Summary
The effect of ipriflavone (IP) on the proliferation and differentiation of rat osteoblast-like (ROB) cells and human periodontal ligament fibroblasts (HPLF) was studied in the presence and absence of estrogen. ROB cells were isolated from newborn rat calvaria by sequential collagenase digestion and HPLF from the outgrowth of human periodontal ligament in culture. The alkaline phosphatase (ALP) activity, employed as a marker of bone cell differentiation, was significantly enhanced by IP in both cell types; however, the concentration at which IP had a maximal effect was lower in ROB cells than in HPLF (10−10 versus 10−7 M, respectively). Cell proliferation judged by DNA content was either constant (ROB cells) or slightly increased (HPLF) by IP up to 10−10 M, and decreased significantly above that concentration. In addition, the dose-dependent effect of estrogen on the growth and differentiation of each cell type in the presence and absence of IP was also tested. At the concentrations of IP which showed maximum effects in the induction of ALP, 10−10 M for ROB cells and 10−7 M for HPLF, IP inhibited DNA increase in an estrogen-independent manner. Estradiol (10−10-10−9 M) itself increased the growth rate of both cell types significantly in a dose-dependent manner. Regardless of the concentrations of estradiol tested, ALP activities of both ROB cells and HPLF were elevated by the addition of IP. The ratio of ALP in the presence and absence of IP was similar over the range of estradiol concentrations tested. Thus, we conclude that IP modulates osteogenic cell differentiation of both ROB cells and HPLF and the effect is estrogen independent.
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References
Riggs BL (1987) Pathogenesis of osteoporosis. Am J Obstet Gynecol 156:1342–1346
Yamazaki I (1986) Effect of ipriflavone on the response of uterus and thyroid to estrogen. Life Sci 38:757–764
Yamazaki I, Kinoshita M (1986) Calcitonin-secreting property of ipriflavone in the presence of estrogen. Life Sci 38:1535–1541
Yamazaki I, Shinn A, Shimizu Y, Tsukuda R, Shirakawa Y, Kinoshita M (1986) Effect of ipriflavone on glucocorticoidinduced osteoporosis in rat. Life Sci 38:951–958
Yamazaki I, Shino A, Tsukuda R (1986) Effect of ipriflavone on osteoporosis induced by ovariectomy in rats. J Bone Miner Metab 3:205–210
Shino A, Matsuo T, Tsuda M, Yamazaki I, Tsukuda R, Kitazaki T, Shiota K, Okada H, Yoshida K (1986) Effect of ipriflavone on bone and mineral metabolism in the streptozotocin diabetic rats. J Bone Miner Metab 3:27–37
Fujita T, Yoshikawa S, Ono K, Inoue T, Orimo H (1986) Usefulness of TC-80 (ipriflavone) tablets in osteoporosis: multicenter, double-blind, placebo-controlled study. Igakunoayumi 138:113–141
Agnusdei D, Zacchei F, Bigazzi S, Cepollaro C, Nardi P, Montagnani M, Gennari C (1989) Metabolic and clinical effects of ipriflavone in established postmenopausal osteoporosis. Drugs Exp Clin Res 15:97–104
Tsuda M, Kitazaki T, Ito T, Fujita T (1987) The effect of ipriflavone (TC-80) on bone resorption in tissue culture. J Bone Miner Res 1:207–211
Benvenuti S, Tanini A, Frediani U, Bianchi S, Masi L, Casano R, Bufalino L, Serio M, Brandi ML (1991) Effect of ipriflavone and its metabolites on a clonal osteoblastic cell line. J Bone Miner Res 6:987–996
Kawase T, Sato S, Miake K, Saito S (1988) Alkaline phosphatase of human periodontal ligament fibroblast-like cells. Adv Dent Res 2(2):234–239
Arceo N, Sauk JJ, Moehring J, Foster RA, Somerman MJ (1991) Human periodontal cells initiate mineral-like nodules in vitro. J Periodontol 62:499–503
Dziak R, Brand JS (1974) Calcium transport in isolated bone cell. I. Bone cell isolation procedure. J Cell Physiol 84:75–83
Brand JS (1982) Cellular biochemistry of bone formation and resorption. In: Parsons JA (ed) Endocrinology of calcium metabolism. Raven Press, New York, pp 423–443
Puzas JE, Goodman DBP (1978) A rapid assay for cellular deoxyribonucleic acid. Anal Biochem 86:50–55
Puzas JE, Brand JS (1985) Bone cell phosphotyrosine phosphatase: characterization and regulation by calcitropic hormones. Endocrinology 116:2463–2468
Pensler JM, Radosevich JA, Higbee R, Langman CB (1990) Osteoclasts isolated from membraneous bone in children exhibit nuclear estrogen and progesterone receptors. J Bone Miner Res 5:797–802
Eriksen EF, Colvard DS, Berg NJ, Graham ML, Mann KG, Spelsberg TC, Riggs BL (1988) Evidence of estrogen receptors in normal human osteoblast-like cells. Science 241:84–86
Komm BS, Terpening CM, Benz DJ, Graeme KA, Gallegos A, Korc M, Greene GL, O'Malley BW, Haussler MR (1988) Estrogen binding, receptor mRNA and biologic response in osteoblast-like osteosarcoma cells. Science 241:81–84
Oursler MJ, Osdoby P, Pyfferoen J, Riggs BL, Spelsberg TC (1991) Avian osteoclasts as estrogen target cells. Proc Natl Acad Sci USA 88:6613–6617
Lindsay R, Tohme JF (1990) Estrogen treatment of patients with established postmenopausal osteoporosis. Obstet Gynecol 72:290–295
Studd J, Savvas M, Watson N, Garnett T, Fogelman I, Cooper D (1990) The relationship between plasma estradiol and the increase in bone density in postmenopausal women after treatment with subcutaneous hormone implants. Am J Obstet Gynecol 163:1474–1479
Cheng E, Story CD, Yoder L, Hale WH (1953) Estrogenic activity of isoflavone derivatives extracted and prepared from soybean oil meal. Science 118:164–165
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Kakai, Y., Kawase, T., Nakano, T. et al. Effect of ipriflavone and estrogen on the differentiation and proliferation of osteogenic cells. Calcif Tissue Int 51 (Suppl 1), S11–S15 (1992). https://doi.org/10.1007/BF02180243
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DOI: https://doi.org/10.1007/BF02180243