Patterned glass surfaces direct cell adhesion and process outgrowth of primary neurons of the central nervous system

Glass surfaces were patterned with cell-adhesive regions of laminin adhesive peptides YIGSR, RGD, and IKVAV, and cell-repulsive regions of poly(ethylene glycol) (PEG). The patterns were created by sputter-coating titanium and then gold onto glass coverslips through electron microscope grids. Gold surfaces were modified with cysteine-terminated peptides to have approximately 450 fmol/ cm2 of peptide incorporated on the glass coverslips as determined with radiolabeled CGYIGSR. Amine-functionalized glass coverslips were prepared using an amine-functionalized silane and then further modified with PEG-aldehyde by a Schiff base reduction. All surfaces were characterized by X-ray photoelectron spectroscopy and water contact angles. Hippocampal neurons, plated from a serum-free medium, adhered preferentially to peptide-functionalized surfaces over PEG-modified surfaces. Cell adhesion and neurite outgrowth were limited to the peptide region, demonstrating that neurite outgrowth could be directed by a combination of cell-adhesive and cell-repulsive cues.