Skip to main content
SpringerLink
Log in

Effects of dexamethasone on expression and maintenance of cartilage in serum-containing cultures of calvaria cells

  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Summary

The effects of dexamethasone on the ability of cells enzymatically isolated from 21-day fetal rat calvaria to produce cartilage in vitro has been investigated. Primary cultures of single-cell suspensions of rat calvaria were grown for up to 28 days in vitro in α-minimal essential medium containing 15% fetal bovine serum, 50 μ/ml ascorbic acid, 10 mM Na β-glycerophosphate and dexamethasone at concentrations of 1 μM to 1 nM. Two types of nodules were present in dexamethasone-containing cultures. One has been characterized previously as bone (Bellows et al. 1986). The second morphologically resembled hyaline cartilage, possessed a strong Alcian blue-positive matrix and contained type-II, but not type-I, collagen. Both bone and cartilaginous nodules were spatially distinct and developed in isolation from each other. Cartilaginous nodules were found in the highest number at a dexamethasone concentration of 100 nM. Time-course experiments revealed that while the number of bone nodules increased continuously at least to day 28, the number of cartilaginous nodules remained constant after cultures had reached confluency. When cells were isolated separately from frontal and parietal bones and suturai regions, the greatest number of cartilaginous nodules developed from parietal bones. Since 21-day fetal rat calvaria contains 2 distinct patches of cartilage at the periphery of the parietal bones, it seems likely that this cartilaginous tissue is the origin of the cartilage cells. The results demonstrate that cultures of rat calvaria cells contain chondrocytes and possibly chondroprogenitor cells that are distinct from osteoprogenitors. Results support previous data that 100 nM dexamethasone permits the expression of and maintains the phenotype of chondrocytes in serum-containing cultures in vitro.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Aubin JE, Heersche JNM, Merrilees MJ, Sodek J (1982) Isolation of bone cell clones with differences in growth, hormone responses and extracellular matrix production. J Cell Biol 192:452–461

    Google Scholar 

  • Baron R, Vignery A, Neff L, Silverglate A, Santa Maria A (1983) Processing of undecalcified bone specimens for bone histomorphometry. In: Recker RR (ed) Bone Histomorphometry, Techniques and Interpretation. CRC Press, Boca Raton, FL, pp 13–35

    Google Scholar 

  • Barrett AJ, Sledge CB, Dingle JT (1966) Effect of cortisol on the synthesis of chondroitin sulphate by embryonic cartilage. Nature 211:83–84

    Google Scholar 

  • Baxter JD, Forsham PJ (1972) Tissue effects of glucocorticoids. Am J Med 19:245–249.

    Google Scholar 

  • Baxter JD, Rousseau GG (1979) Glucocorticoid hormone action. Monogr Endocrinol Vol. 12 Springer, Berlin Heidelberg New York

    Google Scholar 

  • Bellows CG, Aubin JE, Heersche JNM, Antosz ME (1986) Mineralized bone nodules formed in vitro from enzymatically-released calvaria cell populations. Calcif Tissue Int 38:143–154

    Google Scholar 

  • Bellows CG, Aubin JE, Heersche JNM (1987) Physiological concentrations of glucocorticoids stimulate formation of bone nodules from isolated rat calvaria cells in vitro. Endocrinology 121:1985–1994

    Google Scholar 

  • Blanck TJJ, Peterkovsky B (1975) The stimulation of collagen secretion by ascorbate as a result of increased proline hydroxylation in chick embryo fibroblasts. Arch Biochem Biophys 171:250–261

    Google Scholar 

  • Blondelon D, Adolphe M, Zizine L, Léchat P (1980) Evidence for glucocorticoid receptors in cultured rabbit articular chondrocytes. FEBS Lett 117:195–198

    Google Scholar 

  • Burks JK, Peck WA (1978) Bone cells: A serum-free medium supports proliferation in primary culture. Science 199:542–544

    Google Scholar 

  • Burks JK, Peck WA (1979) Growth and differentiation of isolated calvarium cells in serum-free medium. Int Rev Cytol Suppl 10:103–115

    Google Scholar 

  • Cake MH, Litwack G (1975) The glucocorticoid receptor. In: Litwack G (ed) Biochemical actions of hormones, Vol. 3. Academic Press Inc., New York, pp 317–390

    Google Scholar 

  • Calcagno M, Goyena H, Arrambide E, de Urse CA (1970) Action of cortisone and cortisol upon biosynthesis of chondroitin sulfate in femur in vitro cultures of chick embryo. Exp Cell Res 63:131–137

    Google Scholar 

  • Chen WT, Singer SJ (1982) Immunoelectron microscopic studies of the sites of cell-substratum and cell-cell contacts in cultured fibroblasts. J Cell Biol 95:205–222

    Google Scholar 

  • Corvol MT, Malemud CJ, Sokoloff L (1972) A pituitary growthpromoting factor for articular chondrocytes in monolayer culture. Endocrinology 90:262–271

    Google Scholar 

  • Daniel JC, Pauli BU, Kuettner KE (1984) Synthesis of cartilage matrix by mammalian chondrocytes in vitro III Effects of ascorbate. J Cell Biol 99:1960–1969

    Google Scholar 

  • Fessier JH, Fessier LI (1978) Biosynthesis of procollagen. Ann Rev Biochem 47:126–162

    Google Scholar 

  • Hajek AS, Solursh M (1977) The effect of ascorbic acid on growth and synthesis of matrix components by cultured chick embryo chondrocytes. J Exp Zool 200:377–388

    Google Scholar 

  • Hall BK (1983) Tissue interactions and chondrogenesis. In: Hall BK (ed) Cartilage. Vol. 2. Development, Differentiation and Growth. Academic Press, New York, pp 187–222

    Google Scholar 

  • Heersche JNM, Aubin JE, Grigoriadis AE, Moriya Y (1985) Hormone rcsponsiveness of bone cells populations. Searching for answers in vivo and in vitro. In: Butler WT (ed) The Chemistry and Biology of Mineralized Tissues. EBSCO Media, Inc., Birmingham, pp 286–295

    Google Scholar 

  • Heersche JNM, Pitaru S, Aubin JE, Tenenbaum HC (1986) Corticosteroid-induced expression of cartilage phenotype in cultured membrane bone periosteum. In: Cohn DV, Fujita T, Potts JT, Talmage RV (eds) Endocrine Control of Bone and Calcium Metabolism. Elsevier, Amsterdam, pp 147–150

    Google Scholar 

  • Jennings SD, Ham RG (1983) Clonal growth of primary cultures of human hyaline chondrocytes in a defined medium. Cell Biol Int Rep 7:149–159

    Google Scholar 

  • Johnson GD, Davidson RS, McNamee RC, Russell G, Goodwin D, Holborrow EJ (1982) Fading of immunofluorescence during microscopy: a study of the problem and its remedy. J Immunol Methods 55:231–242

    Google Scholar 

  • Kato Y, Gospodarowicz D (1985) Stimulation by glucocorticoid of the synthesis of cartilage matrix proteoglycans by rabbit costal chondrocytes in vitro. J Biol Chem 260:2364–2373

    Google Scholar 

  • Kilgore BS, McNatt ML, Medor S, Lee JA, Hughes ER, Elders MJ (1979) Alteration of cartilage glycosaminoglycan protein acceptor by somatomedin and cortisol. Paediatr Res 13:96–99

    Google Scholar 

  • Lebovitz HE, Eisenbarth GS (1975) Hormonal regulation of cartilage growth and metabolism. Vitam Horm 38:575–648

    Google Scholar 

  • Lee JA, Hicks DC, Hughes ER, Smith D, Elders MJ (1978) Cytoplasmic dexamethasone receptors in embryonic chick cartilage: nature and ontogeny. J Steroid Biochem 9:23–28

    Google Scholar 

  • Lewinson D, Silbermann M (1984) In vitro precocious accumulation of calcium and matrix vesicles formation in young cartilage cells: specific effects of corticosteroids. Calcif Tissue Int 36:702–710

    Google Scholar 

  • Maor G, Silbermann M (1981) In vitro effects of glucocorticoid hormones on the synthesis of DNA in cartilage of neonatal mice. FEBS Lett 129:256–260

    Google Scholar 

  • Martin CE, Cake MH, Hartmann PE, Cook IF (1977) Relationship between foetal corticosteroids, maternal progesterone and parturition in the rat. Acta Endocrinol 84:167–176

    Google Scholar 

  • McLean IW, Nakane PK (1974) Periodate-lysine-paraformaldehyde fixature: a new fixature for immunoelectron microscopy. J Histochem Cytochem 22:1077–1082

    CAS  PubMed  Google Scholar 

  • Meier S, Solursh M (1978) Ultrastructural analysis of the effect of ascorbic acid on secretion and assembly of extracellular matrix by cultured chick embryochondrocytes. J Ultrastruct Res 65:48–59

    Google Scholar 

  • Nefussi J-R, Boy-Lefebre ML, Boulebacke H, Forest N (1985) Mineralization in vitro of matrix formed by osteoblasts isolated by collagenase digestion. Differentiation 29:160–168

    Google Scholar 

  • 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 1233–1241

  • Rao LG, Kalliecharan R, Wang H-M, Heersche JNM, Sodek J (1979) Specific immunohistochemical localization of type I collagen in porcine periodontal tissues using the peroxidase-labelled antibody technique. Histochem J 11:73–82

    Google Scholar 

  • Reynolds JJ (1966) The effect of hydrocortisone on the growth of chick bone rudiments in chemically defined medium. Exp Cell Res 41:174–189

    Google Scholar 

  • Rifas L, Uitto J, Memoli VA, Kuettner KE, Henry RW, Peck W A (1982) Selective emergence of differentiated chondrocytes during serum-free culture of cells derived from fetal rat calvaria. J Cell Biol 92:493–504

    Google Scholar 

  • Rousseau GG (1984) Control of gene expression by glucocorticoid hormones. Biochem J 224:1–12

    Google Scholar 

  • Shen U, Rifas L, Kohler G, Peck WA (1985) Fetal rat chondrocytes sequentially elaborate separate growthand differentiation-promoting peptides during their development in vitro. Endocrinology 116:920–925

    Google Scholar 

  • Silbermann M (1983) Hormones and Cartilage. In: Hall BK, (ed) Cartilage, Vol. 2. Academic Press Inc., pp 327–368

  • Silbermann M, von der Mark K, Maor G, van Menxel M (1987) Dexamethasone impairs growth and collagen synthesis in condylar cartilage in vitro. Bone Mineral 2:87–106

    Google Scholar 

  • Takano T, Takigawa M, Suzuki F (1985) Stimulation by glucocorticoids of the differentiated phenotype of chondrocytes and the proliferation of rabbit costal chondrocytes in culture. J Biochem 97:1093–1100

    Google Scholar 

  • Tenenbaum HC, Heersche JNM (1985) Dexamethasone stimulates osteogenesis in chick periosteum in vitro. Endocrinology 117:2211–2217

    Google Scholar 

  • Tessler RH, Salmon WD (1975) Glucocorticoid inhibition of sulfate incorporation by cartilage of normal rats. Endocrinology 96:898–902

    Google Scholar 

  • Thompson EB, Lippman ME (1974) Mechanism of action of glucocorticoids. Metabolism 23:159–202

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Medical Research Council Group in Periodontal Physiology, Faculty of Dentistry, University of Toronto, Toronto, Ontario, Canada

    C. G. Bellows, J. N. M. Heersche & J. E. Aubin

Authors
  1. C. G. Bellows
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. N. M. Heersche
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. E. Aubin
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bellows, C.G., Heersche, J.N.M. & Aubin, J.E. Effects of dexamethasone on expression and maintenance of cartilage in serum-containing cultures of calvaria cells. Cell Tissue Res. 256, 145–151 (1989). https://doi.org/10.1007/BF00224728

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00224728

Key words

Search

Navigation