The development of biofunctional and bioactive hybrid polymeric scaffolds seek to mitigate the current challenges in the emerging field of tissue engineering. In this paper, we report the fabrication of a biomimetic and biocompatible nanofibrous scaffolds of polyamide-6,6 (PA-6,6) blended with biopolymer chitosan via one step co-electrospinning technique. Different weight percentage of chitosan 10 wt%, 15 wt%, and 20 wt% were blended with PA-6,6, respectively. The nanocomposite electrospun scaffolds mats enabled to provide the osteophilic environment for cells growth and biomineralization. The morphological and physiochemical properties of the resulted scaffolds were studied using field-emission scanning electron microscopy (FE-SEM), X-ray diffraction (XRD), and Fourier transform-infrared (FTIR) spectroscopy. The improvement in hydrophilicity and mechanical strength of the bio-nanocomposite mesh with 20 wt% chitosan embedded, was the desired avenue for adhesion, proliferation and maturation of osteoblast cells as compared to other sample groups and pure PA-6,6 fibrous mat. The biomineralization of the nanocomposite electrospun mats also showed higher ability to nucleate bioactive calcium phosphate (Ca/P) nanoparticles comparing to pristine PA-6,6. Furthermore, the biomimetic nature of scaffolds exhibited the cells viability and regeneration of pre-osteoblast (MC3T3-E1) cells which were assessed via in vitro cell culture test. Collectively, the results suggested that the optimized 20 wt% of chitosan supplemented hybrid electrospun fibrous scaffold has significant effect in biomedical field to create osteogenic capabilities for tissue engineering. (C) 2016 Elsevier Ltd. All rights reserved.