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Targeted cationic poly(D,L-lactic-co-glycolic acid) nanoparticles for gene delivery to cultured cells

Abstract

We developed a new targeted cationic nanoparticulate system composed of poly(D,L-lactic-co-glycolic acid) (PLGA), 1,2-dioleoyl-3-(trimethylammonium) propane (DOTAP) and asialofetuin (AF), and found it to be a highly effective formulation for gene delivery to liver tumor cells. The nanoparticles (NP) were prepared by a modified solvent evaporation process that used two protocols in order to encapsulate (NP1 particles) or adsorb (NP2 particles) plasmid DNA. The final particles are in the nanoscale range. pDNA loaded in PLGA/DOTAP/AF particles with high loading efficiency showed a positive surface charge. Targeted asialofetuin-nanoparticles (AF-NP) carrying genes encoding for luciferase and interleukin-12 (IL-12) resulted in increased transfection efficiencies compared to free DNA and to plain (non-targeted) systems, even in the presence of 60% fetal bovine serum (FBS). The results of transfections performed on HeLa cells, defective in asialoglycoprotein receptors (ASGPr-), confirmed the receptor-mediated endocytosis mechanism. In summary, this is the first time that asialoglycoprotein receptor targeting by PLGA/DOTAP/DNA nanoparticles carrying the therapeutic gene IL-12 has been shown to be efficient in gene delivery to liver cancer cells in the presence of a very high concentration of serum, and this could be a potential system for in vivo application.

Abbreviations

AF:

asialofetuin

ASGPr:

asialoglycoprotein receptor

DMEM-HG:

Dulbecco’s modified Eagle’s medium-high glucose

DOTAP:

1,2-dioleoyl-3-(trimethylammonium propane)

FBS:

fetal bovine serum

IL-12:

interleukin-12

NP:

nanoparticle

PLGA:

poly(D,L-lactic-co-glycolic acid

SEM:

scanning electron microscopy

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Correspondence to Conchita Tros de Ilarduya.

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Díez, S., Miguéliz, I. & Tros de Ilarduya, C. Targeted cationic poly(D,L-lactic-co-glycolic acid) nanoparticles for gene delivery to cultured cells. Cell Mol Biol Lett 14, 347–362 (2009). https://doi.org/10.2478/s11658-009-0003-7

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  • DOI: https://doi.org/10.2478/s11658-009-0003-7

Key words

  • Poly(D,L-lactic-co-glycolic acid) (PLGA)
  • 1,2-dioleoyl-3-(trimethylammonium) propane (DOTAP)
  • Asialofetuin
  • Targeted gene delivery
  • Pharmaceutical nanotechnology