Gene delivery in vitro and in vivo from bioreducible multilayered polyelectrolyte films of plasmid DNA

Jenifer Blacklock, Ye Zi You, Qing Hui Zhou, Guangzhao Mao, David Oupický

Research output: Contribution to journalArticle

98 Citations (Scopus)

Abstract

Layer-by-layer (LbL) films were assembled on flexible stainless steel substrate using plasmid DNA and reducible hyperbranched poly(amido amine) (RHB) polycation. The films were characterized by XPS and their disassembly in reducing conditions confirmed by ellipsometry. Fibroblast and smooth muscle cell attachment and proliferation on DNA/RHB films were indistinguishable from those on control DNA/poly(ethylenimine) (PEI) films. In vitro transfection activity was evaluated using reporter plasmids encoding for secreted alkaline phosphatase (SEAP) and green fluorescent protein (GFP). DNA/RHB films showed higher and longer lasting transfection activity than control DNA/PEI films using SEAP plasmid. It was revealed through the use of GFP plasmid that DNA/RHB films transfected almost the entire cell population growing on the films. In vivo transfection activity was evaluated by subcutaneously implanting a stainless steel substrate coated with the DNA/RHB films containing SEAP plasmid DNA and measuring the levels of SEAP secreted into the blood circulation of rats. It was found that the plasma levels of SEAP peaked at ∼160 ng SEAP/mL five days post-implantation.

Original languageEnglish (US)
Pages (from-to)939-950
Number of pages12
JournalBiomaterials
Volume30
Issue number5
DOIs
StatePublished - Feb 1 2009

Fingerprint

Polyelectrolytes
Plasmids
DNA
Genes
Phosphatases
Alkaline Phosphatase
Transfection
Stainless Steel
Polyetherimides
Green Fluorescent Proteins
Stainless steel
Cells
Proteins
In Vitro Techniques
Blood Circulation
Ellipsometry
Hemodynamics
Substrates
Fibroblasts
Smooth Muscle Myocytes

Keywords

  • DNA
  • In vivo transfection
  • LBL
  • Layer-by-layer
  • Polyelectrolyte films

ASJC Scopus subject areas

  • Bioengineering
  • Ceramics and Composites
  • Biophysics
  • Biomaterials
  • Mechanics of Materials

Cite this

Gene delivery in vitro and in vivo from bioreducible multilayered polyelectrolyte films of plasmid DNA. / Blacklock, Jenifer; You, Ye Zi; Zhou, Qing Hui; Mao, Guangzhao; Oupický, David.

In: Biomaterials, Vol. 30, No. 5, 01.02.2009, p. 939-950.

Research output: Contribution to journalArticle

Blacklock, Jenifer ; You, Ye Zi ; Zhou, Qing Hui ; Mao, Guangzhao ; Oupický, David. / Gene delivery in vitro and in vivo from bioreducible multilayered polyelectrolyte films of plasmid DNA. In: Biomaterials. 2009 ; Vol. 30, No. 5. pp. 939-950.
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