The purpose of this study was to examine whether soybean protein isolate prevents bone loss induced by ovarian hormone deficiency. Thirty-two 95-d-old Sprague-Dawley rats were randomly assigned to four treatment groups [sham-operated (sham); ovariectomized (ovx); ovx+soybean; ovx + 17 beta-estradiol (E2)] and killed after 30 d. Rats in the sham, ovx and ovx + 17 beta-estradiol groups were fed a casein-based diet, and the soybean group was fed soybean protein isolate instead of casein; the diets were otherwise comparable. Rats in the ovx group had significantly lower densities of the right femur (P < 0.001) and the fourth lumbar vertebra (P < 0.05) than rats in the sham group. These lower bone densities were not observed in animals receiving 17 beta-estradiol or fed soybean. The ovx group also had significantly (P < 0.01) greater serum concentrations of 1,25-dihydroxycholecalciferol than the other three groups. Our findings suggest that dietary soybean protein is effective in preventing bone loss due to ovarian hormone deficiency. Because serum activities of both alkaline phosphatase and tartrate-resistant acid phosphatase were significantly greater in the ovx group and in the ovx + soybean group but not in the group receiving 17 beta-estradiol, compared with sham animals, this confirms that ovariectomy enhances and 17 beta-estradiol suppresses the rate of bone turnover. Despite the higher rate of bone turnover in the soybean-fed animals, the vertebral and femoral bone densities of these rats were significantly greater than those of rats in the ovx group, suggesting that formation exceeded resorption. Further studies are needed to clarify whether this protective effect on bone is due to the protein itself or to the presence of isoflavones in soybean protein.