This work introduces a systematic and efficient approach for producing stable AgNPs utilizing Nigella sativa (Ns) seed aqua extract (AE), which exhibit strong antibacterial properties. The characterization of Ns-AgNPs was performed using a UV-visible spectrophotometer (UV-Vis), X-ray diffraction (XRD), Fourier transform infrared (FT-IR) spectroscopy, scanning electronic microscopy (SEM), and transmission electronic microscopy (TEM), and energy-dispersive X-ray spectroscopy (EDX). The Ns-AgNPs did not show aggregation, as shown by the results of STEM and XRD. According to the EDX analysis in this research, it was determined that Ns-AgNPs, gave signals in the silver region (~3 KeV) at 92.3%. Ns-AgNPs showed significant antibacterial performance against Staphylococcus aureus, Escherichia coli and were effective at the low concentrations. Ns-AgNPs may be incorporated into wound dressings, surgical instruments, and medical devices to prevent infections and promote healing.