The bioactivity of coatings on alloy implants plays critical roles in the fast osteointegration and maintenance of a long-term life span of the implants. The aim of this study is to incorporate nanopores and bioactive components into the plasma-sprayed coatings by constructing nanopores and bioactive elements on the surface of coatings and further improve their in vitro bone-forming activity. In this study, mesoporous bioactive glasses (MBG) were used for modifying the surface of ZrO2 coatings on Ti-6Al-4V by spin-coating methods. The result shows that MBG nanolayer has been successfully coated on the surface of ZrO2 coatings. The MBG-modified ZrO2 coatings have improved apatite-mineralization ability and stable bonding strength. In addition, the attachment, proliferation, alkaline phosphate activity, and bone-related gene expression of rabbit bone marrow stromal cells on MBG-ZrO2 coating were also enhanced as compared with conventional bioactive glass-modified ZrO2 coatings and pure ZrO2 coatings. The results indicated that MBG-modified ZrO2 coatings on Ti alloy are promising for orthopedic application.