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Exp Clin Endocrinol Diabetes 2003; 111(2): 66-72
DOI: 10.1055/s-2003-39231
Article

J. A. Barth Verlag in Georg Thieme Verlag Stuttgart · New York

The Effects of Triiodothyronine on Human Osteoblast-Like Cells Metabolism and Interactions with Growth Hormone

C. E. Pepene 1 , 2 , T. Seck 1 , J. Pfeilschifter 1 , L. Gozariu 2 , R. Ziegler 1 , C. H. Kasperk 1
  • 1Department of Internal Medicine I, University of Heidelberg, Heidelberg, Germany
  • 2Department of Endocrinology, University of Medicine and Pharmacy Cluj-Napoca, Cluj-Napoca, Romania
Further Information

Publication History

Received: November 25, 2000 First decision: April 24, 2001

Accepted: July 19, 2002

Publication Date:
14 May 2003 (online)

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Abstract

The expression of thyroid hormones receptors in osteoblasts and osteoclasts has involved these cells as direct targets for triiodothyronine (T3), but thyroid hormones may also interact with other hormones or local growth factors to exert their actions on bone cells. Among these, growth hormone (GH) is recognised as participating in the acquisition and maintenance of bone mass and exerting stimulatory effects on human osteoblastic cells. The aim of this study was to investigate T3 effects on primary human adult osteoblast-like cells (HOB) as well as to test for possible interactions between T3 and GH on bone cell metabolism.

Primary human bone cell cultures were obtained by outgrowth from trabecular bone fragments from the hip and knee. Dose-response studies demonstrated enhanced [3H]-thymidine incorporation for T3 at 10-9, 10-8, 10-7 and 20-7 M, with a maximal response of 162.81 ± 12.97 % with T3 10-8 M, compared to vehicle (p < 0.001). Time-course studies showed an increased osteoblast-like cell proliferation after 24 h, followed by a decrease of cell proliferation by 48 h and 72 h of culture, respectively, when compared to control cells, with a maximal response after 72 h (T3 10-10 M: 45.21 ± 6.97 %, p < 0.01). In addition, T3 markedly increased specific alkaline phosphatase (AP) activity in HOB (10-10 M: 169.86 ± 12.14 % vs. control, p < 0.001), but no significant influence on type I procollagen propeptide (PICP) production was observed. At 10-9 - 10-7 g/ml, GH significantly enhanced HOB proliferation (p < 0.001) however, GH effects were not dose-dependent. Triiodothyronine, at a high concentration (10-7 M), stimulated GH-receptor (GHR) mRNA levels by 165.20 ± 16.54 % after 24 h (p < 0.05). Correspondingly, a synergistic effect of T3 with the same concentration and GH on cell proliferation in human adult osteoblast-like cells was found.

Key words

Thyroid hormone - Somatotropin - Osteoblasts - Somatotropin receptors

References

  • 1 Abu E G, Bord S, Horner A, Chatterjee K K, Compston J E. The expression of thyroid hormone receptors in human bone.  Bone. 1997;  21 137-142
    CrossrefPubMedGoogle Scholar
  • 2 Allain T J, Yen P M, Flanagan A M, McGregor A M. The isoform-specific expression of the tri-iodothyronine receptor in osteoblasts and osteoclasts.  Eur J Clin Invest. 1996;  26 418-425
    CrossrefPubMedGoogle Scholar
  • 3 Amit T, Hertz P, Ish-Shalom S, Lotan R, Luboshitzki R, Youdim M B, Hochberg Z. Effects of hypo or hyper-thyroidism on growth hormone-binding protein.  Clin Endocrinol. 1991;  35 159-162
    PubMedGoogle Scholar
  • 4 Bengtsson B A, Eden S, Lonn L. Treatment of adults with growth hormone (GH) deficiency with recombinant human GH.  J Clin Endocrinol Metab. 1993;  76 309-317
    CrossrefPubMedGoogle Scholar
  • 5 Brameld J M, Weller P A, Saunders J C, Buttery P C, Gilmour R S. Hormonal control of insulin-like growth factor-I and growth hormone receptor RNA expression by porcine hepatocytes in culture.  J Endocrinol. 1995;  146 239-245
    PubMedGoogle Scholar
  • 6 Britto J M, Fenton A J, Holloway W R, Nicholson G C. Osteoblasts mediate thyroid hormone stimulation of osteoclastic bone resorption.  Endocrinology. 1994;  134 169-176
    CrossrefPubMedGoogle Scholar
  • 7 Brixen K, Nielsen H K, Mosekilde L, Flyvbjerg A. A short course of recombinant human growth hormone treatment stimulates osteoblasts and activates bone remodeling in normal human volunteers.  J Bone Miner Res. 1990;  5 609-618
    PubMedGoogle Scholar
  • 8 Calvo M, Eyre D, Gundberg C. Molecular basis and clinical application of biological markers of bone turnover.  Endocr Rev. 1996;  17 333-368
    PubMedGoogle Scholar
  • 9 Chenu C, Valentin-Opran A, Chavassieux P, Saez S, Meunier P J, Delmas P D. Insulin-like growth factor I hormonal regulation by growth hormone and by 1, 25-(OH)2D3 and activity on human osteoblast-like cells in short-term cultures.  Bone. 1990;  11 81-86
    CrossrefPubMedGoogle Scholar
  • 10 Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.  Anal Biochem. 1987;  162 156-159
    PubMedGoogle Scholar
  • 11 Chomczynski P, Soszynski P A, Frohman L A. Stimulatory effect of thyroid hormone on growth hormone gene expression in a human pituitary cell line.  J Clin Endocrinol Metab. 1993;  77 281-285
    CrossrefPubMedGoogle Scholar
  • 12 Cooper D S, Kaplan M M, Ridgway E C, Maloof F, Daniels G H. Alkaline phosphatase isoenzyme patterns in hyperthyroidism.  Ann Intern Med. 1979;  90 164-168
    PubMedGoogle Scholar
  • 13 Degerblad M, Elgindy N, Hall K, Sjoberg H E, Thoren M. Potent effect of recombinant growth hormone on bone mineral density and body composition in adults with panhypopituitarism.  Acta Endocrinol. 1992;  126 387-393
    PubMedGoogle Scholar
  • 14 Eriksen E F, Kassem M, Brixen K. Growth hormone and insulin-like growth factors as anabolic therapies for osteoporosis.  Horm Res. 1993;  40 95-98
    PubMedGoogle Scholar
  • 15 Ernst M, Froesch E R. Triiodothyronine stimulates proliferation of osteoblast-like cells in serum-free culture.  FEBS Lett. 1987;  220 163-166
    CrossrefPubMedGoogle Scholar
  • 16 Garnero P, Vassy V, Bertholin A, Riou J P, Delmas P D. Markers of bone turnover in hyperthyroidism and the effects of treatment.  J Clin Endocrinol Metab. 1994;  78 955-959
    CrossrefPubMedGoogle Scholar
  • 17 Hansen T B, Brixen K, Vahl N, Jorgensen J OO, Christiansen J S, Mosekilde L, Hagen C. Effects of 12 months of growth hormone (GH) treatment on calciotropic hormones, calcium homeostasis and bone metabolism in adults with acquired GH deficiency: a double blind, randomised, placebo-controlled study.  J Clin Endocrinol Metab. 1996;  81 3352-3359
    PubMedGoogle Scholar
  • 18 Kanzaki S, Baxter R C, Knutsen R, Baylink D J, Mohan S. Evidence that human bone cells in culture secrete insulin-like growth factor (IGF)-II and IGF-binding protein-3 but not acid-labile subunit both under basal and regulated conditions.  J Bone Miner Res. 1995;  10 854-858
    PubMedGoogle Scholar
  • 19 Kasperk C H, Fitzsimmons R, Strong D, Mohan S, Jennings J, Werdegal J, Baylink D. Studies of the mechanism by which androgens enhance mitogenesis and differentiation in bone cells.  J Clin Endocrinol Metab. 1990;  71 1322-1329
    CrossrefPubMedGoogle Scholar
  • 20 Kassem M, Blum W, Ristelli J, Mosekilde L, Eriksen E F. Growth hormone stimulates proliferation and differentiation of normal human osteoblast-like cells in vitro.  Calcif Tissue Int. 1993 a;  52 222-226
    CrossrefPubMedGoogle Scholar
  • 21 Kassem M, Mosekilde L, Eriksen E F. Effects of triiodothyronine on DNA synthesis and differentiation markers of normal human osteoblast-like cells in vitro.  Biochem Mol Biol Int. 1993 b;  30 779-788
    PubMedGoogle Scholar
  • 22 Korytko A I, Cuttler L. Thyroid hormone and glucocorticoid regulation of pituitary growth hormone-releasing hormone receptor gene expression.  J Endocrinol. 1997;  152 13-17
    CrossrefPubMedGoogle Scholar
  • 23 Krieger N S, Stappenbeck T S, Stern P H. Characterization of specific thyroid hormone receptors in bone.  J Bone Miner Res. 1988;  3 473-478
    PubMedGoogle Scholar
  • 24 Kung Sutherland M, Rao L G, Sodek J, Wylie J, Ly H, Haussler M R, Murray T M. Regulation of thyroid hormone receptor gene expression in osteoblastic SaOs-2 cells [Abstract No.  62] J Bone Miner Res. 1990;  5 (Suppl) 89
    PubMedGoogle Scholar
  • 25 Lalau J D, Sebert J L, Marie A, Fournier A, Quichaud J. Effect of thyrotoxicosis and its treatment on mineral and bone metabolism.  J Endocrinol Invest. 1986;  491-496
    PubMedGoogle Scholar
  • 26 Lakatos P, Caplice M D, Khanna V, Stern P H. Thyroid hormones increase insulin-like growth factor I content in the medium of rat bone tissue.  J Bone Miner Res. 1993;  8 1475-1481
    PubMedGoogle Scholar
  • 27 LeBron B A, Pekary E A, Mirell C, Hahn T, Hershman J M. Thyroid hormone 5′-deiodinase activity, nuclear binding and effects on mitogenesis in UMR-106 osteoblastic osteosarcoma cells.  J Bone Miner Res. 1989;  4 173-178
    PubMedGoogle Scholar
  • 28 Lee M, Kim S Y, Lee C L, Cho B Y, Lee H K, Koh C S, Min H K. Negative correlation between the change in bone mineral density and serum osteocalcin in patients with hyperthyroidism.  J Clin Endocrinol Metab. 1990;  70 766-770
    PubMedGoogle Scholar
  • 29 Lewinson D, Harel Z, Shenzer P, Silbermann M, Hochberg Z. Effect of thyroid hormone and growth hormone on recovery from hypothyroidism of epiphyseal growth plate cartilage and its adjacent bone.  Endocrinology. 1989;  124 937-945
    PubMedGoogle Scholar
  • 30 Lewinson D, Bialik G M, Hochberg Z. Differential effects of hypothyroidism on the cartilage and the osteogenic process in the mandibular condyle: recovery by growth hormone and thyroxine.  Endocrinology. 1994;  135 1504-1510
    CrossrefPubMedGoogle Scholar
  • 31 Miki N, Ono M, Murata Y, Ohsaki E, Tamitsu K, Ri T, Demura H, Yamada M. Thyroid hormone regulation of gene expression of the pituitary growth-hormone releasing factor receptor.  Biochem Biophys Res Commun. 1995;  217 1087-1093
    CrossrefPubMedGoogle Scholar
  • 32 Milne M, Kang M, Quail J M, Baran D T. Thyroid hormone excess increases insulin-like growth factor I transcripts in bone marrow cell cultures: divergent effects on vertebral and femoral cell cultures.  Endocrinology. 1998;  139 2527-2534
    CrossrefPubMedGoogle Scholar
  • 33 Mohan S, Strong D, Lempert U G, Tremollieres F, Wergedal J E, Baylink D J. Studies on regulation of insulin-like growth factor binding protein (IGFBP)-3 and IGFBP-4 production in human bone cells.  Acta Endocrinol. 1992;  127 555-564
    PubMedGoogle Scholar
  • 34 Mosekilde L, Christensen M S. Decreased parathyroid function in hyperthyroidism: interrelationships between serum parathyroid hormone, calcium-phosphorus metabolism and thyroid function.  Acta Endocrinol. 1977;  84 566-575
    PubMedGoogle Scholar
  • 35 Nilsson A, Swolin D, Enerback S, Ohlsson C. Expression of functional growth hormone receptors in cultured human osteoblast-like cells.  J Clin Endocrinol Metab. 1995;  80 3483-3488
    CrossrefPubMedGoogle Scholar
  • 36 Rizzoli R, Poser J, Burgi U. Nuclear thyroid hormone receptors in cultured bone cells.  Metabolism. 1986;  35 71-74
    PubMedGoogle Scholar
  • 37 Rosen T, Johansson G, Hallgren P, Caidahl K, Bosaeus I, Bengtsson B A. Beneficial effects of 12 months replacement therapy with recombinant human growth hormone to growth hormone deficient adults.  J Clin Endocrinol Metab. 1994;  1 55-66
    PubMedGoogle Scholar
  • 38 Saggesse G, Baroncelli G I, Bertelloni S, Cinquanta L, Di Nero G. Effects of long-term treatment with growth hormone on bone and mineral metabolism in children with growth hormone deficiency.  J Pediatr. 1993;  122 37-45
    PubMedGoogle Scholar
  • 39 Sato K, Han D C, Fuji Y, Tsushima T, Shizume K. Thyroid hormone stimulates alkaline phosphatase activity in cultured rat osteoblastic cells (ROS 17/2. 8) through 3, 5, 3′-triiodo-L-thyronine nuclear receptors.  Endocrinology. 1987;  120 1873-1881
    PubMedGoogle Scholar
  • 40 Scheven B A, Hamilton J, Fakkeldij T M, Duursma S A. Effects of recombinant human insulin-like growth factor I and II (IGF-I/-II) and growth hormone (GH) on the growth of normal adult human osteoblast-like cells and human osteogenic sarcoma cells.  Growth Regulation. 1991;  1 160-167
    PubMedGoogle Scholar
  • 41 Schmid C, Schläpfer I, Futo E, Waldvogel M, Schwander J, Zapf J, Froesch E R. Triiodothyronine (T3) stimulates insulin-like growth factor (IGF)-I and IGF binding protein (IGFBP)-2 production by rat osteoblasts in vitro.  Acta Endocrinol. 1992;  126 467-473
    PubMedGoogle Scholar
  • 42 Shen X Y, Rodriguez-Arnao J, Ross R J. Regulation of hepatic RNA levels for the growth hormone receptor in rats with altered thyroid status.  Growth Regulation. 1995;  5 199-202
    PubMedGoogle Scholar
  • 43 Slootweg M, Ohlsson C, Salles J P, de Vries C P, Netelenbos J. Insulin-like growth factor binding proteins-2 and - 3 stimulate growth hormone receptor binding and mitogenesis in rat osteosarcoma cells.  Endocrinology. 1995;  136 4210-4217
    CrossrefPubMedGoogle Scholar
  • 44 Slootweg M C, Salles J P, Ohlsson C, de Vries C P, Engelbregt M J, Netelenbos J C. Growth hormone binds to a single high affinity receptor site on mouse osteoblasts: modulation by retinoic acids and cell differentiation.  J Endocrinol. 1996;  150 465-472
    CrossrefPubMedGoogle Scholar
  • 45 Slootweg M C, Swolin D, Netelenbos J C, Isaksson O G, Ohlsson C. Estrogen enhances growth hormone receptor expression and growth hormone action in rat osteosarcoma cells and human osteoblast-like cells.  J Endocrinol. 1997;  155 159-164
    CrossrefPubMedGoogle Scholar
  • 46 Slootweg M C, Vangenesen S T, Otte A P, Duursma S A, Kruijer W. Activation of mouse osteoblast growth hormone receptor c-fos oncogene expression independent of phosphoinositide breakdown and cyclic AMP.  J Mol Endocrinol. 1990;  120 1806-1812
    PubMedGoogle Scholar
  • 47 Swollin-Eide D, Nilsson A, Ohlsson C. Cortisol increases growth hormone-receptor expression in human osteoblast-like cells.  J Endocrinol. 1998;  136 99-105
    PubMedGoogle Scholar
  • 48 Tam S P, Lam K S, Srivastava G. Gene expression of hypothalamic somatostatin, growth hormone releasing factor, and their pituitary receptors in hypothyroidism.  Endocrinology. 1996;  137 418-424
    CrossrefPubMedGoogle Scholar
  • 49 Varga F, Rumpler M, Klaushofer K. Thyroid hormones increase insulin-like growth factor mRNA levels in the clonal osteoblastic cell line MC3T3-E1.  FEBS Lett. 1994;  345 67-70
    CrossrefPubMedGoogle Scholar
  • 50 Varga F, Rumpler M, Luegmayr E, Fratzl-Zelman N, Glantschnig H, Klaushofer K. Triiodothyronine, a regulator of osteoblastic differentiation: depression of histone H4, attenuation of c-fos/c-jun and induction of osteocalcin expression.  Calcif Tissue Int. 1997;  61 404-411
    CrossrefPubMedGoogle Scholar
  • 51 Vanderveen E A, Netelenbos J C. Growth hormone (replacement) therapy in adults-bone and calcium metabolism.  Horm Res. 1990;  33 65-68
    PubMedGoogle Scholar
  • 52 Vandeweghe M, Taelman P, Kaufman J M. Short and long-term effects of growth hormone treatment on bone turnover and bone mineral content in adult growth hormone-deficient males.  Clin Endocrinol. 1993;  39 409-415
    PubMedGoogle Scholar

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