In this study, we demonstrated a novel doxorubicin and vincristine-loaded PLGA magnetic microspheres prepared by coaxial electrospray, as a drug delivery system for osteosarcoma treatment. The results showed that microspheres had an minimum mean diameter of 0.32 +/- 0.25m and exhibited a spherical shape. The particle size distribution changed obviously with the different dosing ways. The entrapment efficiency was found to be 65.72, 73.6 and 74.14% for different microspheres. In vitro degradation research showed that circular pores appeared gradually on the surface of particles with the increase in time, which accelerated the degradation of microspheres. CCK-8 test showed that the Fe3O4@PLGA microspheres without drug loading had almost no side effects, the cytotoxicity of Fe3O4@P/(V+D), Fe3O4@(P+D)/V and Fe3O4@(P+V)/D microspheres increased in turn, and Fe3O4@(P+V)/D microspheres with minimum particle size could significantly arrest the growth of osteosarcoma saos-2 cells. Therefore, the drug-loaded magnetic microspheres have good killing effect on osteosarcoma cell lines, which can be used as an ideal targeting treatment for postoperative adjuvant therapy of osteosarcoma.