Visualization of Endothelial Cell Damage Caused by Ultrasonically Induced Microbubble Oscillation Inside a Capillary Phantom
Visualization of Endothelial Cell Damage Caused by Ultrasonically Induced Microbubble Oscillation Inside a Capillary Phantom
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毛细血管体模内超声诱导微泡振荡引起的内皮细胞损伤的可视化
DOI：
10.1109/ius46767.2020.9251785
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
N.
中科院分区：
文献类型：
--
作者：
Shimizu;R.-I.;Suzuki;R.;Kudo;N.
利用三维细胞培养技术建立的毛细血管模型研究了血脑屏障开放的机制。用人脐静脉内皮细胞(HUVECs)悬浮在胶原胶中制作模型。与内皮生长因子孵育一周导致毛细血管腔的形成，并在腔内引入添加FITC-葡聚糖、SYTOX蓝和钙黄素-AM的微泡悬浮液。在中心频率为1.0 MHz、峰值负压为0.4 Mpa的短脉冲超声作用下，用高速摄像机捕捉气泡与细胞的相互作用，并用荧光显微镜观察内腔和内皮细胞的损伤。高速观察显示，气泡的不均匀收缩导致气泡旁的管腔发生明显的局部变形，管腔内强渗出(FITC-葡聚糖)和明显的内皮细胞膜损伤(SYTOX Blue)。致命性损伤也经常被观察到(钙黄绿素)。另一方面，管腔对侧的细胞在气泡扩张过程中受到拉伸力，较小的外渗和细胞损伤保持了内皮细胞的完整性。提示存在不同的血脑屏障开放机制。
Mechanisms of BBB opening were investigated using a capillary model created by a three-dimensional cell culture technique. The model was made using human umbilical vein endothelial cells (HUVECs) suspended in collagen gel. Incubation with endothelial growth factor for one week resulted in the creation of capillary lumens, and a suspension of microbubbles supplemented with FITC-dextran, SYTOX Blue, and Calcein-AM was introduced inside the lumens. Bubble-cell interaction under exposure to short pulsed ultrasound of 1.0 MHz in center frequency and 0.4 MPa in peak negative pressure was captured using a high-speed camera, and lumen and endothelial cell damage was visualized using fluorescence microscopy. High-speed observation showed that non-uniform contraction of a bubble caused significant local deformation of the lumen beside the bubble, and strong extravasation from the lumen (FITC-dextran) and significant membrane damage of the endothelial cells (SYTOX Blue) were confirmed at this location. Lethal damage was also observed frequently (Calcein). On the other hand, cells on the opposite side of the lumen received stretching force during bubble expansion, and smaller extravasation and cell damage maintained the integrity of endothelial cells. These suggested the presence of different mechanisms of BBB opening.