We studied the use of powered ankle-foot orthoses (PAFOs) and walking stability of the wearers, focusing on the ankle joint, which is known to play a critical role in gait stability. Recognizing that the subtalar joint is an important modulator of walking stability, we conducted the walking experiment on a treadmill by applying varying assistance techniques to the 2-degree-of-freedom (DOF) PAFO, which has the subtalar joint as the rotating axis, and the commonly used 1-DOF PAFO. The participants were 8 healthy men (mean±SD: height, 174.8±7.1 cm; weight, 69.8±6.5 kg; and age, 29.1±4.8 years) with no history of gait abnormality. Center of pressure (COP) was measured with an in-shoe pressure sensor, and stability was estimated on the basis of the angular acceleration measured with the inertial measurement unit attached to the trunk. The experimental results of the 2-DOF PAFO, with or without assistance, showed a significantly higher stability than those of the 1-DOF PAFO (up to 23.78%, p<0.0326). With the 1-DOF PAFO, the stability deteriorated with the increase in the degree of assistance provided. With the 2-DOF PAFO, this tendency was not observed. Thus, the importance of the subtalar joint was proven using PAFOs. The mean position analysis of the COP during the stance phase confirmed that the COP highly correlated with stability (Pearson correlation coefficient: -0.6607). Thus, we conclude that only the 2-DOF PAFO can maintain walking stability, regardless of the assistance characteristics, by preserving the COP in the medial position through eversion. Awareness regarding the role of the subtalar joint is necessary during the manufacture or use of PAFOs, as lack of awareness could lead to the degradation of the wearer's gait stability, regardless of effective assistance, and deteriorate the fundamental functionality of PAFO.