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Delivery of an Angiogenic Gene into Ischemic Muscle by Novel Bubble Liposomes Followed by Ultrasound Exposure

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ABSTRACT

Purpose

To develop a safe and efficient gene delivery system into skeletal muscle using the combination of Bubble liposomes (BL) and ultrasound (US) exposure, and to assess the feasibility and the effectiveness of BL for angiogenic gene delivery in clinical use.

Methods

A solution of luciferase-expressing plasmid DNA (pDNA) and BL was injected into the tibialis (TA) muscle, and US was immediately applied to the injection site. The transfection efficiency was estimated by a luciferase assay. The ischemic hindlimb was also treated with BL and US-mediated intramuscular gene transfer of bFGF-expressing plasmid DNA. Capillary vessels were assessed using immunostaining. The blood flow was determined using a laser Doppler blood flow meter.

Results

Highly efficient gene transfer could be achieved in the muscle transfected with BLs, and US mediated the gene transfer. Capillary vessels were enhanced in the treatment groups with this gene transfer method. The blood flow in the treated groups with this gene transfer method quickly recovered compared to other treatment groups (non-treated, bFGF alone, or bFGF+US).

Conclusion

The gene transfer system into skeletal muscle using the combination of BL and US exposure could be an effective means for angiogenic gene therapy in limb ischemia.

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ACKNOWLEDGEMENTS

We are grateful to Dr. Katsuro Tachibana (Department of Anatomy, School of Medicine, Fukuoka University) for technical advice regarding the induction of cavitation with US and to Mr. Yasuhiko Hayakawa, Mr. Takahiro Yamauchi, and Mr. Kosho Suzuki (NEPA GENE CO., LTD.) for technical advice regarding US exposure. This study was supported in part by the Industrial Technology Research Grant Program (04A05010) from New Energy, the Industrial Technology Development Organization (NEDO) of Japan, Grant-in-Aid for Scientific Research (B) (20300179) from the Japan Society of the Promotion of Science, and by a grant for private universities provided by the Promotion and Mutual Aid Corporation for Private Schools of Japan.

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Authors and Affiliations

  1. Department of Drug and Gene Delivery Systems School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo, 192-0392, Japan

    Yoichi Negishi, Keiko Matsuo, Yoko Endo-Takahashi, Kentaro Suzuki, Yuuki Matsuki & Yukihiko Aramaki

  2. Department of Molecular and Cellular Pharmacology School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo, 192-0392, Japan

    Norio Takagi

  3. Department of Pharmaceutics, Teikyo University, 1091-1 Suwarashi, Midori-ku, Sagamihara, Kanagawa, 252-5195, Japan

    Ryo Suzuki & Kazuo Maruyama

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  1. Yoichi Negishi
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  2. Keiko Matsuo
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  3. Yoko Endo-Takahashi
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  4. Kentaro Suzuki
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  5. Yuuki Matsuki
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  6. Norio Takagi
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  7. Ryo Suzuki
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  8. Kazuo Maruyama
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  9. Yukihiko Aramaki
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Correspondence to Yoichi Negishi.

Additional information

Yoichi Negishi and Keiko Matsuo have contributed equally to this work.

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Negishi, Y., Matsuo, K., Endo-Takahashi, Y. et al. Delivery of an Angiogenic Gene into Ischemic Muscle by Novel Bubble Liposomes Followed by Ultrasound Exposure. Pharm Res 28, 712–719 (2011). https://doi.org/10.1007/s11095-010-0286-4

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  • DOI: https://doi.org/10.1007/s11095-010-0286-4

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