title: Ultrasound-triggered release of recombinant tissue-type plasminogen activator from echogenic liposomes.
abstract: Echogenic liposomes (ELIP) were developed as ultrasound-triggered targeted drug or gene delivery vehicles (Lanza et al. 1997; Huang et al. 2001). Recombinant tissue-type plasminogen activator (rt-PA), a thrombolytic, has been loaded into ELIP (Tiukinhoy-Laing et al. 2007). These vesicles have the potential to be used for ultrasound-enhanced thrombolysis in the treatment of acute ischemic stroke, myocardial infarction, deep vein thrombosis or pulmonary embolus. A clinical diagnostic ultrasound scanner (Philips HDI 5000; Philips Medical Systems, Bothell, WA, USA) equipped with a linear array transducer (L12-5) was employed for in vitro studies using rt-PA-loaded ELIP (T-ELIP). The goal of this study was to quantify ultrasound-triggered drug release from rt-PA-loaded echogenic liposomes. T-ELIP samples were exposed to 6.9-MHz B-mode pulses at a low pressure amplitude (600 kPa) to track the echogenicity over time under four experimental conditions: (1) flow alone to monitor gas diffusion from the T-ELIP, (2) pulsed 6.0-MHz color Doppler exposure above the acoustically driven threshold (0.8 MPa) to force gas out of the liposome gently, (3) pulsed 6.0-MHz color Doppler above the rapid fragmentation threshold (2.6 MPa) or (4) Triton X-100 to rupture the T-ELIP chemically as a positive control. Release of rt-PA for each ultrasound exposure protocol was assayed spectrophotometrically. T-ELIP were echogenic in the flow model (5 mL/min) for 30 min. The thrombolytic drug remained associated with the liposome when exposed to low-amplitude B-mode pulses over 60 min and was released when exposed to color Doppler pulses or Triton X-100. The rt-PA released from the liposomes had similar enzymatic activity as the free drug. These T-ELIP are robust and echogenic during continuous fundamental 6.9-MHz B-mode imaging at a low exposure output level (600 kPa). Furthermore, a therapeutic concentration of rt-PA can be released by fragmenting the T-ELIP with pulsed 6.0-MHz color Doppler ultrasound above the rapid fragmentation threshold (1.59 MPa). (E-mail: denise.smith@uc.edu).
authors.affiliation: Department of Biomedical Engineering, University of Cincinnati, Cincinnati, OH, USA. denise.smith@uc.edu
authors.forename: Denise A B
authors.initials: DA
authors.lastname: Smith
authors.forename: Sampada S
authors.initials: SS
authors.lastname: Vaidya
authors.forename: Jonathan A
authors.initials: JA
authors.lastname: Kopechek
authors.forename: Shao-Ling
authors.initials: SL
authors.lastname: Huang
authors.forename: Melvin E
authors.initials: ME
authors.lastname: Klegerman
authors.forename: David D
authors.initials: DD
authors.lastname: McPherson
authors.forename: Christy K
authors.initials: CK
authors.lastname: Holland
mesh_terms: D000818:Animals; D002417:Cattle; D003287:Contrast Media; D004058:Diffusion; D005343:Fibrinolytic Agents / Q000008:administration & dosage*; D008081:Liposomes / Q000737:chemistry; D011994:Recombinant Proteins / Q000008:administration & dosage; D012710:Serum Albumin, Bovine; D015912:Thrombolytic Therapy / Q000379:methods*; D010959:Tissue Plasminogen Activator / Q000008:administration & dosage*; D014465:Ultrasonics; D018615:Ultrasonography, Doppler, Color; D018617:Ultrasonography, Doppler, Pulsed
chemical_list: D003287:Contrast Media; D005343:Fibrinolytic Agents; D008081:Liposomes; D011994:Recombinant Proteins; D012710:Serum Albumin, Bovine; D010959:Tissue Plasminogen Activator