Bone agumentation by gene activated matrix composed of plasmid DNA, BMP4 or Runx2, embedded in atelocollagen

Abstract

Event Abstract Back to Event Bone agumentation by gene activated matrix composed of plasmid DNA, BMP4 or Runx2, embedded in atelocollagen Mayumi Umebayashi1, Yoshinori Sumita1 and Izumi Asahina1 1 Nagasaki University, Oral surgery, Japan Background: To date, therapeutic method for in vivo gene delivery has not been established on bone engineering though their potential usefulness has been suggested. For clinical applications, an effective condition should be developed to transfer the genes in vivo without any of transfection-reagents or virus-vectors. Objective: The aim of this study is to have a new understanding on the usefulness of delivering non-viral GAM without cell transplantation or any transfection reagents/kits for facilitating clinical setting of bone engineering. Methods: In this study, to facilitate the clinical setting of this strategy, we simply investigated whether manufactured gene activated-matrix (GAM) with atelocollagen containing a certain amount of plasmid (p) DNA encoding osteogenic-proteins could augment the cranial bone in rat. GAMs were manufactured by mixing 0.02, 0.1, 0.5 or 1 mg of AcGFP plasmid-vectors harboring cDNA of BMP4 (pBMP4) or Runx2 (pRunx2) with 2% bovine-atelocollagen and β-TCP granules. Before manufactured GAMs, to determine the biological activity of generated pDNAs, we confirmed GFP-expression and increased-level of ALP activities in MC3T3-E1 cells transfected with pBMP4 or pRunx2 during culture. Then, GAMs were lyophilized and transplanted to onlay placement on the cranium. Results: At 2 weeks of transplantation, GFP-expressing cells could be detectable in only GAMs containing 0.5 and 1 mg of AcGFP plasmid vectors. Then, at 4 weeks, significant bone formation was recognized in GAMs containing 0.5 and 1 mg of pDNAs encoding BMP4 or Runx2 but not in 0.02 or 0.1 mg of GAMs. These newly formed bone tissues surrounded by osteocalcin-stained area were augmented markedly until 8 weeks after transplantation. In contrast, minimal bone formation was observed in GAMs without harboring cDNA of osteogenic-proteins. Meanwhile, when GAMs were transplanted to the cranial bone defect, bone formation was detectable in specimens containing 1 mg of pBmp4 or pRunx2 at 8 weeks as well. Conclusion: Thus, atelocollagen-based GAM reliably could form the engineered-bone even for the vertical augmentation when contained a certain amount of plasmid-vectors encoding osteogenic-proteins. This study supports facilitating the clinical application of GAM for bone engineering. Keywords: Bone Regeneration, Tissue Engineering, biomaterial, gene delivery Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: Poster Topic: Biomimetic materials Citation: Umebayashi M, Sumita Y and Asahina I (2016). Bone agumentation by gene activated matrix composed of plasmid DNA, BMP4 or Runx2, embedded in atelocollagen. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.00373 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 27 Mar 2016; Published Online: 30 Mar 2016. Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Mayumi Umebayashi Yoshinori Sumita Izumi Asahina Google Mayumi Umebayashi Yoshinori Sumita Izumi Asahina Google Scholar Mayumi Umebayashi Yoshinori Sumita Izumi Asahina PubMed Mayumi Umebayashi Yoshinori Sumita Izumi Asahina Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.