Abstract: Objective:To clarify the effects of feather keratin-montmorillonite hydrogels on the biological behavior and osteogenic properties of BMSCs. Methods:Keratins were extracted from chicken feathers by the modified Shindai method. The feather keratin-montmorillonite hydrogel was prepared by mixing the montmorillonite and feather keratin by mass ratio（0%, 1%, 3%). The feather keratin was identified and confirmed as coomassie brilliant blue staining and Fourier transform infrared spectroscopy. The morphology of feather keratin-montmorillonite hydrogels was shown by SEM. BMSCs were cultured at the leach liquor of feather keratin-montmorillonite hydrogels in the medium. The cytotoxicity was measured by CCK-8 assay. The BMSCs of ALP activity was also quantified using an ALP assay kit. Mineralization of the extracellular matrix of BMSCs is investigated by alizarin red staining. The expressions of related proteins in BMP/SMAD signaling pathway were detected by Western blot. Results:The results of coomassie brilliant blue staining and Fourier transform infrared spectroscopy demonstrate that we successfully extracted feather keratin from chicken feather. The pores present in feather keratin hydrogels were well-formed and interconnected. The addition of morillonite changes their structure. When 3%MMT was added, the pore size of feather keratin-montmorillonite hydrogel was significantly reduced. The results of the CCK-8 proliferation experiment showed that the leach liquors of the feather keratin+3%MMT group had inhibitory effects on the growth of the cells. The feather keratin+1%MMT group expressed the highest level of ALP activity, ALP staining, and alizarin red staining, and the data indicate that the leach liquors of the feather keratin+1%MMT group may promote the osteogenic differentiation of BMSCs. Protein expression of BMP-2, Wnt3a, p-SMAD1/5/8, RUNX2, OCN, and COL-1 was significantly higher in feather keratin+1%MMT group（P＜0.01）. Conclusions:The feather keratin-montmorillonite hydrogels can potentially promote osteogenic differentiation through BMP/SMAD signaling pathway.