Engineered Fibrillar Fibronectin Networks as Three‐Dimensional Tissue Scaffolds

Extracellular matrix (ECM) proteins, and most prominently, fibronectin (Fn), are routinely used in the form of adsorbed pre‐coatings in an attempt to create a cell‐supporting environment in both two‐ and three‐dimensional cell culture systems. However, these protein coatings are typically deposited in a form which is structurally and functionally distinct from the ECM‐constituting fibrillar protein networks naturally deposited by cells. Here, the cell‐free and scalable synthesis of freely suspended and mechanically robust three‐dimensional (3D) networks of fibrillar fibronectin (fFn) supported by tessellated polymer scaffolds is reported. Hydrodynamically induced Fn fibrillogenesis at the three‐phase contact line between air, an Fn solution, and a tessellated scaffold microstructure yields extended protein networks. Importantly, engineered fFn networks promote cell invasion and proliferation, enable in vitro expansion of primary cancer cells, and induce an epithelial‐to‐mesenchymal transition in cancer cells. Engineered fFn networks support the formation of multicellular cancer structures cells from plural effusions of cancer patients. With further work, engineered fFn networks can have a transformative impact on fundamental cell studies, precision medicine, pharmaceutical testing, and pre‐clinical diagnostics.     

Chemical-vapor-deposition-based polymer substrates for spatially resolved analysis of protein binding by imaging ellipsometry

Biomolecular interactions between proteins and synthetic surfaces impact diverse biomedical fields. Simple, quantitative, label-free technologies for the analysis of protein adsorption and binding of biomolecules are thus needed. Here, we report the use of a novel type of substrate, poly-p-xylylene coatings prepared by chemical vapor deposition (CVD) polymerization, for surface plasmon resonance enhanced ellipsometry (SPREE) studies and assess the reactive coatings as spatially resolved biomolecular sensing arrays. Prior to use in binding studies, reactive coatings were fully characterized by Fourier transform infrared spectroscopy, electrochemical impedance spectroscopy, and ellipsometry. As a result, the chemical structure, thickness, and homogeneous coverage of the substrate surface were confirmed for a series of CVD-coated samples. Subsequent SPREE imaging and fluorescence microscopy indicated that the synthetic substrates supported detectable binding of a cascade of biomolecules. Moreover, analysis revealed a useful thickness range for CVD films in the assessment of protein and/or antigen-antibody binding via SPREE imaging. With a variety of functionalized end groups available for biomolecule immobilization and ease of patterning, CVD thin films are useful substrates for spatially resolved, quantitative binding arrays.     

Engineering, characterization and directional self-assembly of anisotropically modified nanocolloids

Along with traditional attributes such as the size, shape, and chemical structure of polymeric micro-objects, control over material distribution, or selective compartmentalization, appears to be increasingly important for maximizing the functionality and efficacy of biomaterials. The fabrication of tri- and tetracompartmental colloids made from biodegradable poly(lactide-co-glycolide) polymers via electrohydrodynamic co-jetting is demonstrated. The presence of three compartments is confirmed via flow cytometry. Additional chemical functionality is introduced via the incorporation of acetylene-functionalized polymers into individual compartments of the particles. Direct visualization of the spatioselective distribution of acetylene groups is demonstrated by confocal Raman microscopy as well as by reaction of the acetylene groups with azide-biotin via 'click chemistry'. Biotin-streptavidin binding is then utilized for the controlled assembly and orientation of bicompartmental particles onto functionalized, micropatterned substrates prepared via chemical vapor deposition polymerization.     

Synthesis and properties of new thermosensitive polymer systems

The controlled thermal release of aqueous solvent mixtures from polymeric gel particles was investigated. A new type of a polymeric gel consisting of a maleic anhydride/poly(ethylene glycol) condensation product as a crosslinking macromonomer and acrylamides was synthesized by solution or inverse emulsion polymerization for the investigation. Afterward a shell of crosslinked polystyrene was coated to stabilize this new kind of “microcontainer” for the application. This concept was shown as generally useful for various mixtures of organic solvents and water. The cloud point temperature of the polymer gel strongly depended on the following parameters: the type and content of organic cosolvent, the degree of polymerization and constituents of the polyester moiety, and the type and content of the comonomers. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci, 2007     

Biphasic Janus particles with nanoscale anisotropy

Advances in the field of nanotechnology have fuelled the vision of future devices spawned from tiny functional components that are able to assemble according to a master blueprint. In this concept, the controlled distribution of matter or 'patchiness' is important for creating anisotropic building blocks and introduces an extra design parameter--beyond size and shape. Although the reliable and efficient fabrication of building blocks with controllable material distributions will be of interest for many applications in research and technology, their synthesis has been addressed only in a few specialized cases. Here we show the design and synthesis of polymer-based particles with two distinct phases. The biphasic geometry of these Janus particles is induced by the simultaneous electrohydrodynamic jetting of parallel polymer solutions under the influence of an electrical field. The individual phases can be independently loaded with biomolecules or selectively modified with model ligands, as confirmed by confocal microscopy and transmission electron microscopy. The fact that the spatial distribution of matter can be controlled at such small length scales will provide access to unknown anisotropic materials. This type of nanocolloid may enable the design of multicomponent carriers for drug delivery, molecular imaging or guided self-assembly.     

Effective schema for the rigorous modeling of grating diffraction with focused beams

Most modal diffraction methods are formulated for incident plane waves. In practical applications, the probing beam is focused. Usually, this is simulated by means of numerical integration where Gaussian quadrature formulas are most effective. These formulas require smooth integrands, which is not fulfilled for gratings due to Rayleigh singularities and physical resonances. The violation of this condition entails inaccurate integration results, such as kinks and other artifacts. In this paper, a methodology for the efficient treatment of the numerical integration with improved accuracy is presented. It is based on the subdivision of the aperture along the lines of Rayleigh singularities, mapping of these subapertures into unit squares, and separate application of the Gaussian cubature formulas for each subarea.     

Perturbation approach applied to modal diffraction methods

Eigenvalue computation is an important part of many modal diffraction methods, including the rigorous coupled wave approach (RCWA) and the Chandezon method. This procedure is known to be computationally intensive, accounting for a large proportion of the overall run time. However, in many cases, eigenvalue information is already available from previous calculations. Some of the examples include adjacent slices in the RCWA, spectral- or angle-resolved scans in optical scatterometry and parameter derivatives in optimization. In this paper, we present a new technique that provides accurate and highly reliable solutions with significant improvements in computational time. The proposed method takes advantage of known eigensolution information and is based on perturbation method.     

Direct Protein Detection in the Sample Solution by Monitoring Rotational Dynamics of Nickel Nanorods

The feasibility of a recently introduced homogeneous immunodiagnostic approach to directly detect analyte binding by optical observation of the hydrodynamic properties of magnetically rotated nanorods (“PlasMag”) is demonstrated experimentally. Specifically, it is shown that the phase lag of the long axis of nickel nanorods (magnetic core parameters: length 182 nm, diameter 26 nm) with respect to externally applied rotating magnetic fields significantly increases on the adhesion of bovine serum albumin (BSA) protein to their surfaces. To validate these results, the amount of bound protein molecules is independently determined by analysis of the electrophoretic mobility of the nanorods. Furthermore, the data also demonstrate the applicability of recently developed empirical models based on numerical solutions of the Fokker‐Planck equation for describing the dynamics of magnetic nanoparticles in rotating magnetic fields.