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Experimental Studies |
1 From the Molecular Oncology Center (T.L., Motumu Kuroki, Masahide Kuroki) and Department of Anatomy (K.T.), Fukuoka University School of Medicine, Nanakuma 7–45-1, Jonan-ku, Fukuoka 814-0180, Japan. Received April 29, 2002; revision requested July 10; final revision received January 31, 2003; accepted March 10. Supported in part by the Central Research Institute of Fukuoka University and Grant-in-Aid for High-Technology Research and Scientific Research [C] from the Japanese Ministry of Education, Science, Sports and Culture. Address correspondence to K.T.
PURPOSE: To determine if commercially available echo-enhanced microbubble contrast agents could be used to increase gene transfection efficiency by means of relatively low-intensity ultrasound-mediated microbubble destruction in skeletal muscles.
MATERIALS AND METHODS: Three types of ultrasound microbubble contrast agents (0.01 mL of albumin [Albunex] and human albumin [Optison] and 10 mg/mL of SH U 508A [Levovist]) were each separately mixed with the reporter plasmid DNA (25 µg) encoding green fluorescent protein (GFP) prior to intramuscular injection into the quadriceps muscle of a mouse thigh bilaterally (seven mice per contrast agent). One of the muscle sites that was injected with plasmid DNA was irradiated with low-intensity therapeutic ultrasound (1 MHz) at an intensity of 2.0 W/cm2 for 2 minutes. Mice were sacrificed 7 days after ultrasound treatment for gene expression assay. The number of GFP-expressing muscle fibers was counted. Statistical significance was determined with a two-tailed Student t test. P < .05 was considered to indicate statistically significant difference.
RESULTS: Muscle tissue exposed to ultrasound with air-filled Albunex or Levovist microbubbles revealed no difference in the number of GFP-expressing muscle fibers compared with the control non–ultrasound-exposed muscle. Albumin-coated octafluoropropane gas-filled Optison microbubbles showed a 10-fold increase in the number of GFP-expressing fibers (P < .05).
CONCLUSION: Low-intensity ultrasound with echo-enhanced Optison induced efficient gene transfer unlike that with Albunex or Levovist.
© RSNA, 2003
Index terms: Animals • Experimental study • Genes and genetics • Microbubbles • Ultrasound (US), contrast media
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