Cooperative stiffening of flexible high aspect ratio nanostructures impart mechanobactericidal activity to soft substrates

Abstract

Understanding how bacteria interact with surfaces with micrometer and/or sub-micrometer roughness is critical for developing antibiofouling and bactericidal topographies. A primary research focus in this field has been replicating and emulating bioinspired nanostructures on various substrates to investigate their mechanobactericidal potential. Yet, reports on polymer substrates, especially with very high aspect ratios, have been rare, despite their widespread use in our daily lives. Specifically, the role of a decrease in stiffness with an increase in the aspect ratio of nanostructures may be consequential for the mechanobactericidal mechanism, which is biophysical in nature. Therefore, this work reports on generating bioinspired high aspect ratio nanostructures on poly(ethylene terephthalate) (PET) surfaces to study and elucidate their antibacterial and antibiofouling properties. Biomimetic nanotopographies with variable aspect ratios were generated via maskless dry etching of PET in oxygen plasma. It was found that both high and low-aspect ratio structures effectively neutralized Gram-negative bacterial contamination by imparting damage to their membranes but were unable to inactivate Gram-positive cells. Notably, the clustering of the soft, flexible tall nanopillars resulted in cooperative stiffening, as revealed by the nanomechanical behavior of the nanostructures and validated with the help of finite element simulations. Moreover, external capillary forces augmented the killing efficiency by enhancing the strain on the bacterial cell wall. Finally, experimental and computational investigation of the durability of the nanostructured surfaces showed that the structures were robust enough to withstand forces encountered in daily life. Our results demonstrate the potential of the single-step dry etching method for the fabrication of mechanobactericidal topographies and their potential in a wide variety of applications to minimize bacterial colonization of soft substrates like polymers.