Guenther Schuldt Filho1,2,3, Jordi Caballé-Serrano1,2,4, Kosaku Sawada1,5, Dieter D Bosshardt2,6, Marco Aurélio Bianchini3, Daniel Buser2, Reinhard Gruber1,2,7. 1. *Laboratory of Oral Cell Biology, School of Dental Medicine, University of Bern, Bern, Switzerland. 2. †Department of Oral Surgery and Stomatology, School of Dental Medicine, University of Bern. 3. ‡Department of Implant Dentistry, Federal University of Santa Catarina, Florianópolis, Brazil. 4. §Department of Oral and Maxillofacial Surgery, College of Dentistry, Universitat Internacional de Catalunya, Barcelona, Spain. 5. ‖Department of Cranio-Maxillofacial Surgery, Inselspital, University of Bern. 6. ¶Robert K. Schenk Laboratory of Oral Histology, School of Dental Medicine, University of Bern. 7. #Department of Oral Biology, Medical University of Vienna, Vienna, Austria.
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. CONCLUSION: The findings suggest that the DBCM can activate TGF-β signaling in oral fibroblasts.
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. Humangingiva 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 humanDBM modulated the expression of TGF-β target genes. CONCLUSION: The findings suggest that the DBCM can activate TGF-β signaling in oral fibroblasts.
Entities:
Keywords:
Bone and bones; TGF-beta superfamily proteins; bone substitutes; bone transplantation; culture media, conditioned; freeze drying
Authors: David G Armstrong; Dennis P Orgill; Robert D Galiano; Paul Glat; Jarrod Kaufman; Marco Mehr; Charles M Zelen Journal: Plast Reconstr Surg Glob Open Date: 2021-05-21