Reversible Lectin Binding to Glycan-Functionalized Graphene.

The monolayer character of two-dimensional materials predestines them for application as active layers of sensors. However, their inherent high sensitivity is always accompanied by a low selectivity. Chemical functionalization of two-dimensional materials has emerged as a promising way to overcome the selectivity issues. Here, we demonstrate efficient graphene functionalization with carbohydrate ligands-chitooligomers, which bind proteins of the lectin family with high selectivity. Successful grafting of a chitooligomer library was thoroughly characterized, and glycan binding to wheat germ agglutinin was studied by a series of methods. The results demonstrate that the protein quaternary structure remains intact after binding to the functionalized graphene, and that the lectin can be liberated from the surface by the addition of a binding competitor. The chemoenzymatic assay with a horseradish peroxidase conjugate also confirmed the intact catalytic properties of the enzyme. The present approach thus paves the way towards graphene-based sensors for carbohydrate-lectin binding.

Heat treatment dependent cytotoxicity of silicalite-1 films deposited on Ti-6Al-4V alloy evaluated by bone-derived cells.

A silicalite-1 film (SF) deposited on Ti-6Al-4V alloy was investigated in this study as a promising coating for metallic implants. Two forms of SFs were prepared: as-synthesized SFs (SF-RT), and SFs heated up to 500 °C (SF-500) to remove the excess of template species from the SF surface. The SFs were characterized in detail by X-ray photoelectron spectroscopy (XPS), by Fourier transform infrared spectroscopy (FTIR), by scanning electron microscopy (SEM) and water contact angle measurements (WCA). Two types of bone-derived cells (hFOB 1.19 non-tumor fetal osteoblast cell line and U-2 OS osteosarcoma cell line) were used for a biocompatibility assessment. The initial adhesion of hFOB 1.19 cells, evaluated by cell numbers and cell spreading area, was better supported by SF-500 than by SF-RT. While no increase in cell membrane damage, in ROS generation and in TNF-alpha secretion of bone-derived cells grown on both SFs was found, gamma H2AX staining revealed an elevated DNA damage response of U-2 OS cells grown on heat-treated samples (SF-500). This study also discusses differences between osteosarcoma cell lines and non-tumor osteoblastic cells, stressing the importance of choosing the right cell type model.

Silicalite-1 Layers as a Biocompatible Nano- and Micro-Structured Coating: An In Vitro Study on MG-63 Cells.

Silicalite-1 is a purely siliceous form of zeolite, which does not contain potentially harmful aluminum in its structure as opposed to ZSM-5 aluminosilicate types of zeolite. This paper reports on a study of a silicalite-1 film, deposited on a silicon Si(100) substrate, as a potential anti-corrosive and biocompatible coating for orthopaedic implants. Silicalite-1 film was prepared in situ on the surface of Si(100) wafers using a reaction mixture of tetrapropyl-ammonium hydroxide (TPAOH), tetraethyl-orthosilicate (TEOS), and diH2O. The physico-chemical properties of the obtained surface were characterized by means of X-ray photoelectron spectroscopy, water contact angle measurement, atomic force microscopy, and scanning electron microscopy. The biocompatibility was assessed by interaction with the MG-63 cell line (human osteosarcoma) in terms of cell adhesion, morphology, proliferation, and viability. The synthesized silicalite-1 film consisted of two layers (b- and a, b-oriented crystals) creating a combination of micro- and nano-scale surface morphology suitable for cell growth. Despite its hydrophobicity, the silicalite-1 film increased the number of initially adhered human osteoblast-like MG-63 cells and the proliferation rate of these cells. The silicalite-1 film also improved the cell viability in comparison with the reference Si(100) substrate. It is therefore a promising candidate for coating of orthopaedic implants.

The Photodynamic Properties and the Genotoxicity of Heat-Treated Silicalite-1 Films.

We investigated the use of a supported silicalite-1 film (SF) as a promising coating for metallic materials used in the fabrication of prostheses. The role of carbonaceous residua present on high-temperature calcined-SF in generating singlet oxygen for future use as a sterilization method has also been addressed, and the potential genotoxicity of these residua in osteoblast-like cells has been investigated. Calcination of as-synthesized SF induced the appearance of a rather complicated mixture of aliphatic and aromatic species on its outer surface. A series of variously volatile polycyclic aromatic hydrocarbons (PAH), including naphthalene, fluorene, phenanthrene, anthracene, fluoranthene, and pyrene, were identified in micromole concentrations. Irradiation of these PAHs on calcined-SF immersed in air-saturated chloroform led to the formation of very low concentrations of singlet oxygen. However, an increased level of DNA damage was observed on calcined-SF by immunofluorescence staining of phosphorylated histone H2AX analyzed by flow cytometry.

Applications of zeolites in biotechnology and medicine - a review.

Zeolites are microporous tectosilicates of natural or synthetic origin, which have been extensively used in various technological applications, e.g. as catalysts and as molecular sieves, for separating and sorting various molecules, for water and air purification, including removal of radioactive contaminants, for harvesting waste heat and solar heat energy, for adsorption refrigeration, as detergents, etc. These applications of zeolites were typically related with their porous character, their high adsorption capacity, and their ion exchange properties. This review is focused on potential or already practically implemented applications of zeolites in biotechnology and medicine. Zeolites are promising for environment protection, detoxication of animal and human organisms, improvement of the nutrition status and immunity of farm animals, separation of various biomolecules and cells, construction of biosensors and detection of biomarkers of various diseases, controlled drug and gene delivery, radical scavenging, and particularly tissue engineering and biomaterial coating. As components of scaffolds for bone tissue engineering, zeolites can deliver oxygen to cells, can stimulate osteogenic cell differentiation, and can inhibit bone resorption. Zeolites can also act as oxygen reservoirs, and can improve cell performance in vascular and skin tissue engineering and wound healing. When deposited on metallic materials for bone implantation, zeolite films showed anticorrosion effects, and improved the osseointegration of these implants. In our studies, silicalite-1 films deposited on silicon or stainless steel substrates improved the adhesion, growth, viability and osteogenic differentiation of human osteoblast-like Saos-2 cells. Zeolites have been clinically used as components of haemostatics, e.g. in the Advanced Clotting Sponge, as gastroprotective drugs, e.g. Absorbatox® 2.4D, or as antioxidative agents (Klinobind®). Some zeolites are highly cytotoxic and carcinogenic, e.g. erionite. However, in other zeolites, the antiproliferative and pro-apoptotic effects can be used for tumor therapy.

Interaction of human osteoblast-like Saos-2 cells with stainless steel coated by silicalite-1 films.

This paper investigates the interaction of human osteoblast-like Saos-2 cells with stainless steel covered by a film of densely inter-grown silicalite-1 crystals with defined outer and inner surfaces. The chemical composition of this film, labeled as SF(RT), was tuned by heat treatment at 300°C and 500°C (labeled as SF(300) and SF(500), respectively) and characterized by X-ray photoelectron spectroscopy (XPS), water drop contact angle (WCA) measurements and scanning electron microscopy (SEM). The number, the spreading area and the activity of alkaline phosphatase of human osteoblast-like Saos-2 cells in cultures on the silicalite-1 film were affected by the chemical composition of its outer surface and by its micro-porous structure. The number and the spreading area of the adhered osteoblast-like cells on day 1 was highest on the surface of SF(RT) relative to their adhesion and spreading on a glass cover slip due to the SF(RT) topology. However, SF(300) markedly supported cell growth during days 3 and 7 after seeding.

Facile synthesis of CuO nanosheets via the controlled delamination of layered copper hydroxide acetate.

CuO nanosheets were prepared by the controlled delamination of layered copper hydroxide acetate followed by the in situ solvothermal transformation of hydroxide to oxide. The reaction was performed in 1-butanol in order to prevent recrystallization or nanoparticle aggregation. Analyses by small angle X-ray scattering, transmission electron microscopy, and atomic force microscopy revealed that the CuO nanosheets are approximately 1 nm thin, corresponding to three to four stacked CuO6 octahedral layers. The average lateral size is approximately 5 nm. The nanosheets form stable dispersions in 1-butanol that are suitable for the fabrication of transparent and homogeneous CuO thin films by spin-coating or inkjet printing techniques. The present synthesis is a rare example of the top down strategy leading to the nanometric two-dimensional nanosheets of non-layered oxide materials.

Interaction of human osteoblast-like Saos-2 and MG-63 cells with thermally oxidized surfaces of a titanium-niobium alloy.

An investigation was made of the adhesion, growth and differentiation of osteoblast-like MG-63 and Saos-2 cells on titanium (Ti) and niobium (Nb) supports and on TiNb alloy with surfaces oxidized at 165°C under hydrothermal conditions and at 600°C in a stream of air. The oxidation mode and the chemical composition of the samples tuned the morphology, topography and distribution of the charge on their surfaces, which enabled us to evaluate the importance of these material characteristics in the interaction of the cells with the sample surface. Numbers of adhered MG-63 and Saos-2 cells correlated with the number of positively-charged (related with the Nb2O5 phase) and negatively-charged sites (related with the TiO2 phase) on the alloy surface. Proliferation of these cells is correlated with the presence of positively-charged (i.e. basic) sites of the Nb2O5 alloy phase, while cell differentiation is correlated with negatively-charged (acidic) sites of the TiO2 alloy phase. The number of charged sites and adhered cells was substantially higher on the alloy sample oxidized at 600°C than on the hydrothermally treated sample at 165°C. The expression values of osteoblast differentiation markers (collagen type I and osteocalcin) were higher for cells grown on the Ti samples than for those grown on the TiNb samples. This was more particularly apparent in the samples treated at 165°C. No considerable immune activation of murine macrophage-like RAW 264.7 cells on the tested samples was found. The secretion of TNF-α by these cells into the cell culture media was much lower than for either cells grown in the presence of bacterial lipopolysaccharide, or untreated control samples. Thus, oxidized Ti and TiNb are both promising materials for bone implantation; TiNb for applications where bone cell proliferation is desirable, and Ti for induction of osteogenic cell differentiation.

On the role of Nb-related sites of an oxidized ß-TiNb alloy surface in its interaction with osteoblast-like MG-63 cells.

ß-Stabilized titanium (Ti) alloys containing non-toxic elements, particularly niobium (Nb), are promising materials for the construction of bone implants. Their biocompatibility can be further increased by oxidation of their surface. Therefore, in this study, the adhesion, growth and viability of human osteoblast-like MG 63 cells in cultures on oxidized surfaces of a ß-TiNb alloy were investigated and compared with the cell behavior on thermally oxidized Ti, i.e. a metal commonly used for constructing bone implants. Four experimental groups of samples were prepared: Ti or TiNb samples annealed to 600 °C for 60 min in a stream of dry air, and Ti and TiNb samples treated in Piranha solution prior to annealing. We found that on all TiNb-based samples, the cell population densities on days 1, 3 and 7 after seeding were higher than on the corresponding Ti-based samples. As revealed by XPS and Raman spectroscopy, and also by isoelectric point measurements, these results can be attributed to the presence of T-Nb2O5 oxide phase in the surface of the alloy sample, which decreased its negative zeta (ζ)-potential in comparison with zeta (ζ)-potential of the Ti sample at physiological pH. This effect was tentatively explained by the presence of positively charged defects acting as Lewis sites of the surface Nb2O5 phase. Piranha treatment slightly decreases the biocompatibility of the samples, which for the alloy samples may be explained by a decrease in the number of defective sites with this treatment. Thus, the presence of Nb and thermal oxidation of ß-stabilized Ti alloys play a significant role in the increased biocompatibility of TiNb alloys.

Interaction of CO with palladium supported on oxidized tungsten.

A model catalyst system, palladium on tungsten oxide, has been examined by temperature-programmed desorption and photoemission spectroscopy. The samples were prepared by evaporation of palladium onto an oxidized tungsten foil under ultrahigh vacuum conditions. Mostly three-dimensional (3-D) palladium (Pd) clusters were found to be present on oxidized tungsten (WOx) surfaces at room temperature. Upon annealing to 670 K, the palladium clusters are redispersed and decorated by the WOx surface layer. The nature of the WOx phase on top of the palladium clusters is dependent on the mode of oxidation of the tungsten foil prior to palladium deposition. Mainly W(2+) species decorate palladium deposits on tungsten oxidized at room temperature, while mainly W(4+) species are on top of palladium deposits on the surface oxidized at 1300 K. The appearance of a Pd(n+)-O-W(4+) mixed oxide phase with n < 2 was observed on the oxidized tungsten surface. The substantial reduction (relative to nonannealed samples) of molecular CO coverage induced by annealing is discussed in terms of the changes in chemical composition and morphology of the outermost surface.