Osteosynthesis for canine long bones is a complex process requiring knowledge of biology, surgical techniques and (bio)mechanical principles. Subject-specific finite element analysis constitutes a promising tool to evaluate the effect of surgical intervention on the global properties of a bone–implant construct, but suffers from a lack of validation. In this study, the biomechanical behavior of 10 canine humeri was compared before and after creation of a 10 mm bone defect stabilized with an eight-hole locking compression plate (Synthes®) and two locking screws on each fragment. The response under compression of both intact and plated samples was measured experimentally and reproduced with a finite element model. The experimental stiffness ratio between plated and intact bone was equal to 0.39 ± 0.06. A subject-specific finite element analysis including density-dependent elasto-plastic material properties for canine bone and automatic generation of orthopedic implants was then conducted to recover these experimental results. The stiffness of intact and plated samples could be predicted, with no significant differences with experimental data. The simulated stiffness ratio between plated and intact canine bone was equal to 0.43 ± 0.03. This study constitutes a first step toward the building of a virtual database of pre-computed cases, aiming at helping the veterinary surgeons to make decisions regarding the most suited orthopedic solution for a given dog and a given fracture.