Bacteria have the extraordinary ability to adhere to biomaterial surfaces and form multicellular structures known as biofilms, which have a detrimental impact on the performance of medical devices. Herein, an investigation highlighted the effective inhibition of bacteria adhesion and overgrowth on silicone rubber surface by grafting polysaccharide, agarose (AG), to construct hydrophilic and negatively charged surfaces. Because of the strong hydration capacity of agarose, the water contact angle of the modified silicone rubber surfaces was significantly reduced from 107.6 ± 2.7° to 19.3 ± 2.6°, which successfully limited bacterial adherence. Most importantly, the durability and stability of coating were observed after 10 days of simulated dynamic microenvironment in vivo, exhibiting a long service life. This modification method did not compromise biocompatibility of silicone rubber, opening a door to new applications for silicone rubber in the field of biomedical materials.