Chondroitin sulfate is required for follicle epithelial integrity and organ shape maintenance in Drosophila.

Heparan sulfate (HS) and chondroitin sulfate (CS) are evolutionarily conserved glycosaminoglycans that are found in most animal species, including the genetically tractable model organism Drosophila. In contrast to extensive in vivo studies elucidating co-receptor functions of Drosophila HS proteoglycans (PGs), only a limited number of studies have been conducted for those of CSPGs. To investigate the global function of CS in development, we generated mutants for Chondroitin sulfate synthase (Chsy), which encodes the Drosophila homolog of mammalian chondroitin synthase 1, a crucial CS biosynthetic enzyme. Our characterizations of the Chsy mutants indicated that a fraction survive to adult stage, which allowed us to analyze the morphology of the adult organs. In the ovary, Chsy mutants exhibited altered stiffness of the basement membrane and muscle dysfunction, leading to a gradual degradation of the gross organ structure as mutant animals aged. Our observations show that normal CS function is required for the maintenance of the structural integrity of the ECM and gross organ architecture.

Nanoscale Abrasive Wear of Polyethylene: A Novel Approach To Probe Nanoplastic Release at the Single Asperity Level.

There is a growing concern that nanoplastic pollution may pose planetary threats to human and ecosystem health. However, a quantitative and mechanistic understanding of nanoplastic release via nanoscale mechanical degradation of bulk plastics and its interplay with photoweathering remains elusive. We developed a lateral force microscope (LFM)-based nanoscratch method to investigate mechanisms of nanoscale abrasive wear of low-density polyethylene (LDPE) surfaces by a single sand particle (simulated by a 300 nm tip) under environmentally relevant load, sliding motion, and sand size. For virgin LDPE, we found plowing as the dominant wear mechanism (i.e., deformed material pushed around the perimeter of scratch). After UVA-weathering, the wear mechanism of LDPE distinctively shifted to cutting wear (i.e., deformed material detached and pushed to the end of scratch). The shift in the mechanism was quantitatively described by a new parameter, which can be incorporated into calculating the NP release rate. We determined a 10-fold higher wear rate due to UV weathering. We also observed an unexpected resistance to initiate wear for UV-aged LDPE, likely due to nanohardness increase induced by UV. For the first time, we report 0.4-4 × 10-3 µm3/µm sliding distance/µN applied load as an initial approximate nanoplastic release rate for LDPE. Our novel findings reveal nanoplastic release mechanisms in the environment, enabling physics-based prediction of the global environmental inventory of nanoplastics.

Spatial Distribution of PEO-PPO-PEO Block Copolymer and PEO Homopolymer in Lipid Bilayers.

Maintaining the integrity of cell membranes is indispensable for cellular viability. Poloxamer 188 (P188), a poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide) (PEO-PPO-PEO) triblock copolymer with a number-average molecular weight of 8700 g/mol and containing 80% by mass PEO, protects cell membranes from various external injuries and has the potential to be used as a therapeutic agent in diverse applications. The membrane protection mechanism associated with P188 is intimately connected with how this block copolymer interacts with the lipid bilayer, the main component of a cell membrane. Here, we report the distribution of P188 in a model lipid bilayer comprising 1-palmitoyl-2-oleoyl-glycero-3-phosphocholine (POPC) using neutron reflectivity (NR) and atomic force microscopy (AFM). We also investigated the association of a PEO homopolymer (PEO8.4K; Mn = 8400 g/mol) that does not protect living cell membranes. These experiments were conducted following incubation of a 4.5 mmol/L polymer solution in a buffer that mimics physiological conditions with supported POPC bilayer membranes followed by washing with the aqueous medium. In contrast to previous reports, which dealt with P188 and PEO in salt-free solutions, both P188 and PEO8.4K penetrate into the inner portion of the lipid bilayer as revealed by NR, with approximately 30% by volume occupancy across the membrane without loss of bilayer structural integrity. These results indicate that PEO is the chemical moiety that principally drives P188 binding to bilayer membranes. No defects or phase-separated domains were observed in either P188- or PEO8.4K-incubated lipid bilayers when examined by AFM, indicating that polymer chains mingle homogeneously with lipid molecules in the bilayer. Remarkably, the breakthrough force required for penetration of the AFM tip through the bilayer membrane is unaffected by the presence of the large amount of P188 and PEO8.4K.

Mechanical Deformation Distinguishes Tunneling Pathways in Molecular Junctions.

Developing a clearer understanding of electron tunneling through molecules is a central challenge in molecular electronics. Here we demonstrate the use of mechanical stretching to distinguish orbital pathways that facilitate tunneling in molecular junctions. Our experiments employ junctions based on self-assembled monolayers (SAMs) of homologous alkanethiols (C nT) and oligophenylene thiols (OPT n), which serve as prototypical examples of σ-bonded and π-bonded backbones, respectively. Surprisingly, molecular conductances ( Gmolecule) for stretched C nT SAMs have exactly the same length dependence as unstretched C nT SAMs in which molecular length is tuned by the number of CH2 repeat units, n. In contrast, OPT n SAMs exhibit a 10-fold-greater decrease in Gmolecule with molecular length for stretched versus unstretched cases. Experiment and theory show that these divergent results are explained by the dependence of the molecule-electrode electronic coupling Γ on strain and the spatial extent of the principal orbital facilitating tunneling. In particular, differences in the strain sensitivity of Γ versus the repeat-length ( n) sensitivity can be used to distinguish tunneling via delocalized orbitals versus localized orbitals. Angstrom-level tuning of interelectrode separation thus provides a strategy for examining the relationship between orbital localization or delocalization and electronic coupling in molecular junctions and therefore for distinguishing tunneling pathways.

Exceptionally Small Statistical Variations in the Transport Properties of Metal-Molecule-Metal Junctions Composed of 80 Oligophenylene Dithiol Molecules.

Strong stochastic fluctuations witnessed as very broad resistance (R) histograms with widths comparable to or even larger than the most probable values characterize many measurements in the field of molecular electronics, particularly those measurements based on single molecule junctions at room temperature. Here we show that molecular junctions containing 80 oligophenylene dithiol molecules (OPDn, 1 ≤ n ≤ 4) connected in parallel display small relative statistical deviations-δR/R ≈ 25% after only ∼200 independent measurements-and we analyze the sources of these deviations quantitatively. The junctions are made by conducting probe atomic force microscopy (CP-AFM) in which an Au-coated tip contacts a self-assembled monolayer (SAM) of OPDs on Au. Using contact mechanics and direct measurements of the molecular surface coverage, the tip radius, tip-SAM adhesion force (F), and sample elastic modulus (E), we find that the tip-SAM contact area is approximately 25 nm2, corresponding to about 80 molecules in the junction. Supplementing this information with I-V data and an analytic transport model, we are able to quantitatively describe the sources of deviations δR in R: namely, δN (deviations in the number of molecules in the junction), δε (deviations in energetic position of the dominant molecular orbital), and Î´Γ (deviations in molecule-electrode coupling). Our main results are (1) direct determination of N; (2) demonstration that δN/N for CP-AFM junctions is remarkably small (≤2%) and that the largest contributions to δR are δε and δΓ; (3) demonstration that δR/R after only ∼200 measurements is substantially smaller than most reports based on >1000 measurements for single molecule break junctions. Overall, these results highlight the excellent reproducibility of junctions composed of tens of parallel molecules, which may be important for continued efforts to build robust molecular devices.

Growth of Thin, Anisotropic, π-Conjugated Molecular Films by Stepwise "Click" Assembly of Molecular Building Blocks: Characterizing Reaction Yield, Surface Coverage, and Film Thickness versus Addition Step Number.

We report the systematic characterization of anisotropic, π-conjugated oligophenyleneimine (OPI) films synthesized using stepwise imine condensation, or "click" chemistry. Film synthesis began with a self-assembled monolayer (SAM) of 4-formylthiophenol or 4-aminothiophenol on Au, followed by repetitive, alternate addition of terephthalaldehyde (benzene-1,4-dicarbaldehyde) or 1,4-benzenediamine to form π-conjugated films ranging from 0.6-5.5 nm in thickness. By systematically capping the OPI films with a redox or halogen label, we were able to measure the relative surface coverage after each monomer addition via Rutherford backscattering spectrometry, X-ray photoelectron spectroscopy, spectroscopic ellipsometry, reflection-absorption infrared spectroscopy, and cyclic voltammetry. Nuclear reaction analysis was also employed for the first time on a SAM to calculate the surface coverage of carbon atoms after each stepwise addition. These six different analysis methods indicate that the average extent of reaction is 99% for each addition step. The high yield and molecular surface coverage confirm the efficacy of Schiff base chemistry, at least with the terephthalaldehyde and 1,4-benzenediamine monomers, for preparing high-quality molecular films with π conjugation normal to the substrate.

AFM probing of polymer/nanofiller interfacial adhesion and its correlation with bulk mechanical properties in a poly(ethylene terephthalate) nanocomposite.

The interfacial adhesion between polymer and nanofiller plays an important role in affecting the properties of nanocomposites. The detailed relationship between interfacial adhesion and bulk properties, however, is unclear. In this work, we developed an atomic force microscopy (AFM)-based abrasive scanning methodology, as applied to model laminate systems, to probe the strength of interfacial adhesion relevant to poly(ethylene terephthalate) (PET)/graphene or clay nanocomposites. Graphite and mica substrates covered with ∼2 nm thick PET films were abrasively sheared by an AFM tip as a model measurement of interfacial strength between matrix PET and dispersed graphene and clay, respectively. During several abrasive raster-scan cycles, PET was shear-displaced from the scanned region. At temperatures below the PET glass transition, PET on graphite exhibited abrupt delamination (i.e., full adhesive failure), whereas PET on mica did not; rather, it exhibited a degree of cohesive failure within the shear-displaced layer. Moreover, 100-fold higher force scanning procedures were required to abrade through an ultimate "precursor" layer of PET only ∼0.2-0.5 nm thick, which must be largely disentangled from the matrix polymer. Thus, the adhesive interface of relevance to the strength of clay-filler nanocomposites is between matrix polymer and strongly bound polymer. At 90 °C, above the bulk PET glass transition temperature, the PET film exhibited cohesive failure on both graphite and mica. Our results suggest that there is little difference in the strength of the relevant interfacial adhesion in the two nanocomposites within the rubbery dynamic regime. Further, the bulk mechanical properties of melt mixed PET/graphene and PET/clay nanocomposites were evaluated by dynamic mechanical analysis. The glassy dynamic storage modulus of the PET/clay nanocomposite was higher than that of PET/graphene, correlating with the differences in interfacial adhesion probed by AFM.

Crystal-chemical origins of the ultrahigh conductivity of metallic delafossites.

Despite their highly anisotropic complex-oxidic nature, certain delafossite compounds (e.g., PdCoO2, PtCoO2) are the most conductive oxides known, for reasons that remain poorly understood. Their room-temperature conductivity can exceed that of Au, while their low-temperature electronic mean-free-paths reach an astonishing 20 µm. It is widely accepted that these materials must be ultrapure to achieve this, although the methods for their growth (which produce only small crystals) are not typically capable of such. Here, we report a different approach to PdCoO2 crystal growth, using chemical vapor transport methods to achieve order-of-magnitude gains in size, the highest structural qualities yet reported, and record residual resistivity ratios ( > 440). Nevertheless, detailed mass spectrometry measurements on these materials reveal that they are not ultrapure in a general sense, typically harboring 100s-of-parts-per-million impurity levels. Through quantitative crystal-chemical analyses, we resolve this apparent dichotomy, showing that the vast majority of impurities are forced to reside in the Co-O octahedral layers, leaving the conductive Pd sheets highly pure (∼1 ppm impurity concentrations). These purities are shown to be in quantitative agreement with measured residual resistivities. We thus conclude that a sublattice purification mechanism is essential to the ultrahigh low-temperature conductivity and mean-free-path of metallic delafossites.

Ultrathin, Transferred Layers of Silicon Oxynitrides as Tunable Biofluid Barriers for Bioresorbable Electronic Systems.

Bio/ecoresorbable electronic systems create unique opportunities in implantable medical devices that serve a need over a finite time period and then disappear naturally to eliminate the need for extraction surgeries. A critical challenge in the development of this type of technology is in materials that can serve as thin, stable barriers to surrounding ground water or biofluids, yet ultimately dissolve completely to benign end products. This paper describes a class of inorganic material (silicon oxynitride, SiON) that can be formed in thin films by plasma-enhanced chemical vapor deposition for this purpose. In vitro studies suggest that SiON and its dissolution products are biocompatible, indicating the potential for its use in implantable devices. A facile process to fabricate flexible, wafer-scale multilayer films bypasses limitations associated with the mechanical fragility of inorganic thin films. Systematic computational, analytical, and experimental studies highlight the essential materials aspects. Demonstrations in wireless light-emitting diodes both in vitro and in vivo illustrate the practical use of these materials strategies. The ability to select degradation rates and water permeability through fine tuning of chemical compositions and thicknesses provides the opportunity to obtain a range of functional lifetimes to meet different application requirements.

Delivery of RNA to the Blood-Brain Barrier Endothelium Using Cationic Bicelles.

Blood-brain barrier (BBB) dysfunction is prevalent in Alzheimer's disease and other neurological disorders. Restoring normal BBB function through RNA therapy is a potential avenue for addressing cerebrovascular changes in these disorders that may lead to cognitive decline. Although lipid nanoparticles have been traditionally used as drug carriers for RNA, bicelles have been emerging as a better alternative because of their higher cellular uptake and superior transfection capabilities. Cationic bicelles composed of DPPC/DC7PC/DOTAP at molar ratios of 63.8/25.0/11.2 were evaluated for the delivery of RNA in polarized hCMEC/D3 monolayers, a widely used BBB cell culture model. RNA-bicelle complexes were formed at five N/P ratios (1:1 to 5:1) by a thin-film hydration method. The RNA-bicelle complexes at N/P ratios of 3:1 and 4:1 exhibited optimal particle characteristics for cellular delivery. The cellular uptake of cationic bicelles laced with 1 mol% DiI-C18 was confirmed by flow cytometry and confocal microscopy. The ability of cationic bicelles (N/P ratio 4:1) to transfect polarized hCMEC/D3 with FITC-labeled control siRNA was tested vis-a-vis commercially available Lipofectamine RNAiMAX. These studies demonstrated the higher transfection efficiency and greater potential of cationic bicelles for RNA delivery to the BBB endothelium.

Use of Atomic Force Microscopy (AFM) to monitor surface crystallization in caffeine-oxalic acid (CAFOXA) cocrystal compacts.

Our objective was to monitor the surface crystallization in disordered caffeine-oxalic acid (CAFOXA) cocrystals following exposure to elevated water vapor pressure. This was accomplished using atomic force microscopy (AFM). Disorder was induced in the cocrystal particles by the common pharmaceutical unit operations of milling and compaction. The 'activated' solid, upon exposure to elevated water vapor pressure, had a high propensity to sorb water. This led to a rise in molecular mobility and the surface underwent rapid crystallization to form needle shaped crystals of CAFOXA. Using AFM height and phase imaging, we were able to directly visualize phase transformations on the compact surface. The milled compacts exhibited higher processing induced disorder than the unmilled compacts, thereby accelerating the surface recrystallization.

Holey Substrate-Directed Strain Patterning in Bilayer MoS2.

Key properties of two-dimensional (2D) layered materials are highly strain tunable, arising from bond modulation and associated reconfiguration of the energy bands around the Fermi level. Approaches to locally controlling and patterning strain have included both active and passive elastic deformation via sustained loading and templating with nanostructures. Here, by float-capturing ultrathin flakes of single-crystal 2H-MoS2 on amorphous holey silicon nitride substrates, we find that highly symmetric, high-fidelity strain patterns are formed. The hexagonally arranged holes and surface topography combine to generate highly conformal flake-substrate coverage creating patterns that match optimal centroidal Voronoi tessellation in 2D Euclidean space. Using TEM imaging and diffraction, as well as AFM topographic mapping, we determine that the substrate-driven 3D geometry of the flakes over the holes consists of symmetric, out-of-plane bowl-like deformation of up to 35 nm, with in-plane, isotropic tensile strains of up to 1.8% (measured with both selected-area diffraction and AFM). Atomistic and image simulations accurately predict spontaneous formation of the strain patterns, with van der Waals forces and substrate topography as the input parameters. These results show that predictable patterns and 3D topography can be spontaneously induced in 2D materials captured on bare, holey substrates. The method also enables electron scattering studies of precisely aligned, substrate-free strained regions in transmission mode.

Distinguishing elastic shear deformation from friction on the surfaces of molecular crystals.

Elastic deformation on the surfaces of molecular crystals can be imaged using a variant of lateral force microscopy in which the tip is scanned parallel to the cantilever axis. The shear force transverse to this direction has a distinctly different origin than the friction force as determined by the tip velocity and temperature dependence of the cantilever torque. An elastic deformation model for the tip-sample interaction predicts the crystallographic anisotropy of the transverse shear contrast, establishing its connection with the relative magnitude of the in-plane elastic tensor components.

Coexistence of Low Damping and Strong Magnetoelastic Coupling in Epitaxial Spinel Ferrite Thin Films.

Low-loss magnetization dynamics and strong magnetoelastic coupling are generally mutually exclusive properties due to opposing dependencies on spin-orbit interactions. So far, the lack of low-damping, magnetostrictive ferrite films has hindered the development of power-efficient magnetoelectric and acoustic spintronic devices. Here, magnetically soft epitaxial spinel NiZnAl-ferrite thin films with an unusually low Gilbert damping parameter (<3 × 10-3 ), as well as strong magnetoelastic coupling evidenced by a giant strain-induced anisotropy field (≈1 T) and a sizable magnetostriction coefficient (≈10 ppm), are reported. This exceptional combination of low intrinsic damping and substantial magnetostriction arises from the cation chemistry of NiZnAl-ferrite. At the same time, the coherently strained film structure suppresses extrinsic damping, enables soft magnetic behavior, and generates large easy-plane magnetoelastic anisotropy. These findings provide a foundation for a new class of low-loss, magnetoelastic thin film materials that are promising for spin-mechanical devices.

Measuring the Thickness and Potential Profiles of the Space-Charge Layer at Organic/Organic Interfaces under Illumination and in the Dark by Scanning Kelvin Probe Microscopy.

Scanning Kelvin probe microscopy was used to measure band-bending at the model donor/acceptor heterojunction poly(3-hexylthiophene) (P3HT)/fullerene (C60). Specifically, we measured the variation in the surface potential of C60 films with increasing thicknesses grown on P3HT to produce a surface potential profile normal to the substrate both in the dark and under illumination. The results confirm a space-charge carrier region with a thickness of 10 nm, consistent with previous observations. We discuss the possibility that the domain size in bulk heterojunction organic solar cells, which is comparable to the space-charge layer thickness, is actually partly responsible for less than expected electron/hole recombination rates.

Characterization of Pebax angioplasty balloon surfaces with AFM, SEM, TEM, and SAXS.

In the medical device industry, angioplasty balloons have been widely used in the less invasive treatment of heart disease by expanding and relieving clogged structures in various arterial segments. However, new applications using thin coatings on the balloon surface have been explored to enhance therapeutic value in the delivery of pharmaceuticals (drug-elution) or control thermal energy output (RF ablation). In this study, angioplasty balloon materials comprised of poly(ether-block-amide) (Pebax) were investigated via atomic force microscopy (AFM), transmission electron microscopy (TEM), scanning electron microscopy (SEM), and small-angle X-ray scattering (SAXS) to characterize physical properties at the balloon surface that may affect coating adhesion. The soft segment of this Pebax 1074 material is polyethylene oxide (PEO) and the hard segment is nylon-12. The morphology of the hard segments of this block co-polymer are found via AFM stiffness measurements to be (40 ± 20) nm by (300 ± 150) nm and are oriented parallel to the surface of the balloon. SAXS measurements found the lamellar spacing to be (18.5 ± 0.5) nm, and demonstrate a preferential orientation in agreement with TEM and AFM measurements. Fixation of this balloon in resin, followed by cryo-sectioning is shown to provide a novel manner in which to investigate surface characteristics on the balloon such as material or coating thickness as well as uniformity in comparison to the bulk structure. These outputs were deemed critical to improve overall balloon processing such as molding and surface treatment options for robust designs toward better procedural outcomes targeting new therapeutic areas.

Strain effects on the work function of an organic semiconductor.

Establishing fundamental relationships between strain and work function (WF) in organic semiconductors is important not only for understanding electrical properties of organic thin films, which are subject to both intrinsic and extrinsic strains, but also for developing flexible electronic devices. Here we investigate tensile and compressive strain effects on the WF of rubrene single crystals. Mechanical strain induced by thermal expansion mismatch between the substrate and rubrene is quantified by X-ray diffraction. The corresponding WF change is measured by scanning Kelvin probe microscopy. The WF of rubrene increases (decreases) significantly with in-plane tensile (compressive) strain, which agrees qualitatively with density functional theory calculations. An elastic-to-plastic transition, characterized by a steep rise of the WF, occurs at ∼0.05% tensile strain along the rubrene π-stacking direction. The results provide the first concrete link between mechanical strain and WF of an organic semiconductor and have important implications for understanding the connection between structural and electronic disorder in soft organic electronic materials.

Water sorption induced transformations in crystalline solid surfaces: characterization by atomic force microscopy.

The effect of water sorption on the mobility of molecules on the surface of a crystalline anhydrous solid was investigated to understand the mechanism of its transformation to the corresponding hydrate. Theophylline was chosen as the model compound. The transition water activity for anhydrate to hydrate transformation, RH(T), and the deliquescence RH, RH(0), was determined to be 62% and 99%, respectively (25 degrees C). Atomic force microscopy (AFM) was used to study the surface changes of theophylline above and below the transition water activity. Contact-mode AFM showed that the jump-to-contact distance increased appreciably above RH(T), suggesting formation of solution on the surface. At RH(T) < RH < RH(0), using dynamic (AC/"tapping" mode) AFM, the movements of surface steps were visualized. These results from AFM indicated that, below RH(0), the formation of a thin solution film significantly increased surface mobility. Furthermore, when the anhydrate crystal surface was seeded with the hydrate, the propagation of a new hydrate phase was observed by polarized light microscopy. In conclusion, atomic force microscopy provided direct evidence that the phase transformation of anhydrous theophylline to theophylline monohydrate in the solid-state is mediated by a surface solution as a result of water adsorption.

Multimodal dynamic imaging of therapeutic biomedical coatings in aqueous medium.

Drug release from therapeutic biomedical films such as drug-polymer composite coatings on drug eluting stents is a highly complex and poorly understood process. The dynamics of drug release and the evolution of surface morphology during release have direct impact on the performance of the device. This information is not easily accessible, and there have been few systematic studies to investigate drug release from biomedical coatings in real time. In this study, the complementary analytical techniques of confocal Raman microscopy, in-liquid atomic force microscopy, scanning electron microscopy, and high performance liquid chromatography were used to examine real-time mobilization and release of the drug rapamycin from polyisobutylene-block-polystyrene thin films, during immersion in buffered saline for 12 h. Each technique was found to have distinct limitations in either temporal or spatial resolution; in combination, however, the overlapping techniques provided a level of detail that is not available using any single approach.