Micropatterned polymer substrates control alignment of proliferating Schwann cells to direct neuronal regeneration

Microcontact printed polymeric substrates were evaluated for their ability to control Schwann cell attachment and direct proliferation, as Schwann cell guidance is a crucial factor in directing peripheral nerve regeneration. Elastomeric stamps of poly(dimethylsiloxane) were "inked" with laminin, a permissive protein for Schwann cell adhesion, and stamped onto poly(methyl methacrylate) substrates to create patterns of lines and intervals varying from 10 to 50 microm wide. Schwann cells were seeded onto the substrates in serum-free media. After 4h, media was replaced with serum-containing growth media and changed daily thereafter. The addition of growth media to stimulate proliferation initially caused some loss in cell orientation relative to the laminin pattern, but when monolayer formation was complete, a high degree of cell orientation was observed. As both cell-cell contacts and surface coverage were maximized, the Schwann cells achieved an even higher order of orientation than observed during the early stages of proliferation. Significantly, smaller pattern widths increased the degree of orientation, regardless of interval width. Our results indicate that patterned polymeric substrates may enhance peripheral nerve regeneration by creating a highly ordered Schwann cell matrix for guidance of neurons.