Biodegradable cellulose acetate (CA) nanofibers containing Rose Bengal (RB) dye were fabricated by electrospinning technique. RB dye, an anionic photosensitizer, has been used in photodynamic therapy due to its excellent biocompatibility and ability to absorb light to generate reactive oxygen species (ROS), but has a decisive disadvantage of water solubility on infection prevention. Firstly, water-insoluble RB dye was synthesized through complexation with cationic ionic liquid (IL) for antiviral agents. The synthesized water-insoluble RB dyes were embedded into biodegradable CA nanofibers by electrospinning. The electrospun nanofibers passed both antiviral test for φx174 virus under visible light irradiation and biodegradability-test using enzymes. The fabricated RB nanofibers absorbed light and generated ROS to inactivate the virus. As a result, the log reduction (-Log10(N/N0)) of φx174 titer under visible light reached a detection limit of 5.00 within 30 min. Also, the fabricated nanofibers were degraded up to 34 wt % in 9 weeks by lipase and cellulase enzymes compared with non-biodegradable nanofibers.