Abstract
Intracellular calcium ion delivery via sonoporation has been validated to be a substitute for classical chemotherapy. However, the mechanism behind calcium sonoporation remains unclear to this day. To elucidate the role of calcium in the process of sonoporation, we aimed to investigate the influence of different calcium concentration on cell membrane permeabilization and cell viability after sonoporation. In this study, we present experimental evidence that extracellular calcium plays a major role in cell membrane molecular transport after applying ultrasound pulses. Ultrasound-microbubble cavitation in the presence of different calcium concentration affects fundamental cell bio-physio-chemical conditions: cell membrane integrity, metabolic activity, and colony formation. Corresponding vital characteristics were evaluated using three independent viability tests: propidium iodide assay (20 min–3 h), MTT assay (48 h), and cell clonogenic assay (6 d). The results indicate instant cell death, as the level of cell viability was determined to be similar within a 20 min–48 h–6 d period. Inertial cavitation activities have been determined to be directly involved in calcium delivery via sonoporation according to high correlation (R2 > 0.85, p < 0.01) of inertial cavitation dose with change in either cell membrane permeabilization, metabolic activity, and colony formation efficiency. In general, calcium delivery via sonoporation induces rapid cell death, occurring within 20 min after treatment, that is the result of ultrasound mediated microbubble cavitation.
Highlights
Cell sonoporation (SP) is a physical method, developed to attain spatiotemporally regulated intracellular transfer of bioactive compounds
SP process is intrinsically related to microbubble (MB) cavitation engendered by irradiation with ultrasound (US)
Ca intracellular delivery via SP, 2+ concentra‐. The latter phenomenon is directly associated to inertial cavitation acoustic pressure was varied in the range of 0–500 kPa at constant
Summary
Cell sonoporation (SP) is a physical method, developed to attain spatiotemporally regulated intracellular transfer of bioactive compounds. SP process is intrinsically related to microbubble (MB) cavitation engendered by irradiation with ultrasound (US). Simultaneous US–MB interaction leads to temporal increase in cell membrane permeability for exogenous therapeutic agents, such as anticancer drugs, DNA, RNA, etc. Cellular entry of calcium ions (Ca2+ ), facilitated by physical therapeutic agent delivery techniques, has been proposed as an option for conventional chemotherapy [2]. Ca2+ intracellular apportionment inside membrane-bound organelles plays an essential role in cellular response to diverse stress stimuli as well as coordination of fatal mechanisms. Substantial importance of Ca2+ has been demonstrated to facilitate the resealing rate of cell membrane injuries, caused by MB activity after irradiation with US [3,4]
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