Bioactive glass nanoparticles-loaded poly(ɛ-caprolactone) nanofibers (BIOG PCL nanofibers) were synthesized and evaluated as substrates for ocular cells (ARPE-19). BIOG PCL nanofibers were characterized using SEM, FTIR, and DSC, and thein vitrodegradation profile was also investigated. Thein vitroocular biocompatibility of nanofibers was exploited in Müller glial cells (MIO-M1 cells) and in chorioallantoic membrane (CAM); and the proliferative capacity, cytotoxicity, and functionality were evaluated. Finally, ARPE-19 cells were seeded onto BIOG PCL nanofibers and they were investigated as supports forin vitrocell adhesion and proliferation. SEM images revealed the incorporation of BIOG nanoparticles into PCL nanofibers. Nanoparticles did not induce modifications in the chemical structure and semicrystalline nature of PCL in the nanofiber, as shown by FTIR and DSC. MIO-M1 cells exposed to BIOG PCL nanofibers showed viability, and they were able to proliferate and to express GFAP, indicating cellular functionality. Moreover, nanofibers were well tolerated by CAM. These findings suggested thein vitroocular biocompatibility and absence of toxicity of these nanofibers. Finally, the BIOG nanoparticles modulated the protein adsorption, and, subsequently, ARPE-19 cells adhered and proliferated onto the nanostructured supports, establishing cell-substrate interactions. In conclusion, the biodegradable and biocompatible BIOG PCL nanofibers supported the ARPE-19 cells.