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Polysialic acid (PSA) is crucial for the induction and maintenance of nervous system plasticity and repair after injury. In order to exploit the immense therapeutic potential of PSA, previous studies have focused on the identification and development of peptide‐based or synthetic PSA mimetics. 5‐Nonyloxytryptamine (5‐NOT) has been previously reported as a PSA‐mimicking compound for promoting functional recovery after spinal cord injury in mice. In order to explore the neuroregeneration potential of 5‐NOT, the current study was based on a biomaterial approach using collagen–laminin (C/L) scaffolds. In in vitro studies, 5‐NOT was observed to promote neurite outgrowth, migration, and fasciculation in cerebellar neuronal cells, whereas in 3D cell cultures it showed more ramification and complex Sholl profiles. 5‐NOT promoted the survival and neurite length of cortical neurons when cocultured with glutamate‐challenged astrocytes. In in vivo studies, spinal cord compression injury mice were used with immediate application of C/L hydrogels impregnated with 5‐NOT. C/L + 5‐NOT–treated mice demonstrated ∼75% of motor recovery 14 days after injury. Furthermore, this effect was shown to be dependent on the ERK–MAPK pathway and augmentation of cell survival. Thus, based on a biomaterial approach, our current study provides new insight for 5‐NOT–containing hydrogels as a promising candidate to speed up recovery after central nervous system injuries.

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