The microarchitecture and alignment of trabecular bone adapts to the particular mechanical milieu applied to it. Due to this anisotropic mechanical property, measurement orientation has to be taken into consideration when assessing trabecular bone quality and fracture risk prediction. Quantitative ultrasound (QUS) has demonstrated the ability in predicting the principal structural orientation (PSO) of trabecular bone. Although the QUS prediction for PSO is very close to that of μCT, certain angle differences still exist. It remains unknown whether this angle difference can induce significant differences in mechanical properties or not. The objective of this study is to evaluate the mechanical properties in different PSOs predicted using different methods, QUS and μCT, thus to investigate the ability of QUS as a means to predict the PSO of trabecular bone noninvasively. By validating the ability of QUS to predict the PSO of trabecular bone, it is beneficial for future QUS applications because QUS measurements in the PSO can provide information more correlated with the mechanical properties than with other orientations. In this study, seven trabecular bone balls from distal bovine femurs were used to generate finite element models based on the 3-dimensional μCT images. Uniaxial compressive loading was performed on the bone ball models in the finite element analysis (FEA) in 6 different orientations (three anatomical orientations, two PSOs predicted by QUS and the longest vector of mean intercept length (MIL) tensor calculated by μCT). The stiffness was calculated based on the reaction force of the bone balls under loading and the von Mises stress results showed that both the mechanical properties in the PSOs predicted by QUS is significantly higher than the anatomical orientations and comparatively close to the longest vector of MIL tensor. The stiffness in the PSOs predicted by QUS is also highly correlated with the stiffness in the MIL tensor orientation (ATTmax vs. MIL, R2=0.98, p<001; UVmax vs. MIL, R2=0.92, p<001). These results were validated by in vitro mechanical testing on the bone ball samples. This study demonstrates that the PSO of trabecular bone predicted by QUS has an equally strong apparent stiffness with the orientation predicted by μCT.