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Hyaluronic Acid/Chitosan-g-Poly(ethylene glycol) Nanoparticles for Gene Therapy: An Application for pDNA and siRNA Delivery

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ABSTRACT

Purpose

To design hyaluronic acid (HA) and chitosan-g-poly(ethylene glycol) (CS-g-PEG) nanoparticles intended for a broad range of gene delivery applications.

Methods

Nanoparticles formulated at different HA/CS-g-PEG mass ratios were developed to associate either pDNA or siRNA. The physico-chemical characteristics, morphology, association efficiency and nuclease protection ability of the nanocarriers were compared for these two molecules. Their biological performance, including transfection effciency, nanoparticle cellular uptake and citotoxicity, was assesed.

Results

The resulting nanoparticles showed an adequate size (between 130 and 180 nm), and their surface charge could be modulated according to the nanoparticle composition (from +30 mV to −20 mV). All prototypes exhibited a greater association efficiency and nuclease protection for pDNA than for siRNA. However, cell culture experiments evidenced that HA/CS-g-PEG nanoparticles were effective carriers for the delivery of both, siRNA and pDNA, eliciting a biological response with minimal cytotoxicity. Moreover, experiments performed in the HEK-EGFP-Snail1 cell line showed the potential of the HA/CS-g-PEG nanoparticles to silence the expression of the Snail1 transcription factor, an important mediator in tumor progression.

Conclusions

HA/CS-g-PEG nanoparticles can be easily modulated for the delivery of different types of gene molecules, offering great potential for gene therapy applications, as evidenced by their biological performance.

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ACKNOWLEDGEMENTS

The authors gratefully acknowledge valuable discussion with Dr. M. de la Fuente and support from the Ministry of Science and Technology (Spain) (Refs. SAF2004-09230-004-01 and SAF2004-C04-02). The first author also acknowledges the fellowship received from the Spanish Government (FPI).

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Authors and Affiliations

  1. Departamento de Farmacia y Tecnología Farmacéutica, Facultad de Farmacia, Universidad de Santiago de Compostela, Avda. de las Ciencias s/n, 15782, Santiago de Compostela, Spain

    Manuela Raviña, Alejandro Sánchez & María José Alonso

  2. Instituto Nacional de Toxicologia y Ciencias Forenses (INTCF), Servicio de Biología, Las Rozas de Madrid, 28032, Madrid, Spain

    Eva Cubillo

  3. Centro Nacional de Investigaciones Oncologicas (CNIO), Melchor Fernandez, Almagro, 4, 28029, Madrid, Spain

    David Olmeda

  4. Department of Organic Chemistry and Center for Research in Biological Chemistry and Molecular Materials, University of Santiago de Compostela, Jenaro de la Fuente s/n, 15782, Santiago de Compostela, Spain

    Ramón Novoa-Carballal, Eduardo Fernandez-Megia & Ricardo Riguera

  5. Departamento de Bioquímica, Instituto de Investigaciones Biomédicas ‘Alberto Sols’ CSIC-UAM, c/Arturo Duperier, Madrid, Spain

    Amparo Cano

  6. Department of Pharmacy and Pharmaceutical Technology, School of Pharmacy, University of Santiago de Compostela, 15782, Santiago de Compostela, Spain

    María José Alonso

Authors
  1. Manuela Raviña
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  2. Eva Cubillo
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  3. David Olmeda
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  4. Ramón Novoa-Carballal
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  5. Eduardo Fernandez-Megia
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  6. Ricardo Riguera
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  7. Alejandro Sánchez
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  8. Amparo Cano
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  9. María José Alonso
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Corresponding author

Correspondence to María José Alonso.

Elecrtonic Supplementary Material

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Supplementary Figure 1

Expression of EGFP and Snail-EGFP protein in HEK-EGFP and HEK-EGFP-Snail1 cell line by western blotting. WB on whole cell lysates from HEK-EGFP (left) and HEK-EGFP-Snail1 (right) cells was performed as previously described (20) using anti-Snail (upper panel) or anti-EGFP (middle panels). Anti-tubulin antibodies were used as a loading control. (DOC 56 kb)

Supplementary Figure 2

Comparison of EGFP expression in non treated HEK-EGFP cells or after si-Snail1 loaded HA/CS-g-PEG 1/1 nanoparticles treatment (100 nM siRNA/well). (DOC 38 kb)

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Raviña, M., Cubillo, E., Olmeda, D. et al. Hyaluronic Acid/Chitosan-g-Poly(ethylene glycol) Nanoparticles for Gene Therapy: An Application for pDNA and siRNA Delivery. Pharm Res 27, 2544–2555 (2010). https://doi.org/10.1007/s11095-010-0263-y

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  • DOI: https://doi.org/10.1007/s11095-010-0263-y

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