Proliferation and differentiation of osteoblasts and adipocytes in rat bone marrow stromal cell cultures: Effects of dexamethasone and calcitriol

During bone loss, osteoblast population can be replaced by adipose tissue. This apparent reciprocal relationship between decreased bone density and increased fat formation can be explained by an imbalance in the production of bone‐forming and fat‐forming cells in the marrow cavity. Thus, osteoblast and adipocyte pathways seem more closely and inversely related. In the present study, we investigated the effects of dexamethasone (dex) and calcitriol [1,25(OH)2D3] on proliferation and differentiation of osteoblasts and adipocytes in rat bone marrow stromal cell cultures. Stromal cells were grown in primoculture in presence of dex and subcultivated in presence of dex and/or 1,25(OH)2D3. Total cell proliferation, osteoblast and adipocyte‐cells number, and ‐mRNA specific markers were used to study the effects of hormonal treatment on stromal cells. Total cell proliferation was stimulated by dex and inhibited by 1,25(OH)2D3. Dex increased osteoblast and adipocyte cell population whereas calcitriol decreased bone‐forming cell number and increased fat cell population. The presence of both hormones led to a strong decrease in osteoblastic cells and to a strong increase in adipocytic cell number. Dex induced mRNA osteoblastic markers expression like bone sialoprotein (BSP) and osteocalcin (OC) and an adipocyte marker expression, the fatty acid binding protein aP2. Calcitriol decreased the dex‐induced BSP expression but stimulated slightly OC and aP2 mRNA. The effects of both hormones was to increase strongly OC and aP2 mRNA. These results support that, in rat bone marrow, adipocyte proliferation and differentiation are stimulated by glucocorticoids and calcitriol which act synergically, whereas osteoblastic cell proliferation and differentiation are increased by dex and inhibited by 1,25(OH)2D3. J. Cell. Biochem. 89: 364–372, 2003. © 2003 Wiley‐Liss, Inc.

[1]  J. Chen,et al.  Regulation of bone sialoprotein and osteopontin mRNA expression by dexamethasone and 1,25-dihydroxyvitamin D3 in rat bone organ cultures. , 1996, Connective tissue research.

[2]  J. Aubin,et al.  1,25-dihydroxyvitamin D3 stimulates adipocyte differentiation in cultures of fetal rat calvaria cells: comparison with the effects of dexamethasone. , 1994, Endocrinology.

[3]  D. Rowe,et al.  Regulation of collagen synthesis in fetal rat calvaria by 1,25-dihydroxyvitamin D3. , 1982, The Journal of biological chemistry.

[4]  J Denton,et al.  Adipocytic proportion of bone marrow is inversely related to bone formation in osteoporosis , 2002, Journal of clinical pathology.

[5]  J. Aubin Osteoprogenitor cell frequency in rat bone marrow stromal populations: Role for heterotypic cell–cell interactions in osteoblast differentiation , 1999, Journal of cellular biochemistry.

[6]  L. Mosekilde,et al.  Adipocyte tissue volume in bone marrow is increased with aging and in patients with osteoporosis , 2004, Biogerontology.

[7]  D. Prockop Marrow Stromal Cells as Stem Cells for Nonhematopoietic Tissues , 1997, Science.

[8]  M. Jensen,et al.  Leptin acts on human marrow stromal cells to enhance differentiation to osteoblasts and to inhibit differentiation to adipocytes. , 1999, Endocrinology.

[9]  G. Jordan,et al.  High concentrations of dexamethasone suppress the proliferation but not the differentiation or further maturation of human osteoblast precursors in vitro: relevance to glucocorticoid-induced osteoporosis. , 2001, Rheumatology.

[10]  G. Ringold,et al.  Glucocorticoid regulation of adipocyte differentiation: hormonal triggering of the developmental program and induction of a differentiation-dependent gene , 1985, The Journal of cell biology.

[11]  J. Gimble,et al.  Is there a therapeutic opportunity to either prevent or treat osteopenic disorders by inhibiting marrow adipogenesis? , 2000, Bone.

[12]  H. Senoo,et al.  L‐ascorbic acid 2‐phosphate stimulates collagen accumulation, cell proliferation, and formation of a three‐dimensional tissuelike substance by skin fibroblasts , 1989, Journal of cellular physiology.

[13]  E. Véricel,et al.  Role of polyunsaturated fatty acids in the inhibitory effect of human adipocytes on osteoblastic proliferation. , 2002, Bone.

[14]  J. Gimble,et al.  Regulation of bone marrow stromal cell differentiation by cytokines whose receptors share the gp130 Protein , 1994, Journal of cellular biochemistry.

[15]  A. Scutt,et al.  Regulation of Osteogenic Differentiation of Human Bone Marrow Stromal Cells: Interaction Between Transforming Growth Factor-β and 1,25(OH)2 Vitamin D3In Vitro , 1999, Calcified Tissue International.

[16]  J. Gimble,et al.  1,25-Dihydroxy vitamin D3 inhibits adipocyte differentiation and gene expression in murine bone marrow stromal cell clones and primary cultures. , 1998, Endocrinology.

[17]  T. Kawada,et al.  Counteraction of retinoic acid and 1,25-dihydroxyvitamin D3 on up-regulation of adipocyte differentiation with PPARgamma ligand, an antidiabetic thiazolidinedione, in 3T3-L1 cells. , 1998, Life sciences.

[18]  Y. Reyne,et al.  Differentiation of adipocyte precursors in a serum-free medium is influenced by glucocorticoids and endogenously produced insulin-like growth factor-I. , 1993, International journal of obesity and related metabolic disorders : journal of the International Association for the Study of Obesity.

[19]  O. Shinohara,et al.  Enhancement of differentiation of rat adipocyte precursor cells by pertussis toxin. , 1991, Biochemical medicine and metabolic biology.

[20]  Makoto Sato,et al.  Targeted Disruption of Cbfa1 Results in a Complete Lack of Bone Formation owing to Maturational Arrest of Osteoblasts , 1997, Cell.

[21]  B. Cheatham,et al.  Regulation of ob gene expression and leptin secretion by insulin and dexamethasone in rat adipocytes. , 1999, Diabetes.

[22]  F. Casanueva,et al.  Retinoic acid and vitamin D(3) powerfully inhibit in vitro leptin secretion by human adipose tissue. , 2001, The Journal of endocrinology.

[23]  Z. Wu,et al.  Conditional ectopic expression of C/EBP beta in NIH-3T3 cells induces PPAR gamma and stimulates adipogenesis. , 1995, Genes & development.

[24]  P. Marie,et al.  Reciprocal control of osteoblast/chondroblast and osteoblast/adipocyte differentiation of multipotential clonal human marrow stromal F/STRO‐1+ cells , 2001, Journal of cellular biochemistry.

[25]  R. Considine,et al.  The Journal of Clinical Endocrinology & Metabolism Printed in U.S.A. Copyright © 2000 by The Endocrine Society Leptin Production in Adipocytes from Morbidly Obese Subjects: Stimulation by Dexamethasone, Inhibition with Troglitazone, and Influence of Gende , 2022 .

[26]  J. Lehmann,et al.  Printed in U.S.A. Copyright © 2002 by The Endocrine Society Divergent Effects of Selective Peroxisome Proliferator- Activated Receptor-�2 Ligands on Adipocyte Versus Osteoblast Differentiation , 2022 .

[27]  T. Clemens,et al.  1,25-Dihydroxyvitamin D induces lipoprotein lipase expression in 3T3-L1 cells in association with adipocyte differentiation. , 1996, Endocrinology.

[28]  R. Baron,et al.  Murine bone marrow stromally derived BMS2 adipocytes support differentiation and function of osteoclast-like cells in vitro. , 1998, Endocrinology.

[29]  J. Thompson,et al.  Structural properties of the adipocyte lipid binding protein. , 1999, Biochimica et biophysica acta.

[30]  M. Jimenez-Linan,et al.  Peroxisome proliferator-activated receptor gene expression in human tissues. Effects of obesity, weight loss, and regulation by insulin and glucocorticoids. , 1997, The Journal of clinical investigation.

[31]  C. G. Bellows,et al.  The Frequency of Common Progenitors for Adipocytes and Osteoblasts and of Committed and Restricted Adipocyte and Osteoblast Progenitors in Fetal Rat Calvaria Cell Populations * , 2001, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[32]  Toshio Matsumoto,et al.  Estrogen Promotes Early Osteoblast Differentiation and Inhibits Adipocyte Differentiation in Mouse Bone Marrow Stromal Cell Lines That Express Estrogen Receptor (er) ␣ or ␤ , 2022 .

[33]  F. Kraemer,et al.  Human BMP‐7/OP‐1 induces the growth and differentiation of adipocytes and osteoblasts in bone marrow stromal cell cultures * , 2001, Journal of cellular biochemistry.

[34]  B. L. Riggs,et al.  Differentiation of Human Marrow Stromal Precursor Cells: Bone Morphogenetic Protein‐2 Increases OSF2/CBFA1, Enhances Osteoblast Commitment, and Inhibits Late Adipocyte Maturation , 1999, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[35]  J. Aubin,et al.  Dexamethasone alters the subpopulation make‐up of rat bone marrow stromal cell cultures , 1995, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[36]  M. Cooper,et al.  Glucocorticoid activity, inactivity and the osteoblast. , 1999, The Journal of endocrinology.

[37]  G. Stein,et al.  The developmental stages of osteoblast growth and differentiation exhibit selective responses of genes to growth factors (TGF beta 1) and hormones (vitamin D and glucocorticoids). , 1993, The Journal of oral implantology.

[38]  P. Grimaldi,et al.  Cellular and molecular aspects of adipose tissue development. , 1992, Annual review of nutrition.

[39]  Daniel Chappard,et al.  Phenotypic effects of continuous or discontinuous treatment with dexamethasone and/or calcitriol on osteoblasts differentiated from rat bone marrow stromal cells , 2002, Journal of cellular biochemistry.

[40]  R. Oreffo,et al.  Effects of TGFβ and BFGF on the differentiation of human bone marrow stromal fibroblasts , 1999, Cell biology international.

[41]  X. Holy,et al.  Transforming Growth Factor β2 Inhibits Adipocyte Differentiation Induced by Skeletal Unloading in Rat Bone Marrow Stroma , 2002, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[42]  J. Lehmann,et al.  Divergent Effects of Selective Peroxisome Proliferator-Activated Receptor- (cid:1) 2 Ligands on Adipocyte Versus Osteoblast Differentiation , 2002 .

[43]  B. Ganss,et al.  On Behalf Of: International and American Associations for Dental Research , 2022 .

[44]  G. Karsenty,et al.  Osf2/Cbfa1: A Transcriptional Activator of Osteoblast Differentiation , 1997, Cell.

[45]  G. Karsenty,et al.  The osteoblast: a sophisticated fibroblast under central surveillance. , 2000, Science.

[46]  L. Lecoeur,et al.  In vitro induction of osteogenic differentiation from non-osteogenic mesenchymal cells. , 1997, Biomaterials.

[47]  M. Hüfner,et al.  Differential regulation of Cbfa1/Runx2 and osteocalcin gene expression by vitamin‐D3, dexamethasone, and local growth factors in primary human osteoblasts , 2002, Journal of cellular biochemistry.

[48]  C. Devlin,et al.  Evidence for an inverse relationship between the differentiation of adipocytic and osteogenic cells in rat marrow stromal cell cultures. , 1992, Journal of cell science.

[49]  M. Nuttall,et al.  Human Trabecular Bone Cells Are Able to Express Both Osteoblastic and Adipocytic Phenotype: Implications for Osteopenic Disorders , 1998, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[50]  G. Rodan,et al.  Thiazolidinedione effects on glucocorticoid receptor-mediated gene transcription and differentiation in osteoblastic cells. , 1999, Endocrinology.

[51]  J. Triffitt,et al.  In vitro effects of growth factors and dexamethasone on rat marrow stromal cells. , 1995, Clinical orthopaedics and related research.

[52]  G. Stein,et al.  1,25-(OH)2-vitamin D3 suppresses the bone-related Runx2/Cbfa1 gene promoter. , 2002, Experimental cell research.

[53]  J. Aubin,et al.  Characterization of the 1,25-(OH)2D3-induced inhibition of bone nodule formation in long-term cultures of fetal rat calvaria cells. , 1993, Endocrinology.

[54]  Toshio Matsumoto,et al.  Estrogen promotes early osteoblast differentiation and inhibits adipocyte differentiation in mouse bone marrow stromal cell lines that express estrogen receptor (ER) alpha or beta. , 2002, Endocrinology.

[55]  M. Wabitsch,et al.  Promoting effect of glucocorticoids on the differentiation of human adipocyte precursor cells cultured in a chemically defined medium. , 1989, The Journal of clinical investigation.

[56]  G. Stein,et al.  CBFa(AML/PEBP2)‐related elements in the TGF‐β type I receptor promoter and expression with osteoblast differentiation , 1998, Journal of cellular biochemistry.