Journal of Korean Society of Ultrasound in Medicine 2013;32(1): 59-65.
Synthesis of Ultrasound Contrast Agent: Characteristics and Size Distribution Analysis.
Hak Jong Lee, Tae Jong Yoon, Young Il Yoon
1Program in Nano Science and Technology, Department of Transdisciplinary Studies, Seoul National University Graduate School of Convergence Science and Technology, Korea.
2Department of Radiology, Seoul National University College of Medicine and Seoul National University Bundang Hospital, Korea.
3Department of Applied Bioscience, CHA University, Korea.
  Published online: March 31, 2013.
PURPOSE: The purpose of this study is to establish the methodology regarding synthesis of ultrasound contrast agent imaging, and to evaluate the characteristics of the synthesized ultrasound contrast agents, including size or degradation interval and image quality. MATERIALS AND METHODS: The ultrasound contrast agent, composed of liposome and SF6, was synthesized from the mixture solution of 21 micromol DPPC (1, 2-Dihexadecanoyl-sn-glycero-3-phosphocholine, C40H80NO8P), 9 micromol cholesterol, 1.9 micromol of DCP(Dihexadecylphosphate, [CH3(CH2)15O]2P(O)OH), and chloroform. After evaporation in a warm water bath and drying during a period of 12-24 hours, the contrast agent was synthesized by the sonication process by addition of buffer and SF6 gas. The size of the contrast agent was controlled by use of either extruder or sonication methods. After synthesis of contrast agents, analysis of the size distribution of the bubbles was performed using dynamic light scattering measurement methods. The degradation curve was also evaluated by changes in the number of contrast agents via light microscopy immediate, 12 hours, 24 hours, 36 hours, 48 hours, 60 hours, 72 hours, and 84 hours after synthesis. For evaluation of the role as an US contrast agent, the echogenicity of the synthesized microbubble was compared with commercially available microbubbles (SonoVue, Bracco, Milan, Italy) using a clinical ultrasound machine and phantom. RESULTS: The contrast agents were synthesized successfully using an evaporation-drying-sonication method. The majority of bubbles showed a mean size of 154.2 nanometers, and they showed marked degradation 24 hours after synthesis. ANOVA test revealed a significant difference among SonoVue, synthesized contrast agent, and saline (p < 0.001). Although no significant difference was observed between SonoVue and the synthesized contrast agent, difference in echogenicity was observed between synthesized contrast agent and saline (p < 0.01). CONCLUSION: We could synthesize ultrasound contrast agents using an evaporation-drying-sonication method. On the basis of these results, many prospective types of research, such as anticancer drug delivery, gene delivery, including siRNA or microRNA, targeted molecular imaging, and targeted therapy can be performed.
Keywords: Ultrasound; Contrast agent; Synthesis; Drug delivery; Image guided therapy
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