Bacterial viability on chemically modified silicon nanowire arrays

The global threat of antimicrobial resistance is driving an urgent need for novel antimicrobial strategies. Functional surfaces are essential to prevent spreading of infection and reduce surface contamination. In this study we have fabricated and characterized multiscale-functional nanotopographies with three levels of functionalization: (1) nanostructure topography in the form of silicon nanowires, (2) covalent chemical modification with (3-aminopropyl)triethoxysilane, and (3) incorporation of chlorhexidine digluconate. Cell viability assays were carried out on two model microorganisms E. coli and S. aureus over these nanotopographic surfaces. Using SEM we have identified two growth modes producing distinctive multicellular structures, i.e. in plane growth for E. coli and out of plane growth for S. aureus. We have also shown that these chemically modified SiNWs arrays are effective in reducing the number of planktonic and surface-attached microorganisms

Macroporous gel particles as robust macroporous matrices for cell immobilization

A new design of robust matrices for cell immobilization is described. Macroporous gels (MGs) with immobilized microbial cells were prepared at subzero temperatures and were formed inside a plastic core (so-called, protective housing). Due to the protective housing the macroporous gel particles with immobilized cells can be used in well-stirred bioreactors. High retained activity of yeast (77-92%) and Escherichia coli (50-91%) cells immobilized in MGs after drying and storage in the dried state was due to the high structural stability and heterogeneous porous structure of the MGs

Phosphorylation and activation of phosphodiesterase type 3B (PDE3B) in adipocytes in response to serine/threonine phosphatase inhibitors: deactivation of PDE3B in vitro by protein phosphatase type 2A.

Phosphodiesterase type 3B (PDE3B) has been shown to be activated and phosphorylated in response to insulin and hormones that increase cAMP. In order to study serine/threonine protein phosphatases involved in the regulation of rat adipocyte PDE3B, we investigated the phosphorylation and activation of PDE3B in vivo in response to phosphatase inhibitors and the dephosphorylation and deactivation of PDE3B in vitro by phosphatases purified from rat adipocyte homogenates. Okadaic acid and calyculin A induced dose- and time-dependent activation of PDE3B. Maximal effects were obtained after 30 min using 1 microM okadaic acid (1.8-fold activation) and 300 nM calyculin A (4-fold activation), respectively. Tautomycin and cyclosporin A did not induce activation of PDE3B. Incubation of adipocytes with 300 nM calyculin A inhibited protein phosphatase (PP) 1 and PP2A completely. Okadaic acid (1 microM) reduced PP2A activity by approx. 50% but did not affect PP1 activity, and 1 microM tautomycin reduced PP1 activity by approx. 60% but PP2A activity by only 11%. This indicates an important role for PP2A in the regulation of PDE3B. Furthermore, rat adipocyte PDE3B phosphatase activity co-purified with PP2A but not with PP1 during MonoQ chromatography. As compared with insulin, okadaic acid and calyculin A induced phosphorylation of PDE3B by 2.8- and 14-fold respectively, whereas tautomycin and cyclosporin A had no effect. Both calyculin A and okadaic acid induced phosphorylation on serine 302, the site known to be phosphorylated on PDE3B in response to insulin and isoproterenol (isoprenaline), as well as on sites not identified previously. In summary, PP2A seems to be involved in the regulation of PDE3B in vivo and can act as a PDE3B phosphatase in vitro. In comparison with insulin, calyculin A induced a dramatic activation of PDE3B and both calyculin A and okadaic acid induced phosphorylation on additional sites, which could have a role in signalling pathways not yet identified

Strategies applied to modify structured and smooth surfaces: A step closer to reduce bacterial adhesion and biofilm formation

Nearly a century has passed since the discovery of the first antibiotics. With each passing decade, more bacterial strains developed resistance towards existing antibiotics. Alternative methods to reduce contamination by bacteria and biofilms have arisen to reduce the pressure on existing or currently developed antibiotics. This review highlights promising approaches to prevent bacterial contamination of the surface. Special attention is paid to antibiotic-free antibacterial strategies that are not affected by bacterial resistance. The approaches have been divided into four categories: (i) anti-adhesive, (ii) contact active, and (iii) biocide attached/biocide release, which can be integrated with (iv) topographical modification. Anti-adhesive approaches can reduce the adhesion between bacteria and a solid surface to prevent bacteria from contacting and contaminating the surface. Contact active approaches provide antibacterial activity by attachment of antibacterial agents to the substratum. Biocide attached/biocide release integrates contact-release of toxic chemicals to bacteria attached to the surface. Lastly, topographical modification relies on approaches to produce small structural features capable of matching cellular components killing bacteria. Combining one or more antibacterial strategies can lead to a more robust approach to deal with dangerous pathogenic bacterial species. In this case, a way forward is by combining various coatings onto topographically modified surfaces, enabling multifunctionality to reduce adhesion and biofilm formation. A perspective on the current antibacterial surface challenge is provided

Pattern Formation by Staphylococcus epidermidis via Droplet Evaporation on Micropillars Arrays at a Surface

We evaluate the effect of epoxy surface structuring on the evaporation of water droplets containing Staphylococcus epidermidis (S. epidermidis). During evaporation, droplets with S. epidermidis cells yield to complex wetting patterns such as the zipping-wetting1-3 and the coffee-stain effects. Depending on the height of the microstructure, the wetting fronts propagate circularly or in a stepwise manner, leading to the formation of octagonal or square-shaped deposition patterns.4,5 We observed that the shape of the dried droplets has considerable influence on the local spatial distribution of S. epidermidis deposited between micropillars. These changes are attributed to an unexplored interplay between the zipping-wetting1 and the coffee-stain6 effects in polygonally shaped droplets containing S. epidermidis. Induced capillary flows during evaporation of S. epidermidis are modeled with polystyrene particles. Bacterial viability measurements for S. epidermidis show high viability of planktonic cells, but low biomass deposition on the microstructured surfaces. Our findings provide insights into design criteria for the development of microstructured surfaces on which bacterial propagation could be controlled, limiting the use of biocides

Bacterial Footprints in Elastic Pillared Microstructures

Soft substrates decorated with micropillar arrays are known to be sensitive to deflection due to capillary action. In this work, we demonstrate that micropillared epoxy surfaces are sensitive to single drops of bacterial suspensions. The micropillars can show significant deformations upon evaporation, just as capillary action does in soft substrates. The phenomenon has been studied with five bacterial strains: S. epidermidis, L. sakei, P. aeruginosa, E. coli, and B. subtilis. The results reveal that only droplets containing motile microbes with flagella stimulate micropillar bending, which leads to significant distortions and pillar aggregations forming dimers, trimers, and higher order clusters. Such deformation is manifested in characteristic patterns that are left on the microarrayed surface following evaporation and can be easily identified even by the naked eye. Our findings could lay the ground for the design and fabrication of mechanically responsive substrates, sensitive to specific types of microorganisms

Pattern Formation by <i>Staphylococcus epidermidis</i> via Droplet Evaporation on Micropillars Arrays at a Surface

We evaluate the effect of epoxy surface structuring on the evaporation of water droplets containing <i>Staphylococcus epidermidis</i> (<i>S. epidermidis</i>). During evaporation, droplets with <i>S. epidermidis</i> cells yield to complex wetting patterns such as the zipping-wetting− and the coffee-stain effects. Depending on the height of the microstructure, the wetting fronts propagate circularly or in a stepwise manner, leading to the formation of octagonal or square-shaped deposition patterns., We observed that the shape of the dried droplets has considerable influence on the local spatial distribution of <i>S. epidermidis</i> deposited between micropillars. These changes are attributed to an unexplored interplay between the zipping-wetting and the coffee-stain effects in polygonally shaped droplets containing <i>S. epidermidis</i>. Induced capillary flows during evaporation of <i>S. epidermidis</i> are modeled with polystyrene particles. Bacterial viability measurements for <i>S. epidermidis</i> show high viability of planktonic cells, but low biomass deposition on the microstructured surfaces. Our findings provide insights into design criteria for the development of microstructured surfaces on which bacterial propagation could be controlled, limiting the use of biocides