title: Production and action of transforming growth factor-beta in human osteoblast cultures: dependence on cell differentiation and modulation by calcitriol.
abstract: BACKGROUND
  Transforming growth factor beta (TGF-beta) plays an important role in skeletal remodelling. However, few studies have examined its effects on cultured human osteoblasts. Our aim is to characterise the biological effects of TGF-beta1 on human osteoblasts and to examine the interaction between TGF-beta1 and calcitriol.
  
  
  METHODS
  In vitro study employing two models of normal human osteoblasts: human bone marrow stromal cells [hMS/(OB)] containing osteoprogenitor cells and trabecular bone osteoblasts (hOB), which are mature osteoblasts. A reverse-transcriptase-polymerase-chain-reaction assay was employed to measure steady state mRNA levels of TGF-beta(s) isoforms and receptors. Effects of short-term treatment of TGF-beta1 on osteoblast proliferation and differentiation markers were assessed. The effect of cotreatment of calcitriol (10-8 M) and TGF-beta1 on osteoblast differentiation was also determined.
  
  
  RESULTS
  Both hMS(OB) and hOB cells expressed mRNA transcripts of TGF-beta1, TGF-beta2, TGF-beta 3, TGF-beta type I and type II receptors. TGF-beta 1 stimulated osteoblast proliferation in hMS(OB) and in hOB cultures. In hOB cultures, TGF-beta1 stimulated AP production and cotreatment with calcitriol induced a synergistic increase in AP levels to 250 +/- 61% of calcitriol-treated controls. Effects of TGF-beta1 and calcitriol were less pronounced in hMS(OB) cultures. TGF-beta1 inhibited collagen type I production in hMS(OB) cells and these effects were abolished in presence of calcitriol. In presence of calcitriol, TGF-beta1 increased collagen type I production in hOB cells. In both hOB and hMS(OB) cultures, TGF-beta1 inhibited osteocalcin production.
  
  
  CONCLUSIONS
  TGF-beta increases osteoblastic cell proliferation irrespective of the differentiation state. In presence of calcitriol, it initiates osteoblast cell differentiation and matrix formation. As TGF-beta inhibits osteocalcin production, other factors are necessary for inducing terminal differentiation of osteoblasts. The observed effects of TGF-beta on human osteoblasts in vitro may represent important regulatory steps in controlling osteoblast cell proliferation and differentiation in vivo.
authors.affiliation: Danish Centre for Molecular Gerontology, Aarhus Bone and Mineral Research Group, University Department of Endocrinology and Metabolism, Aarhus, Amtssygehus, DK-8000 Aarhus C, Denmark. mkassem@dadlnet.dk
authors.forename: M
authors.initials: M
authors.lastname: Kassem
authors.forename: M
authors.initials: M
authors.lastname: Kveiborg
authors.forename: E F
authors.initials: EF
authors.lastname: Eriksen
mesh_terms: D030201:Activin Receptors, Type I; D000469:Alkaline Phosphatase / Q000096:biosynthesis / Q000378:metabolism; D002117:Calcitriol / Q000494:pharmacology*; D002120:Calcium Channel Agonists / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis; D015870:Gene Expression / Q000502:physiology; D006801:Humans; D007536:Isomerism; D010006:Osteoblasts / Q000737:chemistry / Q000166:cytology* / Q000201:enzymology*; D015675:Osteocalcin / Q000096:biosynthesis; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D000077293:Receptor, Transforming Growth Factor-beta Type I; D000077294:Receptor, Transforming Growth Factor-beta Type II; D018125:Receptors, Transforming Growth Factor beta / Q000235:genetics; D017154:Stromal Cells / Q000737:chemistry / Q000166:cytology / Q000201:enzymology; D016212:Transforming Growth Factor beta / Q000737:chemistry / Q000235:genetics*
chemical_list: D002120:Calcium Channel Agonists; D012333:RNA, Messenger; D018125:Receptors, Transforming Growth Factor beta; D016212:Transforming Growth Factor beta; D015675:Osteocalcin; D003094:Collagen; D017346:Protein Serine-Threonine Kinases; D030201:Activin Receptors, Type I; D000077293:Receptor, Transforming Growth Factor-beta Type I; D000077294:Receptor, Transforming Growth Factor-beta Type II; D000469:Alkaline Phosphatase; D002117:Calcitriol