摘要：SummaryAstrocytes and microglia are critical regulators of inflammatory cascade after spinal cord injury (SCI). Existing glialin vitrostudies do not replicate inflammatory phases associated with SCI. Here, we report anin vitromodel of mixed glial culture where inflammation is induced by the administration of pro-inflammatory cytokines (tumor necrosis factor-α, interleukin-1β, and interleukin-6) to promote pathologically relevant “acute” and “chronic” inflammatory phases. We observed SCI relevant differential modulation of inflammatory pathways, cytokines, chemokines, and growth factors over 21 days. Mitochondrial dysfunction was associated with a cytokine combination treatment. Highly expressed cytokine induced neutrophil chemoattractant (CINC-3) chemokine was used as a biomarker to establish an enzyme-linked immunosorbent assay-based high-throughput screening (HTS) platform. We screened a 786-compound drug library to demonstrate the efficacy of the HTS platform. The developed model is robust and will facilitatein vitroscreening of anti-reactive glial therapeutics for the treatment of SCI.Graphical abstractDisplay OmittedHighlights•Anin vitroMGC model replicates the inflammatory phases associated with SCI•Differential modulation in NF-κB, MAPK, and immunomodulatory pathways over 21 days•Change in mitochondrial bioenergetics over seven days•ELISA-based HTS platform using CINC-3 as a biomarker is establishedMolecular Neuroscience; Cellular Neuroscience; Immunology; Proteomics
