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Ultrasound-based nonviral gene delivery induces bone formation in vivo

Gene Therapy volume 15, pages 257–266 (2008)Cite this article

Abstract

Nonviral gene delivery is a promising, safe, therapeutic tool in regenerative medicine. This study is the first to achieve nonviral, ultrasound-based, osteogenic gene delivery that leads to bone tissue formation, in vivo. We hypothesized that direct in vivo sonoporation of naked DNA encoding for the osteogenic gene, recombinant human bone morphogenetic protein-9 (rhBMP-9) would induce bone formation. A luciferase plasmid (Luc), encoding rhBMP-9 or empty pcDNA3 vector mixed with microbubbles, was injected into the thigh muscles of mice. After injection, noninvasive sonoporation was applied. Luc activity was monitored noninvasively, and quantitatively using bioluminescence imaging in vivo, and found for 14 days with a peak expression on day 7. To examine osteogenesis in vivo, rhBMP-9 plasmid was sonoporated into the thigh muscles of transgenic mice that express the Luc gene under the control of a human osteocalcin promoter. Following rhBMP-9 sonoporation, osteocalcin-dependent Luc expression lasted for 24 days and peaked on day 10. Bone tissue was formed in the site of rhBMP-9 delivery, as was shown by micro-computerized tomography and histology. The sonoporation method was also compared with previously developed electrotransfer-based gene delivery and was found significantly inferior in its efficiency of gene delivery. We conclude that ultrasound-mediated osteogenic gene delivery could serve as a therapeutic solution in conditions requiring bone tissue regeneration after further development that will increase the transfection efficiency.

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Acknowledgements

We thank Rich-Mar Corp. (Inola) for kindly providing the sonoporation Sonitron2000 device for the experiments described in this study.

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Authors and Affiliations

  1. Skeletal Biotech Laboratory, Hebrew University of Jerusalem, Jerusalem, Israel

    D Sheyn, N Kimelman-Bleich, G Pelled, Y Zilberman, D Gazit & Z Gazit

  2. Department of Surgery, International Stem Cell Institute, Cedars Sinai Medical Center, Los Angeles, CA, USA

    D Gazit

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  1. D Sheyn
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Correspondence to Z Gazit.

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Sheyn, D., Kimelman-Bleich, N., Pelled, G. et al. Ultrasound-based nonviral gene delivery induces bone formation in vivo. Gene Ther 15, 257–266 (2008). https://doi.org/10.1038/sj.gt.3303070

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