Pharmaceutical Research

, Volume 27, Issue 12, pp 2544–2555

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

  • Manuela Raviña
  • Eva Cubillo
  • David Olmeda
  • Ramón Novoa-Carballal
  • Eduardo Fernandez-Megia
  • Ricardo Riguera
  • Alejandro Sánchez
  • Amparo Cano
  • María José Alonso
Research Paper

DOI: 10.1007/s11095-010-0263-y

Cite this article as:
Raviña, M., Cubillo, E., Olmeda, D. et al. Pharm Res (2010) 27: 2544. doi:10.1007/s11095-010-0263-y

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.

KEY WORDS

cancer therapychitosangene deliveryhyaluronic acidnanoparticles

Supplementary material

11095_2010_263_MOESM1_ESM.doc (56 kb)
Supplementary Figure 1Expression 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)
11095_2010_263_MOESM2_ESM.doc (38 kb)
Supplementary Figure 2Comparison 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)

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Manuela Raviña
    • 1
  • Eva Cubillo
    • 2
  • David Olmeda
    • 3
  • Ramón Novoa-Carballal
    • 4
  • Eduardo Fernandez-Megia
    • 4
  • Ricardo Riguera
    • 4
  • Alejandro Sánchez
    • 1
  • Amparo Cano
    • 5
  • María José Alonso
    • 1
    • 6
  1. 1.Departamento de Farmacia y Tecnología Farmacéutica, Facultad de FarmaciaUniversidad de Santiago de CompostelaSantiago de CompostelaSpain
  2. 2.Instituto Nacional de Toxicologia y Ciencias Forenses (INTCF)Servicio de BiologíaMadridSpain
  3. 3.Centro Nacional de Investigaciones Oncologicas (CNIO), Melchor FernandezMadridSpain
  4. 4.Department of Organic Chemistry and Center for Research in Biological Chemistry and Molecular MaterialsUniversity of Santiago de CompostelaSantiago de CompostelaSpain
  5. 5.Departamento de BioquímicaInstituto de Investigaciones Biomédicas ‘Alberto Sols’ CSIC-UAMMadridSpain
  6. 6.Department of Pharmacy and Pharmaceutical Technology, School of PharmacyUniversity of Santiago de CompostelaSantiago de CompostelaSpain