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首页> 外文期刊>Advanced Functional Materials >Aqueous Two-Phase System Patterning of Microbubbles: Localized Induction of Apoptosis in Sonoporated Cells
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Aqueous Two-Phase System Patterning of Microbubbles: Localized Induction of Apoptosis in Sonoporated Cells

机译：微泡的水两相系统模式：超声诱导的细胞凋亡的局部诱导。

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Ultrasound-driven microbubbles produce mechanical forces that can disrupt cell membranes (sonoporation). However, it is difficult to control microbubble location with respect to cells. This lack of control leads to low sonoporation efficiencies and variable outcomes. In this study, aqueous two-phase system (ATPS) droplets are used to localize microbubbles in select micro-regions at the surface of living cells. This is achieved by stably partitioning microbubbles in dextran (DEX) droplets, deposited on living adherent cells in medium containing polyethylene glycol (PEG). The interfacial energy at the PEC-DEX interface overcomes microbubble buoyancy and prevents microbubbles from floating away from the cells. Spreading of the small DEX droplets retains microbubbles at the cell surface in defined lateral positions without the need for antibody or cell-binding ligand conjugation. The patterned microbubbles are activated on a cell monolayer exposed to a broadly applied ultrasound field (center frequency 1.25 MHz, active element diameter 0.6 cm, pulse duration 8 μs or 30 s). This system enables efficient testing of different ultrasound conditions for their effects on sonoporation-mediated membrane disruption and cell viability. Regions of cells without patterned microbubbles show no injury or membrane disruption. In microbubble patterned regions, 8 μs ultrasound pulses (0.2-0.6 MPa) produce cell death that is primarily apoptotic. Ultrasound-induced apoptosis increases with higher extracellular calcium concentrations, with cells displaying all of the hallmarks of apoptosis including annexinV labeling, loss of mitochondrial membrane potential, cas-pase activation and changes in nuclear morphology.
机译：超声波驱动的微气泡会产生机械力，从而破坏细胞膜（超音波）。但是，很难控制微泡相对于细胞的位置。缺乏控制会导致声穿孔效率低下和结果可变。在这项研究中，使用含水两相系统（ATPS）液滴将微泡定位在活细胞表面的选定微区域中。这是通过将微气泡稳定地分配在葡聚糖（DEX）小滴中而实现的，这些小滴沉积在包含聚乙二醇（PEG）的培养基中的活细胞上。 PEC-DEX界面处的界面能克服了微气泡的浮力，并防止了微气泡从细胞中漂浮出来。小的DEX液滴的散布将微泡保留在细胞表面的确定的侧向位置，而无需抗体或细胞结合配体的结合。图案化的微气泡在暴露于广泛应用的超声场（中心频率1.25 MHz，有源元件直径0.6 cm，脉冲持续时间8μs或30 s）的细胞单层上被激活。该系统能够有效测试不同的超声条件对超声介导的膜破坏和细胞活力的影响。没有图案化的微泡的细胞区域没有显示出损伤或膜破坏。在微气泡图案区域中，8μs超声脉冲（0.2-0.6 MPa）产生的细胞死亡主要是凋亡。超声诱导的细胞凋亡随着细胞外钙浓度的升高而增加，细胞表现出所有细胞凋亡的标志，包括膜联蛋白V标记，线粒体膜电位丧失，cas-ase激活和核形态改变。

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  • 来源
    《Advanced Functional Materials》 |2013年第27期|3420-3431|共12页
  • 作者

    John P. Fmmpton; Zhenzhen Fan; Arlyne Simon; Di Chen; Cheri X. Deng; Shuichi Takayama;

  • 作者单位

    Department of Biomedical Engineering University of Michigan Ann Arbor, USA;

    Department of Biomedical Engineering University of Michigan Ann Arbor, USA;

    Department of Macromolecular Science and Engineering University of Michigan Ann Arbor, USA;

    Department of Biomedical Engineering University of Michigan Ann Arbor, USA;

    Department of Biomedical Engineering University of Michigan Ann Arbor, USA;

    Department of Biomedical Engineering University of Michigan Ann Arbor, USA;

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