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A novel biological approach is attempted to convert the human urinary waste into a well-designed bionanomaterial. In the present study, biological activated ammonia gas (NH3(g)) mediated synthesis of hydroxyapatite material (B-HAp) and then impregnation of silver nanoparticles (AgNPs) on the B-HAp material surface was performed by photoreduction method and was followed by an evaluation of its antibacterial activity and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. X-ray diffraction spectroscopy (XRD) and Field emission-scanning electron microscopy (FE-SEM) were engaged to analyze the synthesized materials. Analytical studies revealed the morphology of the crystalline B-HAp synthesized by biologically activated NH3(g) as spherical shaped with AgNPs impregnated on over it. Atomic Absorption Spectrometers (AAS) estimated 2–7 ppm of Ag+ ion were released from the 100 ppm of Ag concentration was impregnated with B-HAp material (B-HAp-Ag-10). It was also found to be an excellent performance of antibacterial activity against Pseudomonas sp, E.coli and S. aureus. The cell- material interaction study of the thus synthesised B-HAp-Ag-10 was found to exhibit a minimal cytotoxicity level when incorporated in MG63 osteosarcoma cell lines, thus confirming the prospective biological application of our material in the biomedical field.