title: Aerosolized Non-viral Nucleic Acid Delivery in the Vaginal Tract of Pigs.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Laboratory of General Biochemistry and Physical Pharmacy, Ghent University, Ottergemsesteenweg 460, B-9000, Ghent, Belgium.
authors.forename: Katrien
authors.initials: K
authors.lastname: Remaut
authors.affiliation: Laboratory of Immunology and Animal Biotechnology, Department of Molecular Biotechnology, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, Ghent, B-9000, Belgium.
authors.forename: Evelien
authors.initials: E
authors.lastname: De Clercq
authors.affiliation: Laboratory of Gene Therapy, Department of Nutrition, Genetics and Ethology, Faculty of Veterinary Medicine, Ghent University, Heidestraat 19, B-9820, Merelbeke, Belgium.
authors.forename: Oliwia
authors.initials: O
authors.lastname: Andries
authors.affiliation: Laboratory of General Biochemistry and Physical Pharmacy, Ghent University, Ottergemsesteenweg 460, B-9000, Ghent, Belgium.
authors.forename: Koen
authors.initials: K
authors.lastname: Rombouts
authors.affiliation: Laboratory of Immunology and Animal Biotechnology, Department of Molecular Biotechnology, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, Ghent, B-9000, Belgium.
authors.forename: Matthias
authors.initials: M
authors.lastname: Van Gils
authors.affiliation: Laboratory of Gene Therapy, Department of Nutrition, Genetics and Ethology, Faculty of Veterinary Medicine, Ghent University, Heidestraat 19, B-9820, Merelbeke, Belgium.
authors.forename: Laetitia
authors.initials: L
authors.lastname: Cicchelero
authors.affiliation: Laboratory of General Biochemistry and Physical Pharmacy, Ghent University, Ottergemsesteenweg 460, B-9000, Ghent, Belgium.
authors.forename: Ian
authors.initials: I
authors.lastname: Vandenbussche
authors.affiliation: Laboratory of General Biochemistry and Physical Pharmacy, Ghent University, Ottergemsesteenweg 460, B-9000, Ghent, Belgium.
authors.forename: Sarah
authors.initials: S
authors.lastname: Van Praet
authors.affiliation: Institute for Chemical Research, CSIC, University of Sevilla, Americo Vespucio 49, Isla de la Cartuja, E-41092, Sevilla, Spain.
authors.forename: Juan Manuel
authors.initials: JM
authors.lastname: Benito
authors.affiliation: Institute for Chemical Research, CSIC, University of Sevilla, Americo Vespucio 49, Isla de la Cartuja, E-41092, Sevilla, Spain.
authors.forename: José Manuel Garcia
authors.initials: JM
authors.lastname: Fernandéz
authors.affiliation: Laboratory of Gene Therapy, Department of Nutrition, Genetics and Ethology, Faculty of Veterinary Medicine, Ghent University, Heidestraat 19, B-9820, Merelbeke, Belgium.
authors.forename: Niek
authors.initials: N
authors.lastname: Sanders
authors.affiliation: Laboratory of Immunology and Animal Biotechnology, Department of Molecular Biotechnology, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, Ghent, B-9000, Belgium. Daisy.Vanrompay@ugent.be.|Department of Animal Production, Faculty of Bioscience Engineering, Ghent University, Coupure links, 653, B-9000, Ghent, Belgium. Daisy.Vanrompay@ugent.be.
authors.forename: Daisy
authors.initials: D
authors.lastname: Vanrompay
mesh_terms: D000336:Aerosols / Q000737:chemistry; D000818:Animals; D002460:Cell Line; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics; D004337:Drug Carriers / Q000737:chemistry; D005260:Female; D018014:Gene Transfer Techniques; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D012333:RNA, Messenger / Q000008:administration & dosage* / Q000235:genetics; D013552:Swine; D014162:Transfection; D014621:Vagina / Q000378:metabolism*
chemical_list: D000336:Aerosols; D004337:Drug Carriers; D012333:RNA, Messenger; D004247:DNA
keywords: aerosolization; mRNA; non-viral gene delivery complexes; plasmid DNA; vaginal administration