Folate-Targeted Multifunctional Amino Acid-Chitosan Nanoparticles for Improved Cancer Therapy
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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.
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.
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.
These findings corroborate the improved efficiency of this delivery system and demonstrate its potential for application in cancer therapy.
Key Wordscancer therapy gene delivery targeted nanoparticles 2D co-cultures 3D tumor spheroids
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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