Axonal outgrowth on nano-imprinted patterns

Nanotechnology has provided methods to fabricate surface patterns with features down to a few nm. If cells or cell processes exhibit contact guidance in response to such small patterns is an interesting question and could be pertinent for many applications. In the present study we investigated if axonal outgrowth was affected by nano-printed patterns in polymethylmethacrylate (PMMA)-covered silicon chips. To this end adult mouse sympathetic and sensory ganglia were mounted in Matrigel on the chips close to the nano-patterns. The patterns consisted of parallel grooves with depths of 300 nm and varying widths of 100-400 nm. The distance between two adjacent grooves was 100-1600 nm. The chips were cultured in medium containing 25 ng/ml of nerve growth factor to stimulate axonal outgrowth. After 1 week of incubation, axonal outgrowth was investigated by immunocytochemistry or scanning electron microscopy. Axons displayed contact guidance on all patterns. Furthermore, we found that the nerve cell processes preferred to grow on ridge edges and elevations in the patterns rather than in grooves, a seemingly claustrophobic behavior. We conclude that axons of peripheral neurons might be guided by nanopatterns on PMMA when the lateral features are 100 nm or larger. The present results can be utilized for nerve regenerating scaffolds or the construction of a stable, high-resolution electronic interface to neurons, which is required for future brain machine interfaces.