A large dose of 24R,25(OH)2D3 was administered to the vitamin D-repleted rat to examine its effect on the bone. Male Wistar rats were fed a diet containing 0, 0.025, 1.25, 4.0, and 12.5 ppm 24R, 25(OH)2D3 for 2 years starting at age 6 weeks. The estimated amounts of daily intake of 24R,25(OH)2D3 were 0, 93, 4640, 14680, and 49580 ng/100 g body weight, respectively. No notable difference was found in either the weight or the death rate of the animal. The long-term administration of massive doses of 24R,25(OH)2D3 did not lead to hypercalcemia nor did it affect the blood phosphorus, alkaline-phosphatase, or creatinine levels. Radiographs revealed a striking increase in the bone density on the bones from the animals treated with 1.25 ppm or more 24R,25(OH)2D3. Direct single photon absorptiometry revealed a dose-dependent increase in total bone minerals of both the femur and coccyx. Histological examination revealed a marked increase in the cortical thickness of the femur as well as in the cancellous bone volume of the coccyx. Polarizing microscopy demonstrated the lamellar structure of the bone, and undecalcified sections confirmed the increase of mineralized bone. Ash weight, calcium, phosphorus, and magnesium contents on the tibia and fibula also indicated the ascending dose-dependent increase up to 150% of the control. The parameters of bone size were not altered in any group. These results clearly suggest that 24R,25(OH)2D3 given in massive doses has the pharmacological action of increasing bone volume in the rat without causing remarkable hypercalcemia.