Composite scaffolds are commonly used strategies and materials employed to achieve similar analogs of bone tissue. This study aims to fabricate 10% wt polylactic acid (PLA) composite fiber scaffolds by the air-jet spinning technique (AJS) doped with 0.5 or 0.1 g of zirconium oxide nanoparticles (ZrO2) for guide bone tissue engineering. ZrO2 nanoparticles were obtained by the hydrothermal method and characterized by X-ray diffraction (XRD) and scanning electron microscopy (SEM). SEM and fourier-transform infrared spectroscopy (FTIR) analyzed the synthesized PLA/ZrO2 fiber scaffolds. The in vitro biocompatibility and bioactivity of the PLA/ZrO2 were studied using human fetal osteoblast cells. Our results showed that the hydrothermal technique allowed ZrO2 nanoparticles to be obtained. SEM analysis showed that PLA/ZrO2 composite has a fiber diameter of 395 nm, and the FITR spectra confirmed that the scaffolds’ chemical characteristics are not affected by the synthesized technique. In vitro studies demonstrated that PLA/ZrO2 scaffolds increased cell adhesion, cellular proliferation, and biomineralization of osteoblasts. In conclusion, the PLA/ZrO2 scaffolds are bioactive, improve osteoblasts behavior, and can be used in tissue bone engineering applications.