Display options
Cite
Umebayashi M, Sumita Y, Kawai Y, et al. Gene-Activated Matrix Comprised of Atelocollagen and Plasmid DNA Encoding BMP4 or Runx2 Promotes Rat Cranial Bone Augmentation. Biores Open Access. 2015;4(1):164-74doi: 10.1089/biores.2014.0057.
Umebayashi, M., Sumita, Y., Kawai, Y., Watanabe, S., & Asahina, I. (2015). Gene-Activated Matrix Comprised of Atelocollagen and Plasmid DNA Encoding BMP4 or Runx2 Promotes Rat Cranial Bone Augmentation. BioResearch open access, 4(1), 164-74. https://doi.org/10.1089/biores.2014.0057
Umebayashi, Mayumi, et al. "Gene-Activated Matrix Comprised of Atelocollagen and Plasmid DNA Encoding BMP4 or Runx2 Promotes Rat Cranial Bone Augmentation." BioResearch open access vol. 4,1 (2015): 164-74. doi: https://doi.org/10.1089/biores.2014.0057
Umebayashi M, Sumita Y, Kawai Y, Watanabe S, Asahina I. Gene-Activated Matrix Comprised of Atelocollagen and Plasmid DNA Encoding BMP4 or Runx2 Promotes Rat Cranial Bone Augmentation. Biores Open Access. 2015 Feb 01;4(1):164-74. doi: 10.1089/biores.2014.0057. eCollection 2015. PMID: 26309793; PMCID: PMC4497668.
Copy Download .nbib Format: NLM
Share it on

Biores Open Access. 2015 Feb 01;4(1):164-74. doi: 10.1089/biores.2014.0057. eCollection 2015.

Gene-Activated Matrix Comprised of Atelocollagen and Plasmid DNA Encoding BMP4 or Runx2 Promotes Rat Cranial Bone Augmentation.

BioResearch open access

Mayumi Umebayashi, Yoshinori Sumita, Yousuke Kawai, Sumiko Watanabe, Izumi Asahina

Affiliations

  1. Department of Regenerative Oral Surgery, Unit of Translational Medicine, Graduate School of Biomedical Sciences, Nagasaki University , Nagasaki, Japan .
  2. Division of Molecular and Developmental Biology, Institute of Medical Science, The University of Tokyo , Tokyo, Japan .

PMID: 26309793 PMCID: PMC4497668 DOI: 10.1089/biores.2014.0057

Abstract

To date, therapeutic method for in vivo gene delivery has not been established on bone engineering though its potential usefulness has been suggested. For clinical applications, an effective condition should be developed to transfer the genes in vivo without any transfection reagents or virus vectors. In this study, to facilitate the clinical setting of this strategy, particularly aimed at atrophic bone repair, 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, or 1 mg of AcGFP plasmid vectors harboring cDNA of BMP4 (pBMP4) or Runx2 (pRunx2) with 2% bovine atelocollagen and β-tricalcium phosphate granules. Before manufacturing GAMs, to determine the biological activity of generated pDNAs, we confirmed GFP expression and increased level of alkaline phosphatase activities in MC3T3-E1 cells transfected with pBMP4 or pRunx2 during culture. Then, GAMs were lyophilized and transplanted to onlay placement on the cranium. At 2 weeks of transplantation, GFP-expressing cells could be detectable in only GAMs containing 1 mg of AcGFP plasmid vectors. Then, at 4 weeks, significant bone formation was recognized in GAMs containing 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. Thus, atelocollagen-based GAM reliably could form the engineered bone even for the vertical augmentation when containing a certain amount of plasmid vectors encoding osteogenic proteins. This study supports facilitating the clinical application of GAM for bone engineering.

Keywords: atelocollagen; bmp4; bone regeneration; gene-activated matrix; in vivo; runx2

References

  1. Hypertension. 2004 Aug;44(2):203-9 - PubMed
  2. Biomaterials. 2014 Jan;35(2):737-47 - PubMed
  3. Am J Vet Res. 2004 Sep;65(9):1223-32 - PubMed
  4. J Orthop Res. 2000 Mar;18(2):289-302 - PubMed
  5. Tissue Eng. 2006 Mar;12(3):489-97 - PubMed
  6. Adv Drug Deliv Rev. 2012 Sep;64(12):1331-40 - PubMed
  7. J Dent Res. 2012 Jun;91(6):592-7 - PubMed
  8. Adv Drug Deliv Rev. 2003 Nov 28;55(12):1651-77 - PubMed
  9. Cancer. 2004 Dec 1;101(11):2557-66 - PubMed
  10. Proc Natl Acad Sci U S A. 2005 Aug 23;102(34):12177-82 - PubMed
  11. J Pharmacol Sci. 2005 Feb;97(2):227-33 - PubMed
  12. Arthritis Res Ther. 2008;10(3):110 - PubMed
  13. Biomaterials. 2010 Aug;31(23):5953-65 - PubMed
  14. Yakugaku Zasshi. 2007 May;127(5):807-12 - PubMed
  15. Mol Ther. 2008 Feb;16(2):387-95 - PubMed
  16. J Vasc Surg. 2002 May;35(5):930-6 - PubMed
  17. Tissue Eng Part B Rev. 2014 Jun;20(3):229-32 - PubMed
  18. J Mater Sci Mater Med. 2011 Feb;22(2):397-404 - PubMed
  19. Arthritis Rheum. 2009 Sep;60(9):2677-83 - PubMed
  20. Crit Rev Oral Biol Med. 1992;3(4):333-52 - PubMed
  21. J Bone Miner Res. 2005 Nov;20(11):2028-35 - PubMed
  22. Nat Med. 1999 Jun;5(6):707-10 - PubMed
  23. Adv Healthc Mater. 2015 Jan 28;4(2):223-7 - PubMed
  24. Bone. 2003 May;32(5):502-12 - PubMed
  25. J Periodontol. 2004 Oct;75(10):1364-70 - PubMed
  26. Gene Ther. 2005 Mar;12(5):418-26 - PubMed
  27. Biotechnol Adv. 2013 Dec;31(8):1695-706 - PubMed
  28. Nat Med. 1999 Jul;5(7):753-9 - PubMed
  29. Biochem Biophys Res Commun. 2002 Nov 29;299(2):208-15 - PubMed
  30. Annu Rev Biochem. 2005;74:711-38 - PubMed
  31. Eur Cell Mater. 2011 Mar 15;21:230-42; discussion 242 - PubMed
  32. Mol Pharm. 2011 Jun 6;8(3):913-9 - PubMed
  33. Gene Ther. 2002 Dec;9(24):1647-52 - PubMed
  34. Proc Natl Acad Sci U S A. 1996 Jun 11;93(12):5753-8 - PubMed
  35. Curr Gene Ther. 2001 May;1(1):31-52 - PubMed
  36. J Bone Joint Surg Am. 2002 Jun;84-A(6):1032-44 - PubMed
  37. J Biomed Mater Res B Appl Biomater. 2009 Jul;90(1):101-9 - PubMed
  38. J Tissue Eng Regen Med. 2014 Oct;8(10):763-70 - PubMed
  39. Spine J. 2006 Jul-Aug;6(4):397-403; discussion 404 - PubMed
  40. J Control Release. 2010 Mar 19;142(3):404-10 - PubMed
  41. Clin Oral Implants Res. 2004 Apr;15(2):187-93 - PubMed
  42. Nucleic Acids Res. 2004 Jul 22;32(13):e109 - PubMed
  43. Biomed Mater Eng. 2013;23(3):163-72 - PubMed
  44. Nat Biotechnol. 1999 Jun;17(6):551-4 - PubMed
  45. PLoS One. 2012;7(7):e40833 - PubMed

Publication Types