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Research Article

Transfection of primary human nasal epithelial cells using a biodegradable poly (ester amine) based on polycaprolactone and polyethylenimine as a gene carrier

, , , , , & show all
Pages 684-690 | Received 27 Mar 2007, Accepted 28 Jul 2007, Published online: 08 Oct 2008
 

Abstract

The purpose of this study was to prepare and characterize poly (ester amine) (PEA)/pGL3 complexes and investigate their transfection efficiency in human nasal epithelial (HNE) cells. Particle size, zeta potential, and gel retardation characteristics of PEA /pGL3 complexes were also measured. After treatment of DNase-I, protection and release assay of PEA/pGL3 complexes were performed. To assess the transfection efficiency and cytotoxicity, measurement of relative luciferase activity and MTS assay were performed. PEA/pGL3 complexes showed effective and stable DNA condensation with the particle sizes below 200 nm, implicating their potential for intracellular delivery. PEA/pGL3 complexes successfully transfected into the HNE cells with higher viability of the cells. These results suggested that, the PEA can be used as an efficient cationic polymeric vehicle which provides a versatile platform for further investigation of structure property relationship along with the controlled degradation, significant low cytotoxicity, and high transfection efficiency of the primary HNE cells.

Abbreviations
BEGM=

bronchial epithelial growth medium

CF=

cystic fibrosis

CFTR=

cystic fibrosis transmembrane conductance regulator

EtBr=

ethidium bromide

GPC-MALLS=

gel permeation chromatography with multi angle laser light scattering

HNE cell=

human nasal epithelial cell

1H-NMR=

1H-nuclear magnetic resonance

PCL=

polycaprolactone

PCLDA=

polycaprolactone diacrylate

PEA=

poly (ester amine)

PEI=

polyethylenimine

RLU=

relative light units

SDS=

sodium dodecyl sulfate

TAE=

Tris-acetate

Abbreviations
BEGM=

bronchial epithelial growth medium

CF=

cystic fibrosis

CFTR=

cystic fibrosis transmembrane conductance regulator

EtBr=

ethidium bromide

GPC-MALLS=

gel permeation chromatography with multi angle laser light scattering

HNE cell=

human nasal epithelial cell

1H-NMR=

1H-nuclear magnetic resonance

PCL=

polycaprolactone

PCLDA=

polycaprolactone diacrylate

PEA=

poly (ester amine)

PEI=

polyethylenimine

RLU=

relative light units

SDS=

sodium dodecyl sulfate

TAE=

Tris-acetate

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