Objective To study the effects of prosthetic alignment on internal contact mechanical characteristics of intact knee joint for transfemoral amputees. Methods The gait experiment of transfemoral amputees was performed under different alignment conditions, and the differences of lower limb motion and ground reaction force (GRF) were analyzed and compared with those of the non-amputees. The three-dimensional （30） finite element model of knee joint was build and used to analyze the effect of alignments of socket adduction and abduction on internal contact mechanical characteristics between femur cartilage, tibia cartilage and meniscus. Results For knee joint of the non-amputees, contact force was mainly concentrated on the medial sides at the moment of the first GRF peak, while contact force was mainly concentrated on the lateral sides at the moment of the second GRF peak. However, for intact knee joint of the transfemoral amputees, contact force was mainly concentrated on the medial side at the moment of two GRF peaks. The stress of the medial meniscus, contact force and contact area between the medial meniscus and cartilage all obviously increased under the alignment of 6° socket adduction. Conclusions Compared with non-amputees, the incidence of knee osteoarthritis (OA) in amputees was higher, which was related to the long-term overload of the medial knee joint. The alignment of socket adduction may increase the risk of knee OA in the intact side of transfemoral amputees. In clinic, excessive adduction of the socket should be avoided during prosthetic alignment.