Because of their immense economic, wellness, and remedial value, the seeds of palm (Phoenix dactylifera) were selected with synthesized silver nanoparticles (AgNPs) based on their properties for increasing the antibacterial efficacy of medical cotton. This study aimed to be contingent upon the characterisation of raw cotton fabrics treated by AgNPs with date seed extract (DSE) of P. dactylifera both individually and in combination and to investigate their antibacterial activity against various human pathogens. The prepared cotton materials with the synthesized AgNPs and/or DSE were described by both X-ray diffraction (XRD) and scanning electron microscope (SEM). At the same time, gas chromatography-mass spectrometry (GC-MS) and High-performance liquid chromatography (HPLC) were employed to determine the bioactive components in the aqueous date seed extract. The greater antibacterial activity was recorded by cotton treated with DSE and AgNPs mix, in which inhibition zones of all treatments were against Escherichia coli (8 cm), followed by Staphylococcus aureus (2.33-5.87cm) and Bacillus subtilis (2.17-4.63 cm), respectively. Overall, these findings indicate that treated cotton fabrics with synthesised AgNPs and DSE may be widely applied in various potential biological and medical applications, which could enhance environmental sustainability in closed production and consumption.