In the present study, cotton fabric was exposed to laser exposure at different energy levels and then the silver nanoparticles were coated on untreated and laser treated cotton fabrics. Methylene blue dye was used to detect the presence of carboxylic acid groups (-COO−
) on laser treated cotton and the dye absorption results were determined spectrophotometrically. ICP-OES (Inductively Coupled Plasma Optical Emission Spectroscopy) analysis and antibacterial tests were carried out to investigate the silver ion content and bactericidal properties of silver nanoparticles on cotton fabrics. Infrared spectroscopy (FTIR/ATR) analysis and scanning electron microscopy (SEM) were used to identify chemical changes and to study the morphology of the surface of the fibers. EDAX (Energy Dispersive X-ray Spectroscopy) analysis was calculated for SEM micrographs. The results showed according to the higher uptake of methylene blue dye that the negative charge of the carboxylic acid groups had been created by laser treatment. Although the FTIR spectroscopy results did not show an increase in carboxylic acid groups, the cationic dye absorption increased. The durability of the Ag+
ion particles on repeated laundered laser treated cotton was proven by antibacterial and ICP tests, particularly when the laser energy was increased.