title: In Situ Transfection by Controlled Release of Lipoplexes Using Acoustic Droplet Vaporization.
abstract: Localized delivery of nucleic acids to target sites (e.g., diseased tissue) is critical for safe and efficacious gene therapy. An ultrasound-based technique termed acoustic droplet vaporization (ADV) has been used to spatiotemporally control the release of therapeutic small molecules and proteins contained within sonosensitive emulsions. Here, ADV is used to control the release of lipoplex-containing plasmid DNA encoding an enhanced green fluorescent protein reporter-from a sonosensitive emulsion. Focused ultrasound (3.5 MHz, mechanical index (MI) ≥ 1.5) generates robust release of fluorescein (i.e., surrogate payload) and lipoplex from the emulsion. In situ release of the lipoplex from the emulsion using ADV (MI = 1.5, 30 cycles) yields a 55% release efficiency, resulting in 43% transfection efficiency and 95% viability with C3H/10T1/2 cells. Without exposure to ultrasound, the release and transfection efficiencies are 5% and 7%, respectively, with 99% viability. Lipoplex released by ADV retains its bioactivity while the ADV process does not yield any measureable sonoporative enhancement of transfection. Co-encapsulation of Ficoll PM 400 within the lipoplex-loaded emulsion, and its subsequent release using ADV, yield higher transfection efficiency than the lipoplex alone. The results demonstrate that ADV can have utility in the spatiotemporal control of gene delivery.
authors.affiliation: Department of Radiology, University of Michigan Health System, Ann Arbor, MI, 48109, USA.
authors.forename: Benjamin A
authors.initials: BA
authors.lastname: Juliar
authors.affiliation: Department of Radiology, University of Michigan Health System, Ann Arbor, MI, 48109, USA.
authors.forename: Melissa M
authors.initials: MM
authors.lastname: Bromley
authors.affiliation: Department of Radiology, University of Michigan Health System, Ann Arbor, MI, 48109, USA.|Applied Physics Program, University of Michigan, Ann Arbor, MI, 48109, USA.
authors.forename: Alexander
authors.initials: A
authors.lastname: Moncion
authors.affiliation: Department of Radiology, University of Michigan Health System, Ann Arbor, MI, 48109, USA.
authors.forename: Denise C
authors.initials: DC
authors.lastname: Jones
authors.affiliation: Department of Radiology, University of Michigan Health System, Ann Arbor, MI, 48109, USA.
authors.forename: Eric G
authors.initials: EG
authors.lastname: O'Neill
authors.affiliation: School of Dentistry, University of Michigan, Ann Arbor, MI, 48109, USA.
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Wilson
authors.affiliation: School of Dentistry, University of Michigan, Ann Arbor, MI, 48109, USA.|Department of Biological Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA.|Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA.
authors.forename: Renny T
authors.initials: RT
authors.lastname: Franceschi
authors.affiliation: Department of Radiology, University of Michigan Health System, Ann Arbor, MI, 48109, USA.|Applied Physics Program, University of Michigan, Ann Arbor, MI, 48109, USA.
authors.forename: Mario L
authors.initials: ML
authors.lastname: Fabiilli
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D039902:Multipotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D010957:Plasmids / Q000737:chemistry / Q000235:genetics; D014162:Transfection / Q000379:methods*; D000069453:Ultrasonic Waves
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: acoustic droplet vaporization; emulsion; gene delivery; lipoplex; perfluorocarbon; ultrasound