Rapid and High-Density Antibody Immobilization Using Electropolymerization of Pyrrole for Highly Sensitive Immunoassay.

Polypyrrole (Ppy) is a biologically compatible polymer that is used as a matrix, in which drugs and enzymes can be incorporated by doping. Here, we suggest an inventive application of Ppy as a biorecognition film encapsulated with an antibody (Ab) as an alternative strategy for the on-site multistep functionalization of thiol-based self-assembled monolayers. The fabrication steps of the recognition films were followed by dropping pyrrole and Ab mixed solutions onto the electrode and obtaining a thin film by direct current electropolymerization. The efficiency of Ab immobilization was studied by using fluorescence microscopy and electrochemical (EC) methods. Finally, the Ab density was increased and immobilized in 1 min, and the sensing performance as an EC immunosensor was demonstrated using α-fetoprotein with a limit of detection of 3.13 pg/mL and sensing range from 1 pg/mL to 100 ng/mL. This study demonstrates the potential for electrochemical functionalization of biomolecules with high affinity and rapidity.

Density-controlled electrochemical synthesis of ZnO nanowire arrays using nanotextured cathode.

Zinc oxide (ZnO) nanowires fabricated via wet chemical synthesis on flexible polymer substrates are inherently unstable against mechanical bending stress because of their high density and weak adhesion to the substrate. We introduce a novel method for controlling the density of such ZnO nanowire arrays using a three-dimensional corrugated metal substrate. These metal substrates, featuring extruded and recessed patterns fabricated via nanoimprint lithography, were employed as cathodes during the electrochemical deposition of ZnO nanowire arrays. The ZnO nanowire arrays synthesized on the patterned metal thin film exhibited smaller diameters and lower densities compared to those on non-patterned metal films. This reduction in density can be attributed to aligned nucleation and limited growth on the patterned metal surface. Crucially, ZnO nanowires synthesized on patterned metal substrates displayed remarkable mechanical robustness against external forces, a direct consequence of their reduced density. In contrast, nanowires synthesized on non-patterned metal substrates were broken under mechanical bending. Detailed morphological analyses performed after mechanical bending tests confirm that ZnO nanowires synthesized on nanoimprinted metal electrodes exhibited enhanced mechanical characteristics compared to those on non-patterned metal electrodes. These findings clearly demonstrate the promise of utilizing density-controlled ZnO nanowires in piezoelectric devices.

NanoIEA: A Nanopatterned Interdigitated Electrode Array-Based Impedance Assay for Real-Time Measurement of Aligned Endothelial Cell Barrier Functions.

A nanopatterned interdigitated electrode array (nanoIEA)-based impedance assay is developed for quantitative real-time measurement of aligned endothelial cell (EC) barrier functions in vitro. A bioinspired poly(3,4-dihydroxy-L-phenylalanine) (poly (l-DOPA)) coating is applied to improve the human brain EC adhesion onto the Nafion nanopatterned surfaces. It is found that a poly (l-DOPA)-coated Nafion grooved nanopattern makes the human brain ECs orient along the nanopattern direction. Aligned human brain ECs on Nafion nanopatterns exhibit increased expression of genes encoding tight and adherens junction proteins. Aligned human brain ECs also have enhanced impedance and resistance versus unaligned ones. Treatment with a glycogen synthase kinase-3 inhibitor (GSK3i) further increases impedance and resistance, suggesting synergistic effects occur on the cell-cell tightness of in vitro human brain ECs via a combination of anisotropic matrix nanotopography and GSK3i treatment. It is found that this enhanced cell-cell tightness of the combined approach is accompanied by increased expression of claudin protein. These data demonstrate that the proposed nanoIEA assay integrated with poly (l-DOPA)-coated Nafion nanopatterns and interdigitated electrode arrays can make not only biomimetic aligned ECs, but also enable real-time measurement of the enhanced barrier functions of aligned ECs via tighter cell-cell junctions.

Nanotextured Polymer Substrate for Flexible and Mechanically Robust Metal Electrodes by Nanoimprint Lithography.

Metal thin film electrodes on flexible polymer substrates are inherently unstable against humidity and mechanical stresses because of their poor adhesion properties. We introduce a novel approach for improving the adhesion characteristics of metal-polymer interface based on the nanostructuring of the polymer substrate by using nanoimprint lithography. The adhesion characteristics of metal-polymer interface were measured by accelerated test, cyclic bending test and double cantilever beam (DCB) test. The interface of Au/Ti dual layer thin film and nanoimprinted PMMA substrate shows over 2.03 and 1.95 times higher adhesion energy (G(c)) than that of Au/Ti dual layer thin film and plane PMMA substrate in air and wet environments, respectively. The adhesion energy between metal thin film and polymer substrate was dramatically improved by the increased surface roughness and mechanical interlocking effect of numerous nanoscale anchors at the edges of nanoimprinted surface, which was verified by both experiment and numerical analysis.