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Synthetic scaffold coating with adeno-associated virus encoding BMP2 to promote endogenous bone repair

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Abstract

Biomaterial scaffolds functionalized to stimulate endogenous repair mechanisms via the incorporation of osteogenic cues offer a potential alternative to bone grafting for the treatment of large bone defects. We first quantified the ability of a self-complementary adeno-associated viral vector encoding bone morphogenetic protein 2 (scAAV2.5-BMP2) to enhance human stem cell osteogenic differentiation in vitro. In two-dimensional culture, scAAV2.5-BMP2-transduced human mesenchymal stem cells (hMSCs) displayed significant increases in BMP2 production and alkaline phosphatase activity compared with controls. hMSCs and human amniotic-fluid-derived stem cells (hAFS cells) seeded on scAAV2.5-BMP2-coated three-dimensional porous polymer Poly(ε-caprolactone) (PCL) scaffolds also displayed significant increases in BMP2 production compared with controls during 12 weeks of culture, although only hMSC-seeded scaffolds displayed significantly increased mineral formation. PCL scaffolds coated with scAAV2.5-BMP2 were implanted into critically sized immunocompromised rat femoral defects, both with or without pre-seeding of hMSCs, representing ex vivo and in vivo gene therapy treatments, respectively. After 12 weeks, defects treated with acellular scAAV2.5-BMP2-coated scaffolds displayed increased bony bridging and had significantly higher bone ingrowth and mechanical properties compared with controls, whereas defects treated with scAAV2.5-BMP2 scaffolds pre-seeded with hMSCs failed to display significant differences relative to controls. When pooled, defect treatment with scAAV2.5-BMP2-coated scaffolds, both with or without inclusion of pre-seeded hMSCs, led to significant increases in defect mineral formation at all time points and increased mechanical properties compared with controls. This study thus presents a novel acellular bone-graft-free endogenous repair therapy for orthotopic tissue-engineered bone regeneration.

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Acknowledgements

The authors thank Angela Lin, Mela Johnson, Chris Dosier, Jessica O’Neal Green, Jason Wang, Laura O’Farrell, Alex Peister, and Brent Uhrig for additional assitance during these studies, and Andres Garcia for providing GFOGER.

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Authors and Affiliations

  1. Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, 315 Ferst Drive, Atlanta, GA, 30332, USA

    Kenneth M. Dupont, Joel D. Boerckel, Hazel Y. Stevens, Tamim Diab, Yash M. Kolambkar & Robert E. Guldberg

  2. George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, 315 Ferst Drive, Atlanta, GA, 30332, USA

    Kenneth M. Dupont, Joel D. Boerckel, Hazel Y. Stevens, Tamim Diab & Robert E. Guldberg

  3. Exponent Failure Analysis Associates, 3401 Market Street, Suite 300, Philadelphia, PA, 19104, USA

    Kenneth M. Dupont

  4. Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, 315 Ferst Drive, Atlanta, GA, 30332, USA

    Yash M. Kolambkar & Robert E. Guldberg

  5. Center for Musculoskeletal Research, School of Medicine and Dentistry, University of Rochester, 601 Elmwood Avenue, Box 665, Rochester, NY, 14642, USA

    Masahiko Takahata & Edward M. Schwarz

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  1. Kenneth M. Dupont
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  2. Joel D. Boerckel
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  4. Tamim Diab
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  5. Yash M. Kolambkar
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  6. Masahiko Takahata
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  7. Edward M. Schwarz
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  8. Robert E. Guldberg
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Corresponding author

Correspondence to Robert E. Guldberg.

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Funding was provided by National Institutes of Health Grant R01 AR051336 and PHS Awards DE19902/AR54041 and by a Musculoskeletal Transplant Foundation grant.

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Dupont, K.M., Boerckel, J.D., Stevens, H.Y. et al. Synthetic scaffold coating with adeno-associated virus encoding BMP2 to promote endogenous bone repair. Cell Tissue Res 347, 575–588 (2012). https://doi.org/10.1007/s00441-011-1197-3

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