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An In Vitro Assay for Sonothrombolysis Based on the Spectrophotometric Measurement of Clot Thickness.

Journal of ultrasound in medicine : official journal of the American Institute of Ultrasound in Medicine (2017-02-06)
Zuojun Wang, Yoshikazu Sawaguchi, Hideo Hirose, Kazumasa Ohara, Shinichiro Sakamoto, Hidetaka Mitsumura, Takeki Ogawa, Yasuyuki Iguchi, Masayuki Yokoyama
RÉSUMÉ

For improved thrombolysis therapy based on ultrasound irradiation, researchers and practitioners would strongly benefit from an easy and efficient in vitro assay system of thrombolysis activity involving irradiated ultrasound. For the present study, we designed a new in vitro sonothrombolysis assay system using a sheet-type clot. We designed a cell for clot assay, and we confirmed that this clot cell did not significantly intervene in the acoustic field. Using human plasma, we made a sheet-type clot in the cell. Clot thicknesses at 100 points along 4 directions were measured photometrically at a rate of approximately 4 points/s. The sonothrombolysis effects at 13 levels of ultrasonic intensity were obtained with only one sheet-type clot. With this method, we used a clinically oriented probe at 0.7 and 0.3 W/cm We successfully established a new, easy, and efficient method for conducting in vitro sonothrombolysis assays. This method involves little intervention of either ultrasound reflection or standing waves in the clot cell. We believe that this new assay method is very useful for fundamental analyses of ultrasound's thrombolysis effects.

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Sigma-Aldrich
Thrombine from bovine plasma, lyophilized powder, 40-300 NIH units/mg protein (biuret)
Sigma-Aldrich
Trizma® hydrochloride solution, pH 7.5, BioPerformance Certified, 1 M, suitable for cell culture