Expanded polytetrafluoroethylene ePTFE grafts are mostly employed to replace damaged blood vessels and to restore normal blood flow. However, the dilemma of early thrombosis, inflammation, and development of biofilms after implantation limit ePTFE long-term patency and restrict the patient’s life quality. In this study, poly lactic-co-glycolic acid (PLGA) nanoparticles were covalently immobilized on ePTFE surface for local therapeutic purposes. First, the ePTFE surface was primarily oxidized by H2O2/H2SO4 solution to create hydroxyl groups. Consequently, free amino groups were introduced onto ePTFE surface by an aminolyzation reaction of the activated hydroxyl groups using 3-aminopropyl triethoxysilane. The produced amino groups were further used as anchor sites for covalent immobilization of previously prepared PLGA nanoparticles. The functional groups originated on ePTFE surface were confirmed by FTIR analysis. Furthermore, the scanning electron microscopy visualization evidenced a homogeneous distribution pattern of the immobilized PLGA nanoparticles on the surface. The immobilized PLGA nanoparticles showed stability on ePTFE surface under blood flow mimetic conditions. Additionally, light microscopy observation confirmed the biocompatibility of mouse L929 fibroblasts on the nano-coated ePTFE graft. The cellular adhesion and growth did not reveal remarkable cytotoxicity in the tested modified ePTFE grafts.