There is a need for high performance scaffold in tissue engineering. Keeping this perspective in mind, the present study delineates the preparation and physico-chemical characterization of soluble eggshell protein (SEP) modified silk fibroin (SF)-polyvinyl alcohol (PVA) scaffold and its application in bone tissue engineering. The SF/PVA scaffold were prepared by salt leaching and modified with eggshell protein. Micro-architecture and porosity analysis revealed that all the scaffolds were having desired pore size (230-360 μm), interconnected porous network and 90% porosity. The scaffolds were found with suitable swelling behavior and biodegradability to support cell proliferation till replaces native osseous tissue. In vitro cytocompatibility and differentiation study showed that SEP(SF-PVA) supports viability, proliferation and differentiation of cord blood derived human mesenchymal stem cell. Further, in vivo study in mice model showed that the scaffolds are non-immunogenic and support tissue growth. In conclusion, SEP modified SF-PVA scaffold could be a better option for tissue engineering. © 2017 Trans Tech Publications, Switzerland.