Decoding the extracellular space and interstitial fluid of brain: A forward-looking review via the ECS-ISF functional...

The extracellular space (ECS) of the brain and its associated interstitial fluid (ISF)-treated as a single functional entity-make up an ever-dynamic but nevertheless functionally restricted medium within the brain. This system is critical for the transport of substances, diffusion of extracellular signals and preservation of neural homeostasis, while also being closely involved in neuronal activity regulation, metabolic waste drainage, disease progression and brain disease development. While many relevant studies have been documented in recent years, the microscopic geometric heterogeneity of the ECS, spatiotemporal dynamics in the ISF, and coupled organization of neurons-glia-vasculature across multiple scales are currently lacking from a broad and operable quantitative analysis framework for interstudy evaluation. This limitation also restricts the ability to systematise understanding of the global role of brain extracellular microenvironments. The extracellular space-interstitial fluid (ECS-ISF) functional unit is summarized and operationally defined as a local, parameterized tissue domain based on ECS geometry, diffusion-dominated transport, context-dependent convective/advection terms, and dynamic regulation by cells and vasculature as part of a state-dependent reconstructible and coordinated system. The geometric and physical properties of the brain ECS, as well as mechanisms regulating ISF generation, transport and exchange in this functional unit that governs tissue organization and contributes to metabolic regulation and brain homeostasis under normal versus pathological conditions, are summarized. In addition, recent quantitative research techniques for investigating modifications of the ECS-ISF system at various scales are summarized, including high-resolution imaging approaches, tracking methods, diffusion-convection transport models and multiscale computational simulation analyses, as well as collaborative multiphysics approaches. Their strengths and weaknesses are evaluated with respect to spatio-temporal resolution, parameter interpretability and applied scale. This review proposes the signal propagation efficiency index (SPEI) as a dimensionless reporting vector for signal-carrier propagation around explicitly defined ECS-ISF functional units (E-I units) by combining structural variables, dynamic indices and functional phenotypes, while highlighting that it remains a conceptual and testable framework rather than a validated biomarker. This perspective further outlines future research directions driven by the functional unit- and quantitative framework to advance fundamental principles of complex brain organization and the pathology of prevalent brain disorders.