Nanohydroxyapatite synthesized from calcium oxide and its characterization

Spherical and nano-sized hydroxyapatite (HAp) was synthesized from calcium oxide by a continuous rapid precipitation method without using any secondary route. The maximum particles have a size around 10 nm according to X-ray diffraction (XRD), 35 nm based on scanning electron microscopy (SEM) and 15 nm according to transmission electron microscopy (TEM) studies. The sintering effects on structural properties of HAp were studied by XRD, Fourier transformation infrared (FTIR), ³¹P solid-state nuclear magnetic resonance (NMR) and Raman spectroscopy. The particle size varied from 5 to 70 nm and nanopores from 10 to 450 nm were revealed by SEM and transmission electron microscopy TEM. This result was also strongly supported by XRD study. The combined effect of heating and centrifuging force exerted by vigorous magnetic stirring causes the dispersion of tiny crystals followed by precipitation to form spherical shaped particles. The chemical structure of HAp was established by FTIR, NMR and Raman spectroscopy.     

Effects of the Ar ions pre‐amorphization of Si substrateon interface mixing of Fe/Si bilayers

Ion beam mixing of Fe/Si bilayers, induced by 100 keV ⁴⁰Arions at room temperature was investigated. Rutherford backscattering spectroscopy (RBS), atomic force microscopy (AFM), scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were applied for structural characterization. The main focus of this study was on the influence of the substrate structure on interface mixing. The influence of the substrate structure is due to the two classes of irradiated bilayers, Fe thin films deposited on crystalline or pre‐amorphized Si substrates. An about 76% higher efficiency of atomic transport across the pre‐amorphized Fe/a‐Si interface as compared to that of Fe/c‐Si bilayers was observed.     

The application of heat‐deproteinization to the morphological study of cortical bone: A contribution to the knowledge of the osteonal structure

Observation of heat‐deproteinized cortical bone specimens in incident light enabled the high definition documentation of the osteonal pattern of diaphyseal Haversian bone. This prompted a study to compare these images with those revealed by polarized light microscopy, carried out either on decalcified or thin, undecalcified, resin‐embedded sections. Different bone processing methods can reveal structural aspects of the intercellular matrix, depending on the light diffraction mode: birefringency in decalcified sections can be ascribed to the collagen fibrils orientation alone; in undecalcified sections, to both the ordered layout of collagen and the inorganic phase; in the heat‐deproteinized samples, exclusively to the hydroxyapatite crystals aggregation mode. The elemental chemical analysis documented low content of carbon and hydrogen, no detectable levels of nitrogen and significantly higher content of calcium and phosphorus in heat‐deproteinized samples, as compared with dehydrated controls. In both samples, the X‐ray diffraction (XRD) pattern did not show any significant difference in pattern of hydroxyapatite, with no peaks of any possible decomposition phases. Scanning electron microscopic (SEM) morphology of heat‐deproteinized samples could be documented with the fracturing technique facilitated by the bone brittleness. The structure of crystal aggregates, oriented in parallel and with marks of time periods, was documented. Comparative study of deproteinized and undecalcified samples showed that the matrix inorganic phase did not undergo a coarse grain thermal conversion until it reached 500°C, maintaining the original crystals structure and orientation. Incident light stereomicroscopy, combined with SEM analysis of deproteinized bone fractured surfaces, is a new enforceable technique which can be used in morphometric studies to improve the understanding of the osteonal dynamics. Microsc. Res. Tech. 79:691–699, 2016. © 2016 Wiley Periodicals, Inc.     

Characterization of nanostructured binary molybdenum oxide catalyst precursors for propene oxidation

High‐resolution transmission electron microscopy characterization of molybdenum oxide catalyst precursors is carried out for a better understanding of the mechanisms leading to the active catalytic material. The precipitated material consists of both large‐unit cell crystals and randomly orientated clusters on a 3–5 nm scale. The crystalline clusters are embedded in non‐crystalline material, preventing the formation of large well‐defined inactive orthorhombic MoO₃ crystals during activation of the catalyst material.     

Freestanding lipid bilayers as substrates for electron cryomicroscopy of integral membrane proteins

Phospholipid bilayers, 40 Å thick, were generated as electron microscope substrates by submerging copper grids overlaid with holey plastic through a lipid monolayer on a water surface. Previously formed proteoliposomes containing single‐particle membrane proteins in their bilayers were then fused into the newly formed bilayer substrate. To demonstrate this methodology, multi‐drug resistance protein P‐glycoprotein was incorporated into these bilayers and imaged by fixed beam microscopy and scanning transmission electron microscopy.     

Regenerating titanium ventricular assist device surfaces after gold/palladium coating for scanning electron microscopy

Titanium is one of the most commonly used materials for implantable devices in humans. Scanning electron microscopy (SEM) serves as an important tool for imaging titanium surfaces and analyzing cells and other organic matter adhering to titanium implants. However, high‐vacuum SEM imaging of a nonconductive sample requires a conductive coating on the surface. A gold/palladium coating is commonly used and to date no method has been described to “clean” such gold/palladium covered surfaces for repeated experiments without etching the titanium itself. This constitutes a major problem with titanium‐based implantable devices which are very expensive and thus in short supply. Our objective was to devise a protocol to regenerate titaniumsurfaces after SEM analysis. In a series of experiments, titanium samples from implantable cardiac assist devices were coated with fibronectin, seeded with cells and then coated with gold/palladium for SEM analysis. X‐ray photoelectron spectroscopy spectra were obtained before and after five different cleaning protocols. Treatment with aqua regia (a 1:3 solution of concentrated nitric and hydrochloric acid), with or without ozonolysis, followed by sonication in soap solution and sonication in deionized water, allowed regenerating titanium surfaces to their original state. Atomic force microscopy confirmed that the established protocol did not alter the titanium microstructure. The protocol described herein is applicable to almost all titanium surfaces used in biomedical sciences and because of its short exposure time to aqua regia, will likely work for many titanium alloys as well. Microsc. Res. Tech., 2009. © 2009 Wiley‐Liss, Inc.     

Microstructural characteristics of calcium hydroxyapatite/poly- l-lactide based composites

Besides its high osteoinductive properties, hydroxyapatite (HAp) exhibits a relatively low mechanical strength. In order to improve the mechanical properties and reliability of HAp based composites, the addition of selected polymers is highly recommended. The main objective of this work is to study the microstructural characteristics of HAp/poly- l -lactide (PLLA) composites obtained by cold or hot processing. The composites were prepared from a mixture of a chloroform solution of poly- l -lactide with granulated HAp. After elimination of chloroform by vacuum evaporation, dense compacts were obtained by cold or hot pressing. The pressing pressure ranged from 98.10 to 294.3 MPa for both cold and hot pressing. The hot pressing was performed in the temperature region 293–457 K for a time period of 15–60 min. Depending on the PLLA amount and the pressing procedure it is possible to obtain highly porous or nearly fully dense composites. The scanning electron microscopy examination of fracture as well as of free surfaces revealed that the final porosity and wetting are affected to a great extent by the synthesis conditions and amount of polymer added. An increase in temperature to 457 K for a longer period of time results in fully dense compacts. The formation of a nearly continuous polymer network that leads to the hardening of HAp has also been observed. However, it should be pointed out that some layers of HAp may be free of polymer film since PLLA penetrates more deeply into the porous HAp.     

Potential of a sensitive uric acid biosensor fabricated using hydroxyapatite nanowire/reduced graphene oxide/gold nanoparticle

In this study, a ternary nanocomposite consisting of gold nanoparticles (AuNPs), hydroxyapatite (HAP) nanowires, and reduced graphene oxide (rGO) is synthesized by a simple one‐step hydrothermal method, which is used to modify glassy carbon electrode (GCE) for detecting uric acid. The nanocomposite is characterized through various methods such as scanning electron microscopy, transmission electron microscopy, and X‐ray diffraction. Electrochemical measurements of the modified GCE are performed in a conventional three‐electrode system. Experimental results show that the obtained HAP nanowire and rGO are mixed homogeneously, and the AuNPs are deposited into this matrix. The GCE modified by the nanocomposites have superior electrocatalytic activities for uric acid. The peak current intensities of UAO (uricase)/HAP‐rGO/AuNPs sensing system linearly increase as the uric acid concentration increases substantially in a range of 1.95 × 10^?5 to 6.0 × 10^?3 M (R² = .9943), with a detection limit of 3.9 × 10^?6 M (S/N = 3) and analytical sensitivity of 13.86 mA/M. The biosensor performs well in determining uric acid concentration in human urine samples.     

Microstructural characterization of Ti‐6Al‐4V alloy subjected to the duplex SMAT/plasma nitriding

In this study, microstructural characterization of Ti‐6Al‐4V alloy, subjected to the duplex surface mechanical attrition treatment (SMAT)/nitriding treatment, leading to improve its mechanical properties, was carried out through novel and original samples preparation methods. Instead of acid etching which is limited for morphological characterization by scanning electron microscopy (SEM), an original ion polishing method was developed. Moreover, for structural characterization by transmission electron microscopy (TEM), an ion milling method based with the use of two ions guns was also carried out for cross‐section preparation. To demonstrate the efficiency of the two developed methods, morphological investigations were done by traditional SEM and field emission gun SEM. This was followed by structural investigations through selected area electron diffraction (SAED) coupled with TEM and X‐ray diffraction techniques. The results demonstrated that ionic polishing allowed to reveal a variation of the microstructure according to the surface treatment that could not be observed by acid etching preparation. TEM associated to SAED and X‐ray diffraction provided information regarding the nanostructure compositional changes induced by the duplex SMAT/nitriding process. Microsc. Res. Tech. 76:897–903, 2013. © 2013 Wiley Periodicals, Inc.     

Micromorphology analysis of TiO2 thin films by atomic force microscopy images: The influence of postannealing

This work describes an analysis of titanium dioxide (TiO₂) thin films prepared on silicon substrates by direct current (DC) planar magnetron sputtering system in O₂/Ar atmosphere in correlation with three‐dimensional (3D) surface characterization using atomic force microscopy (AFM). The samples were grown at temperatures 200, 300, and 400°C on silicon substrate using the same deposition time (30 min) and were distributed into four groups: Group I (as‐deposited samples), Group II (samples annealed at 200°C), Group III (samples annealed at 300°C), and Group IV (samples annealed at 400°C). AFM images with a size of 0.95 μm × 0.95 μm were recorded with a scanning resolution of 256 × 256 pixels. Stereometric analysis was carried out on the basis of AFM data, and the surface topography was described according to ISO 25178‐2:2012 and American Society of Mechanical Engineers (ASME) B46.1‐2009 standards. The maximum and minimum root mean square roughnesses were observed in surfaces of Group II (Sq = 7.96 ± 0.1 nm) and Group IV (Sq = 3.87 ± 0.1 nm), respectively.     

Microstructural characterization of mechanicallyactivated ZnO powders

In this paper, changes of microstructural characteristics of disperse systems during mechanical activation of zinc oxide (ZnO) have been investigated. ZnO powder was activated by grinding in a planetary ball mill in a continuous regime in air during 300 min at the basic disc rotation speed of 320 rpm and rotation speed of bowls of 400 rpm but with various balls‐to‐powder mass ratios. During ball milling in a planetary ball mill, initial ZnO powder suffered high‐energy impacts. These impacts are very strong, and large amounts of microstructural and structural defects were introduced in the milled powders. The morphology and dispersivity of particles and agglomerates of all powders were investigated by scanning electron microscopy and scanning transmission electron microscopy. The specific surface area of initial ZnO powder was determined as 3.60 m² g^?1 and it increased to 4.42 m² g^?1 in mechanically activated powders. An increase of the ball‐to‐powder mass ratio led to a decrease of particle dimensions as well as increased the tendency for joining into quite compact agglomerates, that is aggregates, compared with the very loose, soft initial agglomerates. The obtained results pointed out that activation of ZnO powders produces a highly disperse, nano‐scaled mixture of small particles, that is crystallites with sizes in the range of 10–40 nm. Most of these particles are in the form of aggregates with dimensions of 0.3–0.1 μm. The crystallite and aggregate size strongly depend on milling conditions, that is ball‐to‐powder mass ratio, as shown in this investigation.     

Thick rutile layer on titanium for tribological applications

In the present work, efforts have been made to oxidise the titanium surface, followed by very slow cooling to produce a thick and adherent oxide layer. The response of titanium to oxidation at various temperatures and timings has been investigated, in terms of layer thickness, phase evolution, surface morphology, oxide layer–substrate adhesion, hardness and tribological characteristics. A variety of experimental and analytical techniques, including X-ray diffraction, field-emission scanning electron microscopy, scanning electron microscopy, micro-hardness and tribological testing, have been used to characterise the resultant thermal oxidised surfaces. The results showed that a thick oxide layer with rutile TiO₂ and oxygen diffused Ti structure can be produced, which exhibited excellent adhesion with the titanium substrate, low friction and superior wear resistance during pin-on-disc sliding tests and thus good potential for tribological applications.     

A new HPF specimen carrier adapter for the use of high‐pressure freezing with cryoscanning electron microscope: two applications: stearic acid organization in a hydroxypropyl methylcellulose matrix and mice myocardium

Cryogenic transmission electron microscopy of high‐pressure freezing (HPF) samples is a well‐established technique for the analysis of liquid containing specimens. This technique enables observation without removing water or other volatile components. The HPF technique is less used in scanning electron microscopy (SEM) due to the lack of a suitable HPF specimen carrier adapter. The traditional SEM cryotransfer system (PP3000T Quorum Laughton, East Sussex, UK; Alto Gatan, Pleasanton, CA, USA) usually uses nitrogen slush. Unfortunately, and unlike HPF, nitrogen slush produces water crystal artefacts. So, we propose a new HPF specimen carrier adapter for sample transfer from HPF system to cryogenic‐scanning electronic microscope (Cryo‐SEM). The new transfer system is validated using technical two applications, a stearic acid in hydroxypropyl methylcellulose solution and mice myocardium. Preservation of samples is suitable in both cases. Cryo‐SEM examination of HPF samples enables a good correlation between acid stearic liquid concentration and acid stearic occupation surface (only for homogeneous solution). For biological samples as myocardium, cytoplasmic structures of cardiomyocyte are easily recognized with adequate preservation of organelle contacts and inner cell organization. We expect this new HPF specimen carrier adapter would enable more SEM‐studies using HPF.     

Reducing Friction and Wear of a Zinc Substrate by Combining a Stearic Acid Overcoat with a Nanostructured Zinc Oxide Underlying Film: Perspectives to Super-Hydrophobicity

In this study, a simple two-step process was developed to render zinc (Zn) surfaces super-hydrophobic for the purpose of lowering friction and increasing wear resistance. Zn substrates were immersed in an aqueous solution of N,N-dimethylformamide (4%, volume fraction v/v) to fabricate a ZnO film consisting of uniform and well-packed nanorods. A self-assembled monolayer of stearic acid was then prepared on the ZnO-nanorod film to acquire super-hydrophobicity. Scanning electron microscopy, Fourier transform infrared microscopy, and water contact-angle measurements were employed to analyze the morphological features, the chemical composition, and super-hydrophobicity of freshly prepared samples. Moreover, the friction and wear behavior of the organic–inorganic composite film sliding against steel was evaluated in a ball-on-plate configuration using a UMT-3 friction and wear tester. It was found that the stearic acid overcoat on the nanostructured ZnO film led to a large water contact angle of ~155° as well as to significantly decreased friction and greatly extended wear resistance.     

Gloss phenomena and image analysis of atomic force microscopy in molecular and cell biology

Proper sample preparation, scan setup, data collection and image analysis are key factors in successful atomic force microscopy (AFM), which can avoid gloss phenomena effectively from unreasonable manipulations or instrumental defaults. Fresh cleaved mica and newly treated glass cover were checked first as the substrates for all of the sample preparation for AFM. Then, crystals contamination from buffer was studied separately or combined with several biologic samples, and the influence of scanner, scan mode and cantilever to data collection was also discussed intensively using molecular and cellular samples. At last, images treatment and analysis with off‐line software had been focused on standard and biologic samples, and artificial glosses were highly considered for their high probability. SCANNING 31: 49–58, 2009. © 2009 Wiley Periodicals, Inc.     

Mesoporous iron oxide nanoparticles prepared by polyacrylic acid etching and their application in gene delivery to mesenchymal stem cells

Novel monodisperse mesoporous iron oxide nanoparticles (m‐IONPs) were synthesized by a postsynthesis etching approach and characterized by electron microscopy. In this approach, solid iron oxide nanoparticles (s‐IONPs) were first prepared following a solvothermal method, and then etched anisotropically by polyacrylic acid to form the mesoporous nanostructures. MTT cytotoxicity assay demonstrated that the m‐IONPs have good biocompatibility with mesenchymal stem cells (MSCs). Owing to their mesoporous structure and good biocompatibility, these monodisperse m‐IONPs were used as a nonviral vector for the delivery of a gene of vascular endothelial growth factor (VEGF) tagged with a green fluorescence protein (GFP) into the hard‐to‐transfect stem cells. Successful gene delivery and transfection were verified by detecting the GFP fluorescence from MSCs using fluorescence microscopy. Our results illustrated that the m‐IONPs synthesized in this work can serve as a potential nonviral carrier in gene therapy where stem cells should be first transfected and then implanted into disease sites for disease treatment. Microsc. Res. Tech. 76:936–941, 2013. © 2013 Wiley Periodicals, Inc.     

Quantification of metallic nanoparticle morphology on TiO2 using HAADF-STEM tomography

Electron tomography is applied to photocatalytic gold/titanium oxide and gold/silver/titanium oxide samples. In order to obtain a tilt series for the electron tomography measurement, high-angle annular dark-field scanning transmission electron microscopy (HAADF-STEM) is used under cryogenic conditions. Dedicated programs have been developed for measuring volume, surface area, thickness distribution and nearest-neighbour distance of metallic nanoparticles on samples. Using these quantification programs, the 3D morphology of gold and silver nanoparticles is accurately characterized. We paid particular attention to the quantitative measurement of surface area. The measurement error of the method and appropriate magnification are defined using spherical nanoparticle models. We measured the 3D morphology of gold nanoparticles supported on titanium oxide (total volume=6.5×10⁵ [nm³], surface area=1.4×10⁵ [nm²], and average nearest-neighbour distance=40 [nm]).     

Studies of inherent lubricity coatings for low surface roughness galvanised steel for automotive applications

Surface lubricity on TiO₂‐coated galvanised steels can be controlled by solution depositing perfluorooctanoic ( C8 ), lauric ( C12 ) or stearic ( C18 ) acids to avoid lubricating oils/emulsions or substrate pre‐etching to remove surface oxide that add cost and waste. Water contact angles reveal increased surface hydrophobicity on coated samples that correlate with linear friction testing, suggesting water contact angle can be used to screen lubricity compounds. Linear friction testing shows that C12 and C18 lower the coefficient of friction (μ ) by 50–60% compared with uncoated substrates whilst C8 drops μ from 0.31 to 0.22. Surfaces have been characterised by X‐ray photoelectron spectroscopy, scanning electron microscopy and atomic force microscopy, whilst infrared confirms that as‐deposited coatings contain physisorbed and deprotonated acids chemisorbed through esters and thermal gravimetric analysis confirms increasing loadings from C8 to C12 to C18 . Surface washing removes physisorbed material and lowers μ by increasing surface organisation and alkyl chain packing that enhances frictional energy dissipation through steric quenching. Copyright © 2017 John Wiley & Sons, Ltd.     

Preparation of ultra-thin oxide windows on titanium for tem analysis

Using submerged jet electropolishing, extremely thin (<10 nm), continuous, thermal oxide “windows” have been prepared on poly crystalline titanium (Ti). The preparation technique is described in detail. It has allowed a systematic investigation of the structure of thermal surface oxide layers on Ti in the thickness range 6–40 nm, corresponding to oxidation temperatures 100–450°C. Auger electron spectroscopy was used for oxide characterization and for depth profiling to determine oxide thickness. The thinnest oxides, < 10 nm, are amorphous, morphologically homogeneous, and with essentially no contrast in the transmission electron microscopy (TEM) pictures. As the oxide thickness is increased up to 40 nm, a texture corresponding to the grain structure of the oxidized metal becomes gradually more visible. At the same time the oxide becomes increasingly more crystalline. The results are compared with previously published corresponding results for thicker anodic oxides on Ti.