Intracellular trafficking and therapeutic outcome of multiwalled carbon nanotubes modified with cyclodextrins and polyethylenimine

Colloids Surf B Biointerfaces. 2018 Mar 1:163:55-63. doi: 10.1016/j.colsurfb.2017.12.028. Epub 2017 Dec 15.

Abstract

Functionalized carbon nanotubes (CNTs) have been proposed in the last years as vectors for delivery of biomolecules, proteins and DNA into various cells. In this study, a new multiwalled carbon nanotube β-cyclodextrin platform (MWCNT-CD) modified with branched polyethylenimine (PEI) and doped with Rhodamine (Rhod), MWCNT-CD-PEI-Rhod, was designed and investigated as drug delivery system. The drug binding abilities of MWCNT-CD-PEI-Rhod towards Cidofovir (Cid) and DNA plasmid encoding enhanced green fluorescence protein (pCMS-EGFP) were investigated by complementary spectroscopic techniques. MWCNT-CD-PEI-Rhod showed no significative cytotoxicity and an excellent ability to entrap and delivery Cid. The present study broadens the spectrum of biological evaluation by investigating platform-treatment induced cellular response such as antiviral activity, transfection properties, cellular uptake, internalization mechanisms and cellular localization. The mechanism of cellular uptake was elucidated monitoring the dependence of intracellular red fluorescence from the assembly concentration, time and presence of specific uptake inhibitors. The biological results indicated that MWCNT-CD-PEI-Rhod loaded with Cid and/or pCMS-EGFP crossed the cell membrane via clathrin-dependent pathway and co-localized in lysosomal compartment. However, no green fluorescent protein expression of pCMS-EGFP was detected, whereas the efficient escape of Cid from lysosome and the release close to nuclear region prompted the antiviral activity.

Keywords: Cellular response; Cellular uptake; Cidofovir; Multiwalled carbon nanotubes; Polyethylenimine; Spectroscopic properties; Transfection properties; β-cyclodextrin.

MeSH terms

  • Animals
  • Antiviral Agents / chemistry
  • Antiviral Agents / pharmacology*
  • Biological Transport
  • Chlorocebus aethiops
  • Cidofovir
  • Cytomegalovirus / drug effects
  • Cytomegalovirus / growth & development
  • Cytosine / analogs & derivatives*
  • Cytosine / chemistry
  • Cytosine / pharmacology
  • Drug Carriers*
  • Drug Liberation
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Kinetics
  • Lysosomes / drug effects
  • Lysosomes / metabolism
  • Nanotubes, Carbon / chemistry*
  • Organophosphonates / chemistry
  • Organophosphonates / pharmacology*
  • Plasmids / chemistry
  • Plasmids / metabolism
  • Polyethyleneimine / chemistry*
  • Rhodamines / chemistry
  • Rhodamines / metabolism
  • Vero Cells
  • beta-Cyclodextrins / chemistry*

Substances

  • Antiviral Agents
  • Drug Carriers
  • Nanotubes, Carbon
  • Organophosphonates
  • Rhodamines
  • beta-Cyclodextrins
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Cytosine
  • Polyethyleneimine
  • Cidofovir
  • betadex