title: Bone morphogenetic protein 6 stimulates mineralization in human dental follicle cells without dexamethasone.
abstract: OBJECTIVE
  The aim of this study is to investigate the osteogenic differentiation human dental follicle cells (hDFCs) cultured with in osteogenic induction medium (OIM) without dexamethasone (DEX), and to analyze the gene expression profile during osteogenic differentiation.
  
  
  METHODS
  hDFCs, which isolated from dental follicle tissue from impacted third molar teeth, were cultured with OIM with or without DEX. Osteogenic differentiation of hDFCs was examined using Alkaline phosphatase activity and Arizarin red staining. Gene expression analysis was performed by Microarray and real time-PCR.
  
  
  RESULTS
  We showed that hDFCs have the capacity to differentiate into osteogenic lineages in osteogenic induction medium lacking DEX. We also analyzed gene expression profiling of hDFCs during osteogenic differentiation. BMP6 is up-regulated in both the presence and absence of DEX. In addition, BMP6 enhances gene expression levels of DLX-5, Runx2, and Osterix, which are transcription factors associated with osteogenic differentiation. BMP6 also stimulates phosphorylation of Smad1/5/8 which are transcription factors associated with BMP signalling at protein levels. Additionally BMP6 stimulates mineralization of hDFCs monolayers examined by Arizarin red S staining.
  
  
  CONCLUSIONS
  These findings suggest that hDFCs can differentiate to osteogenic lineage cells osteogenic induction medium without DEX, and BMP6 is a key gene in the osteogenic differentiation of hDFCs, and has therapeutic utility for bone regeneration and bone research.
authors: Department of Maxillofacial Surgery, Nihon University School of Dentistry at Matsudo, Matsudo City, Matsudo Prefecture, Japan.
authors: Kosuke
authors: K
authors: Takahashi
authors: Naomi
authors: N
authors: Ogura
authors: Haruna
authors: H
authors: Aonuma
authors: Ko
authors: K
authors: Ito
authors: Daisuke
authors: D
authors: Ishigami
authors: Yoshikazu
authors: Y
authors: Kamino
authors: Toshirou
authors: T
authors: Kondoh
mesh_terms: D000293:Adolescent; D000469:Alkaline Phosphatase / Q000032:analysis; D000880:Anthraquinones; D055418:Bone Morphogenetic Protein 6 / Q000494:pharmacology*; D002113:Calcification, Physiologic / Q000187:drug effects*; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D004396:Coloring Agents; D050659:Core Binding Factor Alpha 1 Subunit / Q000187:drug effects; D003470:Culture Media; D003795:Dental Sac / Q000166:cytology* / Q000187:drug effects; D003907:Dexamethasone / Q000494:pharmacology*; D020869:Gene Expression Profiling; D005938:Glucocorticoids / Q000494:pharmacology*; D018398:Homeodomain Proteins / Q000187:drug effects; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000187:drug effects*; D010012:Osteogenesis / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects; D051898:Smad1 Protein / Q000187:drug effects; D051902:Smad5 Protein / Q000187:drug effects; D051904:Smad8 Protein / Q000187:drug effects; D000076182:Sp7 Transcription Factor; D014157:Transcription Factors / Q000187:drug effects; D016212:Transforming Growth Factor beta / Q000187:drug effects; D015854:Up-Regulation / Q000187:drug effects; D055815:Young Adult
chemical_list: D000880:Anthraquinones; C525816:BMP6 protein, human; D055418:Bone Morphogenetic Protein 6; D004396:Coloring Agents; D050659:Core Binding Factor Alpha 1 Subunit; D003470:Culture Media; C100596:DLX5 protein, human; D005938:Glucocorticoids; D018398:Homeodomain Proteins; C493709:RUNX2 protein, human; C497399:SMAD1 protein, human; C497797:SMAD5 protein, human; C107514:SMAD9 protein, human; D051898:Smad1 Protein; D051902:Smad5 Protein; D051904:Smad8 Protein; D000076182:Sp7 Transcription Factor; C478163:SP7 protein, human; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; C010078:alizarin; D003907:Dexamethasone; D000469:Alkaline Phosphatase