Abstract
The effects of PGE2 on mineralized bone nodule formation were studied in fetal rat calvarial (RC) cells in vitro. Continuous exposure of RC cells to 3x10-8M PGE2 induced a twofold increase in mineralized bone nodule formation and a 1.5-fold increase in alkaline phosphatase (ALPase) activity without affecting RC cell growth. These stimulatory effects were evoked by concentrations of 3x 10-9-3x10-6 M PGE2 and the maximal effect was observed with 3x10-8 M PGE2. The in vitro effects of PGE2 were evident when RC cells were exposed to it on days 8–14 and 8–21, which correspond to the post-confluent culture stage, but no effects were observed when the cells were exposed on days 1–7, the growth stage. The ALPase activity was also higher (1.2–1.4-fold) when 3x10-8 M PGE2 was added during the post-confluent stage. In order to determine the effect of PGE2 during the mineralization phase of bone nodules in the presence of a large population of osteoprogenitor cells, RC cells were exposed to dexamethasone for 7 days before PGE2 was added during the post-confluent stage. A significantly higher percentage of nodules mineralized were observed with 3x10-8-3x10-9 M PGE2 (1.6-and 1.4-fold, respectively), than in control cultures. Analysis of the mineral-related proteins by EDTA extraction of bone nodules followed by electrophoresis and Stains-All staining revealed an increased total amount of osteopontin extracted from the mineralized matrix after PGE2 treatment. The osteopontin content was highest after 3x10-8 M PGE2, with a 73% increase of the densitometric intensity of the bands, although this increase, reflected the increased number of mineralized bone nodules due to PGE2. These findings suggest that PGE2 may increase the proportion of functional osteoblasts able to produce mineralized bone nodules in the population by stimulating differentiation during the postconfluent stage of RC cell culture.
Similar content being viewed by others
References
Raisz LG, Rodan GA (1990) Cellular basis for bone turnover. In: Avioli LV, Krane SM (eds) Metabolic bone disease, 2nd ed. WB Saunders Philadelphia, pp 1–41
Wallach S, Carstens JH, Avioli LV, Feinblatt JD (1992) Physical and biochemical factors in bone metabolism. Calcif Tissue Int 50:1–4
Jee WSS, Ueno K, Deng YP, Woodbury DM (1985) The effects of prostaglandin E2 in growing rats: increased metaphyseal hard tissue and cortico-endosteal bone formation. Calcif Tissue Int 37:148–157
High WB (1988) Effect of orally administered prostaglandin E2 on cortical bone turnover in adult dog: a histomorphometric study. Bone 8:87–90
Norrdin RW, Shih M-S (1988) Systemic effects of prostaglandin E2 on vertebral trabecular remodeling in beagles used in healing study. Calcif Tissue Int 42:363–368
Mori S, Lee WSS, Li XJ (1992) Production of new trabecular bone in osteopenic ovariectomized rats by prostaglandin E2. Calcif Tissue Int 50:80–87
Ueda K, Saito A, Nakano H, Aoshima M, Yokota M, Muraoka R, Iwaya T (1980) Cortical hyperosteosis following long-term administration of prostaglandin E2 in infants with cyanotic congenital heart disease. J Pediatr 97:834–836
Ke H-Z, Li M, Jee WSS (1992) Prostaglandin E2 prevents ovariectomy-induced cancellous bone loss in rat. Bone Miner 19:45–62
Yong S, Chyun YS, Lawrence G, Raisz LG (1984) Stimulation of bone formation by prostaglandin E2. Prostaglandins 27:97–103
Partridge NC, Alcon D, Michelangeli VP, Kemp BE, Ryan GB, Martin TJ (1986) Functional properties of hormonally responsive cultured normal and malignant rat osteoblastic cells. Endocrinology 108:213–219
Farr D, Pochal W, Brown M, Shapiro E, Weinfeld N, Dziak R (1984) Effect of prostaglandins on rat calvarial bone-cell calcium. Arch Oral Biol 29:885–891
Haketa Y, Yoshino T, Nakatani Y, Kurihara N, Ikeda E, Maeda N, Kumegawa M (1985) Prostaglandin E2 stimulates collagen and non-collagen protein synthesis and prolyl hydroxylase activity in osteoblastic clone MC3T3-E1. Biochem Biophys Res Commun 126:340–345
Bellows CG, Aubin JE, Heersche JNM, Antosz ME (1986) Mineralized bone nodules formed in vitro from enzymatically released rat calvaria cell populations. Calcif Tissue Int 38:143–154
Bellows CG, Aubin JE (1989) Determination of numbers of osteoprogenitors present in isolated fetal rat calvaria cells in vitro. Development Biol 133:8–13
Nagata T, Bellows CG, Kasugai S, Butler WT, Sodek J (1991) Biosynthesis of bone proteins [SPP-1 (secreted phosphoprotein-1, osteopontin), BSP (bone sialoprotein) and SPARC (osteonectin)] in association with mineralized-tissue formation by fetal-rat calvarial cells in culture. Biochem J 274:513–520
Ishida H, Bellows CG, Aubin JE, Heersche JNM (1993) Characterization of the 1,25-(OH)2D3-induced inhibition of bone nodule formation in long-term cultures of fetal rat calvaria cells. Endocrinology 132:61–66
Flanagan AM, Chambers TJ (1992) Stimulation of bone nodule formation in vitro by prostaglandins E1 and E2. Endocrinology 130:443–448
Rao LG, Ng B, Brunette DM, Heersche JNM (1977) Parathyroid hormone and prostaglandin E1-response in a selected population of bone cells after repeated subculture and storage at-80°C. Endocrinology 100:1233–1241
Bellows CG Aubin JE, Heersche JNM (1987) Physiological concentrations of glucocorticoids stimulate formation of bone nodules from isolated rat calvaria cells in vitro. Endocrinology 121:1985–1992
Bellows CG, Aubin JE, Heersche JNM (1991) Initiation and progression of mineralization of bone nodules formed in vitro: the role of alkaline phosphatase and organic phosphate. Bone Miner 14:27–40
Kasugai S, Todescan Jr R, Nagata T, Yao K-L Butler WT, Sodek J (1991) Expression of bone matrix protein associated with mineralized tissue formation by adult rat bone marrow cells in vitro: inductive effects of dexamethasone on the osteoblastic phenotype. J Cell Physiol 147:111–120
Kissane JM, Robins E (1958) The fluorometric measurement of deoxyribonucleic acid in animal tissues with special reference to the central nervous system. J Biol Chem 233:184–188
Lowry OH, Roberts NR, Wu M-L, Hixon WS, Crawford EJ (1954) The quantitative histochemistry of brain. II. Enzyme measurement. J Biol Chem 207:19–37
Nagata T, Todescan R, Goldberg HA Zhang Q, Sodek J (1989) Sulphation of secreted phosphoprotein I (SPPI, osteopontin) is associated with mineralized tissue formation. Biochem Biophys Res Commun 165:234–240
Nagata T, Goldberg HA Zhang Q, Domenicucci C, Sodek J (1991) Biosynthesis of bone proteins by fetal porcine calvariae in vitro. Rapid association of sulfated sialoproteins (secreted phosphoprotein-1 and bone sialoprotein) and chondroitin sulfate proteoglycan (CS-PGIII) with bone mineral. Matrix 11:86–100
Campbell KP, MacLennan DH, Jorgensen AO (1983) Staining of the Ca2+-binding proteins, calsequetin, calmodulin, troponin C, and S-100, with the cationic cabocyanine dye “Stains-all”. J Biol Chem 258:11267–11273
Senger DR, Peruzzi CA, Gracey CF, Papadopoulos A, Tenen DG (1988) Secreted phosphoprotein associated with neoplastic transformation. Close homology with plasma proteins cleaved during blood coagulation. Cancer Res 48:5770–5774
Kasugai S, Nagata T, Sodek J (1992) Temporal studies on the tissue compartmentalization of bone sialoprotein (BSP), osteopontin (OPN), and SPARC protein during bone formation in vitro. J Cell Physiol 152:467–477
Ueno K, Kimmel DB, Haba T, Jee WSS (1984) Increased metaphyseal hard tissue mass in growing long bone following prostaglandin E2 administration: In: Cohn DV, Fujita T, Potts JJR JT, Talmage REV (eds) Endocrine control of bone and calcium metabolism. Elsevier, Amsterdam, pp 151–154
Li XJ, Jee WSS, Li YL, Patterson-Buckendahl P (1990) Transient effects of subcutaneously administered prostaglandin E2 on cancellous and cortical bone in young adult dogs. Bone 11: 353–364
Norrdin RW, Shim MS (1988) Systemic effects of prostaglandin E2 on vertebral trabecular remodeling in beagles used in a healing study. Calcif Tissue Int 42:363–368
Kasahara S, Nishikawa S, Ishida H, Nagata T, Yamauchi N, Ohishi K, Wakano Y, Inoue H (1992) The role of 5′-methylthioadenosine on rat calvaria cell differentiation. Biochem Biophys Res Commun 182:817–823
Kubota T, Zhang Q, Wrana JL, Ber R, Aubin JE, Butler WT, Sodek J (1989) Multiple forms of SPPI (secreted phosphoprotein, osteopontin) synthesized by normal and transformed rat bone cell populations: regulation by TGF-β. Biochem Biophys Res Commun 162:1453–1459
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Nagata, T., Kaho, K., Nishikawa, S. et al. Effect of prostaglandin E2 on mineralization of bone nodules formed by fetal rat calvarial cells. Calcif Tissue Int 55, 451–457 (1994). https://doi.org/10.1007/BF00298559
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00298559