The invention provides a constructing method of a femur middle-lower section bone defect model. The constructing method of the bone defect model comprises the following steps: (1) taking preserved skin from posterior limb of an anesthetized animal and disinfecting the preserved skin, and making incisions on middle, lower and front outer sides of femur of the animal to expose the middle-lower section of the femur; and (2) exciding femurs differing in size from the middle-lower section of the femur to form bone defect, irrigating wounds, dressing and stitching so as to obtain the femur middle-lower section bone defect model. On a femur middle-lower section bone defect operative approach, important blood vessels and nerves are avoided, so that secondary injury and bleeding are relieved in a process of establishing the bone defect model to the greatest extent; and the model is good in unity and repeatability, and is conducive to simulating a clinical bone defect repairing effect. The invention also provides a femur middle-lower section bone defect model prepared based on the method and application of the bone defect model.