Electrical stimulation promotes structural plasticity in cortical spheroids by modulating N-cadherin-dependent...

Electrical stimulation (ES) is a promising tool for brain-computer interfaces and therapeutic neuromodulation, yet its cellular effects on neural tissue remain incompletely understood. Here, we examined the mechanobiological remodeling of 3D cortical spheroids in response to DC electrical stimulation. Prolonged stimulation (60 min) produced a duration-dependent phenotypic shift marked by reduced collective spreading, elevated traction forces, and enhanced neurite outgrowth and complexity, along with increased expression of the plasticity-related immediate-early genes Arc and c-Fos. Higher traction was observed near the spheroid periphery, where IBA1-positive microglia were enriched, suggesting a possible role for microglia in restricting spheroid spreading. This response was accompanied by spatial reorganization of N-cadherin from a peripheral cohesive pattern to a dense, heterogeneous clustered distribution. Pharmacological inhibition of N-cadherin with ADH-1 reversed the migration arrest, supporting a functional role for N-cadherin-mediated adhesion in this phenotype. In parallel, integrin β1 and fibronectin were upregulated and spatially reorganized, indicating substantial remodeling of the adhesion- and ECM-related microenvironment. Together, these findings provide a mechanobiological framework for understanding how electrical stimulation reshapes tissue organization and plasticity-related responses in neural systems, and highlight 3D cortical spheroids as a useful platform for investigating stimulation-induced tissue remodeling.

The online version contains supplementary material available at https://doi.org/10.1007/s13534-026-00586-9.