Silk fibroin (SF) based biomaterials have received more attention due to SF's superior biocompatibility, biodegradability, and special properties such as a minimal inflammatory reaction and low immunogenicity especially for the development of bioactive scaffolds in bone regeneration applications. To date, Silk fibroins are blended with a variety of biocompatible polymers and inorganic materials, to enhance their in vitro and in vivo properties. This study includes the detailed protocol of extracting silk fibroin from Bombyx mori silk cocoons followed by the degumming process using aqueous Na2CO3. Hydroxyapatite was synthesized by co-precipitation method using NH4H2PO4 and Ca(NO3)2. 4H2O, later it was blended with extracted silk fibroin and Polyvinyl alcohol (PVA) solution to fabricate desire nanofibers/scaffolds. The viscosity of the blended solution was tuned at an appropriate concentration for the preparation of nanofibrous scaffold by electrospinning and freeze-dried method. The obtained scaffolds were further characterized by FT-IR, XRD, and SEM to confirm the functional groups, phase formation, porosity, and surface morphology. Cell viability was evaluated using MG-63 cell lines and in -vitro bioactivity was further evaluated by immersing in simulated body fluid (SBF) solution. Drug loading and releasing profile of scaffolds were examined using Cephalexin Monohydrate (CEM) in PBS medium. The results inferred that the prepared PVA/HAP/SF based scaffolds showed better osteogenic cell proliferation with prolonged drug release, Hence, this optimized nanofiber may act as an effective material for bone tissue engineering applications. © 2020 Elsevier B.V.