化学学报 ›› 2017, Vol. 75 ›› Issue (11): 1091-1096.DOI: 10.6023/A17070330 上一篇    下一篇

所属专题: 纳米传感分析

研究论文

光致清洁电极可见光在线更新与细胞实时监测

度欢欢, 刘艳玲, 王雅文, 汤匀, 黄卫华   

  1. 生物医学分析化学教育部重点实验室 武汉大学化学与分子科学学院 武汉 430072
  • 收稿日期:2017-07-22 出版日期:2017-11-15 发布日期:2017-09-25
  • 通讯作者: 黄卫华 E-mail:whhuang@whu.edu.cn
  • 基金资助:

    项目受国家自然科学基金(Nos.21375099,21675121)资助.

Photocatalytically Renewable Electrode for On-Line Regeneration under Visible Light Irradiation and Real-Time Monitoring of Living Cells

Duo Huanhuan, Liu Yanling, Wang Yawen, Tang Yun, Huang Weihua   

  1. Key Laboratory of Analytical Chemistry for Biology and Medicine, Ministry of Education, College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072
  • Received:2017-07-22 Online:2017-11-15 Published:2017-09-25
  • Contact: 10.6023/A17070330 E-mail:whhuang@whu.edu.cn
  • Supported by:

    Project supported by the National Natural Science Foundation of China (Nos. 21375099, 21675121).

电化学传感器在用于细胞实时监测过程中,电极界面污染严重影响其检测性能.通过将纳米光催化剂与电化学传感材料复合,构建光致清洁电化学传感器,为电极界面的高效及无损更新提供了新思路.然而光催化产生的活性氧自由基导致细胞损伤,限制了细胞培养及检测过程中电极界面的实时更新.为此,我们在PEDOT@CdS/TiO2/ITO可见光致更新电极表面旋涂明胶薄层,在保持电极良好的光致清洁和电化学传感性能同时,利用明胶薄层阻碍光催化产生的活性氧自由基扩散至细胞表面,显著降低了细胞损伤.此外,明胶优良的生物相容性有利于细胞的黏附及增殖.利用该电极,我们实现了人脐静脉内皮细胞(HUVECs)培养过程中,电极的在线更新以及细胞释放一氧化氮的实时监测.

关键词: 纳米光催化剂, 光致清洁电极, 可见光, 明胶, 人脐静脉内皮细胞, 一氧化氮, 实时监测

Electrode fouling and passivation is an inevitable problem which seriously affects the electrode performance in cell culture and detection. Construction of photocatalytically renewable electrode by combination of nanophotocatalysts with electrochemical sensing materials could provide a promising approach for highly efficient renewal of electrode surface without damaging its micro-or nanostructures. However, the reactive oxygen species generated during photocatalysis always cause damages to cells being adhered or cultured on the electrode surface, which precludes on-line renewal of electrode during cell culture and detection. To address this issue, based on the visible light-induced renewable electrode (poly(3,4-ethylenedioxythiophene) (PEDOT)-modified TiO2/CdS nanocomposites electrode we previously developed, a thin layer of gelatin hydrogel was spin-coated on the electrode in this work to realize efficient electrode renewal under visible light irradiation during the culture and detection of living cells. The optimized thickness (ca. 2 μm) of gelatin hydrogel was obtained by spin-coating under 3000 r/min. Benefitting from the network structure of gelatin hydrogel and the renewable performance of PEDOT@CdS/TiO2 nanocomposites, the gelatin coating efficiently blocked the diffusion of biomacromolecules from the bulk medium to the electrode surface and thus significantly diminished the fouling caused by these macromolecules, while the pollutants derived from small molecules could be efficiently degraded under visible light irradiation. Meanwhile, gelatin coating did not induce obviously decline in detection sensitivity, and a low detection limit of 4.2 nmol/L (S/N=3) could be obtained towards electrochemical detection of nitric oxide (NO). Most importantly, the gelatin layer efficiently blocked the ultrashort-lived but highly reactive oxygen species such as OH·(generated by photocatalytic process) diffusing from the electrode surface to the cells, and the damages to the cells caused by these highly reactive species could be therefore significantly decreased. The results from live/dead cell staining demonstrated that almost all the cells (>95%) cultured on gelatin-coated electrodes maintain their viability when the electrode was irradiated by visible light for 6 h, while a considerable part of cells (>40%) culture on the uncoated electrode lost their viability under the same conditions. These features allowed on-line renewal of the electrode during cell culture and detection as well as real-time monitoring of NO released from the human umbilical vein endothelial cells (HUVECs).

Key words: nanophotocatalysts, photocatalytically renewable electrode, visible light, gelatin, human umbilical vein endothelical cells, nitric oxide, real-time monitoring