This study aims to develop multifunctional pile cotton fabrics by implementing different compositions of lycra yarns with different densities of the cotton fabric under study. Highly dispersed silica nanoparticles (SiO2 NPs) with small sizes—in the range of 10–40 nm—were successfully prepared and were analyzed using scanning electron microscopy (SEM). The particle size distribution of nano silica was determined via dynamic laser scattering (DLS) and measurements of its zeta potential. Cotton/lycra fabrics were treated using prepared SiO2 NPs in presence of ethylenediaminetetraacetic acid (EDTA) as a crosslinking agent. Energy dispersive X-ray (EDX) analysis and scanning electron microscopy (SEM) were used to characterize the nano-treated fabrics and assure homogeneous dispersion of SiO2 NPs on the cotton/lycra composites. Additionally, the nanoparticles were screened for their in vitro antibacterial activity against human pathogens such as Gram-positive Staphylococcus aureus and Bacillus cereus and Gram-negative Escherichia coli and Pseudomonas aeruginosa strains. The functional properties of the new composite pile cotton fabrics include excellent antibacterial, highly self-cleaning, and excellent UV protection factor (UPF) properties