In this study, chitosan/alginate/hydroxyapatite/nanocrystalline cellulose scaffolds were successfully fabricated by the using of freeze-drying method, followed by dicationic crosslinking using CaCl2. The chemical structure and morphology along with mechanical properties of the formed scaffolds respecting to various CNC contents were studied by Fourier-transform infrared spectroscopy (FTIR), X-Ray Diffraction (XRD), Scanning Electron Microscopy (SEM) and mechanical compression test. Chemical interaction and electrostatic attraction between chitosan (CS) and alginate with various CNC ratios were affirmed by FTIR spectroscopy. Results depicted that, scaffolds containing CNC exhibited remarkable improvement in both swelling ratio up to 110% compared without CNC (63%) and compressive strength when compared with other scaffolds. In addition, the average pore size increased, dramatically, with increasing of CNC up to 230 mu m. Porosity was also obeyed the sequence and attainted a maximum value at 93.6%. The growth and cell attachment of fibroblast cells of the selected scaffold were examined prolonging to the cell viability by using Alamar Blue (AB) and then confirmed using SEM. The results indicated that the scaffold comprising CNC has a promising cell growth and cell adherence, and thus expected to have a potent possibility for applications in bone tissue culture.