Guiding of neuronal cells on surfaces is required for the investigation of fundamental aspects of neurobiology, for tissue engineering, and for numerous bioelectronic applications. A modular method to establish nanostructured chemical templates for local deposition of gold nanoparticles is presented. A process comprising nanoimprint lithography, silanization, lift-off, and gold nanoparticle immobilization is used to fabricate the particle patterns. The chemical composition of the surface can be modified by in situ adsorption of cell-binding ligands to locally addressed particles. The versatility of this approach is demonstrated by inverting the binding affinity between rat cortical neurons and nanopatterned surfaces via wet-chemical means and thereby reversing the pattern of guided neurons.
High resolution lithography combined with microcontact printing (µCP) by means of polyolefine polymer (POP) stamps enabled to create protein gradient patterns. By this means, discrete purely biochemical gradients of extracellular matrix proteins were fabricated. It was possible to adjust independently both the size of elements of a protein pattern and the distance between them with sub 100 nm resolution. Adhesion of primary neurons and directed neuronal outgrowth were observed on these protein patterns. Cellular constituents such as filopodia adhere to different printed protein elements of the discontinuous gradient including features as small as 75 nm.
A colorized neurite of a rat cortical neuron is shown growing on a pattern of gold nanoparticles (AuNPs). The background image illustrates a larger area of AuNPs assembled in a pattern predefined by nanoimprint lithography. The AuNPs are equipped with different surface functionalities by D. Mayer and co‐workers , and used to investigate patterned neuron surface adhesion and neurite outgrowth. The neurites can be guided between or along the pattern using bare AuNPs or particles with positively charged ligands. Surface areas are easily switched between being cellattractive/repulsive by simple thiol chemistry.
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