We show for the first time that imaging of osteoarthritic subchondral bone with 99mTc-sestamibi (MIBI) may be a valuable diagnostic tool to assess the severity of subchondral sclerosis. This is a report on a patient who had osteoarthritis of both knees and where MIBI was accumulated in the lesion sites. Due to the accumulation of MIBI in the bone, we believe that excessive stress had been applied to the bone and microfractures had activated bone remodeling. Typically, MIBI accumulation is not observed in the bone, but MIBI is believed to exhibit high accumulation where there is an increased presence of osteoblasts and osteoclasts. Osteoclasts absorb calcium, consequently causing a depolarization of the mitochondrial membrane potential, which may decrease the ability to retain MIBI and enhance washout. Pathological examination of the tissues of this case confirmed an abundance of osteoclasts. This suggests that using MIBI may be useful for determining the increased presence of osteoblasts and osteoclasts. J. Med. Invest. 63: 127-130, February, 2016