Effects of shear stress cultivation on cell membrane disruption and intracellular calcium concentration in sonoporation of endothelial cells.
Effects of shear stress cultivation on cell membrane disruption and intracellular calcium concentration in sonoporation of endothelial cells.
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DOI：
10.1016/j.jbiomech.2010.09.003
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发表时间：
2011-01-04
影响因子：
2.4
通讯作者：
Deng, Cheri X.
中科院分区：
文献类型：
--
作者：
Park, Juyoung;Fan, Zhenzhen;Deng, Cheri X.
微泡超声（microbubble facilitated ultrasound，US）应用可以通过瞬时破坏细胞膜或声致穿孔（sonoporation）来增强静态条件下培养的内皮细胞中药物和基因的细胞内递送。然而，体内不断暴露于血流的内皮细胞可能表现出不同的声致穿孔特性。本研究探讨了剪切应力培养声穿孔的内皮细胞的膜破坏和细胞内钙离子浓度（[Ca 2 +]i）的变化方面的影响。使用在微通道环境中在静态或剪切应力（5达因/cm 2，5天）条件下培养的小鼠脑微血管内皮（bEnd.3）细胞和人脐静脉内皮细胞（HUVEC）进行声致孔实验。将细胞暴露于短的US短纯音（1.25 MHz，8 μs持续时间，0.24 MPa），并在Deeprock ™微泡存在下促进声致穿孔。碘化丙啶（PI）法测定细胞膜的破坏程度，Fura-2AM法测定细胞内[Ca 2 +]i的变化。这项研究的结果表明，剪切应力培养显着降低了超声驱动的微泡活动对内皮细胞的影响。与静态培养的细胞相比，在剪切应力条件下培养的细胞表现出低得多的百分比与膜破裂和[Ca 2 +]i的变化。PI摄取和[Ca ~（2+）]i的最大增加也显着低于切应力培养的细胞。此外，与静态培养的细胞相比，剪切培养的HUVECs中的[Ca 2 +]i波的范围减少。
Microbubble facilitated ultrasound (US) application can enhance intracellular delivery of drugs and genes in endothelial cells cultured in static condition by transiently disrupting the cell membrane, or sonoporation. However, endothelial cells in vivo that are constantly exposed to blood flow may exhibit different sonoporation characteristics. This study investigates the effects of shear stress cultivation on sonoporation of endothelial cells in terms of membrane disruption and changes in the intracellular calcium concentration ([Ca2+]i). Sonoporation experiments were conducted using murine brain microvascular endothelial (bEnd.3) cells and human umbilical vein endothelial cells (HUVECs) cultured under static or shear stress (5 dyne/cm2 for 5 days) condition in a microchannel environment. The cells were exposed to a short US tone burst (1.25 MHz, 8 μs duration, 0.24 MPa) in the presence of Definity™ microbubbles to facilitate sonoporation. Membrane disruption was assessed by propidium iodide (PI) and changes in [Ca2+]i measured by fura-2AM. Results from this study show that shear stress cultivation significantly reduced the impact of ultrasound-driven microbubbles activities on endothelial cells. Cells cultured under shear stress condition exhibited much lower percentage with membrane disruption and changes in [Ca2+]i compared to statically cultured cells. The maximum increases of PI uptake and [Ca2+]i were also significantly lower in the shear stress cultured cells. In addition, the extent of [Ca2+]i waves in shear cultured HUVECs was reduced compared to the statically cultured cells.
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