Calcium Imaging of Sonoporation of Mammalian Cells

Sabens, D.; Aehle, M.; Steyer, Grant J.; Kourennyi, Dmitri E.; Deng, Cheri X.

doi:10.1063/1.2205531

Cited by 5 publications

(5 citation statements)

References 4 publications

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“…In addition, Ca 21 plays an important role in cell recovery after sonoporation (5). Recent studies using calcium imaging show US-induced increases in [Ca 21 ] i in various types of cells (6)(7)(8)(9)(10). In this study we report the observation of [Ca 21 ] i oscillations and waves induced by US in Chinese hamster ovary (CHO) cells in a solution containing microbubbles.…”

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confidence: 90%

“…For our experiments, the cells were grown in culture dishes for 3 days following standard protocols. As described previously (8), the cells were first loaded in darkness with a 5-mM fura-2 AM (Invitrogen, Carlsbad, CA) solution for 60 min. Next, the cells were washed and immersed in a phosphate-buffered solution with [Ca 21 ] o ¼ 0.9 mM at room temperature and placed under the microscope for online measurement.…”

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confidence: 99%

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

Kumon

Aehle

Sabens

et al. 2007

Biophysical Journal

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This study investigated the effects of ultrasound on the intracellular [Ca(2+)] 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 Ca(2+). Calcium imaging of the cells was performed using digital video fluorescence microscopy and Ca(2+)-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 Ca(2+). These cells exhibited a large influx of Ca(2+) over 3-4 s and did not return to their equilibrium levels. Subsequently, some cells exhibited one or more Ca(2+) 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.

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confidence: 90%

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confidence: 99%

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

Kumon

Aehle

Sabens

et al. 2007

Biophysical Journal

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“…We have previously demonstrated the feasibility of employing this imaging technique to observe the effects of sonoporation in individual cells (Kumon et al, 2007b; Sabens et al, 2006) and have shown for the first time that ultrasound could induce Ca 2+ transients and waves in cells in the presence of microbubbles (Kumon et al, 2007a). The current study focuses on the use of the real-time fluorescence imaging technique to obtain spatiotemporal information about Ca 2+ during ultrasound-mediated sonoporation facilitated by microbubbles in the presence or absence of extracellular Ca 2+ .…”

Section: Introductionmentioning

confidence: 99%

Spatiotemporal Effects of Sonoporation Measured by Real-Time Calcium Imaging

Kumon

Aehle

Sabens

et al. 2009

Ultrasound in Medicine & Biology

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To investigate the effects of sonoporation, spatiotemporal evolution of ultrasound-induced changes in intracellular calcium ion concentration ([Ca 2+ ] i ) was determined using real time fura-2AM fluorescence imaging. Monolayers of Chinese hamster ovary (CHO) cells were exposed to 1-MHz ultrasound tone burst (0.2 s, 0.45 MPa) in the presence of Optison ™ microbubbles. At extracellular [Ca 2+ ] o of 0.9 mM, ultrasound application generated both non-oscillating and oscillating (periods 12-30 s) transients (changes of [Ca 2+ ] i in time) with durations of 100-180 s. Immediate [Ca 2+ ] i transients after ultrasound application were induced by ultrasound-mediated microbubble-cell interactions. In some cases, the immediately-affected cells did not return to pre-ultrasound equilibrium [Ca 2+ ] i levels, thereby indicating irreversible membrane damage. Spatial evolution of [Ca 2+ ] i in different cells formed a calcium wave and was observed to propagate outward from the immediately-affected cells at 7-20 μm/s over a distance greater than 200 μm, causing delayed transients in cells to occur sometimes 60 s or more after ultrasound application. In calcium-free solution, ultrasound-affected cells did not recover, consistent with the requirement of extracellular Ca 2+ for cell membrane recovery subsequent to sonoporation. In summary, ultrasound application in the presence of Optison ™ microbubbles can generate transient [Ca 2+ ] i changes and oscillations at a focal site and in surrounding cells via calcium waves that last longer than the ultrasound duration and spread beyond the focal site. These results demonstrate the complexity of downstream effects of sonoporation beyond the initial pore formation and subsequent diffusion-related transport through the cellular membrane.

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“…Recent studies using calcium imaging showed that mechanical stresses on cells due to ultrasound induced intracellular Ca 2+ transients in various types of cells [7,[10][11][12]. The cells showed an increase in Ca 2+ level over 3-8 s and then decayed back to equilibrium over 150-200 s, with a delayed onset of 8-10 s from the US application [8,13].…”

Section: Introductionmentioning

confidence: 97%

Simulations of calcium related signaling in response to ultrasound stimulations

Yang

Sun

Jiang

et al. 2013

2013 6th International Conference on Biomedical Engineering and Informatics

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As an important second messenger implicated in a wide variety of intracellular signaling and regulatory pathways, cytoplasmic calcium transients are likely involved in the recognition and transduction of extracellular signals. In this work, the calcium transients and related signaling induced by low-intensity ultrasound stimulations were mathematically simulated. The results reproduced the characteristics of elevated intracellular calcium ion transients induced by low-intensity ultrasound and made it clear that the peak level of the transients would increase sharply with the intensity above a certain value as 1200 mW/cm 2 because of the limited capabilities of cell extrusion mechanisms, which might cause cell damage.

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Calcium Imaging of Sonoporation of Mammalian Cells

Cited by 5 publications

References 4 publications

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

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

Spatiotemporal Effects of Sonoporation Measured by Real-Time Calcium Imaging

Simulations of calcium related signaling in response to ultrasound stimulations

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