Background Endosteal augmentation enhances the stability of osteoporotic proximal humeral fracture fixation, but the optimal configuration is unknown. The purpose of this study was to compare the biomechanical properties of different lengths of fibula with or without calcar screw in osteoporotic proximal humeral fracture. Methods Three-part proximal humeral fractures with osteopenia were created on 20 pairs of fresh-frozen humeri specimens and allocated to four groups: (1) locking plate with a 6-cm fibular strut allograft, (2) locking plate with a 6-cm fibular strut allograft and additional calcar screws, (3) locking plate with a 12-cm fibular strut allograft,and (4) locking plate with a 12-cm fibular strut allograft and additional calcar screws. Specimens were loaded to simulate the force at 25° abduction. Thereafter, an axial stiffness test and a compound cyclic load to failure test were applied. Structural stiffness, number of cycles loaded to failure and relative displacement values for 5000 cycles at predetermined measurement points were recorded using a testing machine and a synchronized 3D video tracking system. Findings In terms of initial stiffness, number of cycles loaded to failure, and relative displacement values, the groups with 12-cm fibular strut showed obvious improvement compared to the groups with 6-cm fibular strut irrespective of the influence of calcar screw implementation. Further, the groups implemented with calcar screws also showed promising biomechanical stability irrespective of fibular length. Interpretation Lateral locking plate with longer endosteal fibular augmentation and calcar screw can significantly improve biomechanical stability for elderly proximal humeral fractures with posteromedial comminution.