Ultrasound-induced molecular delivery to erythrocytes using a microfluidic system
Ultrasound-induced molecular delivery to erythrocytes using a microfluidic system
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DOI：
10.1063/1.5144617
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发表时间：
2020-03-01
期刊：
影响因子：
3.2
通讯作者：
Kopechek, Jonathan A.
中科院分区：
文献类型：
--
作者：
Centner, Connor S.;Murphy, Emily M.;Kopechek, Jonathan A.
将红细胞保存在干燥状态下储存在环境温度下可以简化严峻环境中的输血，例如农村诊所，遥远的军事行动和太空旅行期间。目前，红细胞的储存受到4 ℃下42天的短保质期的限制，并且长期保存需要复杂的过程，该过程涉及分别在-80 ℃下储存之前和之后从红细胞中添加和去除甘油。天然化合物，如海藻糖，如果它们在细胞内以足够的水平存在，可以保护处于干燥状态的细胞，但哺乳动物细胞膜缺乏这种化合物的转运蛋白。为了促进化合物通过超声和微泡（声致穿孔）加载穿过质膜，开发了基于聚二甲基硅氧烷的微流体装置。在各种条件下测试荧光素向红细胞中的递送，以评估诸如超声压力、超声脉冲间隔、微泡剂量和流速等参数的影响。与对照组相比，超声压力和平均流速的变化分别导致荧光素输送的统计学显著增加高达73 +/- 37%（p < 0.05）和44 +/- 33%（p < 0.01），但超声脉冲间隔的变化没有检测到统计学显著差异。冷冻干燥和再水化后，与对照组相比，超声介导的海藻糖负载后，活红细胞的回收率增加了128 +/- 32%（p < 0.05）。这些结果表明，在微流体通道中的超声介导的分子递送可能是一种可行的方法来处理红细胞，以在环境温度下以干燥状态长期储存。
Preservation of erythrocytes in a desiccated state for storage at ambient temperature could simplify blood transfusions in austere environments, such as rural clinics, far-forward military operations, and during space travel. Currently, storage of erythrocytes is limited by a short shelf-life of 42 days at 4 degrees C, and long-term preservation requires a complex process that involves the addition and removal of glycerol from erythrocytes before and after storage at -80 degrees C, respectively. Natural compounds, such as trehalose, can protect cells in a desiccated state if they are present at sufficient levels inside the cell, but mammalian cell membranes lack transporters for this compound. To facilitate compound loading across the plasma membrane via ultrasound and microbubbles (sonoporation), a polydimethylsiloxane-based microfluidic device was developed. Delivery of fluorescein into erythrocytes was tested at various conditions to assess the effects of parameters such as ultrasound pressure, ultrasound pulse interval, microbubble dose, and flow rate. Changes in ultrasound pressure and mean flow rate caused statistically significant increases in fluorescein delivery of up to 73 +/- 37% (p < 0.05) and 44 +/- 33% (p < 0.01), respectively, compared to control groups, but no statistically significant differences were detected with changes in ultrasound pulse intervals. Following freeze-drying and rehydration, recovery of viable erythrocytes increased by up to 128 +/- 32% after ultrasound-mediated loading of trehalose compared to control groups (p < 0.05). These results suggest that ultrasound-mediated molecular delivery in microfluidic channels may be a viable approach to process erythrocytes for long-term storage in a desiccated state at ambient temperatures.