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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]

The effect of temperature and buffer solutions with different pH (often used in biomedical applications) on the behavior of POEGMA brush coatings, synthesized without incorporated functional groups, was for the first time studied in details using water contact angle (CA) measurements and atomic force microscopy (AFM). Thermal response of grafted brush-coatings based on poly(oligo(ethylene glycol) methacrylate)s (POEGMA)s is driven by lower critical solution temperature (LCST) phenomenon. Obtained CA and AFM results suggest strong impact of the buffer solutions on the values of LCST transition and contact angle ranges, as well as on coatings morphology. In turn, ellipsometry data reflect penetration of salt ions from buffer solutions into brush-coatings. In contrast to "typical" behavior of POEGMA coatings in water, different mechanisms available below LCST in the buffer solutions destroy hydrated layers surrounding POEGMA macromolecules leading to their collapse. [ABSTRACT FROM AUTHOR]

In this paper the combined effect of elasticity and chemical properties of the substrate on cellular behavior is applied to control the spatial arrangement of bladder cancer cells (HCV29). For this purpose regular and isotropic patterns of polystyrene (PS) are fabricated on a soft polydimethylsiloxane (PDMS) matrix. The topography and chemical composition of the patterns are determined using atomic force microscopy (AFM) and time-of-flight secondary ion mass spectrometry (ToF-SIMS), respectively, whereas fluorescence microscopy provides information on cellular location. Obtained results show that cells locate preferentially on the soft PDMS matrix whereas the stiffer and presenting weaker adhesive properties PS regions remain unoccupied. This effect may be used for spatial control of adhesion and proliferation of cells over the large areas. [ABSTRACT FROM AUTHOR]

Many of bioelectronic and biosensor applications are based on poly(3-alkylthiophenes), conducting and solution-processable polymers. The most facile approach for the fabrication of such devices relies on biofunctionalization of P3AT surfaces with antibodies through adsorption. The success of this approach depends critically on antibody orientation that affects its biorecognition. As demonstrated here both these features are controlled by the surface structure of spin-cast P3ATs. In particular, a multi-technique and multivariate study that involved Atomic Force Microscopy, Grazing Incidence X-ray Diffraction, Angle-Resolved X-ray Photoelectron Spectroscopy, Enzyme-Linked ImmunoSorbent Assay, and Time-of-Flight Secondary Ion Mass Spectrometry combined with Principal Component Analysis is conducted in order to deduce the crystalline texture of three P3AT polymers as well as its effect on orientation of adsorbed rabbit immunoglobulin (IgG) molecules. An edge-on crystalline texture is concluded for regioregular poly(3-butylthiophene) (RP3BT) and poly(3-hexylthiophene) (RP3HT), while amorphous morphology is inferred for poly(3-butylthiophene) (P3BT). In addition, end-on and head-on orientations similar for all P3ATs were concluded, based on the amount of adsorbed rabbit IgG molecules. Examination of amino acids characteristic for F(ab’) 2 and Fc fragments, and dominant in the external regions of adsorbed immunoglobulin molecules, points to end-on IgG alignment on RP3BT and RP3HT, but not on P3BT. Moreover, the binding of an anti-rabbit IgG antibody on the absorbed rabbit IgG is higher (up to 71%) when the biorecognition reactions are performed on regioregular rather than regiorandom P3AT surfaces. In particular, the highest biorecognition efficiency and IgG orientational order is observed for the RP3BT surfaces with the more developed crystallinity. [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]

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]

[Display omitted] • Application of poly(4-vinyl pyridine) layers cross-linked with transition metal complexes as active layers in biomedical sensors. • Sensitivity of the coatings to the presence of proteins and cells in different concentrations verified by impedance spectroscopy. • Different detection limits for different transition metals. • High selectivity of the coatings toward the defined analyte confirmed by the specific antigen–antibody immunoreaction. • Conductive response of a bilayer system that mimics Organic Field Effect Transistor. The possibility of application of poly(4-vinyl pyridine) layers cross-linked with transition metal complexes as active layers in biomedical sensors was tested. The successful modification of the P4VP coating with CuBr 2 or ZnBr 2 was verified using time of flight - secondary ion mass spectrometry and X-ray photoelectron spectroscopy. The topography and wettability of the coatings were examined by using atomic force microscopy and water contact angles measurements, respectively. Tests of biological activity of coatings indicated strong protein adsorption, good biocompatibility, and no antimicrobial activity. The potential of the coatings to be used as active layers of biosensors was verified, by systematic impedance-based measurements, which showed the sensitivity of the P4VP:CuBr 2 coatings to the presence of proteins and cells in different concentrations. The high selectivity of the coatings toward the defined analyte was confirmed by the specific antigen–antibody immunoreaction, and the possibility of in situ monitoring of protein adsorption and cell adhesion also for individual cells was presented. Finally, the conductive response of a bilayer system that mimics Organic Field Effect Transistor was shown. These results point to a great potential for both coatings to serve as active layers of sensitive and highly selective biosensors. [ABSTRACT FROM AUTHOR]

[Display omitted] • Architecture of PS-PGL copolymers influences its properties in thin films. • Copolymer with coil-brush structure reorients PGL segments in aqueous solution. • Reorientation of PGL chains to the film surface significantly reduces BSA adsorption. • Protein molecules due to adsorption to polymer film reduce mobility of polymer blocks. The effect of the architecture of amphiphilic polystyrene-b-polyglycidol and polystyrene-b-(polyglycidol-g-polyglycidol) copolymers on their behavior in thin films and interaction with the protein has been studied. The properties of the copolymer films surface were investigated before and after incubation in water and aqueous solution of PBS buffer. Surface sensitive techniques: time-of-flight secondary ion mass spectrometry, atomic force microscopy, and contact angle measurements show the differences in film surfaces behavior depending on the copolymer architecture. It was found that exposure of the coil-brush copolymer to an aqueous solution reoriented the PGL segments, changing the properties of the surface layer. Greater exposure of antifouling PGL chains at the surface of the copolymer film modified the interaction with bovine serum albumin, significantly reducing its adsorption. In contrast, no changes in the orientation of the PGL chains were observed after incubation of the copolymer films in an aqueous solution of protein in PBS buffer. This suggests that protein adsorption to the thin copolymer film blocks the conformational changes of the copolymer chains and reduces their mobility under the protein molecules. [ABSTRACT FROM AUTHOR]

[Display omitted] • Complex polymer structures were grafted on PEEK to increase its biocompatibility. • SI-ATRP was used to synthesize hydrophilic polymer brushes over 300 nm. • High density of functional groups enhanced the formation of 3D HAp layers. • Extensive analysis of product hints its high potential of use in implantology. In the search of the ideal bone implant material, surface initiated atom transfer radical polymerization (SI-ATRP) techniques were successfully employed to modify the polyetheretherketone (PEEK) surface with complex architecture nanolayers featuring hydrophilic poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) or poly(2-hydroxyethyl methacrylate) (PHEMA) branches. As a result, polymer layers with height up to over 300 nm were synthesized to decrease the water contact angle from 92.8° to 23.0°. In-depth analysis of layer topography, structure and elemental composition realized with atomic force microscopy (AFM), attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), time-of-flight secondary ions mass spectroscopy (TOF-SIMS) and X-ray photon spectroscopy (XPS) confirmed successful surface modification. Additionally, to demonstrate the applicability of proposed PEEK functionalization, the biomineralization assay in simulated body fluid (SBF) solution was conducted. The high density of hydroxyl and amine groups in branched polymer brushes structure enhanced the rate of hydroxyapatites (HAp) formation substantially. Compared to untreated PEEK, modification with polymer brushes resulted in the creation of cauliflower-like structures with high specific surface area and Ca/P ratio equal to 1.83–1.87 for PEEK grafted with brushes composed of PHEMA backbone segment and PDMAEMA or PHEMA as block polymer branches (PEEK- g -PHEMA- g -PDMAEMA or PEEK- g -PHEMA- g -PHEMA) after 2 weeks. Finally, the process of the hydroxyapatite formation and increase of the layer thickness in a range of 14 days to 6 weeks of immersion in SBF was thoroughly visualized by scanning electron microscopy (SEM). [ABSTRACT FROM AUTHOR]

We have investigated the formation of surface nanostructures caused by early-stage wear of polystyrene (PS) and poly(n -butyl methacrylate) (PnBMA) thin films as pure materials, or mixed in the form of a 1:1 blend. To this end, atomic force microscopy (AFM) was used to repeatedly scratch the sample surfaces, measure the accompanying friction forces, and image the resulting features. In the very first stage ordered ripples are formed in all cases. As the process goes on, hillocks are nucleated on the crests of the ripples, and progressively released in the form of nanoplastics while the ripples become wider and less regular. On the blend surface the more compliant PnBMA presents more corrugated ripple structures and larger friction oscillations than PS in the beginning, but the scenario becomes again more complex as the wear test is repeated and the original 'rim and hole' geometry of the blend is disrupted. Quite noticeably, the wear damage is reduced if the surfaces are scraped forth and back and not only in one direction. The influence of the scan pattern (distance between scan lines) and of the normal force (well below and above the force threshold of about 50 nN leading to surface ripples in the first stage) have been also addressed. • Evolution of early-stage wear on a polymer blend is characterized on the nanoscale. • Hillocks are nucleated from the surface ripples formed in the very first stage. • As the process goes on the hillocks are released in the form of nanoplastics. • Wear is reduced if the polymer blend is scraped forth and back. [ABSTRACT FROM AUTHOR]

[Display omitted] • pH-response of complex architecture polymer brushes generated via low-ppm SI-ATRP. • Polyelectrolyte nanolayers as complex architecture brushes were grated from Si wafer. • Sequential SI-ATRP in microliter-scale applied as novel tool for surface nanoengineering. • 20 μL/cm2 of reaction mixture resulted in effective synthesis of amphiphilic polymer brushes. The article presents a step-by-step protocol, designed on the basis of principles of green chemistry, for the synthesis of polymer brushes with complex structure via surface-initiated atom transfer radical polymerization (SI-ATRP) approach. The selection of anionic (poly(acrylic acid), PAA) and cationic (poly(2-(dimethylamino)ethyl methacrylate, PDMAEMA) macromolecular chains enabled the formation of ampholytic polymer coating on silicon (Si) substrates. Scanning electron microscopy (SEM), atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS) analyses confirmed changes in the structure of the synthesized nanolayer at each step of the surface modification. Finally, the synthesis protocol was rescaled presenting an extremely economically and environmentally beneficial procedure for the synthesis of Si wafers grafted with brushes composed of poly(2-hydroxyethyl methacrylate) (PHEMA) core and PDMAEMA and PAA as block polymer branches (Si- g -PHEMA- g -(PDMAEMA- b -PAA)) with application of microliter volumes of reactants. [ABSTRACT FROM AUTHOR]

Mass fabrication of integrated biosensors on silicon chips is facilitated by contact pin-printing, applied for biofunctionalization of individual Si 3 N 4 -based transducers at wafer-scale. To optimize the biofunctionalization for immunochemical (competitive) detection of fungicide thiabendazole (TBZ), Si 3 N 4 surfaces are modified with (3-aminopropyl)triethoxysilane and examined after: immobilization of BSA-TBZ conjugate (probe) from solutions with different concentration, blocking with bovine serum albumin (BSA), and immunoreaction with a mouse monoclonal antibody against TBZ. Nanostructure, surface density, probe composition and coverage uniformity of protein layers are evaluated with Atomic Force Microscopy, Spectroscopic Ellipsometry, Time-of-Flight Secondary Ion Mass Spectrometry and X-ray Photoelectron Spectroscopy. Contact pin-printing of overlapping probe spots is compared with hand spotted areas. Contact pin-printing resulted in two-fold increase of immobilized probe surface density as compared to hand spotting. Regarding BSA-TBZ immobilization, an incomplete monolayer develops into a bilayer as the concentration of BSA-TBZ molecules in the printing solution increases from 25 to 100 μg/mL. Upon blocking, however, a complete protein monolayer is formed for all the BSA-TBZ concentrations used. Free surface sites are filled with BSA for low surface coverage with BSA-TBZ, whereas loosely bound BSA-TBZ molecules are removed from the BSA-TBZ bilayer. As a consequence immunoreaction efficiency increases with the printing probe concentration. [ABSTRACT FROM AUTHOR]

Time-of-flight secondary ion mass spectrometry (imaging, micro-analysis) has been employed to evaluate biofunctionalization of the sensing arm areas of Mach-Zehnder interferometers monolithically integrated on silicon chips for the immunochemical (competitive) detection of bovine κ-casein in goat milk. Biosensor surfaces are examined after: modification with (3-aminopropyl)triethoxysilane, application of multiple overlapping spots of κ-casein solutions, blocking with 100-times diluted goat milk, and reaction with monoclonal mouse anti-κ-casein antibodies in blocking solution. The areas spotted with κ-casein solutions of different concentrations are examined and optimum concentration providing homogeneous coverage is determined. Coverage of biosensor surfaces with biomolecules after each of the sequential steps employed in immunodetection is also evaluated with TOF-SIMS, supplemented by Atomic force microscopy and X-ray photoelectron spectroscopy. Uniform molecular distributions are observed on the sensing arm areas after spotting with optimum κ-casein concentration, blocking and immunoreaction. The corresponding biomolecular compositions are determined with a Principal Component Analysis that distinguished between protein amino acids and milk glycerides, as well as between amino acids characteristic for Mabs and κ-casein, respectively. Use of the optimum conditions (κ-casein concentration) for functionalization of chips with arrays of ten Mach-Zehnder interferometers provided on-chips assays with dramatically improved both intra-chip response repeatability and assay detection sensitivity. [ABSTRACT FROM AUTHOR]

The methods of preparation and characterization of core-shell poly(styrene/α- tert -butoxy-ω-vinylbenzyl-polyglycidol) particles arranged in colloidal crystals are described. The particles were prepared via emulsifier free emulsion copolymerization of styrene and α- tert -butoxy-ω-vinylbenzyl-polyglycidol macromonomer in aqueous medium, initiated by potassium persulfate. The individual polymerizations differed in a way of addition of macromonomer to the polymerization mixture. The particles assemblies were characterized by atomic force microscopy, Secondary Ion Mass Spectrometry (SIMS) and X-ray Photoelectron Spectroscopy (XPS). In-depth distribution of chemical states was determined by XPS combined with argon gas cluster ion sputtering (Ar-GCIB). The morphology of microspheres assemblies reflected the composition of the entire particles. It was found that the method of macromonomer addition to the polymerization mixture affects the particles size, surface and overall morphology. The XPS profiles revealed the chemical composition of the particles arranged in multilayers. It was found that distribution of polyglycidol segments at the particles interfacial layer depends on the time and way of addition of macromonomer. The thickest outer layer composed from polystyrene-polyglycidol segments was obtained for particles when macromonomer was added at the beginning of polymerization. [ABSTRACT FROM AUTHOR]

[Display omitted] • Preparation procedures of effective plasmonic probes for TERS measurements of DNA are developed. • Characterization of plasmonic properties of various types of TERS probes is demonstrated. • DNA – a standard sample for nanospectroscopic techniques. Tip-enhanced Raman (TER) spectroscopy combines the nanometric spatial resolution of atomic force microscopy (AFM) and the chemical sensitivity of Raman spectroscopy. Thus, it provides a unique possibility to obtain spectroscopic information on individual, nanometre-size molecules. The enhancement of Raman scattering cross-section requires modification of the AFM tip apex with a plasmonic nanostructure. Despite numerous advances of TERS research, attaining good reproducibility and stable enhancement is still challenging mainly due to the lack of optimized probes and sample preparation procedures. Moreover, current nanospectroscopic standard samples - carbon nanotubes (CNTs) have relatively simple chemical structure, and therefore, they are far from real-life analytes, especially biological samples. In this work we focus on the optimization of TERS technique for efficient DNA measurements, including: a preparation of atomically-flat gold substrates, fixative free deposition of DNA and optimization of TERS probe preparation. Here we demonstrate a comprehensive comparison of the efficacy of several types of TERS probes. Applying the systematic approach, we obtained reliable and reproducible TER spectra of DNA. Thus, we provide preparation procedures of a new standard TERS sample, TERS substrates and TERS probes. Our research provides a solid foundation for further research on DNA and its interaction with other biomolecules upon biologically significant processes such as DNA damage and repair. [ABSTRACT FROM AUTHOR]