Calcium-dependent ultrasound stimulation of secretory events from pancreatic beta cells.

BACKGROUND: Our previous studies have indicated that ultrasound can stimulate the release of insulin from pancreatic beta cells, providing a potential novel treatment for type 2 diabetes. The purpose of this study was to explore the temporal dynamics and Ca2+-dependency of ultrasound-stimulated secretory events from dopamine-loaded pancreatic beta cells in an in vitro setup. METHODS: Carbon fiber amperometry was used to detect secretion from INS-1832/13 beta cells in real time. The levels of released insulin were also measured in response to ultrasound treatment using insulin-specific ELISA kit. Beta cells were exposed to continuous wave 800 kHz ultrasound at intensities of 0.1 W/cm2, 0.5 W/cm2 and 1 W/cm2 for several seconds. Cell viability tests were done with trypan blue dye exclusion test and MTT analysis. RESULTS: Carbon fiber amperometry experiments showed that application of 800 kHz ultrasound at intensities of 0.5 and 1 W/cm2 was capable of stimulating secretory events for durations lasting as long as the duration of the stimulus. Furthermore, the amplitude of the detected peaks was reduced by 64% (p < 0.01) when extracellular Ca2+ was chelated with 10 mM EGTA in cells exposed to ultrasound intensity of 0.5 W/cm2. Measurements of released insulin in response to ultrasound stimulation showed complete inhibition of insulin secretion by chelating extracellular Ca2+ with 10 mM EGTA (p < 0.01). Viability studies showed that 800 kHz, 0.5 W/cm2 ultrasound did not cause any significant effects on viability and metabolic activity in cells exposed to ultrasound as compared to sham-treated cells. CONCLUSIONS: Our results demonstrated that application of ultrasound was capable of stimulating the release of insulin from pancreatic beta cells in a safe, controlled and Ca2+-dependent manner.

Therapeutic Modulation of Calcium Dynamics using Ultrasound and Other Energy-based Techniques.

Ultrasound, along with other types of energy-based methods, has been widely investigated for use in various therapeutic applications because of its ability to stimulate specific biological processes. Many of these processes are mediated by calcium (Ca2+) signaling, thus making modulation of Ca2+ dynamics an evident therapeutic target for energy-based techniques. Various diseases have been associated with abnormal Ca2+ signaling and could therefore benefit from therapeutic approaches trying to regulate the transport of Ca2+ across cell membranes. Here, we review published literature on the use of mechanical, electrical, magnetic and electromagnetic energy in modulating Ca2+ transients with particular emphasis on therapeutic ultrasound. We further provide brief discussions on the role of Ca2+ in living cells and the use of different experimental techniques to determine and measure its contribution to different biological processes. Finally, we explore the benefits, limitations and potential clinical applications of different energy-based modalities that can be utilized in modulation of Ca2+ signaling.