Brandon B. Seo

Responses of Staphylococcus aureus bacterial cells to nanocrystalline nickel nanostructures.

A broad range of human diseases are associated with bacterial infections, often initiated by specific adhesion of a bacterium to the target environment. Despite the significant role of bacterial adhesion in human infectious diseases, details and mechanisms of bacterial adhesion have remained elusive. Herein, we study the physical interactions between Staphylococcus aureus, a type of micro-organism relevant to infections associated with medical implants, and nanocrystalline (nc) nickel nanostructures with various columnar features, including solid core, hollow, x-shaped and c-shaped pillars. Scanning electron microscopy results show the tendency of these bacterial cells to attach to the nickel nanostructures. Moreover, unique single bacterium attachment characteristics were observed on nickel nanostructures with dimensions comparable to the size of a single bacterium.

Cell responses to metallic nanostructure arrays with complex geometries.

Metallic nanopillar/nanowires are emerging as promising platforms for biological applications, as they allow for the direct characterization and regulation of cell function. Herein we study the response of cells to a versatile nanopillar platform. Nanopillar arrays of various shape, size, and spacing and different nanopillar-substrate interfacial strengths were fabricated and interfaced with fibroblasts and several unique cell-nanopillar interactions were observed using high resolution scanning electron microscopy. Nanopillar penetration, engulfment, tilting, lift off and membrane thinning, were observed by manipulating nanopillar material, size, shape and spacing. These unique cell responses to various nanostructures can be employed for a wide range of applications including the design of highly sensitive nano-electrodes for single-cell probing.

Differential Collective- and Single-Cell Behaviors on Silicon Micropillar Arrays

Three-dimensional vertically aligned nano- and micropillars have emerged as promising tools for a variety of biological applications. Despite their increasing usage, the interaction mechanisms of cells with these rigid structures and their effect on single- and collective-cell behaviors are not well understood for different cell types. In the present study, we examine the response of glioma cells to micropillar arrays using a new microfabricated platform consisting of rigid silicon micropillar arrays of various shapes, sizes, and configurations fabricated on a single platform. We compare collective- and single-cell behaviors at micropillar array interfaces and show that glial cells under identical chemical conditions form distinct arrangements on arrays of different shapes and sizes. Tumor-like aggregation and branching of glial cells only occur on arrays with feature diameters greater than 2 μm, and distinct transitions are observed at interfaces between various arrays on the platform. Additionally, despite the same side-to-side spacing and gaps between micropillars, single glial cells interact with the flat silicon surface in the gap between small pillars but sit on top of larger micropillars. Furthermore, micropillars induced local changes in stress fibers and actin-rich filopodia protrusions as the cells conformed to the shape of spatial cues formed by these micropillars.

Bacterial Networks on Hydrophobic Micropillars

Bacteria have evolved as intelligent microorganisms that can colonize and form highly structured and cooperative multicellular communities with sophisticated singular and collective behaviors. The initial stages of colony formation and intercellular communication are particularly important to understand and depend highly on the spatial organization of cells. Controlling the distribution and growth of bacterial cells at the nanoscale is, therefore, of great interest in understanding the mechanisms of cell-cell communication at the initial stages of colony formation. Staphyloccocus aureus, a ubiquitous human pathogen, is of specific clinical importance due to the rise of antibiotic resistant strains of this species, which can cause life-threatening infections. Although several methods have attempted to pattern bacterial cells onto solid surfaces at single cell resolution, no study has truly controlled the 3D architectures of growing colonies. Herein, we present a simple, low-cost method to pattern S. aureus bacterial colonies and control the architecture of their growth. Using the wetting properties of micropatterened poly(dimethyl siloxane) platforms, with help from the physiological activities of the S. aureus cells, we fabricated connected networks of bacterial microcolonies of various sizes. Unlike conventional heterogeneous growth of biofilms on surfaces, the patterned S. aureus microcolonies in this work grow radially from nanostrings of a few bacterial cells, to form micrometer-thick rods when provided with a nutrient rich environment. This simple, efficient, and low-cost method can be used as a platform for studies of cell-cell communication phenomena, such as quorum sensing, horizontal gene transfer, and metabolic cross-feeding especially during initial stages of colony formation.

Adhesion characteristics ofStaphylococcus aureusbacterial cells on funnel-shaped palladium–cobalt alloy nanostructures

ABSTRACT The adhesion properties of Staphylococcus aureus on palladium–cobalt (Pd–Co) alloy nanostructures with various cross-sectional geometries have been characterised. They include solid core, hollow, c-shaped, and x-shaped pillars. These pillars have unique funnel-shaped geometric features on the top surfaces with average included angles between ∼142o and ∼149o. The success rates of cell attachment on these pillar tops were quantified by using field emission scanning electron microscopy techniques. Results show the Staphylococcus aureus attachment rates of Pd–Co solid core, hollow, and x-shaped pillars are statistically indistinguishable with success rate up to 82%. X-shaped pillars have the lowest attachment rate among the four geometries of 46 ± 5%.

Mechanical Contact Characteristics of PC3 Human Prostate Cancer Cells on Complex-Shaped Silicon Micropillars

In this study we investigated the contact characteristics of human prostate cancer cells (PC3) on silicon micropillar arrays with complex shapes by using high-resolution confocal fluorescence microscopy techniques. These arrays consist of micropillars that are of various cross-sectional geometries which produce different deformation profiles in adherent cells. Fluorescence micrographs reveal that some DAPI (4′,6-diamidino-2-phenylindole)-stained nuclei from cells attached to the pillars develop nanometer scale slits and contain low concentrations of DNA. The lengths of these slits, and their frequency of occurrence, were characterized for various cross-sectional geometries. These DNA-depleted features are only observed in locations below the pillar’s top surfaces. Results produced in this study indicate that surface topography can induce unique nanometer scale features in the PC3 cell.

Trapping polystyrene and latex nanospheres inside hollow nanostructures using Staphylococcus aureus cells

A new methodology was developed to trap polystyrene or latex nanospheres in nanocrystalline nickel columnar nanostructures with various cross-sectional geometries. After filling the column interiors with polystyrene or latex nanospheres with diameters of ∼300 and 500 nm, respectively, the top openings of these small metallic nanopillars were capped with Staphylococcus aureus (S. aureus) bacterial cells. High resolution scanning electron microscopy inspection revealed that these bacterial cells were able to cover the top openings of C-shaped pillars within an hour of incubation at 37 °C. Results also demonstrated that capping S. aureus adhesive strengths to the nickel structures are greater than the cell wall cohesive strengths.