Department of Chemistry , University of North Carolina at Chapel Hill , CB 3290 , Chapel Hill , North Carolina 27599 , United States.
ACS Sens. 2019 Jul 26;4(7):1766-1773. doi: 10.1021/acssensors.9b00170. Epub 2019 Jun 20.
Macrophages mediate mammalian inflammation in part by the release of the gasotransmitter, nitric oxide (NO). Electrochemical methods represent the best means of direct, continuous measurement of NO, but monitoring continuous release from immunostimulated macrophages remains analytically challenging. Long release durations necessitate consistent sensor performance (i.e., sensitivity and selectivity for NO) in proteinaceous media. Herein, we describe the fabrication of an electrochemical sensor modified by an electropolymerized 5-amino-1-naphthol (poly(5A1N)) film in conjunction with a fluorinated xerogel topcoat. The unique combination of these membranes ensures selective detection of NO that is maintained over extended periods of use (>24 h) in biological media without performance deterioration. The hydrophobic xerogel topcoat protects the underlying NO-selective poly(5A1N) film from hydration-induced desorption. The bilaminar sensor is then readily adapted for measurement of the temporal NO-release profiles from immunostimulated macrophages.
巨噬细胞通过释放气体递质一氧化氮 (NO) 来介导哺乳动物炎症。电化学方法是直接、连续测量 NO 的最佳手段，但监测免疫刺激的巨噬细胞持续释放仍然具有分析挑战性。较长的释放时间需要在蛋白质介质中保持一致的传感器性能（即对 NO 的灵敏度和选择性）。在此，我们描述了一种电化学传感器的制造方法，该传感器通过电聚合 5-氨基-1-萘酚（聚（5A1N））膜与氟化的气凝胶顶涂层结合而成。这些膜的独特组合确保了对 NO 的选择性检测，并且在生物介质中长时间（>24 小时）使用而不会出现性能下降。疏水性气凝胶顶涂层可防止底层的 NO 选择性聚（5A1N）膜因水合作用引起的解吸。然后，双层传感器可轻松适应测量免疫刺激的巨噬细胞的时间 NO 释放曲线。
