Effects of cell culture scaffold stiffness on cell membrane damage induced by sonoporation
Effects of cell culture scaffold stiffness on cell membrane damage induced by sonoporation
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DOI：
10.1007/s10396-014-0531-2
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发表时间：
2014-10-01
影响因子：
1.8
通讯作者：
Kinoshita, Yuto
中科院分区：
文献类型：
--
作者：
Kudo, Nobuki;Kinoshita, Yuto
目的作为实现体内声致孔的基础研究，利用单层细胞在不同刚度的支架上培养，评价声致孔过程中细胞膜的损伤率。方法采用胶原凝胶、10%和30%丙烯酰胺凝胶和盖玻片构建的4种支架，用于单层细胞的培养。对于凝胶支架，使用原子力显微镜测量的杨氏模量在0.09- 8.6kPa的范围内，而活细胞的杨氏模量为4.5kPa。将粘附有微泡的细胞暴露于8.0/-1.3MPa的峰值正/负压的单次脉冲超声中，持续时间为3、100和10，000 cycles.Results使用碘化丙啶的荧光显微镜观察到细胞膜损伤。3周期超声脉冲无明显影响;然而，100次循环和10次循环造成的损坏率，000-结论实验结果表明，贴壁细胞下层的刚度应被视为声穿孔条件的一个重要参数，最佳声穿孔暴露时间应与贴壁细胞层的刚度和贴壁细胞层的刚度有关。体内声致穿孔的条件应该考虑下面组织的物理性质来确定。
Purpose As basic studies to realize in vivo sonoporation, rates of cell membrane damage during sonoporation were evaluated using monolayer cells cultured on scaffolds with different degrees of stiffness.Methods Four types of scaffolds, constructed using collagen gel, 10 and 30 % acrylamide gels, and a coverslip, were used for cultivation of monolayer cells. Young's moduli measured using an atomic force microscope were in the range 0.09-8.6 kPa for the gel scaffolds, whereas Young's modulus for living cells was 4.5 kPa. Cells with attached microbubbles were exposed to one-shot pulsed ultrasound of 8.0/-1.3 MPa in peak positive/negative pressures with durations of 3, 100, and 10,000 cycles.Results Cell membrane damage was visualized by fluorescence microscopy using propidium iodide. The 3-cycle ultrasound pulse had no significant effect; however, the rates of damage caused by 100-cycle and 10,000-cycle pulses showed a strong tendency for higher rates of damage with a higher Young's modulus.Conclusion The experimental results indicate that the stiffness of the underlying layer of adherent cells should be considered as an essential parameter of the sonoporation condition and that the optimum exposure conditions for in vivo sonoporation should be determined with consideration of the physical properties of underlying tissues.