title: Osteoblastic differentiation of human stem cells derived from bone marrow and periodontal ligament under the effect of enamel matrix derivative and transforming growth factor-beta.
abstract: OBJECTIVE
  To increase the understanding of the applicability of biomaterials and growth factors in enhancing stem cell-based bone regeneration modalities, this study evaluated the effects of enamel matrix derivative (EMD) and recombinant human transforming growth factor-beta (rhTGF-β) on osteoblastic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) as well as human periodontal ligament stem cells (hPDLSCs).
  
  
  METHODS
  hBMSCs and hPDLSCs were obtained, and identification of stem cell surface markers was performed according to the criteria of the International Society for Cellular Therapy. Each group of stem cells was separately treated with a serial dilution of EMD (10, 50, and 100 μg/mL) or rhTGF-β (10 ng/mL). Osteoblastic differentiation was examined through in vitro matrix mineralization by alizarin red staining, and mRNA expression of osteopontin and osteonectin was determined by quantitative reverse-transcriptase polymerase chain reaction. hPDLSCs were further assessed for osteocalcin mRNA expression. Stem cells cultured in osteogenic medium were employed as a standard positive control group.
  
  
  RESULTS
  In none of the experimental groups were bone-related mRNAs detected subsequent to treatment with EMD for 5, 10, and 15 days. Alizarin red staining on day 21 was negative in EMD-treated BMSC and PDLSC cultures. In rhTGF-β-supplemented BMSC culture, expression of osteonectin mRNA was demonstrated on day 15, which was statistically comparable to the positive control group. Nevertheless, extracellular matrix mineralization was inhibited in both groups of stem cells.
  
  
  CONCLUSIONS
  Within the limitations of this study, it could be concluded that EMD with a concentration of 10, 50, or 100 μg/mL has no appreciable effect on osteoblastic differentiation of BMSCs and PDLSCs. Application of rhTGF-β increased osteonectin mRNA expression in BMSCs. This finding corroborates the hypothesis that TGF-β might be involved in early osteoblastic maturation.
authors.forename: Behzad
authors.initials: B
authors.lastname: Houshmand
authors.forename: Hossein
authors.initials: H
authors.lastname: Behnia
authors.forename: Ahad
authors.initials: A
authors.lastname: Khoshzaban
authors.forename: Golnaz
authors.initials: G
authors.lastname: Morad
authors.forename: Gholamreza
authors.initials: G
authors.lastname: Behrouzi
authors.forename: Seyedeh Ghazaleh
authors.initials: SG
authors.lastname: Dashti
authors.forename: Arash
authors.initials: A
authors.lastname: Khojasteh
mesh_terms: D001854:Bone Marrow Cells / Q000166:cytology*; D001861:Bone Regeneration / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects* / Q000502:physiology; D003746:Dental Enamel Proteins / Q000494:pharmacology*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism / Q000494:pharmacology; D010006:Osteoblasts / Q000166:cytology*; D015675:Osteocalcin / Q000235:genetics / Q000378:metabolism; D053495:Osteopontin / Q000235:genetics / Q000378:metabolism; D010513:Periodontal Ligament / Q000166:cytology*; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects*; D016212:Transforming Growth Factor beta / Q000494:pharmacology*
chemical_list: D003746:Dental Enamel Proteins; D036341:Intercellular Signaling Peptides and Proteins; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D015675:Osteocalcin; D053495:Osteopontin