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Enhanced Stability and Knockdown Efficiency of Poly(ethylene glycol)-b-polyphosphoramidate/siRNA Micellar Nanoparticles by Co-condensation with Sodium Triphosphate

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ABSTRACT

Purpose

Polyelectrolyte complex nanoparticles are a promising vehicle for siRNA delivery but suffer from low stability under physiological conditions. An effective stabilization method is essential for the success of polycationic nanoparticle-mediated siRNA delivery. In this study, sodium triphosphate (TPP), an ionic crosslinking agent, is used to stabilize siRNA-containing nanoparticles by co-condensation.

Methods

siRNA and TPP were co-encapsulated into a block copolymer, poly(ethylene glycol)-b-polyphosphoramidate (PEG-b-PPA), to form ternary nanoparticles. Physicochemical characterization was performed by dynamic light scattering and gel electrophoresis. Gene silencing efficiency in cell lines was assessed by dual luciferase assay system.

Results

The PEG-b-PPA/siRNA/TPP ternary nanoparticles exhibited high uniformity with smaller size (80–100 nm) compared with PEG-b-PPA/siRNA nanoparticles and showed increased stability in physiological ionic strength and serum-containing medium, due to the stabilization effect from ionic crosslinks between negatively charged TPP and cationic PPA segment. Transfection and gene silencing efficiency of the TPP-crosslinked nanoparticles were markedly improved over PEG-b-PPA/siRNA complexes in serum-containing medium. No significant difference in cell viability was observed between nanoparticles prepared with and without TPP co-condensation.

Conclusions

These results demonstrated the effectiveness of TPP co-condensation in compacting polycation/siRNA nanoparticles, improving nanoparticle stability and enhancing the transfection and knockdown efficiency in serum-containing medium.

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ACKNOWLEDGMENTS

The authors thank Dr. James F. Dillman III and Dr. Albert L. Ruff at the Cell and Molecular Biology Branch, US Army Medical Research Institute of Chemical Defense, and Dr. Xuan Jiang at Department of Materials Science and Engineering, Johns Hopkins University, for discussions throughout the study. We thank Dr. Noriko Esumi for providing D407 cells. This study was supported by US Army Defense Threat Reduction Agency through the Grant W81XWH-10-2-0053.

Author information

Authors and Affiliations

  1. Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, Maryland, 21218, USA

    Masataka Nakanishi, Rajesh Patil, Yong Ren & Hai-Quan Mao

  2. Translational Tissue Engineering Center, Johns Hopkins University, Baltimore, Maryland, 21218, USA

    Masataka Nakanishi & Hai-Quan Mao

  3. Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland, 21218, USA

    Rishab Shyam

  4. Department of Neuroscience, Johns Hopkins University, Baltimore, Maryland, 21218, USA

    Philip Wong

  5. Whitaker Biomedical Engineering Institute, Johns Hopkins University, Baltimore, Maryland, 21218, USA

    Hai-Quan Mao

  6. 101E Maryland Hall, 3400 North Charles Street, Baltimore, Maryland, 21218, USA

    Hai-Quan Mao

Authors
  1. Masataka Nakanishi
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  2. Rajesh Patil
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  3. Yong Ren
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  4. Rishab Shyam
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  5. Philip Wong
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  6. Hai-Quan Mao
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Corresponding author

Correspondence to Hai-Quan Mao.

Additional information

Masataka Nakanishi and Rajesh Patil contributed equally to this work.

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Nakanishi, M., Patil, R., Ren, Y. et al. Enhanced Stability and Knockdown Efficiency of Poly(ethylene glycol)-b-polyphosphoramidate/siRNA Micellar Nanoparticles by Co-condensation with Sodium Triphosphate. Pharm Res 28, 1723–1732 (2011). https://doi.org/10.1007/s11095-011-0408-7

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  • DOI: https://doi.org/10.1007/s11095-011-0408-7

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