Purpose: The purpose of this study is to compare kinematics and kinetics on the knee joint between stair gait with unstable shoes and barefoot in healthy adult women. Methods: Seventeen healthy adult women were recruited for this study. The subjects performed stair ascent and descent with unstable shoes and barefoot. The experiment was repeated three times for each stair gait with unstable shoes and barefoot. Measurement and analysis of the movements of the knee joint were performed using a three-dimensional analysis system. Results: Statistically significant differences in the knee muscle force of semimembranosus, biceps femoris-long head, biceps femoris-short head and sartorius, patellar ligament, medial gastrocnemius, and lateral gastrocnemius were observed between unstable shoes and barefoot gait during stair ascent. Statistically significant differences in the knee muscle force of sartorius, rectus femoris, medial gastrocnemius, and lateral gastrocnemius were observed between unstable shoes and barefoot gait during stair descent. Statistically significant differences in the knee flexor moment of semitendinosus, biceps femoris-long head, biceps femoris-short head, sartorius, rectus femoris, vastus intermedialis, medial gastrocnemius, and lateral gastrocnemius were observed between unstable shoes and barefoot gait during stair ascent. Conclusion: Therefore, wearing unstable shoes during stair gait in daily life is considered to influence knee joint kinematics and kinetics due to the unstable shoes, and thus suggest the possibility that reducing the risks of pain, and knee osteoarthritis, stabilizing the knee joint caused by changes in the loading of the knee joint.