Endocrinology, Vol 136, 2671-2677, Copyright © 1995 by Endocrine Society

ARTICLES

The effects of bone morphogenetic protein-2, -4, and -6 on differentiation of rat osteoblast cells in vitro

FJ Hughes, J Collyer, M Stanfield and SA Goodman
Department of Periodontology, London Hospital Medical College, United Kingdom.

The effects of bone morphogenetic protein-2 (BMP-2), -4, and -6 were tested on the differentiation of rat osteoprogenitor cells using a bone nodule-forming assay system, and the kinetics of their actions were investigated by double labeling for alkaline phosphatase (ALP) and bromodeoxyuridine (BrdU) uptake in log phase cultures. All BMPs stimulated bone nodule formation, with an optimal concentration of 25 ng/ml resulting in nodule numbers of approximately 250% of controls using BMP-4 and -6. BMP-2 showed reduced potency compared to either BMP- 4 or -6. No evidence of chondrocytic differentiation was found in any of the cultures. The effect of BMPs on nodule formation was seen after only 24 h of exposure to BMPs, but only affected nodule numbers when added to early cultures. Nodule size and number of cells per nodule were increased with BMP6 only. Continuous or 24-h exposure to BMP-2 or - 4 increased the number of postmitotic ALP-positive cells in log phase cultures, whereas BMP-6 increased the number of postmitotic ALP- negative cells. The results demonstrate that BMP-6, like other BMPs, can stimulate osteoblast differentiation independent of any chondrogenic effects and suggest that an early osteoprogenitor cell is an important target cell for the action of BMPs during bone induction. Overall, BMP-2 and -4 showed differences in potency in the assay systems used, but had qualitatively similar effects. In contrast, the qualitative differences found with BMP-6 suggest that BMP-6 may be acting principally on an early stage osteoprogenitor cell.

This article has been cited by other articles:

				J. Dai, C. L. Hall, J. Escara-Wilke, A. Mizokami, J. M. Keller, and E. T. Keller Prostate Cancer Induces Bone Metastasis through Wnt-Induced Bone Morphogenetic Protein-Dependent and Independent Mechanisms Cancer Res., July 15, 2008; 68(14): 5785 - 5794. [Abstract] [Full Text] [PDF]

				M. Yamashita, F. Otsuka, T. Mukai, H. Otani, K. Inagaki, T. Miyoshi, J. Goto, M. Yamamura, and H. Makino Simvastatin antagonizes tumor necrosis factor-{alpha} inhibition of bone morphogenetic proteins-2-induced osteoblast differentiation by regulating Smad signaling and Ras/Rho-mitogen-activated protein kinase pathway J. Endocrinol., March 1, 2008; 196(3): 601 - 613. [Abstract] [Full Text] [PDF]

				N. E. Hatch, M. Hudson, M. L. Seto, M. L. Cunningham, and M. Bothwell Intracellular Retention, Degradation, and Signaling of Glycosylation-deficient FGFR2 and Craniosynostosis Syndrome-associated FGFR2C278F J. Biol. Chem., September 15, 2006; 281(37): 27292 - 27305. [Abstract] [Full Text] [PDF]

				M. Yaar, C. Wu, H.-Y. Park, I. Panova, G. Schutz, and B. A. Gilchrest Bone Morphogenetic Protein-4, a Novel Modulator of Melanogenesis J. Biol. Chem., September 1, 2006; 281(35): 25307 - 25314. [Abstract] [Full Text] [PDF]

				M. P. Valta, T. Hentunen, Q. Qu, E. M. Valve, A. Harjula, J. A. Seppanen, H. K. Vaananen, and P. L. Harkonen Regulation of Osteoblast Differentiation: A Novel Function for Fibroblast Growth Factor 8 Endocrinology, May 1, 2006; 147(5): 2171 - 2182. [Abstract] [Full Text] [PDF]

				V. Deregowski, E. Gazzerro, L. Priest, S. Rydziel, and E. Canalis Notch 1 Overexpression Inhibits Osteoblastogenesis by Suppressing Wnt/beta-Catenin but Not Bone Morphogenetic Protein Signaling J. Biol. Chem., March 10, 2006; 281(10): 6203 - 6210. [Abstract] [Full Text] [PDF]

				M.F. Termaat, F.C. Den Boer, F.C. Bakker, P. Patka, and H.J.Th.M. Haarman Bone Morphogenetic Proteins. Development and Clinical Efficacy in the Treatment of Fractures and Bone Defects J. Bone Joint Surg. Am., June 1, 2005; 87(6): 1367 - 1378. [Abstract] [Full Text] [PDF]

				F. Stossi, D. H. Barnett, J. Frasor, B. Komm, C. R. Lyttle, and B. S. Katzenellenbogen Transcriptional Profiling of Estrogen-Regulated Gene Expression via Estrogen Receptor (ER) {alpha} or ER{beta} in Human Osteosarcoma Cells: Distinct and Common Target Genes for These Receptors Endocrinology, July 1, 2004; 145(7): 3473 - 3486. [Abstract] [Full Text] [PDF]

				T. Akiyoshi, K. Uchida, and S. Tateyama Expression of Bone Morphogenetic Protein-6 and Bone Morphogenetic Protein Receptors in Myoepithelial Cells of Canine Mammary Gland Tumors Vet. Pathol., March 1, 2004; 41(2): 154 - 163. [Abstract] [Full Text] [PDF]

				J. Lemonnier, C. Ghayor, J. Guicheux, and J. Caverzasio Protein Kinase C-independent Activation of Protein Kinase D Is Involved in BMP-2-induced Activation of Stress Mitogen-activated Protein Kinases JNK and p38 and Osteoblastic Cell Differentiation J. Biol. Chem., January 2, 2004; 279(1): 259 - 264. [Abstract] [Full Text] [PDF]

				M. Nagano, B.-Y. Ryu, C. J. Brinster, M. R. Avarbock, and R. L. Brinster Maintenance of Mouse Male Germ Line Stem Cells In Vitro Biol Reprod, June 1, 2003; 68(6): 2207 - 2214. [Abstract] [Full Text] [PDF]

				R. D. Devlin, Z. Du, R. C. Pereira, R. B. Kimble, A. N. Economides, V. Jorgetti, and E. Canalis Skeletal Overexpression of Noggin Results in Osteopenia and Reduced Bone Formation Endocrinology, May 1, 2003; 144(5): 1972 - 1978. [Abstract] [Full Text] [PDF]

				E. Canalis, A. N. Economides, and E. Gazzerro Bone Morphogenetic Proteins, Their Antagonists, and the Skeleton Endocr. Rev., April 1, 2003; 24(2): 218 - 235. [Abstract] [Full Text] [PDF]

				T. Maeda, T. Kawane, and N. Horiuchi Statins Augment Vascular Endothelial Growth Factor Expression in Osteoblastic Cells via Inhibition of Protein Prenylation Endocrinology, February 1, 2003; 144(2): 681 - 692. [Abstract] [Full Text] [PDF]

				H. Tanaka, E. Nagai, H. Murata, T. Tsubone, Y. Shirakura, T. Sugiyama, T. Taguchi, and S. Kawai Involvement of bone morphogenic protein-2 (BMP-2) in the pathological ossification process of the spinal ligament Rheumatology, October 1, 2001; 40(10): 1163 - 1168. [Abstract] [Full Text] [PDF]

				R. C. Pereira, A. N. Economides, and E. Canalis Bone Morphogenetic Proteins Induce Gremlin, a Protein That Limits Their Activity in Osteoblasts Endocrinology, December 1, 2000; 141(12): 4558 - 4563. [Abstract] [Full Text] [PDF]

				A. Yamaguchi, T. Komori, and T. Suda Regulation of Osteoblast Differentiation Mediated by Bone Morphogenetic Proteins, Hedgehogs, and Cbfa1 Endocr. Rev., August 1, 2000; 21(4): 393 - 411. [Abstract] [Full Text] [PDF]

				T. L. McCarthy, C. Ji, Y. Chen, K. Kim, and M. Centrella Time- and Dose-Related Interactions between Glucocorticoid and Cyclic Adenosine 3',5'-Monophosphate on CCAAT/Enhancer-Binding Protein-Dependent Insulin-Like Growth Factor I Expression by Osteoblasts Endocrinology, January 1, 2000; 141(1): 127 - 137. [Abstract] [Full Text] [PDF]

				E. Gazzerro, S. Rydziel, and E. Canalis Skeletal Bone Morphogenetic Proteins Suppress the Expression of Collagenase-3 by Rat Osteoblasts Endocrinology, February 1, 1999; 140(2): 562 - 567. [Abstract] [Full Text]

				S. D. Boden, G. Hair, L. Titus, M. Racine, K. McCuaig, J. M. Wozney, and M. S. Nanes Glucocorticoid-Induced Differentiation of Fetal Rat Calvarial Osteoblasts Is Mediated by Bone Morphogenetic Protein-6 Endocrinology, July 1, 1997; 138(7): 2820 - 2828. [Abstract] [Full Text] [PDF]

				X. Gu, B.-H. Shin, Y. Akbarali, A. Weiss, J. Boltax, P. Oettgen, and T. A. Libermann Tel-2 Is a Novel Transcriptional Repressor Related to the Ets Factor Tel/ETV-6 J. Biol. Chem., March 16, 2001; 276(12): 9421 - 9436. [Abstract] [Full Text] [PDF]