The response of fibroblasts to hexagonal nanotopography fabricated by electron beam lithography

Abstract

It has been known for many years that cells will react to the shape of their microenvironment. It is more recently becoming clear that cells can alter their morphology, adhesions, and cytoskeleton in response to their nanoenvironment. A few studies have gone further and measured cellular response to high-adhesion nanomaterials. There have, however, been practical difficulties associated with genomic studies focusing on low-adhesion nanotopographies. Because of advancement in fabrication techniques allowing the production of large area of structure and the ability to amplify mRNA prior to microarray hybridization, these difficulties can be overcome. Here, electron beam lithography has been used to fabricate arrays of pits with 120 nm diameters, 100 nm depth and 300 nm center to center spacing in hexagonal arrangement. Electron and fluorescent microscopies have been used to observe morphological changes in fibroblasts cultured on the pits. 1.7k gene microarray was used to gauge genomic response to the pits. The results show reduction in cellular adhesion, decrease in spreading, and a broad genomic down-regulation. Also noted was an increase in endocytotic activity in cells on the pits.