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Bioreducible Crosslinked Polyelectrolyte Complexes for MMP-2 siRNA Delivery into Human Vascular Smooth Muscle Cells

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ABSTRACT

Purpose

Bioreducible crosslinked polyplexes were prepared via disulfide bond formation after siRNA condensation with polyethylenimine-modified by deoxycholic acid (PEI-DA) to stabilize polyplex structure in an extracellular environment and to promote transfection efficiency in human smooth muscle cells (hSMCs).

Methods

The PEI-DA/siRNA polyplexes were further modified by crosslinking the primary amines of PEI with thiol-cleavable crosslinkers. The effect of disulfide crosslinked PEI-DA/siRNA (Cr PEI-DA/siRNA) polyplexes on target gene silencing was investigated by transfecting hSMCs with matrix metalloproteinase-2 (MMP-2) siRNA under serum conditions. The MMP-2 levels in the conditioned medium were examined using gelatin zymography.

Results

The Cr PEI-DA/siRNA polyplexes showed increased stability against heparin exchange reactions, while their disulfide linkages were successfully cleaved under reducing conditions. The polyplex crosslinking reaction led to a slight decrease in MMP-2 gene silencing activity in hSMCs due to the insufficient redox potential. However, the gene silencing efficiency of the Cr PEI-DA/siRNA polypexes was gradually improved in response to increasing intracellular reduction potential. The increased serum stability of the Cr PEI-DA/siRNA polyplexes resulted in significant enhancement of the intracellular delivery efficiency especially under serum conditions.

Conclusion

The Cr PEI-DA/siRNA polyplex formulation may be a promising siRNA delivery system for the treatment of incurable genetic disorders.

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ACKNOWLEDGMENTS & DISCLOSURES

This work was supported by a grant from the Korea Health Technology R&D Project, Ministry of Health & Welfare, Republic of Korea (A110879).

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Authors and Affiliations

  1. Severance Integrative Research Institute for Cerebral and Cardiovascular Disease, Yonsei University Health System, 250 Seongsanno, Seodaemun-gu, Seoul, 120-752, South Korea

    Dokyoung Lee, Dongkyu Kim, Donghoon Choi & Sun Hwa Kim

  2. Department of Bioscience and Biotechnology, Konkuk University, 120 Neungdong-ro, Gwangji-gu, Seoul, 143-701, South Korea

    Hyejung Mok

  3. School of Pharmacy, Sungkyunkwan University, Suwon, South Korea

    Ji Hoon Jeong

  4. Center for Theragnosis, Biomedical Research Center, Korea Institute of Science and Technology (KIST), Hwarangno 14-gil 6, Seongbuk-gu, Seoul, 136-791, South Korea

    Sun Hwa Kim

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  1. Dokyoung Lee
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  2. Dongkyu Kim
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  3. Hyejung Mok
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  4. Ji Hoon Jeong
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  5. Donghoon Choi
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Correspondence to Donghoon Choi or Sun Hwa Kim.

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Lee, D., Kim, D., Mok, H. et al. Bioreducible Crosslinked Polyelectrolyte Complexes for MMP-2 siRNA Delivery into Human Vascular Smooth Muscle Cells. Pharm Res 29, 2213–2224 (2012). https://doi.org/10.1007/s11095-012-0750-4

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  • DOI: https://doi.org/10.1007/s11095-012-0750-4

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