Fu Yuning, Okitsu Ryota, Nashimoto Yuji, Shinoda Yasuhiko, Utagawa Yoshinobu, Yamazaki Masateru, Nakaya Koki, Ino Kosuke, Kaji Hirokazu
Department of Diagnostic and Therapeutic Systems Engineering, Laboratory for Biomaterials and Bioengineering (LBB), Institute of Integrated Research (IIR), Institute of Science Tokyo, Meguro, Tokyo, Japan.
Graduate School of Medical and Dental Sciences, Institute of Science Tokyo, Meguro, Tokyo, Japan.
Anal Sci. 2025 Jul 17. doi: 10.1007/s44211-025-00825-6.
Endothelial metabolism is closely linked to vascular function and is often altered in response to pathological stimuli. Although vascular microphysiological systems (MPS) offer a promising in vitro platform for modeling vascular environments, the integration of real-time metabolic monitoring remains technically challenging. Enzyme-based electrochemical sensors are well-suited for detecting metabolites, such as glucose and lactate; however, their direct incorporation into culture systems is limited by the inactivation of enzymes under culture conditions. In this study, we investigated a vascular MPS with a sensor-compatible design to support future integration of printed enzyme-based biosensors. The device features a stacked open-bottom architecture that enables the integration of biosensors beneath the endothelial layer after monolayer formation, thus minimizing sensor exposure during culture. We validated the formation of an endothelial monolayer on a porous polyurethane membrane with a mortar-like structure and confirmed its compatibility with a model sensor substrate. As a preliminary step toward sensor-based metabolic analysis, we quantified glucose consumption and lactate production during endothelial monolayer formation and upon exposure to Minoxidil and Hydralazine, drugs known to induce vascular injury. These findings demonstrate the feasibility of sensor-compatible vascular MPS and provide foundational data to support the future development of integrated platforms for real-time metabolic monitoring in drug toxicity studies.
内皮细胞代谢与血管功能密切相关，并且常常会因病理刺激而发生改变。尽管血管微生理系统（MPS）为模拟血管环境提供了一个很有前景的体外平台，但实时代谢监测的整合在技术上仍然具有挑战性。基于酶的电化学传感器非常适合检测代谢物，如葡萄糖和乳酸；然而，将它们直接整合到培养系统中受到培养条件下酶失活的限制。在本研究中，我们研究了一种具有传感器兼容设计的血管MPS，以支持未来基于印刷酶的生物传感器的整合。该装置具有堆叠式底部开口结构，能够在单层形成后将生物传感器整合到内皮细胞层下方，从而在培养过程中尽量减少传感器的暴露。我们验证了在具有研钵状结构的多孔聚氨酯膜上形成内皮单层，并确认了其与模型传感器基板的兼容性。作为基于传感器的代谢分析的初步步骤，我们对内皮单层形成过程中以及暴露于米诺地尔和肼屈嗪（已知可诱导血管损伤的药物）时的葡萄糖消耗和乳酸生成进行了量化。这些发现证明了传感器兼容的血管MPS的可行性，并提供了基础数据，以支持药物毒性研究中实时代谢监测集成平台的未来发展。
