ITEFI-CSIC, Institute of Physics and Communication Technologies, 28006 Madrid, Spain.
ICTP-CSIC, Institute of Polymer Science and Technology, 28006 Madrid, Spain.
Sensors (Basel). 2021 Jul 25;21(15):5040. doi: 10.3390/s21155040.
Despite the use of therapeutic ultrasound in the treatment of soft tissue pathologies, there remains some controversy regarding its efficacy. In order to develop new treatment protocols, it is a common practice to carry out in vitro studies in cell cultures before conducting animal tests. The lack of reproducibility of the experimental results observed in the literature concerning in vitro experiments motivated us to establish a methodology for characterizing the acoustic field in culture plate wells. In this work, such acoustic fields are fully characterized in a real experimental configuration, with the transducer being placed in contact with the surface of a standard 12-well culture plate. To study the non-thermal effects of ultrasound on fibroblasts, two different treatment protocols are proposed: long pulse (200 cycles) signals, which give rise to a standing wave in the well with the presence of cavitation (I = 19.25 W/cm), and a short pulse (five cycles) of high acoustic pressure, which produces a number of echoes in the cavity (I = 33.1 W/cm, with P = 1.01 MPa). The influence of the acoustic intensity, the number of pulses, and the pulse repetition frequency was studied. We further analyzed the correlation of these acoustic parameters with cell viability, population, occupied surface, and cell morphology. Lytic effects when cavitation was present, as well as mechanotransduction reactions, were observed.
尽管在软组织病变的治疗中已经使用了治疗性超声波，但它的疗效仍存在一些争议。为了开发新的治疗方案，在进行动物试验之前，在细胞培养中进行体外研究是一种常见的做法。文献中观察到的体外实验结果的重复性缺乏促使我们建立了一种在培养板孔中描述声场的方法。在这项工作中，在与标准 12 孔培养板表面接触的实际实验配置中，对这些声场进行了全面描述。为了研究超声对成纤维细胞的非热效应，提出了两种不同的治疗方案：长脉冲（200 个周期）信号，在井中产生驻波并伴有空化（I = 19.25 W/cm），以及短脉冲（五个周期）的高声压，在腔内产生多个回声（I = 33.1 W/cm，P = 1.01 MPa）。研究了声强、脉冲数和脉冲重复频率的影响。我们进一步分析了这些声学参数与细胞活力、群体、占据表面和细胞形态之间的相关性。观察到有空化时的溶胞效应以及机械转导反应。
