在这项工作中，通过激光组装直接生产出具有纳米酶特征的纳米结构导电薄膜，并将其集成到完整的硝化纤维素传感器中；纤维素基质允许容纳活细胞，而纳米结构薄膜纳米酶活性确保了对活细胞释放的过氧化氢（H2O2）的无酶实时检测。具体来说，使用CO2激光绘图仪通过氧化石墨烯和铂阳离子的同时还原和图案化，生产了装饰有裸露铂纳米立方体的高度剥离的还原氧化石墨烯3D薄膜；将纳米结构薄膜集成到硝基纤维素基板中，并使用经济实惠的半自动印刷方法制造完整的传感器。直接 H2O2 测定的线性范围为 0.5-80 μM (R2 = 0.9943)，检测限为 0.2 μM。活细胞测量是通过将传感器放置在培养基中，确保它们粘附在传感器表面来实现的；两种细胞系分别用作非致瘤（Vero 细胞）和致瘤（SKBR3 细胞）模型。实时检测佛波酯刺激细胞释放的H2O2；硝化纤维传感器返回释放的 H2O2 的现场实时定量信息，证明了有用的灵敏度和选择性，可以区分致瘤细胞。所提出的策略允许使用简单的台式仪器低成本连续半自动生产纸质护理点设备，为轻松且经济地监测癌细胞的细胞病理学状态铺平道路。版权所有 © 2024作者。由 Elsevier B.V. 出版。保留所有权利。

In this work, a nanostructured conductive film possessing nanozyme features was straightforwardly produced via laser-assembling and integrated into complete nitrocellulose sensors; the cellulosic substrate allows to host live cells, while the nanostructured film nanozyme activity ensures the enzyme-free real-time detection of hydrogen peroxide (H2O2) released by the sames. In detail, a highly exfoliated reduced graphene oxide 3D film decorated with naked platinum nanocubes was produced using a CO2-laser plotter via the simultaneous reduction and patterning of graphene oxide and platinum cations; the nanostructured film was integrated into a nitrocellulose substrate and the complete sensor was manufactured using an affordable semi-automatic printing approach. The linear range for the direct H2O2 determination was 0.5-80 μM (R2 = 0.9943), with a limit of detection of 0.2 μM. Live cell measurements were achieved by placing the sensor in the culture medium, ensuring their adhesion on the sensors' surface; two cell lines were used as non-tumorigenic (Vero cells) and tumorigenic (SKBR3 cells) models, respectively. Real-time detection of H2O2 released by cells upon stimulation with phorbol ester was carried out; the nitrocellulose sensor returned on-site and real-time quantitative information on the H2O2 released proving useful sensitivity and selectivity, allowing to distinguish tumorigenic cells. The proposed strategy allows low-cost in-series semi-automatic production of paper-based point-of-care devices using simple benchtop instrumentation, paving the way for the easy and affordable monitoring of the cytopathology state of cancer cells.Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.