Microfluidic platforms for probing spontaneous functional recovery in hierarchically modular neuronal networks

Inherent capacity to flexibly reorganize after injury is a hallmark of brain networks, and recent studies suggest that functional consequences of focal damage are strongly influenced by the networks non-random connectivity. Although many of these insights have been derived from animal models and computational simulations, experimental platforms that enable bottom-up investigations of the structure-function relationships underlying damage and recovery processes remain limited. In this study, we used polydimethylsiloxane microfluidic devices to construct hierarchically modular neuronal networks that mimic the architectural features of the mammalian cortex. Laser microdissection was employed to selectively sever intermodular connections, enabling controlled damage to either hub or peripheral connections. Damage to hub connections led to delayed recovery, requiring more than three days for correlations to re-emerge. In contrast, peripheral damage resulted in faster recovery. Experiments that induced repeated injury to neuronal networks further demonstrated that recovery primarily occurred through the formation of alternative pathways rather than restoration of the original connections. These findings highlight how topological features of neuronal networks shape their response to injury and subsequent reorganization, providing mechanistic insights into the intrinsic self-repair capacity of biological systems.