Label-free Detection of Protein Released during Platelet Activation by CNT-Enhanced Love Mode SAW Sensors.

Huiyan Wu, Hongfei Zu, Qing-Ming Wang, Gangyi Zhao, Jamesu H-C Wang
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Abstract

Platelet-rich plasma (PRP) has been applied in a series of clinical treatments. PRP contains high-concentrated platelets, which, when activated, could secret a variety of growth factors and cytokines, to promote and/or enhance healing of injured tissues. Non-activated platelets suspension could be prepared by an isolation method of centrifugation and washing currently. However, it is not clear whether platelets, if any, are already activated during this process and there is no simple method to monitor their activation accordingly. Shear-Horizontal Surface Acoustic Wave sensors (SH-SAW, Love Mode) are promising in fundamental biology as well as biomedical engineering, detecting cell behaviors in liquid in a non-invasive, simple and quantitative manner. In this study, Love mode sensors are adopted for the label-free detection of protein secreted by platelets. Carbon nanotube (CNT) is reported as an advisable platform of both non-specific protein adsorption and specific protein binding. For further improvement of Love mode sensor performance, novel CNT -coated parylene-C film is prepared on its surface as both the acoustic-wave-guiding layer and bio-interface layer. The S21 loss curves of Love mode sensors were recorded and the corresponding resonance frequencies were extracted. The results showed that the CNT-enhanced sensor possessed an increased resonance frequency shift when compared to normal sensor with single parylene-C film under identical collagen concentrations. Then, the modified sensor is used for label-free detection of protein released by various concentrations of platelets. The results revealed high sensitivity and consistency, indicating the potential of CNT-enhanced Love mode sensors in cell-based applications.

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利用 CNT 增强型爱模式声表面波传感器对血小板活化过程中释放的蛋白质进行无标记检测。
富血小板血浆(PRP)已被应用于一系列临床治疗中。血小板富集血浆含有高浓度血小板,激活后可分泌多种生长因子和细胞因子,促进和/或增强损伤组织的愈合。非活化血小板悬浮液目前可通过离心和洗涤的分离方法制备。不过,目前还不清楚在此过程中是否有血小板被激活,也没有简单的方法来监测其激活情况。剪切-水平表面声波传感器(SH-SAW,Love 模式)在基础生物学和生物医学工程中大有可为,它能以非侵入性、简单和定量的方式检测液体中的细胞行为。本研究采用爱模式传感器对血小板分泌的蛋白质进行无标记检测。据报道,碳纳米管(CNT)是非特异性蛋白质吸附和特异性蛋白质结合的理想平台。为了进一步提高爱模式传感器的性能,在其表面制备了新型 CNT 涂层对二甲苯-C 薄膜,作为声波导向层和生物界面层。记录了爱模式传感器的 S21 损耗曲线,并提取了相应的共振频率。结果表明,在胶原蛋白浓度相同的情况下,CNT 增强型传感器的共振频率偏移与单层对二甲苯-C 薄膜的普通传感器相比有所增加。然后,将改进后的传感器用于无标记检测不同浓度血小板释放的蛋白质。结果表明,这种传感器具有高灵敏度和高一致性,表明 CNT 增强爱模式传感器在基于细胞的应用中具有巨大潜力。
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