Pharmaceutical Research

, Volume 33, Issue 2, pp 384–394 | Cite as

Aerosolized Non-viral Nucleic Acid Delivery in the Vaginal Tract of Pigs

  • Katrien Remaut
  • Evelien De Clercq
  • Oliwia Andries
  • Koen Rombouts
  • Matthias Van Gils
  • Laetitia Cicchelero
  • Ian Vandenbussche
  • Sarah Van Praet
  • Juan Manuel Benito
  • José Manuel Garcia Fernandéz
  • Niek Sanders
  • Daisy Vanrompay
Research Paper



The human pathogen Chlamydia trachomatis is worldwide the leading cause of bacterial sexually transmitted disease. Nasal or vaginal nucleic acid vaccination is a promising strategy for controlling genital Chlamydia trachomatis infections. Since naked nucleic acids are generally not efficiently taken up by cells, they are often complexed with carriers that facilitate their intracellular delivery.


In the current study, we screened a variety of commonly used non-viral gene delivery carriers for their ability to transfect newborn pig tracheal cells. The effect of aerosolization on the physicochemical properties and transfection efficiency of the complexes was also evaluated in vitro. Subsequently, a pilot experiment was performed in which the selected complexes were aerosolized in the vaginal tract of pigs.


Both mRNA and pDNA containing lipofectamine and ADM70 complexes showed promise for protein expression in vitro, before and after aerosolization. In vivo, only lipofectamine/pDNA complexes resulted in high protein expression levels 24 h following aerosolization. This correlates to the unexpected observation that the presence of vaginal mucus increases the efficiency of lipofectamine/pDNA complexes 3-fold, while the efficiency of lipofectamine/mRNA complexes and ADM70/mRNA and ADM70/pDNA complexes decreased.


As aerosolization was an easy and effective method to deliver complexes to the vaginal tract of pigs, we believe this application technique has future potential for both vaginal and perhaps nasal vaccination using non-viral gene delivery vectors.

Key words

plasmid DNA mRNA aerosolization vaginal administration non-viral gene delivery complexes 





Dynamic light scattering


Dulbecco’s modified Eagle’s medium










Fetal calf serum


Green fluorescent protein


Genzyme lipid 67




3-(4,5-dimethylthiazole-2yl)-2,5-diphenyl tetrazolium bromide


Newborn pig tracheal


Polyethylene glycol




Regions of interest

Supplementary material

11095_2015_1796_MOESM1_ESM.jpg (1.1 mb)
Suppl Fig 1Cell viability. NPTr cells were treated with different mRNA/carrier complexes as depicted in the x-axis. Blanc represents non-transfected cells and DMSO served as positive control. * Significantly different from non-treated cells (blanc) (P ≤ 0.05). (JPG 1.12 mb)


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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Katrien Remaut
    • 1
  • Evelien De Clercq
    • 2
  • Oliwia Andries
    • 3
  • Koen Rombouts
    • 1
  • Matthias Van Gils
    • 2
  • Laetitia Cicchelero
    • 3
  • Ian Vandenbussche
    • 1
  • Sarah Van Praet
    • 1
  • Juan Manuel Benito
    • 4
  • José Manuel Garcia Fernandéz
    • 4
  • Niek Sanders
    • 3
  • Daisy Vanrompay
    • 2
    • 5
  1. 1.Laboratory of General Biochemistry and Physical PharmacyGhent UniversityGhentBelgium
  2. 2.Laboratory of Immunology and Animal Biotechnology, Department of Molecular Biotechnology, Faculty of Bioscience EngineeringGhent UniversityGhentBelgium
  3. 3.Laboratory of Gene Therapy, Department of Nutrition, Genetics and Ethology, Faculty of Veterinary MedicineGhent UniversityMerelbekeBelgium
  4. 4.Institute for Chemical Research, CSICUniversity of SevillaSevillaSpain
  5. 5.Department of Animal Production, Faculty of Bioscience EngineeringGhent UniversityGhentBelgium

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