Enhancement of vascular permeability with low-frequency contrast-enhanced ultrasound in the chorioallantoic membrane model.
OBJECTIVE
To characterize the effect of low-frequency contrast material-enhanced ultrasound on the vascular endothelium and to determine the parameters and techniques required to deliver a therapeutic agent by using the chorioallantoic membrane (CAM) model.


METHODS
All in vivo animal procedures were conducted with institutional Animal Care and Use Committee approval. Extravasation of 8.5-nm-diameter fluorescein isothiocyanate-labeled dextran was evaluated in the vasculature of a chick CAM model. Intravital microscopy was performed during contrast-enhanced ultrasound exposure (1.00 or 2.25 MHz); results were compared with results of electron microscopy of the insonated regions. Data acquired after insonation with greater mechanical stress (n = 30 animals) (mechanical index [MI] > 1.3) and with lower mechanical stress (n = 86 animals) (MI < 1.13) were compared with measurements in control conditions (n = 46 animals). The diameter of affected vessels; number of extravasation sites; extravasation rate, area, and location; and changes in endothelial cells and basement membrane were evaluated. Differences were tested with analysis of variance or the Student t test.


RESULTS
After ultrasound application, convective transport of the model drug was observed through micron-sized openings with a mean fluid velocity of 188.6 microm/sec in the low-stress class and 362.5 microm/sec in the high-stress class. Electron microscopy revealed micron-sized focal endothelial gaps and disseminated blebs, vacuoles, and filopodia extending across tens of microns. The threshold pressure for extravasation was 0.5 MPa for a transmitted center frequency of 1.00 MHz (MI = 0.5) and 1.6 MPa for a frequency of 2.25 MHz (MI = 1.06); thus, the frequency dependence of the threshold was not predicted simply by the MI.


CONCLUSIONS
Low-frequency contrast-enhanced ultrasound can increase vascular permeability and result in convective extravasation of an 8.5-nm-diameter model drug.
Stieger
Susanne M
SM
Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California, Davis, CA 95616, USA. smstieger@ucdavis.edu
Caskey
Charles F
CF
Adamson
Roger H
RH
Qin
Shengping
S
Curry
Fitz-Roy E
FR
Wisner
Erik R
ER
Ferrara
Katherine W
KW
D000818:Animals; D001692:Biological Transport; D002199:Capillary Permeability / Q000502:physiology; D002642:Chick Embryo; D049033:Chorioallantoic Membrane / Q000098:blood supply / Q000000981:diagnostic imaging*; D003287:Contrast Media / Q000493:pharmacokinetics* / Q000494:pharmacology; D003911:Dextrans / Q000493:pharmacokinetics* / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000000981:diagnostic imaging* / Q000187:drug effects; D005119:Extravasation of Diagnostic and Therapeutic Materials; D020342:Finite Element Analysis; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives* / Q000493:pharmacokinetics / Q000494:pharmacology; D045423:Microbubbles; D008854:Microscopy, Electron; D023421:Models, Animal; D011312:Pressure; D013314:Stress, Mechanical; D014463:Ultrasonography / Q000009:adverse effects
D003287:Contrast Media; D003911:Dextrans; C015219:fluorescein isothiocyanate dextran; D016650:Fluorescein-5-isothiocyanate