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Pharmaceutical Research

, 36:14 | Cite as

Delivery of pDNA Polyplexes to Bronchial and Alveolar Epithelial Cells Using a Mesh Nebulizer

  • Larissa Gomes dos Reis
  • Maree Svolos
  • Lyn M Moir
  • Rima Jaber
  • Norbert Windhab
  • Paul M Young
  • Daniela Traini
Research Paper
  • 179 Downloads

Abstract

Purpose

In this study, a cell penetrating peptide was used as an uptake enhancer for pDNA delivery to the lungs.

Methods

Polyplexes were prepared between pDNA and CPP. Intracellular delivery of pDNA was assessed in both alveolar (A549) and bronchial (Calu-3) epithelial cells. Aerosol delivery was investigated using a mesh nebulizer.

Results

Efficient intracellular delivery of pDNA occurs in both A549 and Calu-3 cells when delivered as polyplexes. Protection against nucleases and endosomal escape mechanism occurs when pDNA is formulated within the polyplexes. For aerosol delivery, 1% (w/v) mannitol was able to protect naked DNA structure during nebulization with a significant increase in fine particle fraction (particles <5 μm). The structure of polyplexes when delivered via a mesh nebulizer using 1% (w/v) mannitol could partially withstand the shear forces involved in aerosolization. Although some loss in functionality occurred after nebulization, membrane-associated fluorescence was observed in A549 cells. In Calu-3 cells mucus entrapment was a limiting factor for polyplex delivery.

Conclusions

The presence of CPP is essential for efficient intracellular delivery of pDNA. The polyplexes can be delivered to lung epithelial cells using mesh nebulizer. The use of different excipients is essential for further optimization of these delivery systems.

Key Words

aerosol cell-penetrating peptide gene delivery lung nebulization 

Abbreviations

CPP

Cell-penetrating peptide

DLS

Dynamic light scattering

dsDNA

Double stranded DNA

ED

Emitted dose

eGFP

Enhanced green fluorescent protein

EMSA

Electrophoretic Mobility Shift Assay

FPF

Fine particle fraction

N:P

Nitrogen:phosphate

NGI

Next Generation Impactor

PBS

Phosphate buffered saline

PDI

Polydispersity index

pDNA

Plasmid DNA

ROI

Regions of Interest

scDNA

Supercoiled DNA

SEM

Scanning electron microscopy

TEER

Transepithelial Electrical Resistance

TSI

Twin Stage Impinger

Notes

Acknowledgments and Disclosures

This work was supported by an Australian Research Council Linkage Grant LP140100181. Larissa Gomes dos Reis is financially supported by the Brazilian Research Funding CAPES.

Supplementary material

11095_2018_2542_MOESM1_ESM.docx (19 kb)
ESM 1 (DOCX 19.2 kb)
11095_2018_2542_MOESM2_ESM.docx (21 kb)
ESM 2 (DOCX 20.6 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Larissa Gomes dos Reis
    • 1
  • Maree Svolos
    • 1
  • Lyn M Moir
    • 1
  • Rima Jaber
    • 2
  • Norbert Windhab
    • 2
  • Paul M Young
    • 1
  • Daniela Traini
    • 1
  1. 1.Respiratory Technology, Woolcock Institute of Medical Research and Discipline of Pharmacology, Sydney Medical SchoolSydney UniversityCamperdownAustralia
  2. 2.Evonik Industries AGDarmstadtGermany

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