Histochemical and fine structural study of bone of ipriflavone-treated rats

Bone labeling, histochemical, and fine structural studies were performed in order to clarify the effects of ipriflavone (IP) on rat bone tissue in vivo and in vitro. Labeling experiments showed a slight increase in bone formation during 3 days' administration. It was also noted that many osteoclasts detached from the bone surface at 1, 2, and 6 hours after administration in vivo. In addition, irregular localization of tartrate-resistant acid phosphatase (TRACPase) activity was observed in osteoclasts. Fine structurally, IP-treated osteoclasts exhibited irregularity in their ruffled borders, as reported in calcitonin administration, and many enlarged rough endoplasmic reticuli and vacuoles were observed. However, osteoclasts at 12 hours after administration, as well as the control, indicated recovery features from the effect of IP. Osteoblast proliferation and differentiation led to increasing alkaline phosphatase activity (ALPase) with time as well as the development of rough endoplasmic reticuli and Golgi apparatus with well-developed fine structure. These findings imply active synthesis of bone matrix. In our in vitro experiment, osteoclasts and osteoblasts displayed histochemical and fine structural characteristics similar to those observed in our in vivo experiment. Moreover, fewer TRACP-positive mononuclear cells were observed after 24-hour culture with IP than with the control. These results suggest that IP inhibits directly and/or indirectly differentiation and activity of osteoclasts and also promotes differentiation of osteoblast-lineage cells and their bone-forming activity.