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A simple and efficient method for the synthesis of biodegradable, highly branched polycaprolactone (PCL) is presented. The solvent-free (bulk) reaction was carried out via ring opening polymerization (ROP), catalyzed by tin octanoate Sn(Oct)2, and it employed hyperbranched polyamide (HPPA) as a macro-initiator. The core–shell structure of the obtained products (PCL-HPPA), with the hyperbranched HPPA core and linear PCL chains as shell, was in the focus of the product characterization. 1H nuclear magnetic resonance (1H NMR) and elemental analysis confirmed the covalent incorporation of the HPPA in the products, as well as a high degree of grafting conversion of its amino functional groups. Confocal Raman Micro spectroscopy, and especially Time-of-Flight Secondary Ion Mass Spectrometry, further supported the existence of a core–shell structure in the products. Direct observation of macromolecules by means of cryogenic transmission electron microscopy, as well as gel permeation chromatography (GPC), suggested the existence of a minor 'aggregated' product fraction with multiple HPPA cores, which was attributed to transesterification reactions. Differential scanning calorimetry, as well as X-ray diffraction, demonstrated that the PCL-HPPA polymers displayed a similar degree of crystallinity to linear neat PCL, but that the branched products possessed smaller and less regular crystallites. [ABSTRACT FROM AUTHOR]

The unique features of poly[2‐(dimethylamino)ethyl methacrylate] (PDMAEMA), such as its sensitivity to external stimuli like pH and the presence of tertiary amine groups that can be easily quaternized to introduce antibacterial properties, make it a promising platform for biomedical applications. In this contribution, a facile, cost‐effective, and ecological procedure for controlled grafting of PDMAEMA brushes from a glass surface, both in mL and µL scale, is developed. This concept involves utilizing an aqueous solution of sunflower honey as a source of reducing sugars to accelerate surface‐initiated activators regenerated by electron transfer atom transfer radical polymerization. The PDMAEMA chains covalently grafted to the glass surface are then quaternized to form an antibacterial film. The thickness of the polymeric brush layer is determined by atomic force microscopy, while the chemical composition is analyzed by time of flight secondary ion mass spectrometry. Water contact angle measurements demonstrate the pH‐sensitivity of PDMAEMA pointing out the potential application of the prepared material as smart surfaces. Furthermore, the antibacterial tests against Gram‐positive and Gram‐negative bacteria strains are performed. The protein adsorption is used to evaluate the biocompatibility of the prepared surfaces. The resulting glass materials can serve as multifunctional surfaces for various purposes. [ABSTRACT FROM AUTHOR]

Fluorinated polymers have unique wettability and protein adsorption properties. The site-specific alteration of these properties could expand their application to different research areas. In this work, a fluorinated homopolymer and two of its copolymers with 4-vinylbenzyl glycidyl ether (VBGE) are synthesized by free radical polymerization. The produced polymers are then used to develop resist formulations by the addition of a photoacid generator. Films of these formulations are exposed to ultraviolet radiation through a binary mask and heated to create the pattern. It is found that the water contact angle values of the exposed films areas are reduced compared to those of the unexposed ones, with the exception of pentafluorophenyl methacrylate (PFMA) homopolymer film. This is attributed to the reaction of the epoxy groups creating x-links and producing hydroxyl groups and the cleavage of the pentafluorophenyl group from the ester group leading to carboxylic acid groups. Both modifications on the exposed areas are verified by FTIR spectroscopy and ToF-SIMS analysis. In addition, the biomolecules adsorption ability of the exposed area is increasing 10–15 times compared to the unexposed one for the PFMA homopolymer and the PFMA/VBGE 1:1 copolymer. Thus, the proposed polymers and patterning procedure could find application to spatially directed immobilization of biomolecules and/or cells onto a surface for both biosensing and tissue engineering purposes. [ABSTRACT FROM AUTHOR]

The fibrotic fibroblasts derived from idiopathic pulmonary fibrosis (IPF) and nonspecific interstitial pneumonia (NSIP) are surrounded by specific environments, characterized by increased stiffness, aberrant extracellular matrix (ECM) composition, and altered lung architecture. The presented research was aimed at investigating the effect of biological, physical, and topographical modification of the substrate on the properties of IPF- and NSIP-derived fibroblasts, and searching for the parameters enabling their identification. Soft and stiff polydimethylsiloxane (PDMS) was chosen for the basic substrates, the properties of which were subsequently tuned. To obtain the biological modification of the substrates, they were covered with ECM proteins, laminin, fibronectin, and collagen. The substrates that mimicked the 3D structure of the lungs were prepared using two approaches, resulting in porous structures that resemble natural lung architecture and honeycomb patterns, typical of IPF tissue. The growth of cells on soft and stiff PDMS covered with proteins, traced using fluorescence microscopy, confirmed an altered behavior of healthy and IPF- and NSIP-derived fibroblasts in response to the modified substrate properties, enabling their identification. In turn, differences in the mechanical properties of healthy and fibrotic fibroblasts, determined using atomic force microscopy working in force spectroscopy mode, as well as their growth on 3D-patterned substrates were not sufficient to discriminate between cell lines. [ABSTRACT FROM AUTHOR]

In the present work, three types of grafted brush coatings [P4VP, POEGMA246, and P(4VP-co-POEGMA246)] were successfully fabricated using graft polymerization of monomers "from the surface." The composition, thickness, and morphology of the grafted brush coatings were analyzed by TOF-SIMS, ellipsometry, and AFM, respectively. The chemical nature of the polymer surface plays a crucial role in the growth and development of the cow granulosa cells and, therefore, also oocyte-cumulus complexes. In comparison with other coatings, the P(4VP-co-POEGMA246) copolymer coating enables the formation of dispersed and small but numerous cell conglomerates and high cumulus expansion in oocyte-cumulus complexes with highly homogeneous cumulus layers surrounding the oocytes. Moreover, the cellular oxygen uptake for this coating in the presence of NaF (inhibitor glycolysis) was stimulated. This new (4VP-co-POEGMA246) copolymer nanostructured coating is a promising material for granulosa cell and oocyte-cumulus complex cultivation and possibly will have great potential for applications in veterinary and reproductive medicine. [ABSTRACT FROM AUTHOR]

Abstract: The origin and formation of the dipole moment at the interface of metal electrode and thin organic film is crucial for the fabrication of high‐performance organic electronic devices. In this work, model polymer multilayer systems are studied in which interfaces are modified heterogeneously with thiol‐based self‐assembled monolayers (SAM). Due to the presence of polar terminal group (‐COOH) in SAM and polar pendant groups (‐COOH, ‐C(O)OCH3) within studied polymers, it is pointed out that the resultant interface energetics differ from the expected ones as one would expect from the chemical character of the self‐assembled molecules used to tune junction properties. This is discussed in terms of inter‐association and depolarization effects between polymer and thiol modified substrate. The presented model systems may serve as an interaction reference for SAMs and semiconducting polymers containing similar moieties, utilized for the fabrication of organic electronic devices. [ABSTRACT FROM AUTHOR]

Poly(n-butyl methacrylate) (PBMA) grafted polymer brushes attached to glass were fabricated in a three-step process involving surface initiated atom transfer radical polymerization. The surface properties of the coatings after subsequent fabrication steps were confirmed using ToF-SIMS and ellipsometry. Measurements of water contact angle and AFM revealed temperature-induced changes in the hydrophobicity and morphology of the coating. The glass transition temperatures (Tg) of the PBMA coatings with different thicknesses were determined from the AFM measurements. For the PBMA grafted brush coatings with thicknesses less than 62 nm, Tg increases sharply with increasing thickness. The PBMA grafted coatings of thickness equal to 86 nm and 43 nm as well as control glass substrates were used as substrates for culturing a urinary bladder cancer HTB-5 cell line. After 144 h of culturing, a well-developed monocellular layer may be observed on the PBMA coating of thickness equal to 86 nm. In turn, the cells incubated on thinner (43 nm) PBMA coatings as well as on a control glass sample only start to form a confluent layer. [ABSTRACT FROM AUTHOR]

Tuning the transition between stable hydrophilicity and high etching-rate and simultaneous hydrophilization of polymethylmethacrylate (PMMA) films was achieved in an open-air environment simply by adjusting the O2 content in He gas injection through the showerhead-type electrode of a novel atmospheric pressure dielectric barrier discharge source. A comparative study was performed for He and He-O2 plasma treatment of PMMA. Electrical and optical measurements indicated severe alteration of plasma characteristics when O2 is added in gas feed. The enhancement of oxygen emission band intensities indicates an enhanced concentration of atomic oxygen, resulting in very high PMMA etching rates of ~320 nm/min, ~four times higher compared to pure He. In contrast, the He plasma showed higher nitrogen emission band intensities, translated into a higher nitrogen content on the PMMA surface according to x-ray photoelectron spectroscopy and time-of-flight secondary ion mass spectrometry. This enhanced nitrogen content due to the partial substitution of ester groups of the PMMA side chain with amide groups was responsible for the more efficient hydrophilization of the PMMA surface. A water contact angle as low as ~28#176; was achieved, stabilizing at ~40° after 10 days and remained constant at least for 45 days, significantly lower than that for the untreated PMMA (~69°) [ABSTRACT FROM AUTHOR]

Time-of-flight-secondary ion mass spectrometry (TOF-SIMS) mass spectra measurements combined with an appropriate sample preparation protocol are the powerful tools to obtain unique information about the chemical composition of biological materials. In our studies, two questions were addressed, i.e., whether it is possible to develop a fixative-based sample preparation protocol and whether it allows one to distinguish between cells originating from various stages of cancer progression. Therefore, four human bladder cancer cell lines (with distinct malignancy degree) have been investigated. A chemical fixation protocol has been used for TOF-SIMS measurements, and mass spectra were obtained using a Bi3+ primary ion beam. The principal component analysis (PCA) has been applied to analyze the whole range of mass spectra (without preselection of any particular masses) using two approaches of data preprocessing, namely, mean centering and autoscaling. The PC3 versus PC2 plot has showed significant differences between nonmalignant cancer cells and the cancerous ones for both of preprocessing approaches. The analysis of mass spectra of human bladder cells allows one tofind a list of mass peaks with intensities significantly larger in cancerous bladder cells compared to nonmalignant cell cancer of the ureter (HCV29 cells). These findings show that TOF-SIMS in combination with PCA can be used to identify reference, human bladder cells from cancerous ones. [ABSTRACT FROM AUTHOR]

Three multi-step multi-molecular approaches using the biotin–streptavidin system to contact-print DNA arrays on SiO2 surfaces modified with (3-glycidoxypropyl)trimethoxysilane are examined after each deposition/reaction step by atomic force microscopy, X-ray photoelectron spectroscopy and time of flight secondary ion mass spectrometry. Surface modification involves the spotting of preformed conjugates of biotinylated oligonucleotides with streptavidin onto surfaces coated with biotinylated bovine serum albumin b-BSA (approach I) or the spotting of biotinylated oligonucleotides onto a streptavidin coating, the latter prepared through a reaction with immobilized b-BSA (approach II) or direct adsorption (approach III). AFM micrographs, quantified by autocorrelation and height histogram parameters (e.g. roughness), reveal uniform coverage after each modification step with distinct nanostructures after the reaction of biotinylated BSA with streptavidin or of a streptavidin conjugate with biotinylated oligonucleotides. XPS relates the immobilization of biomolecules with covalent binding to the epoxy-silanized surface. Protein coverage, estimated from photoelectron attenuation, shows that regarding streptavidin the highest and the lowest immobilization efficiency is achieved by following approaches I and III, respectively, as confirmed by TOF-SIMS microanalysis. The size of the DNA spot reflects the contact radius of the printed droplet and increases with protein coverage (and roughness) prior to the spotting, as epoxy-silanized surfaces are hardly hydrophilic. Representative TOF-SIMS images show sub-millimeter spots: uniform for approach I, doughnut-like (with a small non-zero minimum) for approach II, both with coffee-rings or peak-shaped for approach III. Spot features, originating from pinned contact lines and DNA surface binding and revealed by complementary molecular distributions (all material, DNA, streptavidin, BSA, epoxy, SiO2), indicate two modes of droplet evaporation depending on the details of each applied approach. [ABSTRACT FROM AUTHOR]

Versatility of solution-processing strategy based on the simultaneous rather than additive deposition of different functional molecules is discussed. It is shown that spin-cast polymer blends result in films with domains that could form elements with complementary functions of (i) solar cells, (ii) electronic circuitries, and (iii) test plates for protein micro-arrays: Alternating layers, rich in electron-donating polyfluorene and electron-accepting fullerene derivative, result in optimized solar power conversion. Surface patterns, made by soft lithography, align conductive paths of conjugated poly(3-alkylthiophene) in dielectric polystyrene. Proteins, preserving their biologically activity, are adsorbed to hydrophobic domains of polystyrene in hydrophilic matrix of poly(ethylene oxide). The authors report the research progress on structure formation in three polymer blend families, resulting in films with complementary elements for electronics and biotechnology. Blend film structures are determined with secondary ion mass spectrometry, atomic force microscopy, and fluorescence microscopy. In addition, the authors present recent results on (i) structure formation in fullerene derivative/poly(3-alkylthiophene) blends intended for solar cells, (ii) 3-dimensional SIMS imaging of conductive paths of poly(3-alkylthiophene) in dielectric polystyrene, (iii) test plates for multiprotein micro-arrays fabricated with blend films of hydrophobic polystyrene and thermoresponsive poly( N-isopropylacrylamide). © 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012 [ABSTRACT FROM AUTHOR]

Background: The aim of the research presented here was to find a set of parameters enabling discrimination between three types of fibroblasts, i.e., healthy ones and those derived from two disorders mimicking each other: idiopathic pulmonary fibrosis (IPF), and nonspecific interstitial pneumonia (NSIP). Methods: The morphology and growth of cells were traced using fluorescence microscopy and analyzed quantitatively using cell proliferation and substrate cytotoxicity indices. The viability of cells was recorded using MTS assays, and their stiffness was examined using atomic force microscopy (AFM) working in force spectroscopy (FS) mode. To enhance any possible difference in the examined parameters, experiments were performed with cells cultured on substrates of different elasticities. Moreover, the chemical composition of cells was determined using time-of-flight secondary ion mass spectrometry (ToF-SIMS), combined with sophisticated analytical tools, i.e., Multivariate Curve Resolution (MCR) and Principal Component Analysis (PCA). Results: The obtained results demonstrate that discrimination between cell lines derived from healthy and diseased patients is possible based on the analysis of the growth of cells, as well as their physical and chemical properties. In turn, the comparative analysis of the cellular response to altered stiffness of the substrates enables the identification of each cell line, including distinguishing between IPF- and NSIP-derived fibroblasts. [ABSTRACT FROM AUTHOR]

In the present work, we have successfully prepared and characterized novel nanocomposite material exhibiting temperature-dependent surface wettability changes, based on grafted brush coatings of non-fouling poly(di(ethylene glycol)methyl ether methacrylate) (POEGMA) with the embedded CaCO3 nanoparticles. Grafted polymer brushes attached to the glass surface were prepared in a three-step process using atom transfer radical polymerization (ATRP). Subsequently, uniform CaCO3 nanoparticles (NPs) embedded in POEGMA-grafted brush coatings were synthesized using biomineralized precipitation from solutions of CaCl2 and Na2CO3. An impact of the low concentration of the embedded CaCO3 NPs on cell adhesion and growth depends strongly on the type of studied cell line: keratinocytes (HaCaT), melanoma (WM35) and osteoblastic (MC3T3-e1). Based on the temperature-responsive properties of grafted brush coatings and CaCO3 NPs acting as biologically active substrate, we hope that our research will lead to a new platform for tissue engineering with modified growth of the cells due to the release of biologically active substances from CaCO3 NPs and the ability to detach the cells in a controlled manner using temperature-induced changes of the brush. [ABSTRACT FROM AUTHOR]

The presented research aims to verify whether physicochemical properties of lung fibroblasts, modified by substrate stiffness, can be used to discriminate between normal and fibrotic cells from idiopathic pulmonary fibrosis (IPF). The impact of polydimethylsiloxane (PDMS) substrate stiffness on the physicochemical properties of normal (LL24) and IPF-derived lung fibroblasts (LL97A) was examined in detail. The growth and elasticity of cells were assessed using fluorescence microscopy and atomic force microscopy working in force spectroscopy mode, respectively. The number of fibroblasts, as well as their shape and the arrangement, strongly depends on the mechanical properties of the substrate. Moreover, normal fibroblasts remain more rigid as compared to their fibrotic counterparts, which may indicate the impairments of IPF-derived fibroblasts induced by the fibrosis process. The chemical properties of normal and IPF-derived lung fibroblasts inspected using time-of-flight secondary ion mass spectrometry, and analyzed complexly with principal component analysis (PCA), show a significant difference in the distribution of cholesterol and phospholipids. Based on the observed distinctions between healthy and fibrotic cells, the mechanical properties of cells may serve as prospective diagnostic biomarkers enabling fast and reliable identification of idiopathic pulmonary fibrosis (IPF). [ABSTRACT FROM AUTHOR]

Time-of-flight secondary ion mass spectrometry has been employed to examine, with biomolecular discrimination, sensing arm areas (20 μm × 600 μm) of integrated onto silicon chips Mach-Zehnder interferometers aiming to optimize their biofunctionalization with regard to indirect immunochemical (competitive) detection of ochratoxin A. Sensing areas are examined after: modification with (3-aminopropyl)triethoxysilane, spotting of OTA-ovalbumin conjugate (probe) from solutions with different concentration, blocking with bovine serum albumin, reaction with OTA-specific mouse monoclonal antibody followed by goat anti-mouse IgG secondary antibody. Component mass loadings of all proteins involved in immunodetection are determined from TOF-SIMS micro-analysis combined with ellipsometry of planar surfaces. These data show that partial desorption of surface-bound probe and blocking protein takes place upon primary immunoreaction to a degree that depends on probe concentration in spotting solution. Taking into account this desorption, apparent binding stoichiometry of both antibodies in immune complexes formed onto chip surface is determined more accurately than the respective evaluation based on real-time sensor response. In addition, mass loadings for probe and secondary antibody is observed to saturate for optimum probe concentrations. Also, principal component analysis of TOF-SIMS data could resolve both immunoreactions and biofunctionalization and discriminate surfaces prepared with optimum probe concentrations from those prepared using suboptimum ones. [ABSTRACT FROM AUTHOR]