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一种模拟微重力导向的 AIE 探针-ECM 水凝胶集成芯片,用于细胞培养和超氧阴离子自由基检测。

A simulated microgravity-oriented AIE probe-ECM hydrogel-integrated chip for cell culture and superoxide anion radical detection.

机构信息

Institute of Engineering Medicine, School of Medical Technology, Beijing Institute of Technology, Beijing, 100081, China; Beijing Key Laboratory for Separation and Analysis in Biomedicine and Pharmaceuticals, Beijing, 100081, China.

Department of Neurosurgery, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, Jinan, Shandong, 250014, China.

出版信息

Biosens Bioelectron. 2024 Nov 15;264:116656. doi: 10.1016/j.bios.2024.116656. Epub 2024 Aug 9.

Abstract

Human space activities have been continuously increasing. Astronauts experiencing spaceflight are faced with health problems caused by special space environments such as microgravity, and the investigation of cell injury is fundamental. The development of a platform capable of cell culture and injury detection is the prerequisite for the investigation. Constructing a platform suitable for special conditions in space life science research is the key issue. The ground-based investigation is an indispensable part of the research. Accordingly, a simulated microgravity (SMG)-oriented integrated chip platform capable of 3D cell culture and in situ visual detection of superoxide anion radical (O) is developed. SMG can cause oxidative stress in human cells, and O is one of the signaling molecules. Thus, a O-responsive aggregation-induced emission (AIE) probe is designed, which shows high selectivity and sensitivity to O. Moreover, the probe exhibits abilities of long-term and wash-free staining to cells due to the AIE behavior, which is precious for space cell imaging. Meanwhile, a chip with a high-aspect-ratio chamber for adequate medium storage for the lack of the perfusion system during the SMG experiment and a cell culture chamber which can integrate the extracellular matrix (ECM) hydrogel for the bioinspired 3D cell culture is fabricated. In addition, a porous membrane is introduced between the chambers to prevent the hydrogel from separating during the SMG experiment. The afforded AIE probe-ECM hydrogel-integrated chip can achieve 3D culturing of U87-MG cells and in situ fluorescent detection of endogenous O in the cells after long-term staining under SMG. The chip provides a powerful and potential platform for ground-based investigation in space life science and biomedical research.

摘要

人类的空间活动一直在不断增加。经历太空飞行的宇航员面临着特殊空间环境(如微重力)引起的健康问题,细胞损伤的调查是基础。开发能够进行细胞培养和损伤检测的平台是调查的前提。构建适合空间生命科学研究特殊条件的平台是关键问题。地面调查是研究不可或缺的一部分。因此,开发了一种模拟微重力(SMG)导向的集成芯片平台,能够进行 3D 细胞培养和超氧阴离子自由基(O)的原位可视化检测。SMG 会导致人类细胞氧化应激,而 O 是信号分子之一。因此,设计了一种对 O 具有高选择性和高灵敏度的 O 响应聚集诱导发射(AIE)探针。此外,由于 AIE 行为,探针对细胞具有长期和免洗染色能力,这对于空间细胞成像非常宝贵。同时,制造了一种带有高纵横比腔室的芯片,用于在 SMG 实验中缺乏灌注系统时储存足够的培养基,以及一个细胞培养腔室,该腔室可以集成用于生物启发 3D 细胞培养的细胞外基质(ECM)水凝胶。此外,在腔室之间引入了多孔膜,以防止水凝胶在 SMG 实验中分离。提供的 AIE 探针-ECM 水凝胶集成芯片可以在 SMG 下进行 U87-MG 细胞的 3D 培养,并对细胞内内源性 O 进行原位荧光检测,经过长期染色后。该芯片为空间生命科学和生物医学研究的地面调查提供了一个强大且有潜力的平台。

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