Integrated Workflow for Electromechanical Stimulation of Cells in Active Scaffolds: From Live Cell Imaging to Functional Assays

In vivo, cells are exposed to a dynamic extracellular
matrix (ECM) that delivers biochemical, biophysical, and electrical
cues essential for regulating cellular behavior. However, most existing in vitro culture systems remain static and fail to reproduce
the time-dependent mechanical and electrical signals that characterize
native tissues. Developing biomimetic and dynamic platforms that bridge
this gap is therefore critical for advancing mechanobiology, tissue
engineering, or drug screening studies. Here, we report a 4D cell
culture platform based on polyHIPE-PEDOT scaffolds, combining the
structural versatility of polyHIPE architectures synthesized from
high internal phase emulsion with the electroactive behavior of the
conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT). The resulting
electroactive scaffolds exhibit a highly interconnected porous structure
with tunable morphology and demonstrate reversible pore deformation
under electrical stimulation. To facilitate a broad range of experimental
applications, we designed two complementary devices: one optimized
for in vitro cell culture under standard incubator
conditions, and another tailored for real-time live-cell imaging.
To illustrate this integrated workflow, we provide examples with fibroblast
cells cultured under electromechanical stimulation using our two devices.
The objective of this workflow is to facilitate novel insights into
the study of time-resolved cell–matrix interactions and to
develop responsive biomimetic microenvironments for advanced biomedical
research.

体内环境中，细胞所处的细胞外基质（extracellular matrix, ECM）为动态结构，可提供调控细胞行为所必需的生化、生物物理与电信号。然而，当前绝大多数体外培养体系仍为静态模式，无法复现天然组织所具有的时变力学与电信号特征。因此，开发能够填补这一空白的仿生动态平台，对于推动力学生物学、组织工程与药物筛选等领域的研究至关重要。

本研究报道了一种基于polyHIPE-PEDOT支架的4D细胞培养平台，该平台整合了由高内相乳液（high internal phase emulsion, HIPE）合成的高内相乳液聚合多孔材料（polyHIPE）结构的结构可调性，以及导电聚合物聚(3,4-乙撑二氧噻吩)（PEDOT）的电活性特性。所制备的电活性支架具有高度连通的多孔结构，形貌可调控，且在电刺激下可发生可逆的孔隙形变。

为适配多样化的实验应用需求，本研究设计了两款互补的装置：一款针对标准培养箱环境下的体外细胞培养进行了优化，另一款则专为实时活细胞成像打造。为展示这套整合式实验流程，我们使用两款装置开展了成纤维细胞在机电刺激下培养的示例实验。本实验流程的目标是为时间分辨的细胞-基质相互作用研究提供全新的研究视角，并为先进生物医学研究构建响应型仿生微环境。