Decorating Nanostructured Surfaces with Antimicrobial Peptides to Efficiently Fight Bacteria.

With conventional antibiotic therapies being increasingly ineffective, bacterial infections with subsequent biofilm formation represent a global threat to human health. Here, an active and a passive strategy based on polymeric micelles were combined to fight bacterial growth. The passive strategy involved covalent immobilization of polymeric micelles through Michael addition between exposed maleimide and thiol functionalized surfaces. Compared to the bare surface, micelle-decorated surfaces showed reduced adherence and survival of bacteria. To extend this passive defense against bacteria with an active strategy, the immobilized micelles were equipped with the antimicrobial peptide KYE28 (KYEITTIHNLFRKLTHRLFRRNFGYTLR). The peptide interacted nonspecifically with the immobilized micelles where it retained its antimicrobial property. The successful surface decoration with KYE28 was demonstrated by a combination of X-ray photoelectron spectroscopy and quartz crystal microbalance with dissipation monitoring. The initial antimicrobial activity of the nanostructured surfaces against Escherichia coli was found to be increased by the presence of KYE28. The combination of the active and passive strategy represents a straightforward modular approach that can easily be adapted, for example, by exchanging the antimicrobial peptide to optimize potency against challenging bacterial strains, and/or to simultaneously achieve antimicrobial and anti-infection properties.

Brushing the surface: cascade reactions between immobilized nanoreactors.

Functionalization of hard or soft surfaces with, for example, ligands, enzymes or proteins, is an effective and practical methodology for the development of new applications. We report the assembly of two types of nanoreactors based upon poly(dimethylsiloxane)-block-poly(2-methyl-2-oxazoline) (PDMS-b-PMOXA) diblock copolymers as scaffold, uricase and lactoperoxidase as bio-catalysts located within the nanoreactors, and melittin as the biopores inserted into the hydrophobic shell. The nanoreactors were immobilized on poly(2-hydroxyethyl methacrylate)-co-poly(2-aminoethyl methacrylate hydrochloride) (PHEMA-co-P(2-AEMA·HCl) brushes-grafted wafer surfaces by utilizing the strong supramolecular interactions between biotin and streptavidin. The (PHEMA-co-P(2-AEMA·HCl) brushes on silicon surfaces were prepared by a surface initiating atom transfer radical polymerization (ATRP) "graft-from" technique. Cascade reactions between different surface-anchored nanoreactors were demonstrated by converting Amplex® Red to the fluorescent probe resorufin by using the H2O2 produced from uric acid and H2O. The detailed properties of the nanoreactors on the functionalized surface including the binding behaviours and cascade reactions were investigated using emission spectroscopy, transmission electron microscopy (TEM), light scattering (LS), atomic force microscopy (AFM) and a quartz crystal microbalance (QCM-D). The results are proof-of-principle for the preparation of catalytically functional engineered surface materials and lay the foundation for applying this advanced functional surface material in biosensing, implanting and antimicrobial materials preparation.

Enzymatic reactions in polymeric compartments: nanotechnology meets nature.

One of the main features of living matter is compartmentalization, that is the temporal and spatial division of biological reactions and containment of the cellular components. Nanotechnology aims to replicate this, separating tiny environments from the exterior into nano-sized and micro-sized self-assembled compartments. Those synthetic compartments can perform reactions, be tracked and act in vivo. Here, an overview of the techniques to fabricate vesicular, polymer-based catalytic compartments and the parameters affecting their architecture is presented. How communication can be ensured across their membranes, recent developments in the enzymes that have been loaded into them and the latest advances in biological applications are discussed. This review highlights the characteristics that make polymers an enticing choice, the protection they offer, and their applications in compartmentalizing biologically relevant reactions.

Probing membrane asymmetry of ABC polymersomes.

We report the sensitivity of the membrane asymmetry of ABC (PEO-b-PCL-b-PMOXA) polymersomes towards the end-group modification of a shorter C block. While a non-modified ABC polymer formed polymersomes with the A block outside and the C block inside, a mixture of ABC and ABC-biotin formed polymersomes with the C block outside.

Surfaces with Dual Functionality through Specific Coimmobilization of Self-Assembled Polymeric Nanostructures.

Coimmobilization of functional, nanosized assemblies broadens the possibility to engineer dually functionalized active surfaces with a nanostructured texture. Surfaces decorated with different nanoassemblies, such as micelles, polymersomes, or nanoparticles are in high demand for various applications ranging from catalysis, biosensing up to antimicrobial surfaces. Here, we present a combination of bio-orthogonal and catalyst-free strain-promoted azide-alkyne click (SPAAC) and thiol-ene reactions to simultaneously coimmobilize various nanoassemblies; we selected polymersome-polymersome and polymersome-micelle assemblies. For the first time, the immobilization method using SPAAC reaction was studied in detail to attach soft, polymeric assemblies on a solid support. Together, the SPAAC and thiol-ene reactions successfully coimmobilized two unique self-assembled structures on the surfaces. Additionally, poly(dimethylsiloxane) (PDMS)-based polymersomes were used as "ink" for direct immobilization from a PDMS-based microstamp onto a surface creating locally defined patterns. Combining immobilization reactions has the advantage to attach any kind of nanoassembly pairs, resulting in surfaces with "desired" interfacial properties. Different nanoassemblies that encapsulate multiple active compounds coimmobilized on a surface will pave the way for the development of multifunctional surfaces with controlled properties and efficiency.

Nanoscience-Based Strategies to Engineer Antimicrobial Surfaces.

Microbial contamination and biofilm formation of medical devices is a major issue associated with medical complications and increased costs. Consequently, there is a growing need for novel strategies and exploitation of nanoscience-based technologies to reduce the interaction of bacteria and microbes with synthetic surfaces. This article focuses on surfaces that are nanostructured, have functional coatings, and generate or release antimicrobial compounds, including "smart surfaces" producing antibiotics on demand. Key requirements for successful antimicrobial surfaces including biocompatibility, mechanical stability, durability, and efficiency are discussed and illustrated with examples of the recent literature. Various nanoscience-based technologies are described along with new concepts, their advantages, and remaining open questions. Although at an early stage of research, nanoscience-based strategies for creating antimicrobial surfaces have the advantage of acting at the molecular level, potentially making them more efficient under specific conditions. Moreover, the interface can be fine tuned and specific interactions that depend on the location of the device can be addressed. Finally, remaining important challenges are identified: improvement of the efficacy for long-term use, extension of the application range to a large spectrum of bacteria, standardized evaluation assays, and combination of passive and active approaches in a single surface to produce multifunctional surfaces.

Exploiting exciton coupling of ligand radical intervalence charge transfer transitions to tune NIR absorption.

We detail the rational design of a series of bimetallic bis-ligand radical Ni salen complexes in which the relative orientation of the ligand radical chromophores provides a mechanism to tune the energy of intense intervalence charge transfer (IVCT) bands in the near infrared (NIR) region. Through a suite of experimental (electrochemistry, electron paramagnetic resonance spectroscopy, UV-vis-NIR spectroscopy) and theoretical (density functional theory) techniques, we demonstrate that bimetallic Ni salen complexes form bis-ligand radicals upon two-electron oxidation, whose NIR absorption energies depend on the geometry imposed in the bis-ligand radical complex. Relative to the oxidized monomer [1˙]+ (E = 4500 cm-1, ε = 27 700 M-1 cm-1), oxidation of the cofacially constrained analogue 2 to [2˙˙]2+ results in a blue-shifted NIR band (E = 4830 cm-1, ε = 42 900 M-1 cm-1), while oxidation of 5 to [5˙˙]2+, with parallel arrangement of chromophores, results in a red-shifted NIR band (E = 4150 cm-1, ε = 46 600 M-1 cm-1); the NIR bands exhibit double the intensity in comparison to the monomer. Oxidation of the intermediate orientations results in band splitting for [3˙˙]2+ (E = 4890 and 4200 cm-1; ε = 26 500 and 21 100 M-1 cm-1), and a red-shift for [4˙˙]2+ using ortho- and meta-phenylene linkers, respectively. This study demonstrates for the first time, the applicability of exciton coupling to ligand radical systems absorbing in the NIR region and shows that by simple geometry changes, it is possible to tune the energy of intense low energy absorption by nearly 400 nm.

Does Membrane Thickness Affect the Transport of Selective Ions Mediated by Ionophores in Synthetic Membranes?

Biomimetic polymer nanocompartments (polymersomes) with preserved architecture and ion-selective membrane permeability represent cutting-edge mimics of cellular compartmentalization. Here it is studied whether the membrane thickness affects the functionality of ionophores in respect to the transport of Ca2+ ions in synthetic membranes of polymersomes, which are up to 2.6 times thicker than lipid membranes (5 nm). Selective permeability toward calcium ions is achieved by proper insertion of ionomycin, and demonstrated by using specific fluorescence markers encapsulated in their inner cavities. Preservation of polymersome architecture is shown by a combination of light scattering, transmission electron microscopy, and fluorescence spectroscopy. By using a combination of stopped-flow and fluorescence spectroscopy, it is shown that ionomycin can function and transport calcium ions across polymer membranes with thicknesses in the range 10.7-13.4 nm (7.1-8.9 times larger than the size of the ionophore). Thicker membranes induce a decrease in transport, but do not block it due to the intrinsic flexibility of these synthetic membranes. The design of ion selective biomimetic nanocompartments represents a new path toward the development of cellular ion nanosensors and nano-reactors, in which calcium sensitive biomacromolecules can be triggered for specific biological functions.