Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Adenoviral vector-mediated overexpression of osteoprotegerin accelerates osteointegration of titanium implants in ovariectomized rats

Gene Therapy volume 22, pages 636–644 (2015)Cite this article

Subjects

Abstract

This study investigated the efficacy of human osteoprotegerin (hOPG) transgene to accelerate osteointegration of titanium implant in ovariectomized (OVX) rats. Bone marrow stromal cells transduced with Ad-hOPG-EGFP could sustainedly express hOPG. Osteoclast precursor RAW264.7 cells treated by the hOPG were examined by tartrate-resistant acid phosphatase (TRAP) staining and bone slice resorption assay. The results showed differentiation and function of osteoclasts were significantly suppressed by hOPG in vitro. Ad-hOPG-EGFP was locally administered to the bone defect prior to implant placement in OVX and sham rats. After 3, 7, 28 days of implantation, the femurs were harvested for molecular and histological analyses. Successful transgene expression was confirmed by western blot and cryosectioning. A significant reduction in TRAP+ numbers was detected in Ad-hOPG-EGFP group. Real-time reverse transcriptase-PCR examination revealed that hOPG transgene markedly diminished the expression of cathepsin K and receptor activator for nuclear factor-κ B ligand in vivo. The transgene hOPG modification revealed a marked increasing osteointegration and restored implant stability in OVX rats (P<0.01), compared with the control groups (Ad-EGFP or sterilized phosphate-buffered saline) 28 days after implantation. In conclusion, hOPG via direct adenovirus-mediated gene transfer could accelerate osteointegration of titanium implants in OVX rats. Osteoprotegerin gene therapy may be an effective strategy to osteointegration of implants under osteoporotic conditions.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

References

  1. Rizzoli R, Burlet N, Cahall D, Delmas PD, Eriksen EF, Felsenberg D et al. Osteonecrosis of the jaw and bisphosphonate treatment for osteoporosis. Bone 2008; 42: 841–847.

    Article  CAS  Google Scholar 

  2. Anner R, Grossmann Y, Anner Y, Levin L . Smoking, diabetes mellitus, periodontitis, and supportive periodontal treatment as factors associated with dental implant survival: a long-term retrospective evaluation of patients followed for up to 10 years. Implant Dent 2010; 19: 57–64.

    Article  Google Scholar 

  3. Alsaadi G, Quirynen M, Komarek A, van Steenberghe D . Impact of local and systemic factors on the incidence of oral implant failures, up to abutment connection. J Clin Periodontol 2007; 34: 610–617.

    Article  Google Scholar 

  4. Fini M, Giavaresi G, Torricelli P, Corsari V, Giardino R, Nicolini A et al. Osteoporosis and biomaterial osteointegration. Biomed Pharmacother 2004; 58: 487–493.

    Article  CAS  Google Scholar 

  5. Takayanagi H, Ogasawara K, Hida S, Chiba T, Murata S, Sato K et al. T-cellmediated regulation of osteoclastogenesis by signalling cross-talk between RANKL and IFN-gamma. Nature 2000; 408: 600–605.

    Article  CAS  Google Scholar 

  6. Ho TY, Santora K, Chen JC, Frankshun AL, Bagnell CA . Effects of relaxin and estrogens on bone remodeling markers, receptor activator of NF-κB ligand(RANKL) and osteoprotegerin(OPG), in rat adjuvant-induced arthritis. Bone 2011; 48: 1346–1353.

    Article  CAS  Google Scholar 

  7. Kostenuik PJ, Bolon B, Morony S, Daris M, Geng Z, Carter C et al. Gene therapy with human recombinant osteoprotegerin reverses established osteopenia in ovariectomized mice. Bone 2004; 34: 656–664.

    Article  CAS  Google Scholar 

  8. Bolon B, Carter C, Daris M, Morony S, Capparelli C, Hsieh A et al. Adenoviral delivery of recombinant osteoprotegerin (OPG) ameliorates bone resorption in a mouse ovariectomy model of osteoporosis. Mol Ther 2001; 3: 179–205.

    Article  Google Scholar 

  9. Luo E, Hu J, Bao C, Li Y, Tu Q, Murray D et al. Sustained release of adiponectin improves osteogenesis around hydroxyapatite implants by suppressing osteoclast activity in ovariectomized rabbits. Acta Biomater 2012; 8: 734–743.

    Article  CAS  Google Scholar 

  10. Lugero GG, de Falco Caparbo V, Guzzo ML, Konig Jr B, Jorgetti V . Histomorphometric evaluation of titanium implants in osteoporotic rabbits. Implant Dent 2000; 9: 303–309.

    Article  CAS  Google Scholar 

  11. Kurth AH, Eberhardt C, Muller S, Steinacker M, Schwarz M, Bauss F . The bisphosphonate ibandronate improves implant integration in osteopenic ovariectomized rats. Bone 2005; 37: 204–210.

    Article  CAS  Google Scholar 

  12. Saito T, Kin Y, Koshino T . Osteogenic response of hydroxyapatite cement implanted into the femur of rats with experimentally induced osteoporosis. Biomaterials 2002; 23: 2711–2716.

    Article  CAS  Google Scholar 

  13. Flick LM, Weaver JM, Ulrich-Vinther M, Abuzzahab F, Zhang X, Dougall WC et al. Effects of receptor activator of NFKB (RANK) signaling blockade on fracture healing. J Orthop Res 2003; 21: 676–684.

    Article  CAS  Google Scholar 

  14. Ominsky MS, Stouch B, Schroeder J, Pyrah L, Stolina M, Smith SY et al. Denosumab,a fully human RANKL antibody, reduced bone turnover markers and increased trabecular and cortical bone mass, density,and strength in ovariectomized cynomolgus monkeys. Bone 2011; 49: 162–173.

    Article  CAS  Google Scholar 

  15. Horowitz MC, Xi Y, Wilson K, Kacena MA . Control of osteoclastogenesis and bone resorption by members of the TNF family of receptors and ligands. Cytokine Growth Factor Rev 2001; 12: 9–18.

    Article  CAS  Google Scholar 

  16. Boyce BF, Xing L . Biology of RANK, RANKL, and osteoprotegerin. Arthritis Res Ther 2007; 9: S1.

    Article  Google Scholar 

  17. Boyce BF, Xing L . Functions of RANKL/RANK/OPG in bone modeling and remodeling. Arch Biochem Biophys 2008; 473: 139–146.

    Article  CAS  Google Scholar 

  18. Wang H, Jia TH, Zacharias N, Gong W, Du HX, Wooley PH et al. Combination gene therapy targeting on interleukin-1 β and RANKL for wear debris-induced aseptic loosening. Gene Ther 2013; 20: 128–135.

    Article  CAS  Google Scholar 

  19. Zhang L, Jia TH, Chong AC, Bai L, Yu H, Gong W et al. Cell-based osteoprotegerin therapy for debris-induced aseptic prosthetic loosening on a murine model. Gene Ther 2010; 17: 1262–1269.

    Article  CAS  Google Scholar 

  20. Kearns AE, Khosla S, Kostenuik PJ . Receptor activator of nuclear factorB ligand and osteoprotegerin regulation of bone remodeling in health and disease. Endocr Rev 2008; 29: 155–192.

    Article  CAS  Google Scholar 

  21. Wang FS, Ko JY, Lin CL, Wu HL, Ke HJ, Tai PJ . Knocking down dickkopf-1 alleviates estrogen deficiency induction of bone loss: a histomorphological study in ovariectomized rats. Bone 2007; 40: 485–492.

    Article  CAS  Google Scholar 

  22. Ominsky MS, Li X, Asuncion FJ, Barrero M, Warmington KS, Dwyer D et al. RANKL inhibition with OPG preserves bone density and trabecular architecture in ovariectomized rats. J Bone Miner Res 2008; 23: 672–682.

    Article  CAS  Google Scholar 

  23. Christ M, Lusky M, Stoeckel F, Dreyer D, Dieterle A, Michou AI et al. Gene therapy with recombinant adenovirus vectors: evaluation of the host immune response. Immunol Lett 1997; 57: 19–25.

    Article  CAS  Google Scholar 

  24. Molinier-Frenkel V, Gahery-Segard H, Mehtali M, Le Boulaire C, Ribault S, Boulanger P et al. Immune response to recombinant adenovirus in humans: capsid components from viral input are targets for vector-specific cytotoxic T lymphocytes. J Virol 2000; 74: 7678–7682.

    Article  CAS  Google Scholar 

  25. Davis AR, Wivel NA, Palladino JL, Tao L, Wilson JM . Construction of adenoviral vectors. Mol Biotechnol 2001; 18: 63–70.

    Article  CAS  Google Scholar 

  26. Sun S, Chen G, Xu M, Qiao Y, Zheng S . Differentiation and Migration of Bone Marrow Mesenchymal Stem Cells Transplanted through the Spleen in Rats with Portal Hypertension. Plos One 2013; 8: e83523.

    Article  Google Scholar 

  27. Nakajima A, Sanjay A, Chiusaroli R, Adapala NS, Neff L, Itzsteink C et al. Loss of Cbl-b increases osteoclast bone-resorbing activity and induces osteopenia. J Bone Miner Res 2009; 24: 1162–1172.

    Article  CAS  Google Scholar 

  28. Valverde P, Tu Q, Chen J . BSP and RANKL induce osteoclastogenesis and bone resorption synergistically. J Bone Miner Res 2005; 20: 1669–1679.

    Article  CAS  Google Scholar 

  29. Varkey M, Kucharski C, Doschak MR, Winn SR, Brochmann EJ, Murray S et al. Osteogenic response of bone marrow stromal cells from normal and ovariectomized rats treated with a low dose of basic fibroblast growth factor. Tissue Eng 2007; 13: 809–817.

    Article  CAS  Google Scholar 

  30. Xu B, Zhang J, Brewer E, Tu Q, Yu L, Tang J et al. Osterix enhances BMSC-associated osseointegration of implants. J Dent Res 2009; 88: 1003–1007.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This study was supported by grants from the financial support of NSFC 81321002, ‘Key Technologies Research and Development Program of Science and Technology Department of Sichuan Province (2014SZ0019)’, and National Basic Research Program of China (973 Program, No. 2012CB933902).

Author information

Authors and Affiliations

  1. State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, China

    G Yin, H Wang, Q Chen, Y Lin, J Hu & E Luo

  2. Division of Oral Biology, Department of General Dentistry, Tufts University School of Dental Medicine, Boston, MA, USA

    J Chen & E Luo

  3. Department of Oral and Maxillofacial Surgery, Laboratory of Interdisciplinary Studies, School and Hospital of Stomatology, Peking University, Beijing, China

    S Wei

Authors
  1. G Yin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S Wei
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. H Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Q Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Y Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. J Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. E Luo
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to J Hu or E Luo.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yin, G., Chen, J., Wei, S. et al. Adenoviral vector-mediated overexpression of osteoprotegerin accelerates osteointegration of titanium implants in ovariectomized rats. Gene Ther 22, 636–644 (2015). https://doi.org/10.1038/gt.2015.34

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/gt.2015.34

This article is cited by

Search

Advanced search

Quick links