Maciulevičius Martynas, Navickaitė Diana, Chopra Sonam, Jakštys Baltramiejus, Šatkauskas Saulius
Biophysical Research Group, Faculty of Natural Sciences, Vytautas Magnus University, Vileikos st. 8, LT-44404 Kaunas, Lithuania.
Biomedicines. 2021 Jan 4;9(1):32. doi: 10.3390/biomedicines9010032.
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 (R > 0.85, < 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.
通过声孔效应进行细胞内钙离子递送已被证实可替代传统化疗。然而，钙离子声孔效应背后的机制至今仍不清楚。为了阐明钙离子在声孔效应过程中的作用，我们旨在研究不同钙离子浓度对声孔效应后细胞膜通透性和细胞活力的影响。在本研究中，我们提供了实验证据，表明在施加超声脉冲后，细胞外钙离子在细胞膜分子转运中起主要作用。在不同钙离子浓度存在下的超声微泡空化会影响细胞的基本生物物理化学条件：细胞膜完整性、代谢活性和集落形成。使用三种独立的活力测试评估相应的重要特征：碘化丙啶检测（20分钟 - 3小时）、MTT检测（48小时）和细胞克隆形成检测（6天）。结果表明细胞立即死亡，因为在20分钟 - 48小时 - 6天的时间段内细胞活力水平被确定为相似。根据惯性空化剂量与细胞膜通透性、代谢活性和集落形成效率变化之间的高度相关性（R > 0.85，P < 0.01），已确定惯性空化活动直接参与通过声孔效应进行的钙离子递送。一般来说，通过声孔效应进行的钙离子递送会诱导快速细胞死亡，在处理后20分钟内发生，这是超声介导的微泡空化的结果。
