Nanoimprinting for High-Throughput Fabrication of Geometrically Precise Pillars to Regulate Cell Behavior

Abstract

Developing high-throughput nanopatterning techniques that also allow for precise control over the dimensions of the fabricated features is essential for the study of cell-nanopattern interactions. Here, we developed a process that fulfills both of these criteria. Firstly, we used electron-beam lithography (EBL) to fabricate precisely controlled arrays of high aspect ratio submicron pillars with varying values of interspacing on a large area (3 × 5 mm2) of fused silica substrates. Two types of etching procedures with two different systems were developed to etch the fused silica and create the final desired height. We then studied the interactions of preosteoblasts (MC3T3-E1) with these pillars. Varying interspacing was observed to significantly affect the morphological characteristics of the cell, the organization of actin fibers, and the formation of focal adhesions. The expression of osteopontin (OPN) significantly increased on the patterns, indicating the potential of the pillars for inducing osteogenic differentiation. The EBL arrays of pillars were thereafter used as master molds in two subsequent processing steps, namely soft lithography (using hybrid PDMS) and thermal nanoimprint lithography for high-fidelity replication of the pillars on the substrates of interest. The molding parameters were optimized to maximize the fidelity of the generated patterns and minimize the wear and tear of the master mold. Comparing the replicated feature with those present on the original mold confirmed that the geometry and dimensions of the replicated pillars closely resemble those of the original ones (i.e., 1% deviation in pillar interspacing, < 20% deviation in pillar diameter). The method proposed in this study, therefore, enables the precise fabrication of submicron- and nanopatterns on a wide variety of substrate materials that are relevant for systematic cell studies.