Graphene oxide functionalized with chitosan based nanoparticles as a carrier of siRNA in regulating Bcl-2 expression on Saos-2 & MG-63 cancer cells and its inflammatory response on bone marrow derived cells from mice

Mater Sci Eng C Mater Biol Appl. 2019 Jun:99:1459-1468. doi: 10.1016/j.msec.2019.02.047. Epub 2019 Feb 16.

Abstract

Presently, quite a lot of research that are being carried out to find a potential cure for cancer and many had made to clinical trial stage as well. In the present study, we focus on use of a novel graphene oxide functionalized chitosan nanoparticle targeting Saos-2 and MG-63 osteosarcoma cells. The graphene oxide chitosan nanoparticles were loaded with siRNA, studied for in vitro release with varying concentration & pH, and fitted to peppas model. MTT & ROS assay was used to evaluate biocompatibility of carrier and qPCR to study the inflammatory responses in particular checking gene expression of IL-6, TGF-ß, TNF-α in both RAW 264.7 and bone marrow derived macrophages. The results of study showed that release of siRNA were in a controlled fashion and effective at acidic pH that prevails on tumor site. The material was biocompatible and effective in case of Saos-2 osteosarcoma cells with a viability of 0.4 ± 0.43 and 0.49 ± 0.53 in case of MG-63 cells when treated with highest concentration of 100 μl siRNA compared to untreated cells that were in range of 0.64 ± 0.67 in Saos-2 and 0.61 ± 0.63 in MG-63 cells. The results of expression of inflammatory cytokines IL-6, TGF-β & TNF-α showed negligible amount compared to control group serving the purpose of an effective carrier targeting tumor cells.

Keywords: Bone marrow derived macrophages; Chitosan; Graphene oxide; Inflammation; Osteosarcoma; RAW macrophages; siRNA.

MeSH terms

  • Animals
  • Bone Marrow / pathology*
  • Cell Death / drug effects
  • Cell Line, Tumor
  • Chitosan / chemistry*
  • Drug Liberation
  • Fluorescence
  • Gene Expression Regulation
  • Graphite / chemistry*
  • Humans
  • Hydrogen-Ion Concentration
  • Inflammation / pathology*
  • Kinetics
  • Macrophages / metabolism
  • Mice
  • Nanoparticles / chemistry*
  • Nanoparticles / ultrastructure
  • Proto-Oncogene Proteins c-bcl-2 / metabolism*
  • RNA, Small Interfering / metabolism*
  • Spectrum Analysis, Raman

Substances

  • BCL2 protein, human
  • Proto-Oncogene Proteins c-bcl-2
  • RNA, Small Interfering
  • graphene oxide
  • Graphite
  • Chitosan