Murine models are widely employed in biomechanics research for different preclinical applications, such as evaluating the effectiveness of bone drugs, studying the effect of genetic mutations on bone mechanical properties, and more. Finite Element (FE) models can be used, after appropriate validation, for the non-destructive assessment of bone mechanical properties, thus reducing the number of animals and refining preclinical experimentation. The aim of this project was to develop and validate FE models of the mouse femur to predict stiffness and failure load in four-point bending configuration. Experimental measurements of femur mechanical properties were conducted at the Mechanical Testing Laboratory of Istituto Ortopedico Rizzoli (IOR, Bologna) and used to evaluate the accuracy of FE predictions.