Site-specific sonoporation of human melanoma cells at the cellular level using high lateral-resolution ultrasonic micro-transducer arrays.
Site-specific sonoporation of human melanoma cells at the cellular level using high lateral-resolution ultrasonic micro-transducer arrays.
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DOI：
10.1016/j.bios.2011.05.026
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发表时间：
2011-09-15
影响因子：
12.6
通讯作者：
Xu, Jian
中科院分区：
文献类型：
--
作者：
Thein, Myo;Cheng, An;Khanna, Payal;Zhang, Chunfeng;Park, Eun-Joo;Ahmed, Daniel;Goodrich, Christopher J.;Asphahani, Fareid;Wu, Fengbing;Smith, Nadine B.;Dong, Cheng;Jiang, Xiaoning;Zhang, Miqin;Xu, Jian
关键词：
我们开发了一种新的仪器方法，通过该方法，人黑色素瘤细胞（LU 1205）通过高横向分辨率超声微换能器阵列（UMTA）产生的高度受限的超声波施加的辐射压力进行sonoporated。该方法能够在细胞单层内由于UMTA而实现细胞水平的位点特异性声致穿孔，并且可以适用于在细胞测定中递送药物和基因产物。在该方法中，将细胞接种在采用UMTA的生物芯片上，以获得高空间分辨率和特异性。UMTA由30 MHz正弦信号驱动，并且所产生的辐射压力在靶细胞中诱导声致穿孔。通过对被动转运到细胞中的羧酸衍生的CdSe/ZnS量子点进行荧光显微镜和分析，确定UMTA的声致孔度和有效横向分辨率。开发了代表换能器产生的超声辐射压力、超声造成的细胞膜伤口和通过伤口的跨膜运输的模型，以确定超声压力依赖性伤口大小和增强的纳米颗粒细胞摄取。基于模型的计算表明，纳米颗粒的有效伤口尺寸和细胞摄取随着超声压力的增加而线性增加（即，在0.21、0.29和0.40MPa的辐射压力下，超声诱导的初始有效伤口半径分别为150、460和650 nm，声致孔后细胞内量子点浓度分别为7.8、22.8和29.9nM），并且在LU 1205细胞中诱导声致孔所需的阈值压力为10.12MPa。
We developed a new instrumental method by which human melanoma cells (LU1205) are sonoporated via radiation pressures exerted by highly-confined ultrasonic waves produced by high lateral-resolution ultrasonic micro-transducer arrays (UMTAs). The method enables cellular-level site-specific sonoporation within the cell monolayer due to UMTAs and can be applicable in the delivery of drugs and gene products in cellular assays. In this method, cells are seeded on the biochip that employs UMTAs for high spatial resolution and specificity. UMTAs are driven by 30-MHz sinusoidal signals and the resulting radiation pressures induce sonoporation in the targeted cells. The sonoporation degree and the effective lateral resolution of UMTAs are determined by performing fluorescent microscopy and analysis of carboxylic-acid-derivatized CdSe/ZnS quantum dots passively transported into the cells. Models representing the transducer-generated ultrasound radiation pressure, the ultrasound-inflicted cell membrane wound, and the transmembrane transport through the wound are developed to determine the ultrasound-pressure-dependent wound size and enhanced cellular uptake of nanoparticles. Model-based calculations show that the effective wound size and cellular uptake of nanoparticles increase linearly with increasing ultrasound pressure (i.e., at applied radiation pressures of 0.21, 0.29, and 0.40MPa, the ultrasound-induced initial effective wound radii are 150, 460, and 650nm, respectively, and the post-sonoporation intracellular quantum-dot concentrations are 7.8, 22.8, and 29.9nM, respectively) and the threshold pressure required to induce sonoporation in LU1205 cells is ∼0.12MPa.
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