Nanostructured materials possess unique structural and functional properties that play a crucial position in tissue engineering applications. Present investigation is aimed to synthesize chitosan–sodium alginate (CS) nanocomposite using hydrothermally prepared zirconia nanoparticles. In this, three different weight percentages of (0.5, 1, and 1.5) zirconia nanoparticles are utilized for the preparation of biomimetic nanocomposite scaffolds (CSZ) employing 4 wt% of CS by a solvent casting technique. Physico‐chemical and thermal behavior of the prepared nanoparticles and their CSZ scaffolds are comprehensively characterized. Bioactivity of the prepared zirconia nanoparticles and CSZ scaffolds are explored in terms of in vitro biocompatibility, protein absorption in simulated body fluid (SBF), and phosphate buffered saline (PBS). Agar disc diffusion method is employed to identify the antibacterial property against Staphylococcus aureus and Escherichia coli. In vitro cytotoxicity of zirconia nanoparticles and CSZ scaffolds is identified against human urothelial carcinoma (UC6) and osteosarcoma (MG‐63) cells. These studies explore that zirconia nanoparticles are suitable for biomedical applications while it is interacted with chitosan and sodium alginate (CS) due to their promising biocompatibility. Biomimetically obtained chitosan/sodium alginate scaffold contain 1 wt% zirconia nanoparticles show higher biocompatibility amenable for tissue engineering applications.
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