title: Conditioned medium of demineralized freeze-dried bone activates gene expression in periodontal fibroblasts in vitro.
abstract: BACKGROUND
  Demineralized bone matrix (DBM) is used for the treatment of osseous defects. Conditioned medium from native bone chips can activate transforming growth factor (TGF)-β signaling in mesenchymal cells. The aim of this study is to determine whether processing of native bone into DBM affects the activity of the conditioned medium.
  
  
  METHODS
  Porcine cortical bone blocks were subjected to defatting, different concentrations of hydrochloric acid, and various temperatures. DBM was lyophilized, ground, and placed into culture medium. Human gingiva and periodontal fibroblasts were exposed to the respective conditioned medium obtained from DBM (DBCM). Changes in the expression of TGF-β target genes were determined.
  
  
  RESULTS
  DBCM altered the expression of TGF-β target genes (e.g., adrenomedullin, pentraxin 3, KN motif and ankyrin repeat domains 4, interleukin 11, NADPH oxidase 4, and BTB [POZ] domain containing 11) by at least five-fold. The response was observed in fibroblasts from both sources. Defatting lowered the activity of DBCM. The TGF-β receptor type I kinase inhibitor SB431542 [4-(4-(benzo[d][1,3]dioxol-5-yl)-5-(pyridin-2-yl)-1H-imidazol-2-yl)benzamide] but not the inhibitor of bone morphogenetic protein receptor dorsomorphin, blocked the effects of DBCM on gene expression. Moreover, conditioned medium obtained from commercial human DBM modulated the expression of TGF-β target genes.
  
  
  CONCLUSIONS
  The findings suggest that the DBCM can activate TGF-β signaling in oral fibroblasts.
authors: *Laboratory of Oral Cell Biology, School of Dental Medicine, University of Bern, Bern, Switzerland.|†Department of Oral Surgery and Stomatology, School of Dental Medicine, University of Bern.|‡Department of Implant Dentistry, Federal University of Santa Catarina, Florianópolis, Brazil.
authors: Guenther Schuldt
authors: GS
authors: Filho
authors: *Laboratory of Oral Cell Biology, School of Dental Medicine, University of Bern, Bern, Switzerland.|†Department of Oral Surgery and Stomatology, School of Dental Medicine, University of Bern.|§Department of Oral and Maxillofacial Surgery, College of Dentistry, Universitat Internacional de Catalunya, Barcelona, Spain.
authors: Jordi
authors: J
authors: Caballé-Serrano
authors: *Laboratory of Oral Cell Biology, School of Dental Medicine, University of Bern, Bern, Switzerland.|‖Department of Cranio-Maxillofacial Surgery, Inselspital, University of Bern.
authors: Kosaku
authors: K
authors: Sawada
authors: †Department of Oral Surgery and Stomatology, School of Dental Medicine, University of Bern.|¶Robert K. Schenk Laboratory of Oral Histology, School of Dental Medicine, University of Bern.
authors: Dieter D
authors: DD
authors: Bosshardt
authors: ‡Department of Implant Dentistry, Federal University of Santa Catarina, Florianópolis, Brazil.
authors: Marco Aurélio
authors: MA
authors: Bianchini
authors: †Department of Oral Surgery and Stomatology, School of Dental Medicine, University of Bern.
authors: Daniel
authors: D
authors: Buser
authors: *Laboratory of Oral Cell Biology, School of Dental Medicine, University of Bern, Bern, Switzerland.|†Department of Oral Surgery and Stomatology, School of Dental Medicine, University of Bern.|#Department of Oral Biology, Medical University of Vienna, Vienna, Austria.
authors: Reinhard
authors: R
authors: Gruber
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000037:antagonists & inhibitors; D000209:Acute-Phase Proteins / Q000032:analysis; D053607:Adrenomedullin / Q000032:analysis; D000818:Animals; D017089:Ankyrin Repeat / Q000235:genetics; D001549:Benzamides / Q000494:pharmacology; D018775:Bone Demineralization Technique; D001857:Bone Matrix / Q000187:drug effects / Q000502:physiology*; D002097:C-Reactive Protein / Q000032:analysis; D002460:Cell Line; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D004149:Dioxoles / Q000494:pharmacology; D005347:Fibroblasts / Q000378:metabolism; D005612:Freeze Drying; D005881:Gingiva / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D006851:Hydrochloric Acid / Q000737:chemistry; D017370:Interleukin-11 / Q000032:analysis; D059630:Mesenchymal Stem Cells / Q000378:metabolism; D051379:Mice; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases / Q000032:analysis; D009687:Nuclear Proteins / Q000032:analysis; D010519:Periodontium / Q000166:cytology / Q000378:metabolism*; D011720:Pyrazoles / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D018125:Receptors, Transforming Growth Factor beta / Q000037:antagonists & inhibitors; D000683:Serum Amyloid P-Component / Q000032:analysis; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D013552:Swine; D014021:Tissue Preservation / Q000379:methods*; D016212:Transforming Growth Factor beta / Q000187:drug effects / Q000378:metabolism*; D000099153:Pentraxins
chemical_list: D055372:AMP-Activated Protein Kinases; D000209:Acute-Phase Proteins; D053607:Adrenomedullin; D001549:Benzamides; D002097:C-Reactive Protein; D017077:Culture Media, Conditioned; D004149:Dioxoles; D006851:Hydrochloric Acid; D017370:Interleukin-11; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases; D009687:Nuclear Proteins; D011720:Pyrazoles; D011743:Pyrimidines; D018125:Receptors, Transforming Growth Factor beta; D000683:Serum Amyloid P-Component; D016212:Transforming Growth Factor beta; D000099153:Pentraxins; C459179:4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide; C508611:IL11 protein, human; C516138:dorsomorphin; C498910:NOX4 protein, human
keywords: Bone and bones; TGF-beta superfamily proteins; bone substitutes; bone transplantation; culture media, conditioned; freeze drying