title: Shock waves and DNA-cationic lipid assemblies: a synergistic approach to express exogenous genes in human cells.
abstract: Cationic lipid/DNA complexes (lipoplexes) represent a powerful tool for cell transfection; however, their use is still limited by important concerns, including toxicity and poor internalization into deep tissues. In this work, we investigated the use of shock wave-induced acoustic cavitation in vitro for the transfection of lipoplexes in human embryo kidney 293 cells. We selected shock waves with the ability to internalize 10-kDa fluorescein isothiocyanate-dextran into cells while maintaining survival rates above 50%. Cell transfection was tested using the green fluorescent protein-encoding plasmid pCX::GFPGPI2. Confocal microscopy and fluorescence-assisted cell sorting analyses revealed successful transfection after treatments ranging from 1 to 3 min using 60 to 180 shock waves at peak amplitudes of 12.3 ± 1.5 MPa. Interestingly, the combination of shock waves and lipoplexes induced a 3.1- and 3.8-fold increase in the expression of the reporter gene compared with the use of lipoplexes or shock waves alone, respectively. These results indicate that cationic DNA assembly and shock waves act in a synergistic manner to promote transfection of human cells, revealing a potential approach for non-invasive site-specific gene therapy.
authors.affiliation: Centro de Física Aplicada y Tecnología Avanzada, Universidad Nacional Autónoma de México, Campus Juriquilla, Querétaro, México.
authors.forename: Blanca
authors.initials: B
authors.lastname: Millán-Chiu
authors.affiliation: Centro de Física Aplicada y Tecnología Avanzada, Universidad Nacional Autónoma de México, Campus Juriquilla, Querétaro, México.
authors.forename: Giselle
authors.initials: G
authors.lastname: Camacho
authors.affiliation: Instituto de Neurobiología, Universidad Nacional Autónoma de México, Campus Juriquilla, Querétaro, México.
authors.forename: Alfredo
authors.initials: A
authors.lastname: Varela-Echavarría
authors.affiliation: Instituto de Neurobiología, Universidad Nacional Autónoma de México, Campus Juriquilla, Querétaro, México.
authors.forename: Elisa
authors.initials: E
authors.lastname: Tamariz
authors.affiliation: Centro de Física Aplicada y Tecnología Avanzada, Universidad Nacional Autónoma de México, Campus Juriquilla, Querétaro, México.
authors.forename: Francisco
authors.initials: F
authors.lastname: Fernández
authors.affiliation: Centro de Física Aplicada y Tecnología Avanzada, Universidad Nacional Autónoma de México, Campus Juriquilla, Querétaro, México. Electronic address: lmlm@unam.mx.
authors.forename: Luz M
authors.initials: LM
authors.lastname: López-Marín
authors.affiliation: Centro de Física Aplicada y Tecnología Avanzada, Universidad Nacional Autónoma de México, Campus Juriquilla, Querétaro, México.
authors.forename: Achim M
authors.initials: AM
authors.lastname: Loske
mesh_terms: D002412:Cations; D002463:Cell Membrane Permeability / Q000528:radiation effects*; D004247:DNA / Q000008:administration & dosage / Q000235:genetics*; D018274:Electroporation / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000008:administration & dosage / Q000235:genetics*; D057809:HEK293 Cells; D019220:High-Energy Shock Waves; D006801:Humans; D008081:Liposomes / Q000737:chemistry* / Q000528:radiation effects*; D013010:Sonication / Q000379:methods; D014162:Transfection / Q000379:methods*
chemical_list: D002412:Cations; D008081:Liposomes; D049452:Green Fluorescent Proteins; D004247:DNA
keywords: Acoustic cavitation; Cell transfection; HEK 293 cells; Lipoplexes; Shock waves