, a technique for preparing nanofibers without using chemicals has been gaining attention from the environmental impact and safety perspectives. This study focuses on the structure observation of ground silk fibroin (GF) prepared using a grinding method, which is a physical nanofibrillation method. The fabricated nanofiber samples were examined in detail using the X-ray diffraction (XRD), differential scanning calorimetry (DSC), micro Raman spectroscopy, and atomic force microscopy (AFM) techniques. The nanofibrillated structures were observed in both GF and regenerated silk fibroin (RF) samples prepared using the conventional method. As results, AFM images showed that the nanofibril diameter of GF was about 1.64 nm and that of RF was about 0.32 nm. Methanol treatment induced a structural transition from a random coil to a β-sheet for the RF film, but it had no effect on the GF film. Thus, it is suggested that the grinding method provides not only ultrafine silk fibroin nanofibers without using toxic reagents but also resistance to reagents such as methanol.