Understanding the mechanism of protamine in solid lipid nanoparticle-based lipofection: the importance of the entry pathway

Eur J Pharm Biopharm. 2011 Nov;79(3):495-502. doi: 10.1016/j.ejpb.2011.06.005. Epub 2011 Jun 25.

Abstract

The aim of our study was to evaluate the effect of protamine on the transfection capacity of solid lipid nanoparticles (SLNs) by correlating it to the internalization mechanisms and intracellular trafficking of the vectors. Vectors were prepared with SLN, DNA, and protamine. ARPE-19 and HEK-293 cells were used for the evaluation of the formulations. Protamine induced a 6-fold increase in the transfection of SLNs in retinal cells due to the presence of nuclear localization signals (NLS), its protection capacity, and a shift in the internalization mechanism from caveolae/raft-mediated to clathrin-mediated endocytosis. However, protamine produced an almost complete inhibition of transfection in HEK-293 cells. In spite of the high DNA condensation capacity of protamine and its content in NLS, this does not always lead to an improvement in cell transfection since it may impair some of the limiting steps of the transfection processes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Survival / drug effects
  • DNA / administration & dosage
  • DNA / genetics
  • DNA / metabolism
  • Drug Carriers / chemistry*
  • Drug Carriers / toxicity
  • Electrophoresis, Polyacrylamide Gel
  • Endocytosis
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism
  • Flow Cytometry
  • Green Fluorescent Proteins / genetics
  • HEK293 Cells
  • Humans
  • Lipids / chemistry*
  • Lipids / toxicity
  • Microscopy, Fluorescence
  • Nanoparticles* / chemistry
  • Particle Size
  • Plasmids
  • Protamines* / chemistry
  • Protamines* / metabolism
  • Protein Binding
  • Retinal Pigment Epithelium / cytology
  • Retinal Pigment Epithelium / drug effects
  • Retinal Pigment Epithelium / metabolism
  • Surface Properties
  • Transfection / methods*

Substances

  • Drug Carriers
  • Lipids
  • Protamines
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • DNA