Abstract

As a crucial biological material, the extracellular matrix (ECM) constitutes a major part of the extracellular microenvironment, offering both structural support and essential biological signals to surrounding cells. Research into ECM properties not only deepens our understanding of cellular behavior and metabolism but also opens new avenues for studying disease mechanisms and therapies. In this work, we report a previously unrecognized function of ECM by leveraging a three-dimensional spatial cavity structure. It is termed “Non-contact Transspatial Regulatory” (nCTR), referring to the ECM that remotely regulates cells without traditional direct physical contact. Systematic investigation and validation of key factors reveal that nCTR strongly depends on the 3D spatial distance between ECM and cells, governed by both physical spatial confinement and chemical signaling exchange. The process involves multiple components, pathways, and mechanisms, with macropinocytosis playing a dominant role. This newly identified function of the ECM offers a fresh perspective on cellular behavior and will undoubtedly open a new frontier in the cellular microenvironment studies. Highlights A custom-designed 3D study model utilizing two biomaterials with reverse thermal properties is developed to investigate non-physical contact interactions between the extracellular matrix (ECM) and cells. The Non-Contact Transspatial Regulation (nCTR) function of the ECM is introduced for the first time, defined as the ECM's capacity to remotely modulate cell behaviors across three-dimensional space without requiring direct physical contact. This regulatory function demonstrates strong spatial dependence on the distance between the ECM and target cells, and is jointly constrained by physical barriers and chemical exchanges. Mechanism analysis reveals that nCTR engages multiple signaling pathways and molecular components, with macropinocytosis serving as a potential mediator. The proposed nCTR will br