Benedicta E Beck-Broichsitter1, Anneke N Werk2, Ralf Smeets3, Alexander Gröbe3, Max Heiland3, Ingolf Cascorbi2, Jörg Wiltfang4, Robert Häsler5, Stephan T Becker4. 1. University Medical Center Hamburg-Eppendorf, Department of Oral and Maxillofacial Surgery, Martinistraße 52, Campus Forschung Gebäude N27, 20246 Hamburg, Germany. Electronic address: b.beck-broichsitter@uke.de. 2. Schleswig-Holstein University Hospital, Institute of Clinical and Experimental Pharmacology, Arnold-Heller-Straße 3, Haus 30, 24105 Kiel, Germany. 3. University Medical Center Hamburg-Eppendorf, Department of Oral and Maxillofacial Surgery, Martinistraße 52, Campus Forschung Gebäude N27, 20246 Hamburg, Germany. 4. Schleswig-Holstein University Hospital, Department of Oral and Maxillofacial Surgery (Head: J. Wiltfang, MD, DMD, PhD), Arnold-Heller-Straße 3, Haus 26, 24105 Kiel, Germany. 5. Institute of Clinical Molecular Biology, Center for Molecular Biosciences, Christian Albrechts University of Kiel, Am Botanischen Garten 11, 24118 Kiel, Germany.
Abstract
PURPOSE: Although bone tissue engineering techniques have become more and more sophisticated than in the past, natural bone healing mechanisms have not been sufficiently considered for further improvement of these techniques so far. We used an established animal model with transcriptome analysis to generate an unbiased picture of early bone healing to support tissue engineering concepts. MATERIAL AND METHODS: In 30 Wistar rats, a 3-mm bone defect was created in the mandibular angle. Tissue was sampled at 5, 10, and 15 days, and the former defect area was excised to undergo transcriptome analysis after RNA extraction. Five differentially expressed genes were further evaluated with reverse transcription-polymerase chain reaction (rt-PCR). RESULTS: Transcriptome analysis revealed 2467 significantly over- and under-expressed transcripts after 5 days and 2265 after 15 days of bone healing, respectively. Validation via rt-PCR confirmed overexpression of osteoactivin, angiopoietin-like factor-4, and metallomatrix proteinase-9 and underexpression of mastcellprotease-10 and proteoglycane-2 in comparison to values in the control group. CONCLUSION: This systematic genome-wide transcriptome analysis helps to decipher the physiological mechanisms behind physiological bone healing. The exemplary depiction of 5 genes demonstrates the great complexity of metabolic processes during early bone healing. Here, BMP-2 signaling pathways and local hypoxia play decisive roles in bone formation.
PURPOSE: Although bone tissue engineering techniques have become more and more sophisticated than in the past, natural bone healing mechanisms have not been sufficiently considered for further improvement of these techniques so far. We used an established animal model with transcriptome analysis to generate an unbiased picture of early bone healing to support tissue engineering concepts. MATERIAL AND METHODS: In 30 Wistar rats, a 3-mm bone defect was created in the mandibular angle. Tissue was sampled at 5, 10, and 15 days, and the former defect area was excised to undergo transcriptome analysis after RNA extraction. Five differentially expressed genes were further evaluated with reverse transcription-polymerase chain reaction (rt-PCR). RESULTS: Transcriptome analysis revealed 2467 significantly over- and under-expressed transcripts after 5 days and 2265 after 15 days of bone healing, respectively. Validation via rt-PCR confirmed overexpression of osteoactivin, angiopoietin-like factor-4, and metallomatrix proteinase-9 and underexpression of mastcellprotease-10 and proteoglycane-2 in comparison to values in the control group. CONCLUSION: This systematic genome-wide transcriptome analysis helps to decipher the physiological mechanisms behind physiological bone healing. The exemplary depiction of 5 genes demonstrates the great complexity of metabolic processes during early bone healing. Here, BMP-2 signaling pathways and local hypoxia play decisive roles in bone formation.
Authors: Klara Janjić; Alwina Schellner; Alexander Engenhart; Kurt Kernstock; Barbara Schädl; Andreas Moritz; Hermann Agis Journal: J Periodontal Res Date: 2019-03-20 Impact factor: 4.419