In Vitro Evaluation of the Link Between Cell Activation State and Its Rheological Impact on the Microscale Flow of Neutrophil Suspensions
In Vitro Evaluation of the Link Between Cell Activation State and Its Rheological Impact on the Microscale Flow of Neutrophil Suspensions
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DOI：
10.1115/1.4030824
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发表时间：
2015-09-01
影响因子：
1.7
通讯作者：
Shin，Hainsworth Y.
中科院分区：
文献类型：
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作者：
Akenhead，Michael L.;Horrall，Nolan M.;Shin，Hainsworth Y.
据报道，活化的中性粒细胞会影响外周阻力，例如，通过堵塞毛细血管或粘附于微血管。体内和离体数据表明，血液中循环的活化中性粒细胞也影响外周阻力。我们使用粘度计和微血管模拟物进行体外证实。使用锥板流变仪（450 s-1剪切速率）定量用10 nM fMet-Leu-Phe（fMLP）刺激的分化嗜中性粒细胞样HL-60早幼粒细胞（dHL 60 s）或人中性粒细胞悬液的流变学影响。为了评估它们对微尺度流动阻力的影响，我们使用10 μm Isopore®膜来模拟毛细血管，以及单个200 × 50μm微通道和20个20 × 50μm微流体通道网络来模拟非毛细血管微血管。刺激dHL 60和中性粒细胞群体显着改变了他们的流动行为证明了他们对悬浮液粘度的影响。值得注意的是，血细胞比容消除了白细胞活化对血细胞悬浮液粘度的影响。在生物膜过滤器中，活化的细胞悬浮液增强了流动阻力。红细胞的存在进一步增强了这种效果。我们的非毛细血管微血管模拟流动的活化中性粒细胞悬浮液的阻力显着增加，只有红细胞压积。值得注意的是，与单通道室相比，微网络室中的浓度升高程度更高。总的来说，我们的研究结果提供了支持性证据，即通过微循环的活化中性粒细胞可能会改变血流动力学阻力，由于其在非毛细血管微血管系统的流变学改变。这种效应是由于慢性炎症引起的嗜中性粒细胞活化的另一种方式，其可能至少部分地导致与心血管疾病相关的血液动力学阻力升高（例如，高血压和高胆固醇血症）。
Activated neutrophils have been reported to affect peripheral resistance, for example, by plugging capillaries or adhering to the microvasculature. In vivo and ex vivo data indicate that activated neutrophils circulating in the blood also influence peripheral resistance. We used viscometry and microvascular mimics for in vitro corroboration. The rheological impact of differentiated neutrophil-like HL-60 promyelocytes (dHL60s) or human neutrophil suspensions stimulated with 10 nM fMet-Leu-Phe (fMLP) was quantified using a cone-plate rheometer (450 s−1shear rate). To evaluate their impact on microscale flow resistance, we used 10-μm Isopore®membranes to model capillaries as well as single 200 × 50μm microchannels and networks of twenty 20 × 50μm microfluidic channels to mimic noncapillary microvasculature. Stimulation of dHL60 and neutrophil populations significantly altered their flow behavior as evidenced by their impact on suspension viscosity. Notably, hematocrit abrogated the impact of leukocyte activation on blood cell suspension viscosity. In micropore filters, activated cell suspensions enhanced flow resistance. This effect was further enhanced by the presence of erythrocytes. The resistance of our noncapillary microvascular mimics to flow of activated neutrophil suspensions was significantly increased only with hematocrit. Notably, it was elevated to a higher extent within the micronetwork chambers compared to the single-channel chambers. Collectively, our findings provide supportive evidence that activated neutrophils passing through the microcirculation may alter hemodynamic resistance due to their altered rheology in the noncapillary microvasculature. This effect is another way neutrophil activation due to chronic inflammation may, at least in part, contribute to the elevated hemodynamic resistance associated with cardiovascular diseases (e.g., hypertension and hypercholesterolemia).