Quantification of Endothelial αvβ3 Expression with High-Frequency Ultrasound and Targeted Microbubbles: In Vitro and In Vivo Studies.
Angiogenesis is a critical feature of plaque development in atherosclerosis and might play a key role in both the initiation and later rupture of plaques. The precursory molecular or cellular pro-angiogenic events that initiate plaque growth and that ultimately contribute to plaque instability, however, cannot be detected directly with any current diagnostic modality. This study was designed to investigate the feasibility of ultrasound molecular imaging of endothelial αvβ3 expression in vitro and in vivo using αvβ3-targeted ultrasound contrast agents (UCAs). In the in vitro study, αvβ3 expression was confirmed by immunofluorescence in a murine endothelial cell line and detected using the targeted UCA and ultrasound imaging at 18-MHz transmit frequency. In the in vivo study, expression of endothelial αvβ3 integrin in murine carotid artery vessels and microvessels of the salivary gland was quantified using targeted UCA and high-frequency ultrasound in seven animals. Our results indicated that endothelial αvβ3 expression was significantly higher in the carotid arterial wall containing atherosclerotic lesions than in arterial segments without any lesions. We also found that the salivary gland can be used as an internal positive control for successful binding of targeted UCA to αvβ3 integrin. In conclusion, αvβ3-targeted UCA allows non-invasive assessment of the expression levels of αvβ3 on the vascular endothelium and may provide potential insights into early atherosclerotic plaque detection and treatment monitoring.
Biomedical Engineering, Thorax Center, Erasmus MC, Rotterdam, The Netherlands. Electronic address: v.daeichin@erasmusmc.nl.
Verya
V
Daeichin
Biomedical Engineering, Thorax Center, Erasmus MC, Rotterdam, The Netherlands.
Klazina
K
Kooiman
Biomedical Engineering, Thorax Center, Erasmus MC, Rotterdam, The Netherlands.
Ilya
I
Skachkov
Biomedical Engineering, Thorax Center, Erasmus MC, Rotterdam, The Netherlands.
Johan G
JG
Bosch
Department of Pathology, Maastricht University Medical Center, Maastricht, The Netherlands.
Thomas L
TL
Theelen
Institute of Pathology, Technical University of Munich, Munich, Germany.
Katja
K
Steiger
VisualSonics Inc., Toronto, Ontario, Canada.
Andrew
A
Needles
Department of Pharmacology, MUMC, Maastricht, The Netherlands.
Ben J
BJ
Janssen
Department of Pathology, Academic Medical Center, Amsterdam, The Netherlands.
Mat J A P
MJ
Daemen
Biomedical Engineering, Thorax Center, Erasmus MC, Rotterdam, The Netherlands; Laboratory of Acoustical Wavefield Imaging, Faculty of Applied Sciences, Delft University of Technology, Delft, The Netherlands; Shenzhen Institute of Advanced Technologies, Shenzhen, China.
Antonius F W
AF
van der Steen
Biomedical Engineering, Thorax Center, Erasmus MC, Rotterdam, The Netherlands; Laboratory of Acoustical Wavefield Imaging, Faculty of Applied Sciences, Delft University of Technology, Delft, The Netherlands; Netherlands Heart Institute, Utrecht, The Netherlands.
Nico
N
de Jong
Department of Pathology, Maastricht University Medical Center, Maastricht, The Netherlands.
Judith C
JC
Sluimer
D000818:Animals; D003287:Contrast Media; D004730:Endothelium, Vascular / Q000000981:diagnostic imaging*; D007089:Image Enhancement / Q000379:methods*; D066298:In Vitro Techniques; D039302:Integrin alphaVbeta3 / Q000235:genetics / Q000378:metabolism*; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D045423:Microbubbles; D023421:Models, Animal; D014463:Ultrasonography / Q000379:methods*
D003287:Contrast Media; D039302:Integrin alphaVbeta3
Endothelial cell; High-frequency ultrasound; In vitro; In vivo; Molecular imaging; Murine atherosclerosis; Targeted microbubble; α(v)β(3) integrin