Calcination has been widely demonstrated as a favorable protocol for producing various inorganic nanomaterials for tumor therapy. However, little attention has been paid to its effect on the biotherapeutic efficacy of inorganic nanomaterials. Herein, we compare the effects of different calcination atmospheres on the therapeutic efficacy of Fe-V-O (FVO) nanomaterials. We find that compared with FVO nanomaterials synthesized by calcination in air, those prepared by argon calcination have a lower metallic valence state and a higher near-infrared light absorption capacity, hence resulting in significantly better biosafety and higher chemodynamic therapy (CDT)/photothermal therapy (PTT) efficacy. This study demonstrates that the therapeutic efficacy of inorganic nanomaterials can be optimized by employing different thermal treatment atmospheres, which provides new insights into the development of efficient anti-tumor agents.