Although vanadium compounds are well recognized for their ability to change from insulator to metal, they may also be used therapeutically to address significant medical issues. In this study, we used vanadium oxide thin films synthesized by the pulsed laser deposition (PLD) technique to examine human stem cells generated from bone marrow. According to x-ray reflectivity (XRR) measurements, the films’ thickness ranged from 6 to 26 nm. The water contact angle method has been employed to probe the surface energy and wettability of the films, which influence the cell behavior significantly. We also used a variety of techniques, such as differentiation staining, phase contrast microscopy, and real-time reverse transcription-polymerase chain reaction (RT-PCR), to examine the growth, adhesion, proliferation, and differentiation of human bone marrow mesenchymal stem cells (hBMMSCs) on these oxide films over time. Our results indicated that vanadium oxide films alter hBMMSCs adhesion and growth and affect their differentiation. The application of VOx films in biological and medical materials, as well as future research on cells, is all made possible by these findings, which also improve our understanding of the biological actions of vanadium compounds.