Abstract
To determine the effects of transforming growth factor-beta (TGF-beta) on the different cell types that exist in bone, cell populations (I- IV), progressively enriched in osteoblastic cells relative to fibroblastic cells, were prepared from fetal rat calvaria using timed collagenase digestions. TGF-beta did not induce anchorage-independent growth of these cells, nor was anchorage-dependent growth stimulated in most populations studied, despite a two- to threefold increase in the synthesis of cellular proteins. In all populations the synthesis of secreted proteins increased 2-3.5-fold. In particular, collagen, fibronectin, and plasminogen activator inhibitor synthesis was stimulated. However, different degrees of stimulation of individual proteins were observed both within and between cell populations. A marked preferential stimulation of plasminogen activator inhibitor was observed in each population, together with a slight preferential stimulation of collagen; the effect on collagen expression being directed primarily at type I collagen. In contrast, the synthesis of SPARC (secreted protein acidic rich in cysteine/osteonectin was stimulated approximately two-fold by TGF-beta, but only in fibroblastic populations. Collectively, these results demonstrate that TGF-beta stimulates matrix production by bone cells and, through differential effects on individual matrix components, may also influence the nature of the matrix formed by different bone cell populations. In the presence of TGF-beta, osteoblastic cells lost their polygonal morphology and alkaline phosphatase activity was decreased, reflecting a suppression of osteoblastic features. The differential effects of TGF- beta on bone cell populations are likely to be important in bone remodeling and fracture repair.
Full Text
The Full Text of this article is available as a PDF (2.4 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Assoian R. K., Komoriya A., Meyers C. A., Miller D. M., Sporn M. B. Transforming growth factor-beta in human platelets. Identification of a major storage site, purification, and characterization. J Biol Chem. 1983 Jun 10;258(11):7155–7160. [PubMed] [Google Scholar]
- Aubin J. E., Heersche J. N., Merrilees M. J., Sodek J. Isolation of bone cell clones with differences in growth, hormone responses, and extracellular matrix production. J Cell Biol. 1982 Feb;92(2):452–461. doi: 10.1083/jcb.92.2.452. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Centrella M., Massagué J., Canalis E. Human platelet-derived transforming growth factor-beta stimulates parameters of bone growth in fetal rat calvariae. Endocrinology. 1986 Nov;119(5):2306–2312. doi: 10.1210/endo-119-5-2306. [DOI] [PubMed] [Google Scholar]
- Centrella M., McCarthy T. L., Canalis E. Transforming growth factor beta is a bifunctional regulator of replication and collagen synthesis in osteoblast-enriched cell cultures from fetal rat bone. J Biol Chem. 1987 Feb 25;262(6):2869–2874. [PubMed] [Google Scholar]
- Cheifetz S., Weatherbee J. A., Tsang M. L., Anderson J. K., Mole J. E., Lucas R., Massagué J. The transforming growth factor-beta system, a complex pattern of cross-reactive ligands and receptors. Cell. 1987 Feb 13;48(3):409–415. doi: 10.1016/0092-8674(87)90192-9. [DOI] [PubMed] [Google Scholar]
- Fine A., Goldstein R. H. The effect of transforming growth factor-beta on cell proliferation and collagen formation by lung fibroblasts. J Biol Chem. 1987 Mar 15;262(8):3897–3902. [PubMed] [Google Scholar]
- Hughes R. C., Taylor A., Sage H., Hogan B. L. Distinct patterns of glycosylation of colligin, a collagen-binding glycoprotein, and SPARC (osteonectin), a secreted Ca2+-binding glycoprotein. Evidence for the localisation of colligin in the endoplasmic reticulum. Eur J Biochem. 1987 Feb 16;163(1):57–65. doi: 10.1111/j.1432-1033.1987.tb10736.x. [DOI] [PubMed] [Google Scholar]
- Ignotz R. A., Massagué J. Transforming growth factor-beta stimulates the expression of fibronectin and collagen and their incorporation into the extracellular matrix. J Biol Chem. 1986 Mar 25;261(9):4337–4345. [PubMed] [Google Scholar]
- Kao W. W., Berg R. A., Prockop D. J. Kinetics for the secretion of procollagen by freshly isolated tendon cells. J Biol Chem. 1977 Dec 10;252(23):8391–8397. [PubMed] [Google Scholar]
- Laiho M., Saksela O., Andreasen P. A., Keski-Oja J. Enhanced production and extracellular deposition of the endothelial-type plasminogen activator inhibitor in cultured human lung fibroblasts by transforming growth factor-beta. J Cell Biol. 1986 Dec;103(6 Pt 1):2403–2410. doi: 10.1083/jcb.103.6.2403. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Limeback H. F., Sodek J. Procollagen synthesis and processing in periodontal ligament in vivo and in vitro. A comparative study using slab-gel fluorography. Eur J Biochem. 1979 Oct 15;100(2):541–550. doi: 10.1111/j.1432-1033.1979.tb04200.x. [DOI] [PubMed] [Google Scholar]
- Lund L. R., Riccio A., Andreasen P. A., Nielsen L. S., Kristensen P., Laiho M., Saksela O., Blasi F., Danø K. Transforming growth factor-beta is a strong and fast acting positive regulator of the level of type-1 plasminogen activator inhibitor mRNA in WI-38 human lung fibroblasts. EMBO J. 1987 May;6(5):1281–1286. doi: 10.1002/j.1460-2075.1987.tb02365.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mann K., Deutzmann R., Paulsson M., Timpl R. Solubilization of protein BM-40 from a basement membrane tumor with chelating agents and evidence for its identity with osteonectin and SPARC. FEBS Lett. 1987 Jun 22;218(1):167–172. doi: 10.1016/0014-5793(87)81040-2. [DOI] [PubMed] [Google Scholar]
- Mason I. J., Murphy D., Münke M., Francke U., Elliott R. W., Hogan B. L. Developmental and transformation-sensitive expression of the Sparc gene on mouse chromosome 11. EMBO J. 1986 Aug;5(8):1831–1837. doi: 10.1002/j.1460-2075.1986.tb04434.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mason I. J., Taylor A., Williams J. G., Sage H., Hogan B. L. Evidence from molecular cloning that SPARC, a major product of mouse embryo parietal endoderm, is related to an endothelial cell 'culture shock' glycoprotein of Mr 43,000. EMBO J. 1986 Jul;5(7):1465–1472. doi: 10.1002/j.1460-2075.1986.tb04383.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Noda M., Rodan G. A. Type-beta transforming growth factor inhibits proliferation and expression of alkaline phosphatase in murine osteoblast-like cells. Biochem Biophys Res Commun. 1986 Oct 15;140(1):56–65. doi: 10.1016/0006-291x(86)91057-0. [DOI] [PubMed] [Google Scholar]
- Odermatt E., Tamkun J. W., Hynes R. O. Repeating modular structure of the fibronectin gene: relationship to protein structure and subunit variation. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6571–6575. doi: 10.1073/pnas.82.19.6571. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Otsuka K., Yao K. L., Wasi S., Tung P. S., Aubin J. E., Sodek J., Termine J. D. Biosynthesis of osteonectin by fetal porcine calvarial cells in vitro. J Biol Chem. 1984 Aug 10;259(15):9805–9812. [PubMed] [Google Scholar]
- Pfeilschifter J., Mundy G. R. Modulation of type beta transforming growth factor activity in bone cultures by osteotropic hormones. Proc Natl Acad Sci U S A. 1987 Apr;84(7):2024–2028. doi: 10.1073/pnas.84.7.2024. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rao L. G., Ng B., Brunette D. M., Heersche J. N. Parathyroid hormone- and prostaglandin E1-response in a selected population of bone cells after repeated subculture and storage at -80C. Endocrinology. 1977 May;100(5):1233–1241. doi: 10.1210/endo-100-5-1233. [DOI] [PubMed] [Google Scholar]
- Roberts A. B., Anzano M. A., Meyers C. A., Wideman J., Blacher R., Pan Y. C., Stein S., Lehrman S. R., Smith J. M., Lamb L. C. Purification and properties of a type beta transforming growth factor from bovine kidney. Biochemistry. 1983 Dec 6;22(25):5692–5698. doi: 10.1021/bi00294a002. [DOI] [PubMed] [Google Scholar]
- Roberts A. B., Sporn M. B., Assoian R. K., Smith J. M., Roche N. S., Wakefield L. M., Heine U. I., Liotta L. A., Falanga V., Kehrl J. H. Transforming growth factor type beta: rapid induction of fibrosis and angiogenesis in vivo and stimulation of collagen formation in vitro. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4167–4171. doi: 10.1073/pnas.83.12.4167. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sage H., Johnson C., Bornstein P. Characterization of a novel serum albumin-binding glycoprotein secreted by endothelial cells in culture. J Biol Chem. 1984 Mar 25;259(6):3993–4007. [PubMed] [Google Scholar]
- Sage H., Tupper J., Bramson R. Endothelial cell injury in vitro is associated with increased secretion of an Mr 43,000 glycoprotein ligand. J Cell Physiol. 1986 Jun;127(3):373–387. doi: 10.1002/jcp.1041270305. [DOI] [PubMed] [Google Scholar]
- Seyedin S. M., Segarini P. R., Rosen D. M., Thompson A. Y., Bentz H., Graycar J. Cartilage-inducing factor-B is a unique protein structurally and functionally related to transforming growth factor-beta. J Biol Chem. 1987 Feb 15;262(5):1946–1949. [PubMed] [Google Scholar]
- Seyedin S. M., Thompson A. Y., Bentz H., Rosen D. M., McPherson J. M., Conti A., Siegel N. R., Galluppi G. R., Piez K. A. Cartilage-inducing factor-A. Apparent identity to transforming growth factor-beta. J Biol Chem. 1986 May 5;261(13):5693–5695. [PubMed] [Google Scholar]
- Sodek J., Berkman F. A. Bone cell cultures. Methods Enzymol. 1987;145:303–324. doi: 10.1016/0076-6879(87)45018-0. [DOI] [PubMed] [Google Scholar]
- Sporn M. B., Roberts A. B., Shull J. H., Smith J. M., Ward J. M., Sodek J. Polypeptide transforming growth factors isolated from bovine sources and used for wound healing in vivo. Science. 1983 Mar 18;219(4590):1329–1331. doi: 10.1126/science.6572416. [DOI] [PubMed] [Google Scholar]
- Sporn M. B., Roberts A. B., Wakefield L. M., Assoian R. K. Transforming growth factor-beta: biological function and chemical structure. Science. 1986 Aug 1;233(4763):532–534. doi: 10.1126/science.3487831. [DOI] [PubMed] [Google Scholar]
- Tashjian A. H., Jr, Voelkel E. F., Lazzaro M., Singer F. R., Roberts A. B., Derynck R., Winkler M. E., Levine L. Alpha and beta human transforming growth factors stimulate prostaglandin production and bone resorption in cultured mouse calvaria. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4535–4538. doi: 10.1073/pnas.82.13.4535. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thalacker F. W., Nilsen-Hamilton M. Specific induction of secreted proteins by transforming growth factor-beta and 12-O-tetradecanoylphorbol-13-acetate. Relationship with an inhibitor of plasminogen activator. J Biol Chem. 1987 Feb 15;262(5):2283–2290. [PubMed] [Google Scholar]
- Tung P. S., Domenicucci C., Wasi S., Sodek J. Specific immunohistochemical localization of osteonectin and collagen types I and III in fetal and adult porcine dental tissues. J Histochem Cytochem. 1985 Jun;33(6):531–540. doi: 10.1177/33.6.3889139. [DOI] [PubMed] [Google Scholar]
- Uchida N., Smilowitz H., Tanzer M. L. Monovalent ionophores inhibit secretion of procollagen and fibronectin from cultured human fibroblasts. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1868–1872. doi: 10.1073/pnas.76.4.1868. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wasi S., Otsuka K., Yao K. L., Tung P. S., Aubin J. E., Sodek J., Termine J. D. An osteonectinlike protein in porcine periodontal ligament and its synthesis by periodontal ligament fibroblasts. Can J Biochem Cell Biol. 1984 Jun;62(6):470–478. doi: 10.1139/o84-064. [DOI] [PubMed] [Google Scholar]
- Wong G. L., Cohn D. V. Target cells in bone for parathormone and calcitonin are different: enrichment for each cell type by sequential digestion of mouse calvaria and selective adhesion to polymeric surfaces. Proc Natl Acad Sci U S A. 1975 Aug;72(8):3167–3171. doi: 10.1073/pnas.72.8.3167. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wrana J. L., Sodek J., Ber R. L., Bellows C. G. The effects of platelet-derived transforming growth factor beta on normal human diploid gingival fibroblasts. Eur J Biochem. 1986 Aug 15;159(1):69–76. doi: 10.1111/j.1432-1033.1986.tb09834.x. [DOI] [PubMed] [Google Scholar]
- Zung P., Domenicucci C., Wasi S., Kuwata F., Sodek J. Osteonectin is a minor component of mineralized connective tissues in rat. Biochem Cell Biol. 1986 Apr;64(4):356–362. doi: 10.1139/o86-049. [DOI] [PubMed] [Google Scholar]
- de Larco J. E., Todaro G. J. Growth factors from murine sarcoma virus-transformed cells. Proc Natl Acad Sci U S A. 1978 Aug;75(8):4001–4005. doi: 10.1073/pnas.75.8.4001. [DOI] [PMC free article] [PubMed] [Google Scholar]