Targeted Smart pH and Thermoresponsive N,O-Carboxymethyl Chitosan Conjugated Nanogels for Enhanced Therapeutic Efficacy of Doxorubicin in MCF-7 Breast Cancer Cells

Bioconjug Chem. 2016 Nov 16;27(11):2605-2619. doi: 10.1021/acs.bioconjchem.6b00366. Epub 2016 Oct 25.

Abstract

In cancer treatment, developing ideal anticancer drug delivery systems to target tumor microenvironment by circumventing various physiological barriers still remains a daunting challenge. Here, in our work, a series of pH- and temperature-responsive nanogels based on poly(N-isopropylacrylamide-co-1-propene-2-3-dicarboxylate-co-2-acrylamido-2-methyl-1-propanesulfonate [poly(NIPAAm-IA-AMPS)] cross-linked by ethylene glycol dimethacrylate (EGDMA) were synthesized by random copolymerization. The molar ratio between monomer-comonomers-cross-linker was varied to fine-tune the optimum responsiveness of the nanogels. These optimized nanogels were further coupled to N,O-carboxymethyl chitosan (NOCC) stoichiometrically using EDC-NHS coupling chemistry to enhance the swelling behavior at lower pH. Interestingly, these NOCC-g-nanogels, when dispersed in aqueous media under sonication, attain nanosize and retain their high water-retention capacity with conspicuous pH and temperature responsiveness (viz. nanogel shrinkage in size beyond 35 °C and swelled at acidic pH) in vitro, as reflected by dynamic light scattering data. Doxorubicin (DOX), a potent anticancer drug, was loaded into these nanogels using the physical entrapment method. These drug-loaded nanogels exhibited a slow and sustained DOX release profile at physiological temperature and cytosolic pH. Furthermore, confocal and TEM results demonstrate that these nanogels were swiftly internalized by MCF-7 cells, and cell viability data showed preferential heightened cytotoxicity toward cancer cells (MCF-7 and MDA-MB231) compared to the MCF10A cells (human breast epithelial cell). Furthermore, intracellular DNA damage and cell cycle arrest assays suggest a mitochondrial mediated apoptosis in MCF-7 cells. This study substantiates our NOCC-g-nanogel platform as an excellent modality for passive diffusive loading and targeted release of entrapped drug(s) at physiological conditions in a controlled way for the improved therapeutic efficacy of the drug in anticancer treatment.

MeSH terms

  • Acrylic Resins / chemistry
  • Apoptosis / drug effects
  • Biological Transport
  • Cell Cycle Checkpoints / drug effects
  • Cell Survival / drug effects
  • Chitosan / chemistry*
  • DNA Damage
  • Doxorubicin / chemistry*
  • Doxorubicin / pharmacology*
  • Drug Carriers / chemistry*
  • Drug Carriers / metabolism
  • Gels
  • Humans
  • Hydrogen-Ion Concentration
  • MCF-7 Cells
  • Membrane Potential, Mitochondrial / drug effects
  • Nanostructures / chemistry*
  • Oxidative Stress / drug effects
  • Temperature

Substances

  • Acrylic Resins
  • Drug Carriers
  • Gels
  • O,N-carboxymethylchitosan
  • poly-N-isopropylacrylamide
  • Doxorubicin
  • Chitosan