Biotechnological approach to induce human fibroblast apoptosis using superparamagnetic iron oxide nanoparticles

J Inorg Biochem. 2020 May:206:111017. doi: 10.1016/j.jinorgbio.2020.111017. Epub 2020 Feb 1.

Abstract

Cancer-Associated Fibroblasts (CAFs) contribute to tumour progression and have received significant attention as a therapeutic target. These cells produce growth factors, cytokines and chemokines, stimulating cancer cell proliferation and inhibiting their apoptosis. Recent advances in drug delivery have demonstrated a significant promise of iron oxide nanoparticles in clinics as theranostic agents, mainly due to their magnetic properties. Here, we designed superparamagnetic iron oxide nanoparticles (SPIONs) to induce apoptosis of human fibroblasts. SPIONs were synthesized via co-precipitation method and coated with sodium citrate (SPION_Cit). We assessed the intracellular uptake of SPIONs by human fibroblast cells, as well as their cytotoxicity and ability to induce thermal effects under the magnetic field. The efficiency and time of nanoparticle internalization were assessed by Prussian Blue staining, flow cytometry and transmission electron microscopy. SPIONs_Cit were detected in the cytoplasm of human fibroblasts 15 min after in vitro exposure, entering into cells mainly via endocytosis. Analyses through Cell Titer Blue assay, AnnexinV-fluorescein isothiocyanate (FITC) and propidium iodide (PI) cellular staining demonstrated that concentrations below 8 × 10-2 mg/mL of SPIONs_Cit did not alter cell viability of human fibroblast. Furthermore, it was also demonstrated that SPIONs_Cit associated with alternating current magnetic field were able to induce hyperthermia and human fibroblast cell death in vitro, mainly through apoptosis (83.5%), activating caspase 8 (extrinsic apoptotic via) after a short exposure period. Collectively these findings suggest that our nanoplatform is biocompatible and can be used for therapeutic purposes in human biological systems, such as inducing apoptosis of CAFs.

Keywords: Cancer; Fibroblast cells; Hyperthermia-induced apoptosis; Nanoparticle internalization; Nanotoxicity; SPIONs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / drug effects*
  • Cancer-Associated Fibroblasts / drug effects
  • Cell Death / drug effects
  • Cell Line
  • Cell Survival / drug effects
  • Citric Acid / chemistry
  • Endocytosis
  • Ferric Compounds / chemistry
  • Ferric Compounds / pharmacology*
  • Fibroblasts / drug effects*
  • Flow Cytometry
  • Humans
  • Hyperthermia, Induced
  • Magnetic Iron Oxide Nanoparticles / administration & dosage*
  • Magnetic Iron Oxide Nanoparticles / chemistry
  • Microscopy, Electron, Transmission
  • Neoplasms / metabolism
  • Neoplasms / pathology

Substances

  • Ferric Compounds
  • Citric Acid