Chemokine-Receptor Modulation Shapes Neuroinflammatory and Biomechanical Responses in Organotypic Hippocampal Cultures After Oxygen-Glucose Deprivation.

Ischemic stroke is a life-threatening neurological condition that frequently leads to severe brain damage and long-term disability. Current therapeutic options remain limited and are largely confined to anticoagulant-based interventions. Despite extensive evidence implicating inflammation in stroke pathogenesis, effective therapies targeting neuroinflammatory mechanisms are still missing. Given that chemokines and their receptors have been successfully targeted in clinical contexts outside stroke, we identified these signaling axes as promising candidates for therapeutic modulation in ischemic injury. We tested the hypothesis that pharmacological modulation of selected chemokine-receptor pathways alters the neuroinflammatory response and tissue viability following ischemia. Using organotypic hippocampal cultures subjected to oxygen-glucose deprivation, we demonstrate that inhibition of CX3CR1, CCR2, and CXCR4 differentially affects tissue viability under ischemic conditions. Owing to the pronounced biological effects observed upon CCR2 inhibition, we further investigated its mechanistic basis using atomic force microscopy. These analyses revealed that Irbesartan treatment is associated with cytoskeletal reorganization and significant alterations in the mechanical properties of ischemic brain tissue. In summary, our study establishes the organotypic hippocampal culture-oxygen-glucose deprivation model as a robust platform for investigating neuroinflammatory processes in ischemic stroke. We identify key chemokine-receptor circuits that modulate ischemic injury and uncover a previously underappreciated contribution of tissue biomechanics to the pathophysiological response to ischemia.