Molecular Imaging with Targeted Ultrasound Contrast Microbubbles

  • A. L. Klibanov
Part of the Ernst Schering Research Foundation Workshop book series (SCHERING FOUND, volume 49)

10.6 Conclusions

Microbubbles were prepared from insoluble perfluorocarbon gas, stabilized with a monolayer of phospholipid with a grafted PEG brush and decorated with targeting ligands, such as monoclonal antibodies or peptides on a flexible PEG spacer arm. Such targeted bubbles selectively and firmly bind to the surfaces coated with the specific receptors in vitro. Bubble deposition areas can be detected by ultrasound imaging. Single bubble detection sensitivity is possible.

Targeting of microbubbles to areas of inflammation, ischemia-reperfusion injury, and angiogenesis (including tumors) was achieved in vivo in experimental animal studies, by the use of microbubbles directed towards P-selectin or α v/β 3, respectively. Selective accumulation of ligand-carrying bubbles in the tissues of interest was confirmed by fluorescence microscopy (intravital and confocal) and ultrasound imaging.


Bubble Surface Ultrasound Contrast Agent Myocardial Contrast Echocardiography Ultrasound Contrast Material Contrast Particle 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


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  1. Alkan-Onyuksel H, Demos SM, Lanza GM, Vonesh MJ, Klegerman ME, Kane BJ, Kuszak J, McPherson DD (1996) Development of inherently echogenic liposomes as an ultrasonic contrast agent. J Pharm Sci 85:486–490PubMedCrossRefGoogle Scholar
  2. Andre MP, Steinbach G, Mattrey RF (1993) Enhancement of the echogenicity of flowing blood by the contrast agent perflubron. Invest Radiol 28:502–506PubMedCrossRefGoogle Scholar
  3. Borden MA, Longo ML (2002) Dissolution behavior of lipid-monolayer-coated, air-filled microbubbles: effect of lipid hydrophobic chain length. Langmuir 18:9225–9233CrossRefGoogle Scholar
  4. Brooks PC, Clark RA, Cheresh DA (1994) Requirement of vascular integrin alpha(v)beta3 for angiogenesis. Science 264:569–571PubMedCrossRefGoogle Scholar
  5. Christiansen JP, Leong-Poi H, Klibanov AL, Kaul S, Lindner JR (2002) Noninvasive imaging of myocardial reperfusion injury using leukocyte-targeted contrast echocardiography. Circulation 105:1764–1767PubMedCrossRefGoogle Scholar
  6. Dill-Macky MJ, Burns PN, Khalili K, Wilson SR (2002) Focal hepatic masses: enhancement patterns with SH U 508A and pulse-inversion US. Radiology 222:95–102PubMedCrossRefGoogle Scholar
  7. Ellegala DB, Leong-Poi H, Carpenter JE, Klibanov AL, Kaul S, Shaffrey ME, Sklenar J, Lindner JR (2003) Imaging tumor angiogenesis with contrast ultrasound and microbubbles targeted to alpha(v)beta3. Circulation 108:336–341PubMedCrossRefGoogle Scholar
  8. Epstein PS, Plesset MS (1950) On the stability of gas bubbles in liquid-gas solutions. J Chem Phys 18:1505–1509CrossRefGoogle Scholar
  9. Fisher NG, Christiansen JP, Klibanov AL, Taylor RP, Kaul S, Lindner JR (2002) Influence of microbubble surface charge on capillary transit and myocardial contrast enhancement. J Am Coll Cardiol 40:811–819PubMedCrossRefGoogle Scholar
  10. Fritz TA, Unger EC, Sutherland G, Sahn D (1997) Phase I clinical trials of MRX-115, a new ultrasound contrast agent. Invest Radiol 32:735–740PubMedCrossRefGoogle Scholar
  11. Goldberg BB, Raichlen JS, Forsberg S (eds) (2001) Ultrasound contrast agents: basic principles and clinical applications. Martin Dunitz, LondonGoogle Scholar
  12. Gramiak R, Shah PM (1968) Echocardiography of the aortic root. Invest Radiol 3:356–366PubMedCrossRefGoogle Scholar
  13. Keller MW, Segal SS, Kaul S, Duling B (1989) The behavior of sonicated albumin microbubbles within the microcirculation: a basis for their use during myocardial contrast echocardiography. Circ Res 65:458–467PubMedGoogle Scholar
  14. Kim DH, Klibanov AL, Needham D (2000) The influence of tiered layers of surface-grafted poly(ethylene glycol) on receptor-ligand-mediated adhesion between phospholipid monolayer-stabilized microbubbles and coated glass beads. Langmuir 16:2808–2817CrossRefGoogle Scholar
  15. Kirpotin D, Park JW, Hong K, Zalipsky S, Li WL, Carter P, Benz CC, Papahadjopoulos D (1997) Sterically stabilized anti-HER2 immunoliposomes: design and targeting to human breast cancer cells in vitro. Biochemistry 36:66–75PubMedCrossRefGoogle Scholar
  16. Klibanov AL, Hughes MS, Marsh JN, Hall CS, Miller JG, Wible JH, Brandenburger GH (1997) Targeting of ultrasound contrast material. An in vitro feasibility study. Acta Radiol Suppl 412:113–120PubMedGoogle Scholar
  17. Klibanov AL, Gu H, Wojdyla JK, Wible JH, Kim DH, Needham D, Villanueva FS, Brandenburger GH (1999 a) Attachment of ligands to gas-filled microbubbles via PEG spacer and lipid residues anchored at the interface. In: Proceedings of the 26th international symposium on controlled release of bioactive materials. Controlled release society, Boston, pp 124–125Google Scholar
  18. Klibanov AL, Hughes MS, Villanueva FS, Jankowski RJ, Wagner WR, Wojdyla JK, Wible JH, Brandenburger GH (1999b) Targeting and ultrasound imaging of microbubble-based contrast agents. MAGMA 8:177–184PubMedCrossRefGoogle Scholar
  19. Klibanov AL, Rasche PT, Hughes MS, Wojdyla JK, Galen KP, Wible JH Jr, Brandenburger GH (2002) Detection of individual microbubbles of an ultrasound contrast agent: fundamental and pulse inversion imaging. Acad Radiol 2:S279–S281CrossRefGoogle Scholar
  20. Klibanov AL, Rasche PT, Hughes MS, Wojdyla JK, Galen KP, Wible JH Jr, Brandenburger GH (2004) Detection of individual microbubbles of ultrasound contrast agents: imaging of free-floating and targeted bubbles. Invest Radiol 39:187–195PubMedCrossRefGoogle Scholar
  21. Lanza GM, Wallace KD, Scott MJ, Cacheris WP, Abendschein DR, Christy DH, Sharkey AM, Miller JG, Gaffney PJ, Wickline SA (1996) A novel site-targeted ultrasonic contrast agent with broad biomedical application. Circulation 94:3334–3340PubMedGoogle Scholar
  22. Lawrence MB, Mclntire LV, Eskin SG (1987) Effect of flow on polymorphonuclear leukocyte/endothelial cell adhesion. Blood 70:1284–1290PubMedGoogle Scholar
  23. Lee S, Kim DH, Needham D (2001) Equilibrium and dynamic interfacial tension measurements at microscopic interfaces using a micropipet technique. 2. Dynamics of phospholipid monolayer formation and equilibrium tensions at the water-air interface. Langmuir 17:5544–5550CrossRefGoogle Scholar
  24. Leighton TG (1997) The acoustic bubble. Academic Press, NYGoogle Scholar
  25. Leong-Poi H, Christiansen J, Klibanov AL, Kaul S, Lindner JR (2003) Non-invasive assessment of angiogenesis by ultrasound and microbubbles targeted to alpha(v)-integrins. Circulation 107:455–460PubMedCrossRefGoogle Scholar
  26. Lindner JR, Wei K, Kaul S (1999) Imaging of myocardial perfusion with SonoVue in patients with a prior myocardial infarction. Echocardiography 16:753–760PubMedCrossRefGoogle Scholar
  27. Lindner JR, Coggins MP, Kaul S, Klibanov AL, Brandenburger GH, Ley K (2000) Microbubble persistence in the microcirculation during ischemia/ reperfusion and inflammation is caused by integrin-and complement-mediated adherence to activated leukocytes. Circulation 101:668–675PubMedGoogle Scholar
  28. Lindner JR, Song J, Christiansen J, Klibanov AL, Xu F, Ley K (2001) Ultrasound assessment of inflammation and renal tissue injury with microbubbles targeted to P-selectin. Circulation 104:2107–2112PubMedCrossRefGoogle Scholar
  29. Pither R (2003) PET and the role of in vivo molecular imaging in personalized medicine. Expert Rev Mol Diagn 3:703–713PubMedCrossRefGoogle Scholar
  30. Schumann PA, Christiansen JP, Quigley RM, McCreery TP, Sweitzer RH, Unger EC, Lindner JR, Matsunaga TO (2002) Targeted-microbubble binding selectively to GPIIbIIIa receptors of platelet thrombi. Invest Radiol 37:587–593PubMedCrossRefGoogle Scholar
  31. Schutt EG, Klein DH, Mattrey RM, Riess JG (2003) Injectable microbubbles as contrast agents for diagnostic ultrasound imaging: the key role of perfluorochemicals. Angew Chem Int Ed Engl 42:3218–3235PubMedCrossRefGoogle Scholar
  32. Sipkins DA, Cheresh DA, Kazemi MR, Nevin LM, Bednarski MD, Li KC (1998) Detection of tumor angiogenesis in vivo by alphaVbeta3-targeted magnetic resonance imaging. Nat Med 4:623–626PubMedCrossRefGoogle Scholar
  33. Skyba DM, Camarano G, Goodman NC, Price RJ, Skalak TC, Kaul S (1996) Hemodynamic characteristics, myocardial kinetics and microvascular rheology of FS-069, a second-generation echocardiographic contrast agent capable of producing myocardial opacification from a venous injection. J Am Coll Cardiol 28:1292–1300PubMedCrossRefGoogle Scholar
  34. Villanueva FS, Jankowski RJ, Klibanov S, Pina ML, Alber SM, Watkins SC, Brandenburger GH, Wagner WR (1998) Microbubbles targeted to intercellular adhesion molecule-1 bind to activated coronary artery endothelial cells. Circulation 98:1–5PubMedGoogle Scholar
  35. Villanueva FS, Gertz EW, Csikari M, Pulido G, Fisher D, Sklenar J (2001) Detection of coronary artery stenosis with power Doppler imaging. Circulation 103:2624–2630PubMedGoogle Scholar
  36. Weller GE, Lu E, Csikari MM, Klibanov AL, Fischer D, Wagner WR, Villanueva FS (2003) Ultrasound imaging of acute cardiac transplant rejection with microbubbles targeted to intercellular adhesion molecule-1. Circulation 108:218–224PubMedCrossRefGoogle Scholar
  37. Winter PM, Caruthers SD, Kassner A, Harris TD, Chinen LK, Allen JS, Lacy EK, Zhang H, Robertson JD, Wickline SA, Lanza GM (2003) Molecular imaging of angiogenesis in nascent Vx-2 rabbit tumors using a novel alpha(V)beta3-targeted nanoparticle and 1.5 tesla magnetic resonance imaging. Cancer Res 63:5838–5843PubMedGoogle Scholar
  38. Yahalom D, Wittelsberger A, Mierke DF, Rosenblatt M, Alexander JM, Chorev M (2002) Identification of the principal binding site for RGD-containing ligands in the alphaVbeta3 integrin: a photoaffinity cross-linking study. Biochemistry 41:8321–8331PubMedCrossRefGoogle Scholar
  39. Yao J, Takeuchi M, Teupe C, Sheahan M, Connolly R, Walovitch RC, Fetterman RC, Church CC, Udelson JE, Pandian NG (2002) Evaluation of a new ultrasound contrast agent (AI-700) using two-dimensional and three-dimensional imaging during acute ischemia. J Am Soc Echocardiogr 15:686–694PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2005

Authors and Affiliations

  • A. L. Klibanov
    • 1
  1. 1.Cardiovascular DivisionUniversity of Virginia Medical CenterCharlottesvilleUSA

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