Biocompatibility enhancement via post-processing of microporous scaffolds made by optical 3D printer.

Providing a 3D environment that mimics the native extracellular matrix is becoming increasingly important for various applications such as cell function studies, regenerative medicine, and drug discovery. Among the most critical parameters to consider are the scaffold's complicated micro-scale geometry and material properties. Therefore, stereolithography based on photopolymerization is an emerging technique because of its ability to selectively form volumetric structures from liquid resin through localized polymerization reactions. However, one of the most important parameters of the scaffold is biocompatibility, which depends not only on the material but also on the exposure conditions and post-processing, which is currently underestimated. To investigate this systematically, microporous scaffolds with pore sizes of 0.05 mm3 corresponding to a porosity of 16,4% were fabricated using the stereolithography printer Asiga PICO2 39 UV from the widely used resins FormLabs Clear and Flexible. The use of various polymers is usually limited for cells because, after wet chemical development, the non-negligible amount of remaining monomers intertwined in the photopolymerized structures is significantly toxic to cells. Therefore, the aim of this research was to find the best method to remove monomers from the 3D scaffold by additional UV exposure. For this purpose, a Soxhlet extractor was used for the first time, and the monomers were immersed in different alcohols. A Raman microspectroscopy was also used to investigate whether different post-processing methods affect DC (cross-linking) to find out if this specifically affects the biocompatibility of the scaffolds. Finally, mesenchymal stem cells from rat dental pulp were examined to confirm the increased biocompatibility of the scaffolds and their ability to support cell differentiation into bone tissue cells.

Stable and Reusable Lace-like Black Silicon Nanostructures Coated with Nanometer-Thick Gold Films for SERS-Based Sensing.

We propose a simple, fast, and low-cost method for producing Au-coated black Si-based SERS-active substrates with a proven enhancement factor of 106. Room temperature reactive ion etching of silicon wafer followed by nanometer-thin gold sputtering allows the formation of a highly developed lace-type Si surface covered with homogeneously distributed gold islands. The mosaic structure of deposited gold allows the use of Au-uncovered Si domains for Raman peak intensity normalization. The fabricated SERS substrates have prominent uniformity (with less than 6% SERS signal variations over large areas, 100 × 100 µm2). It has been found that the storage of SERS-active substrates in an ambient environment reduces the SERS signal by less than 3% in 1 month and not more than 40% in 20 months. We showed that Au-coated black Si-based SERS-active substrates can be reused after oxygen plasma cleaning and developed relevant protocols for removing covalently bonded and electrostatically attached molecules. Experiments revealed that the Raman signal of 4-MBA molecules covalently bonded to the Au coating measured after the 10th cycle was just 4 times lower than that observed for the virgin substrate. A case study of the reusability of the black Si-based substrate was conducted for the subsequent detection of 10-5 M doxorubicin, a widely used anticancer drug, after the reuse cycle. The obtained SERS spectra of doxorubicin were highly reproducible. We demonstrated that the fabricated substrate permits not only qualitative but also quantitative monitoring of analytes and is suitable for the determination of concentrations of doxorubicin in the range of 10-9-10-4 M. Reusable, stable, reliable, durable, low-cost Au-coated black Si-based SERS-active substrates are promising tools for routine laboratory research in different areas of science and healthcare.

Nanodiamond surface as a photoluminescent pH sensor.

A systematic spectroscopic characterization of highly homogeneous water suspensions of 'buckydiamonds' comprising sp3cubic nanodiamond (ND) core covered with disordered sp2shell densely decorated with oxygen-containing groups demonstrates the excitation-wavelength-dependent photoluminescence (PL) given by at least four types of specific structures on the ND surface (hydroxyl, C=O containing ketones, carboxylic anhydrides, and carboxyl groups). PL properties of NDs suspensions possess concentration-dependent behavior revealing tendency of NDs to agglomerate. PL of NDs has been found to be strongly sensitive to pH of the environment in wide range of pH values, i.e. 2-11. We disclosed the mechanisms of pH sensitivity of the 'buckydiamond' and proved that it can serve as all-optical sensor of tiny pH variations suitable for further exploitation for pH sensing locally in the area where NDs have been delivered for any purpose, e.g. bioimaging or therapeutic needs.

Terahertz structured light: nonparaxial Airy imaging using silicon diffractive optics.

Structured light - electromagnetic waves with a strong spatial inhomogeneity of amplitude, phase, and polarization - has occupied far-reaching positions in both optical research and applications. Terahertz (THz) waves, due to recent innovations in photonics and nanotechnology, became so robust that it was not only implemented in a wide variety of applications such as communications, spectroscopic analysis, and non-destructive imaging, but also served as a low-cost and easily implementable experimental platform for novel concept illustration. In this work, we show that structured nonparaxial THz light in the form of Airy, Bessel, and Gaussian beams can be generated in a compact way using exclusively silicon diffractive optics prepared by femtosecond laser ablation technology. The accelerating nature of the generated structured light is demonstrated via THz imaging of objects partially obscured by an opaque beam block. Unlike conventional paraxial approaches, when a combination of a lens and a cubic phase (or amplitude) mask creates a nondiffracting Airy beam, we demonstrate simultaneous lensless nonparaxial THz Airy beam generation and its application in imaging system. Images of single objects, imaging with a controllable placed obstacle, and imaging of stacked graphene layers are presented, revealing hence potential of the approach to inspect quality of 2D materials. Structured nonparaxial THz illumination is investigated both theoretically and experimentally with appropriate extensive benchmarks. The structured THz illumination consistently outperforms the conventional one in resolution and contrast, thus opening new frontiers of structured light applications in imaging and inverse scattering problems, as it enables sophisticated estimates of optical properties of the investigated structures.

Fragmented graphene synthesized on a dielectric substrate for THz applications.

Fragmented multi-layered graphene films were directly synthesized via chemical vapor deposition (CVD) on dielectric substrates with a pre-deposited copper catalyst. We demonstrate that the thickness of the sacrificial copper film, process temperature, and growth time essentially influence the integrity, quality, and disorder of the synthesized graphene. Atomic force microscopy and Kelvin probe force microscopy measurements revealed the presence of nano-agglomerates and charge puddles. The potential gradients measured over the sample surface confirmed that the deposited graphene film possessed a multilayered structure, which was modelled as an ensemble of randomly oriented conductive prolate ellipsoids. THz time domain spectroscopy measurements gave theacconductivity of the graphene flakes and homogenized graphitic films as being around 1200 S cm-1and 1000 S cm-1, respectively. Our approach offers a scalable fabrication of graphene structures composed of graphene flakes, which have effective conductivity sufficient for a wide variety of THz applications.

Visualizing hypochlorous acid production by human neutrophils with fluorescent graphene quantum dots.

In living organisms, redox reactions play a crucial role in the progression of disorders accompanied by the overproduction of reactive oxygen and reactive chlorine species, such as hydrogen peroxide and hypochlorous acid, respectively. We demonstrate that green fluorescence graphene quantum dots (GQDs) can be employed for revealing the presence of the hypochlorous acid in aqueous solutions and cellular systems. Hypochlorous acid modifies the oxygen-containing groups of the GQD, predominantly opens epoxide ring C-O-C, forms excessive C=O bonds and damages the carbonic core of GQDs. These changes, which depend on the concentration of the hypochlorous acid and exposure time, manifest themselves in the absorbance and fluorescence spectra of the GQD, and in the fluorescence lifetime. We also show that the GQD fluorescence is not affected by hydrogen peroxide. This finding makes GQDs a promising sensing agent for selective detecting reactive chlorine species produced by neutrophils. Neutrophils actively accumulate GQDs allowing to visualize cells and to examine the redox processes via GQDs fluorescence. At high concentrations GQDs induce neutrophil activation and myeloperoxidase release, leading to the disruption of GQD structure by the produced hypochlorous acid. This makes the GQDs a biodegradable material suitable for various biomedical applications.

Reflection Absorption Infrared Spectroscopy Characterization of SAM Formation from 8-Mercapto-N-(phenethyl)octanamide Thiols with Phe Ring and Amide Groups.

Multifunctional amide-containing self-assembled monolayers (SAMs) provide prospects for the construction of interfaces with required physicochemical properties and distinctive stability. In this study, we report the synthesis of amide-containing thiols with terminal phenylalanine (Phe) ring functionality (HS(CH2)7CONH(CH2)2C6H5) and the characterization of the formation of SAMs from these thiols on gold by reflection absorption infrared spectroscopy (RAIRS). For reliable assignments of vibrational bands, ring deuterated analogs were synthesized and studied as well. Adsorption time induced changes in Amide-II band frequency and relative intensity of Amide-II/Amide-I bands revealed two-state sigmoidal form dependence with a transition inflection points at 2.2 ± 0.5 and 4.7 ± 0.5 min, respectively. The transition from initial (disordered) to final (hydrogen-bonded, ordered) structure resulted in increased Amide-II frequency from 1548 to 1557 cm-1, which is diagnostic for a strongly hydrogen-bonded amide network in trans conformation. However, the lateral interactions between the alkyl chains were found to be somewhat reduced when compared with well-ordered alkane thiol monolayers.

Surface-Enhanced Raman Spectroscopy of Organic Molecules and Living Cells with Gold-Plated Black Silicon.

Black silicon (bSi) refers to an etched silicon surface comprising arrays of microcones that effectively suppress reflection from UV to near-infrared (NIR) while simultaneously enhancing the scattering and absorption of light. This makes bSi covered with a nm-thin layer of plasmonic metal, i.e., gold, an attractive substrate material for sensing of bio-macromolecules and living cells using surface-enhanced Raman spectroscopy (SERS). The performed Raman measurements accompanied with finite element numerical simulation and density functional theory analysis revealed that at the 785 nm excitation wavelength, the SERS enhancement factor of the bSi/Au substrate is as high as 108 due to a combination of electromagnetic and chemical mechanisms. This finding makes the SERS-active bSi/Au substrate suitable for detecting trace amounts of organic molecules. We demonstrate the outstanding performance of this substrate by highly sensitive and specific detection of a small organic molecule of 4-mercaptobenzoic acid and living C6 rat glioma cell nucleic acids/proteins/lipids. Specifically, the bSi/Au SERS-active substrate offers a unique opportunity to investigate the living cells' malignant transformation using characteristic protein disulfide Raman bands as a marker. Our findings evidence that bSi/Au provides a pathway to the highly sensitive and selective, scalable, and low-cost substrate for lab-on-a-chip SERS biosensors that can be integrated into silicon-based photonics devices.

Macro-, Micro- and Nano-Roughness of Carbon-Based Interface with the Living Cells: Towards a Versatile Bio-Sensing Platform.

Integration of living cells with nonbiological surfaces (substrates) of sensors, scaffolds, and implants implies severe restrictions on the interface quality and properties, which broadly cover all elements of the interaction between the living and artificial systems (materials, surface modifications, drug-eluting coatings, etc.). Substrate materials must support cellular viability, preserve sterility, and at the same time allow real-time analysis and control of cellular activity. We have compared new substrates based on graphene and pyrolytic carbon (PyC) for the cultivation of living cells. These are PyC films of nanometer thickness deposited on SiO2 and black silicon and graphene nanowall films composed of graphene flakes oriented perpendicular to the Si substrate. The structure, morphology, and interface properties of these substrates are analyzed in terms of their biocompatibility. The PyC demonstrates interface biocompatibility, promising for controlling cell proliferation and directional intercellular contact formation while as-grown graphene walls possess high hydrophobicity and poor biocompatibility. By performing experiments with C6 glioma cells we discovered that PyC is a cell-friendly coating that can be used without poly-l-lysine or other biopolymers for controlling cell adhesion. Thus, the opportunity to easily control the physical/chemical properties and nanotopography makes the PyC films a perfect candidate for the development of biosensors and 3D bioscaffolds.

Shell-isolated nanoparticle-enhanced Raman spectroscopy for characterization of living yeast cells.

Studying the biochemistry of yeast cells has enabled scientists to understand many essential cellular processes in human cells. Further development of biotechnological and medical progress requires revealing surface chemistry in living cells by using a non-destructive and molecular structure sensitive technique. In this study shell-isolated nanoparticle-enhanced Raman spectroscopy (SHINERS) was applied for probing the molecular structure of Metschnikowia pulcherrima yeast cells. Important function of studied cells is the ability to eliminate iron from growth media by precipitating the insoluble pigment pulcherrimin. Comparative SERS and SHINERS analysis of the yeast cells in combination with bare Au and shell-isolated Au@SiO2 nanoparticles were performed. It was observed that additional bands, such as adenine ring-related vibrational modes appear due to interaction with bare Au nanoparticles; the registered spectra do not coincide with the spectra where Au@SiO2 nanoparticles were used. SHINERS spectra of M. pulcherrima were significantly enhanced comparing to the Raman spectra. Based on first-principles calculations and 830-nm excited Raman analysis of pulcherrimin, the SHINERS signatures of iron pigment in yeast cells were revealed. Being protected from direct interaction of metal with adsorbate, Au@SiO2 nanoparticles yield reproducible and reliable vibrational signatures of yeast cell wall constituents.

On the Molecular Interactions in Lipid Bilayer-Water Assemblies of Different Curvatures.

This work concerns the importance of intermolecular interactions present in aqueous lipid assembly systems depending on the type of aggregates they form. We have studied aqueous mixtures of diglycerol monooleate, Capmul glycerol monoleate (GMO-50) and polyoxyethylene (20) sorbitan monooleate (Polysorbate 80, P80) using small-angle X-ray scattering (SAXS) measurements to reveal the structure of liquid crystalline phases. On the basis of the SAXS data, a phase diagram was constructed. We discuss the effect of curvature changes of the lipid-aqueous interface obtained by changing the water content and the temperature. The results are related to the intermolecular interactions, as revealed by Raman spectroscopy, with a focus on the bilayer type of system of different curvature and bilayer flexibility, namely, the lamellar phase, bicontinuous cubic phase, and sponge phase. All phases show large similarities in their chain conformation and head group interactions as revealed by the Raman spectra, arising from the fact that all three structures are formed by lipid bilayers. However, subtle differences in the molecular organization of the sponge phase were revealed by employing Raman difference spectroscopy and by analysis of key spectroscopic indicators, which show a less dense hydrocarbon chain packing compared to the inverse bicontinuous cubic or lamellar phase.

Synthesis of Reduced Graphene Oxide with Adjustable Microstructure Using Regioselective Reduction in the Melt of Boric Acid: Relationship Between Structural Properties and Electrochemical Performance.

The melt of H3BO3 was used to reach a controllable reduced graphene oxide (rGO) synthesis protocol using a graphene oxide (GO) precursor. Thermogravimetric analysis and differential scanning calorimetry (TG/DSC) investigation and scanning electron microscopy (SEM) images have shown that different from GO powder, reduction of GO in the melt of H3BO3 leads to the formation of less disordered structure of basal graphene planes. Threefold coordinated boron atom acts as a scavenger of oxygen atoms during the process of GO reduction. Fourier-transform infrared (FTIR) spectra of synthesized products have shown that the complex of glycerol and H3BO3 acts as a regioselective catalyst in epoxide ring-opening reaction and suppress the formation of ketone C=O functional groups at vacancy sites. Thermal treatment at 800 °C leads to the increased concentration of point defects in the backbone structure of rGO. Synthesized materials were tested electrochemically. The electrochemical performance of these materials essentially differs depending on the preparation protocol. The highest charge/discharge rate and double-layer capacitance were found for a sample synthesized in the melt of H3BO3 in the presence of glycerol and treated at 800 °C. The effect of optimal porosity and high electrical conductivity on the electrochemical performance of prepared materials also were studied.

Hindered Amine Light Stabilizers Increase the Stability of Methylammonium Lead Iodide Perovskite Against Light and Oxygen.

Methylammonium lead iodide perovskite (MAPI) is a promising material for highly efficient photovoltaic devices. However, it suffers from photooxidation, which imposes strict requirements for its protection from oxygen during processing and operation. A hindered amine light stabilizer (HALS) has been found to exert a stabilization effect on methylammonium iodide (MAI) and MAPI against photooxidation. The HALS prevents the degradation of MAI by inhibiting the oxidation of iodide to iodine. Chemical modification of HALS allows its incorporation in MAPI films, which extends the resistivity of MAPI against photodegradation in ambient air from a couple of hours to several days, while causing no significant changes in key properties, such as optical absorption and charge transport. These results represent an important advance in the stabilization of MAPI against decomposition and demonstrate for the first time that antioxidants improve the stability of MAPI.

Stimuli-controlled self-assembly of diverse tubular aggregates from one single small monomer.

The design and synthesis of new stimuli-responsive hydrogen-bonding monomers that display a diversity of self-assembly pathways is of central importance in supramolecular chemistry. Here we describe the aggregation properties of a simple, intrinsically C2-symmetric enantiopure bicyclic cavity compound bearing a terminally unsubstituted ureidopyrimidinone fragment fused with a pyrrole moiety in different solvents and in the absence and presence of C60 and C70 guests. The tetrameric cyclic aggregate is selectively obtained in chlorinated solvents, where only part of the available hydrogen bonding sites are utilized, whereas in toluene or upon addition of C70 guests, further aggregation into tubular supramolecular polymers is achieved. The open-end cyclic assemblies rearrange into a closed-shell capsule upon introduction of C60 with an accompanied symmetry breaking of the monomer. Our study demonstrates that a C60 switch can be used to simultaneously control the topology and occupancy of tubular assemblies resulting from the aggregation of small monomers.

Interaction of 4-imidazolemethanol with a copper electrode revealed by isotope-edited SERS and theoretical modeling.

Adsorption of 4-imidazolemethanol (ImMeOH) on a copper electrode has been investigated by in situ isotope-edited (H/D and (63)Cu/(65)Cu) surface enhanced Raman spectroscopy (SERS) in aqueous solutions at physiological pH (7.0) in a potential window from -0.500 to -1.100 V. Theoretical modeling by DFT calculations at the B3LYP/6-311++G(d,p) level for light atoms and LANL2DZ with ECP for copper atoms have been employed for the interpretation of experimental data. The copper surface was modeled by a cluster of 6 atoms. It was found that the imidazole ring adopts Tautomer-I form in the adsorbed state and coordinates with the Cu surface through the N3 atom. Linear potential-dependence of ν(C4=C5) mode with the slope of (15 ± 1) cm(-1) V(-1) was experimentally observed. The imidazole ring mode near 1492 cm(-1) primarily due to ν(C2-N3) + ß(C2H) vibration has also showed a considerable decrease in frequency at more negative electrode potentials with the slope of (9 ± 2) cm(-1) V(-1). Both modes can be used as sensitive probes for analysis of interaction of the imidazole ring with the metal surface. In agreement with experimental data theoretical modeling has predicted higher stability of surface bound Tautomer-I compared with Tautomer-II. The formation of a covalent bond between the metal and adsorbate was experimentally evidenced by metal isotopic ((63)Cu/(65)Cu) frequency shift of ν(Cu-N) mode at 222 cm(-1), combined with theoretical modeling of the surface complex.

Molecular structure of mercury(II) thiocyanate complexes based on DFT calculations and experimental UV-electron spectroscopy and Raman studies.

In this work, we report a combined experimental and theoretical study on molecular structure, vibrational and electronic spectra of [Hg(SCN)n](2-)(n) complexes (where n=2, 3, 4) in the aqueous solution. Molecular modeling of the mercury(II) complexes were done by the density functional theory (DFT) method using B3LYP functional with Stuttgart relativistic ECP 78MWB basis set for Hg and 6-311++G(d,p) basis set for all other atoms. The effect of different solvation models with explicit (ligand) and/or implicit water environment upon its geometry, vibrational frequencies and UV spectrum have been studied. The influence of H2O/D2O exchange on the experimental and calculated vibrational frequencies of studied complexes has been established. The double-peak character of the νHgS vibrational mode of the all analyzed mercury complexes and νCN mode of [Hg(SCN)3H2O](-) complex, respectively, were proposed here for the first time. The formation of four-coordinated Hg(II) complexes with thiocyanate and (or) water ligands was verified.

Surface-enhanced Raman spectroscopy for detection of toxic amyloid ß oligomers adsorbed on self-assembled monolayers.

Surface-enhanced Raman spectroscopy (SERS) was used to detect different spectral features of small (1-2 nm) and large (5-10 nm) synthetic amyloid Aß-42 oligomers, exhibiting high and no detectable neurotoxicities, respectively. Adsorption of peptides at self-assembled monolayers (SAM) terminated by methyl and pyridinium groups was employed to differentiate toxic and non-toxic oligomers. Three SAMs were analyzed: hydrophobic heptanethiol (HT) and octadecanethiol (ODT) as well as positively charged N-(6-mercapto)hexylpyridinium (MHP) chloride. SERS study revealed twofold adsorption effect, changes in the monolayer structure and appearance of new bands associated with the adsorbed peptides. A band at 1387 cm(-1), observed as a result of the SAM and Aß-42 interaction, is tentatively assigned to the peptide symmetric stretching vibration of carboxylate groups, and appears to be the most prominent spectral feature distinguishing toxic oligomers from the non-toxic Aß-42 forms. This band was identified in the spectra of Aß-42 adsorption on heptanethiol and MHP monolayers, while no clear perturbations were observed in the case of ODT monolayer.