Cellulose is a natural polymer that is widely used in daily life, but it is susceptible to microorganism growth. In this study, a simple sol-gel technique was utilized to incorporate the cellulose scaffold with Ag/TiO2 nanoparticles. The morphology and crystal structure of the as-prepared Ag/TiO2/cellulose composite film were characterized using scanning electron microscopy (SEM) and X-ray diffraction (XRD) methods. Antibacterial tests involving the use of Escherichia coli (E. coli) were carried out under dark and UV-light conditions to evaluate the efficiency of the Ag/TiO2/cellulose composite film in comparison with pristine cellulose paper and TiO2/cellulose composite film. The results indicated that the antibacterial activity of the Ag/TiO2/cellulose composite film outperformed all other samples, where the Ag content of 0.030 wt% could inhibit more than 99% of E. coli. This study suggests that finely dispersed nanocale Ag/TiO2 particles in the cellulose scaffold were effective at slowing down bacterial growth, and the mechanisms of this are also discussed.