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Ultrasound-Induced Calcium Oscillations and Waves in Chinese Hamster Ovary Cells in the Presence of Microbubbles

R. E. Kumon ^* , M. Aehle , D. Sabens , P. Parikh , D. Kourennyi  and C. X. Deng ^* 

^* Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan; and Department of Biomedical Engineering, Case Western Reserve University, Cleveland, Ohio

Correspondence: Address reprint requests and inquiries to Cheri X. Deng, Tel: 734-936-2855; Fax: 734-936-1905; E-mail: cxdeng{at}umich.edu.

This study investigated the effects of ultrasound on the intracellular [Ca²⁺] of Chinese hamster ovary cells in the presence of albumin-encapsulated Optison microbubbles. Cells were exposed to 1 MHz ultrasound (tone burst of 0.2 s duration, 0.45 MPa peak pressure) while immersed in solution of 0.9 mM Ca²⁺. Calcium imaging of the cells was performed using digital video fluorescence microscopy and Ca²⁺-indicator dye fura-2AM. Experimental evidence indicated that ultrasound caused a direct microbubble-cell interaction resulting in the breaking and eventual dissolution of the microbubble and concomitant permeabilization of the cells to Ca²⁺. These cells exhibited a large influx of Ca²⁺ over 3–4 s and did not return to their equilibrium levels. Subsequently, some cells exhibited one or more Ca²⁺ oscillations with the onset of oscillations delayed by 10–80 s after the ultrasound pulse. A variety of oscillations were observed including decaying oscillations returning to the baseline value over 35–100 s, oscillations superimposed on a more gradual recovery over 150–200 s, and oscillations continued with increased amplitude caused by a second ultrasound tone burst. The delays in onset appeared to result from calcium waves that propagated across the cells after the application of the ultrasound pulse.