Ultrastructural and cytochemical studies on cell death of osteoclasts induced by bisphosphonate treatment.

The process of apoptosis and fate of osteoclasts are not well elucidated because dying osteoclasts are rarely seen in normal bone. Histological, cytochemical, and ultrastructural features of osteoclasts undergoing apoptosis were studied in the femur and tibia of rats treated with a third-generation bisphosphonate (disodium dihydrogen (cycloheptylamino)-methylene-1, 1-bisphosphonate). After the bisphosphonate administration, osteoclasts decreased significantly in number. Initially, they became devoid of ruffled borders and detached from the bone surface. In such osteoclasts, the Golgi apparatus was degraded, or dispersed in the cytoplasm. Later, osteoclasts revealed typical features of apoptosis, with pyknotic nuclei showing condensation and margination of heterochromatins and DNA fragmentation. They were often convoluted to give rise to apoptotic bodies. In addition, enlargement and fusion of nuclear envelopes and subsequent disruption leading to leakage of nuclear contents into the cytoplasm were observed in osteoclasts in the late stage of apoptosis. These osteoclasts as well as apoptotic bodies were surrounded by cytoplasmic processes of macrophages, which often contained degenerated cytoplasmic fragments of osteoclasts. Apoptotic osteoclasts migrating into or present in capillaries were also observed in some areas. In conclusion, bisphosphonate induces apoptosis of osteoclasts, which was characterized by ultrastructural changes of the nucleus typical of apoptosis accompanied by degradation of cell organelles. The majority of them are eliminated by macrophages, but there are some that escape into blood vessels.