Journal of Periodontal and Implant Science

Korean Academy of Periodontology (대한치주과학회)
권호 표지

Osteogenic effects of polyethyleneimine-condensed BMP-2 genes in vitro and in vivo

Polyethyleneimine-응축 BMP-2 발현 유전자를 이용한 골형성 효과

  • Cheong, Hee-Sun (Department of Periodontology, School of Dentistry, Seoul National University) ;
  • Kim, Kyoung-Hwa (Department of Periodontology, School of Dentistry, Seoul National University, BK21 Craniomaxillofacial Life Science) ;
  • Park, Yoon-Jeong (Craniomaxillofacial Reconstructive Science, School of Dentistry, Seoul National University) ;
  • Kim, Tae-Il (Department of Periodontology, School of Dentistry, Seoul National University) ;
  • Lee, Yong-Moo (Department of Periodontology, School of Dentistry, Seoul National University) ;
  • Ku, Young (Department of Periodontology, School of Dentistry, Seoul National University) ;
  • Rhyu, In-Chul (Department of Periodontology, School of Dentistry, Seoul National University) ;
  • Lee, Dong-Soo (Department of Nuclear Medicine, College of Medicine, Seoul National University) ;
  • Lee, Seung-Jin (College of Pharmacy, Ewha Womans University) ;
  • Chung, Chong-Pyoung (Department of Periodontology, School of Dentistry, Seoul National University) ;
  • Han, Soo-Boo (Department of Periodontology, School of Dentistry, Seoul National University) ;
  • Seol, Yang-Jo (Department of Periodontology, School of Dentistry, Seoul National University)
  • 정희선 (서울대학교 치의학전문대학원 치주과학교실) ;
  • 김경화 (서울대학교 치의학전문대학원 치주과학교실, BK21 치의학생명과학사업단) ;
  • 박윤정 (서울대학교 치과대학 두 개악안면재건과학교실) ;
  • 김태일 (서울대학교 치의학전문대학원 치주과학교실) ;
  • 이용무 (서울대학교 치의학전문대학원 치주과학교실) ;
  • 구영 (서울대학교 치의학전문대학원 치주과학교실) ;
  • 류인철 (서울대학교 치의학전문대학원 치주과학교실) ;
  • 이동수 (서울대학교 의과대학 핵의학과) ;
  • 이승진 (이화여자대학교 약학대학 물리약학교실) ;
  • 정종평 (서울대학교 치의학전문대학원 치주과학교실) ;
  • 한수부 (서울대학교 치의학전문대학원 치주과학교실) ;
  • 설양조 (서울대학교 치의학전문대학원 치주과학교실)
  • Published : 2007.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

Naked DNA and standard vectors have been previously used for gene delivery. Among these, PEI can efficiently condense DNA and has high intrinsic endosomal activities. The aim of this study is to investigate whether the cationic polycation PEI could increase the transfection efficiency of BMP expressing DNA using a vector-loaded collagen sponge model. BMP-2/pcDNA3.1 plasmid was constructed by subcloning human BMP-2 cDNA into the pcDNA3.1 plasmid vector. PEI/DNA complexes were prepared by mixing PEI and BMP-2/pcDNA3.1 and the constructed complexes were loaded into the collagen sponges. In vitro studies, BMSCs were transfected with the PEI/BMP-2/pcDNA3.1 complexes from collgen sponge. The level of secreted BMP-2 and alkaline phosphatase activities of transfected BMSCs were significantly higher in PEI/BMP-2/pcDNA3.1 group than in BMP-2/pcDNA3.1 group (p<0.05). Transfected BMSCs were cultured and mineralization was observed only in cells treated with PEI/BMP-2/pcDNA3.1 complexes. In vivo studies, PEI/BMP-2/pcDNA3.1/collagen, BMP-2/pcDNA3.1/collagen and blank collagen were grafted in skeletal muscle of nude mice. Ectopic bone formation was shown in PEI/BMP-2/pcDNA3.1/collagen grafted mouse 4 weeks postimplantation, while not in BMP-2/pcDNA3.1 grafted tissue. This study suggests that PEI-condensed DNA encoding for BMP-2 is capable of inducing bone formation in ectopic site and might increase the transfection rate of BMP-2/pcDNA3.1. As a non-viral vector, PEI offers the potential in gene therapy for bone engineering.

Keywords

References

  1. Urist MR. Bone: Formation by autoinduction. Science 1965;150:893-899 https://doi.org/10.1126/science.150.3698.893
  2. Wang EA, Rosen V, D' Alessandro JS et al. Recombinant human bone morphogenetic protein induces bone formation. Proc Natl Acad Sci 1990;87:2220-2224 https://doi.org/10.1073/pnas.87.6.2220
  3. Lee YM, Nam SH, Seol YJ el al. Enhanced bone augmentation by controlled release of recombinant human bone morphogenetic protein-2 from bioabsorbable membranes. J Periodontol 2003;74:865-872 https://doi.org/10.1902/jop.2003.74.6.865
  4. Wozney JM. The potential role of bone morphogenetic proteins in periodontal reconstruction. J Periodontol 1995;66:50-6510
  5. Wang EA. Bone morphogenetic proteins (BMPs) : Therapeutic potential in healing bony defects. Trends Biotechnol 1993;11:379-383 https://doi.org/10.1016/0167-7799(93)90096-R
  6. Ripamonti U, Reddi A H. Tissue engineering, morphogenesis, and regeneration of the periodontal tissues by bone morphogenetic proteins. Crit Rev Oral Biol Med 1997;8:154-163 https://doi.org/10.1177/10454411970080020401
  7. Ripamonti U, Heliotis M, Rueger DC, Sampath TK. Induction of cementogenesis by recombinant human osteogenic protein-1 (hop-1/bmp-7) in the baboon (Papio ursinus). Arch Oral Biol 1996;41:121-126 https://doi.org/10.1016/0003-9969(95)00110-7
  8. Higuchi T, Kinoshita A, Takahashi K, Oda S, Ishikawa I. Bone regeneration by recombinant human bone morphogenetic protein- 2 in rat mandibular defects. An experimental model of defect filling. J Periodontol 1999;70:1026-1031 https://doi.org/10.1902/jop.1999.70.9.1026
  9. Cochran DL, Jones AA, Lilly LC, Fiorellini JP, Howell H. Evaluation of recombinant human bone morphogenetic protein-2 in oral applications including the use of endosseous implants: 3-year results of a pilot study in humans. J Periodontol 2000;71:1241-1257 https://doi.org/10.1902/jop.2000.71.8.1241
  10. Cochran DL, Nummikoski PV, Jones AA et al. Radiographic analysis of regenerated bone around endosseous implants in the canine using reeombinant human bone morphogenetic protein. Int J Oral Maxillofac Implants 1997;12:739-748
  11. Barboza EP, Duarte ME, Geolas L et al. Ridge augmentation following implantation of recombinant human bone morphogenetic protein-2 in the dog. J Periodontol 2000;71:488-496 https://doi.org/10.1902/jop.2000.71.3.488
  12. Howell TH, Fiorellini J, Jones A et al. A feasibility study evaluating rhBMP-2/absorvavle collagen sponge device for local alveolar ridge preservation or augmentation. Int J Periodontics Restorative Dent 1997;17:124-39
  13. Terheyden H, Jepsen S, Moller B, Tucker MM, Rueger DC. Sinus floor augmentation with simultaneous placement of dental implants using a combination of deproteinized bone xenografts and recombinant human osteogenic protein-1. A histometric study in miniature pigs. Clin Oral Implants Res 1999;10:510-521 https://doi.org/10.1034/j.1600-0501.1999.100609.x
  14. Murphy WL, Mooney DJ. Controlled delivery of inductive proteins, plasmid DNA and cells from tissue engineering matrices. J Periodontal Res 1999;34:413-419 https://doi.org/10.1111/j.1600-0765.1999.tb02275.x
  15. Seeherman H, Wozney J, Rebecca L. Bone morphogenetic protein delivery systems. Spine 2002;278:816-823
  16. Winn SR, Chen JC, Gong X et al. Non-viral- mediated gene therapy approaches for bone repair. Orthod Craniofacial Res 2005;8:183-190 https://doi.org/10.1111/j.1601-6343.2005.00332.x
  17. Alden TD, Varady P, Kallmes DF, Jane JA Jr, Helm GA. Bone morphogenetic protein gene therapy. Spine 2002;15:S87-93 https://doi.org/10.1097/00007632-199002000-00007
  18. Gardlik R, Palffy R, Hodosy Julius et al. Vectors and delivery systems in gene therapy. Med Sci Monit 2005;11:RA110-121
  19. Nishikawa M, Huang L. Nonviral Vectors in the New Millennium : Delivery Barriers in Gene Transfer. Hum Gene Ther 2001;12:861-870 https://doi.org/10.1089/104303401750195836
  20. Herweijer H, Wolff JA. Progress and prospects: naked DNA gene transfer and therapy. Gene Ther 2003;10:453-458 https://doi.org/10.1038/sj.gt.3301983
  21. Fang J, Zhu YY, Smiley E et al. Stimulation of new bone formation by direct transfer of osteogenic plasmid genes. Proc Natl Acad Sci 1996;93:5753-5758 https://doi.org/10.1073/pnas.93.12.5753
  22. Bonadio J, Smiley E, Patil P, Goldstein S. Localized, direct plasmid gene delivery in vivo: prolonged therapy results in reproducible tissue regeneration. Nat Med 1999;5:753-759 https://doi.org/10.1038/10473
  23. Goldstein SA, Bonadio J. Potential role for direct gene transfer in the enhancement of fracture healing. Clin Orthop 1998;3558:S154-162
  24. Pack DW, Hoffman AS, Pun S, Stayton PS. Designing and development of polymers for gene delivery. Nature Reviews Drug Disc 2005;4:581-593 https://doi.org/10.1038/nrd1775
  25. Park TG, Jeong JH, Kim SW. Current status of polymeric gene delivery systems. Adv Drug Del Rev 2006;58:467-486 https://doi.org/10.1016/j.addr.2006.03.007
  26. Boussif O, Lezoualc' h F, Zanta MA et al. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. Proc Natl Acad Sci 1995;92:7297-7301 https://doi.org/10.1073/pnas.92.16.7297
  27. Scherer F, Schillinger U, Putz U, Stemberger A, Plank C. Nonviral vector loaded collagen sponges for sustained gene delivery in vitro and in vivo. J Gene Med 2002;4:634-643 https://doi.org/10.1002/jgm.298
  28. Huang YC, Simmons C, Kaigler D, Rice KG, Mooney DJ. Bone regeneration in a rat cranial defect with delivery of PEI-condensed plasmid DNA encoding for bone morphogenic protein-4 (BMP-4). Gene Ther 2005;12:418-426 https://doi.org/10.1038/sj.gt.3302439
  29. Huang YC, Riddle K, Rice KG, Mooney DJ. Long-Term in vivo gene expression via delivery of PEI-DNA condensates from porous polymer scaffolds. Hum Gene Ther 2005;16:609-617 https://doi.org/10.1089/hum.2005.16.609
  30. Wightman L, Kircheis R, Rossler V et al. Different behavior of branched and linear polyethylenimin for gene delivery in vitro and in vivo. J Gene Med 2001;3:362-372 https://doi.org/10.1002/jgm.187
  31. Fischer D, Bieber T, Li Y, Elsasser HP, Kissel T. A novel non-viral vector for DNA delivery based on low molecular weight, branched polyethyleneimine: Effect of molecular weight on transfection efficiency and cytotoxicity. Pharm Res 1999;16:1273-1279 https://doi.org/10.1023/A:1014861900478
  32. Kunath K, Harpe AV, Fischer D et al. Low-molecular-weight polyethylenimine as a non-viral vector for DNA delivery: comparison of physicochemical properties, transfection efficiency and in vivo distribution with high-molecular-weight polyethylenimine. J Cont Rel 2003;89:113-125 https://doi.org/10.1016/S0168-3659(03)00076-2
  33. Lieberman JR, Daluiski A, Stevenson S et al. The effect of regional gene therapy with bone morphogenetic protein 2 producing bone marrow cells on the repair of segmental femoral defects in rats. J Bone Joint Surg (Am) 1999;81:905-917 https://doi.org/10.2106/00004623-199907000-00002
  34. Jin QM, Anusaksathien O, Webb SA, Rutherford RB, Giannobile WV. Gene therapy of bone morphogenetic protein for periodontal tissue engineering. J Periodontol 2003;74:202-13 https://doi.org/10.1902/jop.2003.74.2.202
  35. Dunn CA, Jin Q, Taba M Jr et al. BMP gene delivery for alveolar bone engineering at dental implant defects. Mol Ther 2005;11:294-9 https://doi.org/10.1016/j.ymthe.2004.10.005
  36. Krebsbach PH, Gu K, Franceschi RT, Rutherford RB. Gene therapy-directed osteogenesis: BMP-7-transduced human fibroblasts form bone in vivo. Hum Gene Ther 2000;11:1201-1210 https://doi.org/10.1089/10430340050015248
  37. Rundle CH, Miyakoshi N, Kasukawa Y et al. In vivo bone formation in fracture re.pair induced by direct retroviral-based gene therapy with bone morphogenetic protein-4. Bone 2003;32:591-601 https://doi.org/10.1016/S8756-3282(03)00096-6
  38. Cohen-Sacks H, Elazar V, Gao J, Golomb A. Delivery and expression of pDNA embedded in collagen matrices. J Cont Release 2004;5;95:309-320
  39. Friess W. Collagen-biomaterial for drug delivery. Eur J Pharm Biopharm 1998;45:113-136 https://doi.org/10.1016/S0939-6411(98)00017-4
  40. Katz JM, Roth CM, Dunn MG. Factors that influence transgene expression and cell viability on DNA-PEI-seeded collagen films. Tissue Engineering 2005;11:1398-1406 https://doi.org/10.1089/ten.2005.11.1398
  41. Southwood LL, Frisbie DD, Kawcak CE, McIlwraith CW. Delivery of growth factors using gene therapy to enhance bone healing. Vet Surg 2004;33:565-578 https://doi.org/10.1111/j.1532-950x.2004.04080.x