In this study, the focus was on Pseudomonas aeruginosa, a common bacterial agent in keratitis that exhibits antibiotic resistance. The aim was to isolate, identify, and assess the antimicrobial potential of a bacteriophage sample for treating keratitis caused by P. aeruginosa. The bacteriophage was isolated from wastewater at Baqiyatallah Hospital in Tehran, Iran, and identified using TEM imaging. Various methods, including plaque assays and host range determination, were employed to confirm the presence of the phage. The study evaluated the phage's minimum inhibitory concentration (MIC), biofilm degradation, cytotoxicity, and the safety of phage-containing eye drops on New Zealand rabbits, as well as their effectiveness in a mouse model. The results indicated that the isolated phage belonged to the Myoviridae family and exhibited anti-biofilm activity against clinical isolates. The MIC and IC50 values for the phage were determined to be 10-8 and 10-1, respectively. Notably, the combination treatment of ciprofloxacin eye drops with phage drops proved more effective in improving keratitis than either treatment alone. In conclusion, while the therapeutic effect of the isolated phage on clinical strains of P. aeruginosa was demonstrated, the combination of antibiotic and phage treatment showed synergistic effects.