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Folate-Targeted Multifunctional Amino Acid-Chitosan Nanoparticles for Improved Cancer Therapy

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Abstract

Purpose

Tumor targeting nanomaterials have potential for improving the efficiency of anti-tumoral therapeutics. However, the evaluation of their biological performance remains highly challenging. In this study we describe the synthesis of multifunctional nanoparticles decorated with folic acid-PEG and dual amino acid-modified chitosan (CM-PFA) complexed with DNA and their evaluation in organotypic 2D co-cultures of cancer-normal cells and also on 3D multicellular tumor spheroids models.

Methods

The physicochemical characterization of CM-PFA multifunctional carriers was performed by FTIR, 1H NMR and DLS. 2D co-culture models were established by using a 1:2 cancer-to-normal cell ratio. 3D organotypic tumor spheroids were assembled using micromolding technology for high throughput screening. Nanoparticle efficiency was evaluated by flow cytometry and confocal microscopy.

Results

The CM-PFA nanocarriers (126–176 nm) showed hemocompatibility and were internalized by target cells, achieving a 3.7 fold increase in gene expression. In vivo-mimicking 2D co-cultures confirmed a real affinity towards cancer cells and a negligible uptake in normal cells. The targeted nanoparticles penetrated into 3D spheroids to a higher extent than non-targeted nanocarriers. Also, CM-PFA-mediated delivery of p53 tumor suppressor promoted a decrease in tumor-spheroids volume.

Conclusion

These findings corroborate the improved efficiency of this delivery system and demonstrate its potential for application in cancer therapy.

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Acknowledgments

The authors would like to acknowledge Eng. Ana Paula for her help with the acquisition of SEM images. This work was supported by the Portuguese Foundation for Science and Technology (FCT), (PTDC/EBB-BIO/114320/2009 and PEst-C/SAU/UI0709/2011). Vítor M. Gaspar is grateful for the PhD fellowship from FCT (SFRH/BD/80402/2011). All the authors do not disclose any conflict of interest.

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Authors and Affiliations

  1. CICS-UBI – Health Sciences Research Centre, Universidade da Beira Interior, Avenida Infante D. Henrique, 6200-506, Covilhã, Portugal

    Vítor M. Gaspar, Elisabete C. Costa, João A. Queiroz, Fani Sousa & Ilídio J. Correia

  2. Centre de Biophysique Moléculaire, CNRS UPR4301, Inserm and University of Orléans, 45071 cedex 02, Orléans, France

    Chantal Pichon

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  1. Vítor M. Gaspar
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  6. Ilídio J. Correia
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Correspondence to Ilídio J. Correia.

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Gaspar, V.M., Costa, E.C., Queiroz, J.A. et al. Folate-Targeted Multifunctional Amino Acid-Chitosan Nanoparticles for Improved Cancer Therapy. Pharm Res 32, 562–577 (2015). https://doi.org/10.1007/s11095-014-1486-0

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  • DOI: https://doi.org/10.1007/s11095-014-1486-0

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