Bacteria that adhere to the surfaces of implanted medical devices can cause catastrophic infection. Since chemical modifications of materials' surfaces have poor long-term performance in preventing bacterial buildup, approaches using bactericidal physical surface topography have been investigated. The authors used Nanoimprint Lithography was used to fabricate a library of biomimetic nanopillars on the surfaces of poly(methyl methacrylate) (PMMA) films. After incubation of Escherichia coli (E. coli) on the structured PMMA surfaces, pillared surfaces were found to have lower densities of adherent cells compared to flat films (67%-91% of densities on flat films). Moreover, of the E. coli that did adhere a greater fraction of them were dead on pillared surfaces (16%-141% higher dead fraction than on flat films). Smaller more closely spaced nanopillars had better performance. The smallest most closely spaced nanopillars were found to reduce the bacterial load in contaminated aqueous suspensions by 50% over a 24-h period compared to flat controls. Through quantitative analysis of cell orientation data, it was determined that the minimum threshold for optimal nanopillar spacing is between 130 and 380 nm. Measurements of bacterial cell length indicate that nanopillars adversely affect E. coli morphology, eliciting a filamentous response. Taken together, this work shows that imprinted polymer nanostructures with precisely defined geometries can kill bacteria without any chemical modifications. These results effectively translate bactericidal nanopillar topographies to PMMA, an important polymer used for medical devices.
Selective cell adhesion is desirable to control cell growth and migration on biomedical implants. Mesenchymal cell migration is regulated through focal adhesions (FAs) and can be modulated by their microenvironment, including changes in surface topography. We use the Number and Molecular Brightness (N&B) imaging analysis to provide a unique perspective on FA assembly and disassembly. This imaging analysis generates a map of real-time fluctuations of protein monomers, dimers, and higher order aggregates of FA proteins, such as paxillin during assembly and disassembly. We show a dynamic view of how nanostructured surfaces (nanoline gratings or nanopillars) regulate single molecular dynamics. In particular, we report that the smallest nanopillars (100 nm spacing) gave rise to a low percentage population of disassembly adhesion cluster size of ~2 paxillin proteins/cluster whereas the larger nanopillars (380 nm spacing) gave rise to a much larger population of larger disassembling cluster of ~3–5 paxillin proteins. Cells were more motile on the smaller nanopillars (spaced 100–130 nm apart) compared to all other surfaces studied. Thus, physical nanotopography influences cell motility, adhesion size, and adhesion assembly and disassembly. We report for the first time, with single molecular detection, how nanotopography influences cell motility and protein reorganization in adhesions.
Nanoimprinting of polymer surfaces is a way to functionalize surfaces by changing surface topography alone. The authors have demonstrated a method for the transfer of nanostructured coatings onto a curved polymer substrate by adapting reversal imprint lithography, an alternative to hot embossing nanoimprint lithography, which is best suited for flat planar surfaces and thermoplastics. The curved surface was made antireflective and hydrophobic without any chemical changes—solely by the addition of a nanostructured surface coating. In the future, this technique could be applied to a variety of materials and to surfaces with even more complex curvatures.
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