Clarifying the differences of mechanical properties in different anatomical regions of bone can provide an important basis for constructing the finite element model of the bone with high biofidelity. One sample was pre? pared from each of the anterior，posterior，medial，and lateral anatomical regions of the bovine femoral shaft. Each sample was tested at 18 points utilizing a Berkovich nanoindenter tip，and the time history of the loading force and in? dentation depth was recorded to obtain the indentation modulus and hardness. The indentation modulus of the ante? rior，posterior，medial and lateral regions of the long bone were 20.78 ± 2.66 GPa，18.66 ± 2.57 GPa，16.39 ± 2.29 GPa，21.57 ± 2.19 GPa respectively，and hardness were 0.65 ± 0.79 GPa，0.58 ± 0.08 GPa，0.44 ± 0.06 GPa，0.61 ± 0.15 GPa respectively. Variance analysis showed that anatomical region had a significant effect on indentation modulus and hardness（p < 0.001）. Multiple comparisons between groups showed that the indentation modulus and hardness of the anterior specimen were significantly higher than those of the lateral specimen，the indentation modu? lus and hardness of the medial specimen were significantly higher than those of the lateral specimen，the hardness of the posterior specimen was significantly higher than that of the lateral specimen，and the indentation modulus of the medial specimen was significantly higher than that of the lateral specimen. Therefore，the use of heterogeneous mate? rials can help to improve the biological fidelity of the finite element model of the long bone.