Jiang Donglei, Xu Yang, Jiang Hui, Xiang Xinyue, Wang Lifeng
College of Food Science and Engineering, Collaborative Innovation Center for Modern Grain Circulation and Safety, Key Laboratory of Grains and Oils Quality Control and Processing, Nanjing University of Finance and Economics, Nanjing, Jiangsu 210023, PR China.
Nanjing Institute for Food and Drug Control, Nanjing, Jiangsu 211198, PR China.
Bioelectrochemistry. 2025 Feb;161:108805. doi: 10.1016/j.bioelechem.2024.108805. Epub 2024 Sep 3.
In this paper, a biomimetic skin microtissue biosensor was developed based on three-dimensional (3D) bioprinting to precisely and accurately determine fish parvalbumin (FV). Based on the principle that allergens stimulate cells to produce ONOO (peroxynitrite anion), a screen-printed electrode for the detection nanomolar level ONOO was innovatively prepared to indirectly detect FV based on the level of ONOO release. Gelatin methacryloyl (GelMA), RBL-2H3 cells, and MS1 cells were used as bio-ink for 3D bioprinting. The high-throughput and standardized preparation of skin microtissue was achieved using stereolithography 3D bioprinting technology. The printed skin microtissues were put into the self-designed 3D platform that integrated cell culture and electrochemical detection. The experimental results showed that the sensor could effectively detect FV when the optimized ratio of RBL-2H3 to MS1 cells and allergen stimulation time were 2:8 and 2 h, respectively. The linear detection range was 0.125-3.0 μg/mL, and the calculated lowest detection limit was 0.122 μg/mL. In addition, the sensor had excellent selectivity, specificity, stability, and reliability. Thus, this study successfully constructed a biomimetic skin microtissue electrochemical sensor for PV detection.
在本文中，基于三维（3D）生物打印技术开发了一种仿生皮肤微组织生物传感器，用于精确、准确地测定鱼小清蛋白（FV）。基于过敏原刺激细胞产生过氧亚硝酸根阴离子（ONOO）的原理，创新性地制备了一种用于检测纳摩尔水平ONOO的丝网印刷电极，以基于ONOO的释放水平间接检测FV。甲基丙烯酸明胶（GelMA）、RBL-2H3细胞和MS1细胞用作3D生物打印的生物墨水。利用立体光刻3D生物打印技术实现了皮肤微组织的高通量标准化制备。将打印好的皮肤微组织放入自行设计的集成细胞培养和电化学检测的3D平台中。实验结果表明，当RBL-2H3与MS1细胞的优化比例为2:8且过敏原刺激时间为2小时时，该传感器能够有效检测FV。线性检测范围为0.125 - 3.0μg/mL，计算得出的最低检测限为0.122μg/mL。此外，该传感器具有优异的选择性、特异性、稳定性和可靠性。因此，本研究成功构建了一种用于检测PV的仿生皮肤微组织电化学传感器。
