Toxicity of mercury on in vitro development of parthenogenetic eggs of a freshwater cladoceran Daphnia carinata

Chronic toxicity test duration of 21 days for daphnid is time consuming and expensive. Therefore, the developmental stages of Daphnia carinata eggs that could be used as potential endpoints for sublethal and chronic toxicity tests have been investigated and defined. Daphnid egg test is simple, easy to conduct and handle in the laboratory, and cost-effective. The 72 h ‘egg arrest’ bioassay system could be an alternative to the classic 21-day chronic test with neonates of daphnid. The main aims of the study were to establish easy to identify stages of D. carinata egg that could be used as potential endpoints for toxicity tests with in vitro cultures of daphnid parthenogenetic eggs. Commonly available Indian freshwater cladoceran Daphnia carinata parthenogenetic eggs in vitro were exposed to water borne mercury concentrations, ranging from 0.1 to 32 μg l⁻¹. Adult female cladoceran D. carinata have eight main developmental stages of parthenogenetic reproduction based on the release of external and internal membranes, formation of cephalic and body regions, appearance of secondary antennae, presence of two pink eyes, than a single black eye, and finally caudal or shell spine separation and finally free-swimming neonate within 65–72 h. At 1, 3.2 and 10 μg l⁻¹ of Hg concentrations; the 25, 50 and 70% embryonic developmental arrests were observed. The lower concentrations of Hg (0.32, 1, and 3.2 μg l⁻¹) tested in the present study are not generally harmful to the neonates and adults daphnid species, but the same are highly toxic to the embryos of D. carinata. The 48 h and 72 h EC₅₀s and their 95% confidence limits for survival and hatchability were lower than previously reported 48 h EC₅₀s for Daphnia magna immobilization assay. The egg of D. carinata turned out to be a suitable alternative model for ecotoxicological and water quality assessment studies.     

Preparation and characterization of an Hf-doped zinc silicate long-lasting phosphorescent material

Hf⁴⁺-doped Zn₂SiO₄ phosphor emitting long-lasting cyan light was prepared by the conventional high temperature solid-state technique. The emission spectrum of the Hf⁴⁺-doped phosphor exhibits one broad band peaking at 471 nm in the visible region excited by 254 nm. The cyan-light afterglow can last about 40 min in darkness after being irradiated with 254 nm UV lamp for 10 min (250 mW/cm²). The afterglow decay curve can be fitted into a second-order exponential curve. The thermoluminescence (TL) curve shows two glow bands centered at about 386 K (0.61 eV) and 440 K (1.56 eV), respectively, with the lower trap energy level being responsible for the long-lasting afterglow emission. A possible mechanism of the long-lasting phosphorescence based on the experimental results is proposed.     

In vitro evaluation of gentamicin release from a bioactive tricalcium silicate bone cement

Tricalcium silicate (Ca₃SiO₅) cement, a novel self-setting biomaterial, has been shown to exhibit good hydraulic properties and excellent bioactivity. In this study, gentamicin sulfate (GS) was integrated into cement pastes and in vitro release of GS from the Ca₃SiO₅ cement was performed in deionized water, phosphate buffer saline (PBS) and HCl solutions with different pH at 37 °C, respectively. The results showed that the initial fast release of GS was restricted to a low level and prolonged release of drugs was achieved in water and PBS. The prolonged GS release is attributed to the interaction of GS with the calcium silicate hydrate network and the formation of unique nano-to-micro porous structure after hydration. Furthermore, GS release from milled powders of the hydrated cement suggested that the constrained GS could be released at low pH environment or during the degradation of the cement. When the samples were soaked in PBS, a nano-structured apatite layer was formed on the surface of the cement, which resulted in a relatively lower GS release rate as compared to that in water. The results suggest that Ca₃SiO₅ cement might be used as bioactive bone implant materials with drug loading and prolonged release properties.     

Order-disorder transformation in Fe-Pd alloy nanoparticles studied by in situ transmission electron microscopy

We have studied order-disorder transformation in Fe-Pd alloy nanoparticles by in situ transmission electron microscopy (TEM) and electron diffraction. The transformation is size-dependent, and the transformation temperatures are lower than those of the bulk alloys. The transformation proceeds continuously but rather steeply as the temperature increases, which differs from the first-order transformation observed in a bulk alloy or gradual transformation predicted by simulations for nanoparticles. Experimental results indicated that the continuous nature can be attributed to the distribution of the transformation temperature due to the distributions of both particle size and alloy composition. Quantitative intensity analyses of nanobeam electron diffraction (NBD) patterns indicated the existence of short-range order (SRO) inside disordered nanoparticles. The SRO as well as particle size distribution are responsible for the remaining weak superlattice reflections above the transformation temperature. In situ high-resolution TEM (HRTEM) observation revealed the existence of the SRO, which was consistent with the results obtained by NBD. We show that the disorder may not necessarily proceed continuously from the surface toward the center of the nanoparticle. Ordering from the disordered phase upon cooling was also observed by in situ HRTEM, which can be attributed to growth of the SRO.     

Processing, physico-chemical characterisation and in vitro evaluation of silicon containing β-tricalcium phosphate ceramics

For bone grafting applications, the elaboration of silicon containing beta-tricalcium phosphate (β-TCP) was studied. The synthesis was performed using a wet precipitation method according to the hypothetical theoretical formula Ca_3 − x(PO₄)_2 − 2x(SiO₄)_x. Two silicon loaded materials (0.46 wt.% and 0.95 wt.%) were investigated and compared to a pure β-TCP. The maturation time of the synthesis required in order to obtain β-TCP decreased with the amount of silicon. Only restrictive synthesis conditions allow preparing silicon containing β-TCP with controlled composition. To obtain dense ceramics, the sintering behaviour of the powders was evaluated. The addition of silicon slowed the densification process and decreased the grain size of the dense ceramics. Rietveld refinement may indicate a partial incorporation of silicon in the β-TCP lattice. X-ray photoelectron spectroscopy and transmission electron microscopy analyses revealed that the remaining silicon formed amorphous clusters of silicon rich phase. The in vitro biological behaviour was investigated with MC3T3-E1 osteoblast-like cells. After the addition of silicon, the ceramics remained cytocompatible, highlighting the high potential of silicon containing β-TCP as optimised bone graft material.     

Microstructural characterization and wear behavior of in situ TiC/7075 composites synthesized by displacement reactions and spray forming

A novel in situ reaction technique is developed to prepare TiC/7075 composites. This technique provides a new approach overcoming the problems of loss and agglomeration of reinforcement particles when they are in situ formed in a molten metal first and then injected into the spray cone of molten droplets during the spray forming process. Experimental results have shown that the presence of strip or rectangular-like Al₃Ti, which is detrimental not only to the fracture toughness, but also to the stability of the microstructure, can be avoided completely from the final product by using a proper Ti:C molar ratio in the Ti-C-Al performs. The mechanisms of formation or absence of Al₃Ti phase in the TiC/7075 composites are explained based on thermodynamics of the system. The modification of the microstructure of the spray-formed 7075 alloy can be understood in the light of atomic diffusion. The wear results showed that the wear rates of the spray-formed 7075 alloy and its composites increased with applied loads. At higher applied loads, the 7075 alloy exhibited superior wear resistance than that of the composites. This is attributed to increased microcracking tendency of the composites than the matrix alloy.     

Silicon surface passivation by hot-wire CVD Si thin films studied by in situ surface spectroscopy

Silicon thin films can provide an excellent surface passivation of crystalline silicon (c-Si) which is of importance for high efficiency heterojunction solar cells or diffused emitter solar cells with well-passivated rear surfaces. Hot-wire chemical vapor deposition (hot-wire CVD) is an attractive method to synthesize Si thin films for these applications as the method is ion-bombardment free yielding good quality films over a wide range of deposition rates. The properties of the interface between hot-wire CVD Si thin films and H-terminated c-Si substrates have been studied during film growth by three complementary in situ techniques. Spectroscopic ellipsometry has been used to determine the optical properties and thickness of the films, whereas information on the H-bonding modes and H-depth profile has been obtained by attenuated total reflection infrared spectroscopy. Second-harmonic generation (SHG), a nonlinear optical technique sensitive to surface and interface states, has been used to probe two-photon resonances related to modified Si-Si bonds at the interface. By correlating the observations with ex situ lifetime spectroscopy experiments the growth and surface passivation mechanism of the Si films are discussed.     

Dielectric properties of sol-gel derived high-k titanium silicate thin films

High-k dielectric titanium silicate (Ti_xSi_1 − xO₂) thin films have been deposited by means of an optimized sol-gel process. At the optimal firing temperature of 600 °C, the Ti_0.5Si_0.5O₂ films are shown to exhibit not only a dielectric constant (k) as high as ∼ 23 but more importantly the lowest leakage current and dielectric losses. Fourier transform infrared spectroscopy shows an absorbance peak at 930 cm^− 1, which is a clear signature of the formation of Ti-O-Si bondings in all the silicate films. The developed sol-gel process offers the required latitude to grow Ti_xSi_1 − xO₂ with any composition (x) in the whole 0 ≤ x ≤ 1 range. Thus, the k value of the Ti_xSi_1 − xO₂ films can be tuned at any value between that of SiO₂ (3.8) to that of TiO₂ (k ∼ 60) by simply controlling the TiO₂ content of the films. The composition dependence of the dielectric constant of the Ti_xSi_1 − xO₂ films is analyzed in the light of existing models for dielectric composites.     

Stress evolution in magnetron sputtered Ti-Zr-N and Ti-Ta-N films studied by in situ wafer curvature: Role of energetic particles

Stress evolution during reactive magnetron sputtering of binary TiN, ZrN and TaN thin films as well as ternary Ti-Zr-N and Ti-Ta-N solid-solutions was studied using real-time wafer curvature measurements. The energy of the incoming particles (sputtered atoms, backscattered Ar, ions) was tuned by changing either the metal target (M_Ti = 47.9, M_Zr = 91.2 and M_Ta = 180.9 g/mol), the plasma conditions (effect of pressure, substrate bias or magnetron configuration) for a given target or by combining different metal targets during co-sputtering. Experimental results were discussed using the average energy of the incoming species, as calculated using Monte-Carlo simulations (SRIM code). In the early stage of growth, a rapid evolution to compressive stress states is noticed for all films. A reversal towards tensile stress is observed with increasing thickness at low energetic deposition conditions, revealing the presence of stress gradients. The tensile stress is ascribed to the development of a ‘zone T’ columnar growth with intercolumnar voids and rough surface. At higher energetic deposition conditions, the atomic peening mechanism is predominant: the stress remains largely compressive and dense films with more globular microstructure and smooth surface are obtained.     

Fabrication and biocompatibility in vitro of potassium titanate biological thin film/titanium alloy biological composite

A potassium titanate biological thin film/titanium alloy biological composite was fabricated by way of bionic chemistry. The biocompatibility in vitro of Ti-15Mo-3Nb and the potassium titanate biological thin film/titanium alloy was studied using simulated body fluid cultivation, kinetic clotting of blood and osteoblast cell cultivation experiments in vitro. By comparing the biological properties of both materials, the following conclusions can be obtained: (1) The deposition of a calcium phosphate layer was not found on the surface of Ti-15Mo-3Nb, so it was bioinert. Because the network of potassium titanate biological thin film could induce the deposition of a calcium phosphate layer, this showed that it had excellent bioactivity. (2) According to the values of kinetic clotting, the blood coagulation time of the potassium titanate biological thin film was more than that of Ti-15Mo-3Nb. It was obvious that the potassium titanate biological thin film possessed good hemocompatibility. (3) The cell compatibility of both materials was very good. However, the growth trend and multiplication of osteoblast cells on the surface of potassium titanate biological thin film was better, which made for the concrescence of wounds during the earlier period. As a result, the potassium titanate biological thin film/titanium alloy showed better biocompatibility and bioactivity. Translated from Journal of Functional Materials, 2006, 37(10): 1,638–1,642 (in Chinese)     

Microstructure effect on mechanical properties of in situ synthesized titanium matrix composites reinforced with TiB and La2O3

High temperature titanium matrix composites (TMCs) with different volume fraction of reinforcements were insitu synthesized by casting and hot forging. An effort was made to investigate the mechanical properties as a function of the microstructure of composites. Tensile tests were performed at room temperature, 600 °C, 650 °C and 700 °C respectively. Creep behavior at 650 °C was characterized in the stress range of 200-300 MPa. Results indicated that the composite with 2.11 vol.% reinforcements had the highest tensile strength and lowest steady state creep rate. Morphology of TiB whiskers was critical to mechanical properties of TMCs. TiB whiskers fracture and debonding acted as the dominant failure modes.     

Preparation of H-bar cross-sectional specimen for in situ TEM straining experiments: A FIB-based method applied to a nitrided Ti-6Al-4V alloy

The in situ tensile straining of cross-sectional specimens inside a TEM is intrinsically very difficult to perform despite its obvious interest to study interfaces of surface treated materials. We have combined a FIB-based method to produce H-bar specimens of a nitrided Ti-6Al-4V alloy and in situ TEM straining stage, to successfully study the plastic deformation mechanisms that are activated close to the nitrided surface in the Ti-based alloy.     

Influence of fluid circulation on in vitro reactivity of bioactive glass particles

The influence of circulation of simulated body fluid (SBF) on the in vitro behavior of glass particles (500–800 μm) was studied for seven glasses by measuring in situ pH inside the particle beds at 37 °C. Reaction layers on particle surfaces were characterized with SEM/EDXA. Differences in the tendency to form reaction layers on surfaces at different locations in the particle beds were determined using samples with interconnected porosity prepared by sintering the particles. Circulation of the solution above the particle bed affected strongly the in vitro behavior of the particles. Due to poor diffusion, the static conditions led to high pH in the SBF within the particle beds and uneven layer formation on the particles. Fluid circulation above the particle beds resulted in a homogenous environment lacking the pronounced pH gradients seen in the static system. The reaction layers on the particles were uniform but thin in the circulating conditions.     

Effects of extrusion and heat treatment on the microstructure and tensile properties of in situ TiBw/Ti6Al4V composite with a network architecture

In situ TiB whisker reinforced Ti6Al4V (TiBw/Ti64) composites with a network architecture were extruded and heat treated in order to further improve their mechanical properties. The microstructure results show that the equiaxed network architecture was extruded to column network architecture and TiB whisker to alignment distribution. The transformed β phase is formed and the residual stress generated during extrusion obviously decreases after water quenching and aging processes. The tensile test results show that the strength, elastic modulus and ductility of the composites can be significantly improved by the subsequent extrusion, and then, the strength can be further improved by water quenching and aging processes after hot extrusion deformation. The elastic modulus of the as-sintered composites with a novel network microstructure follows the upper bound of Hashin-Shtrikman (H-S) theory before extrusion, while that of the as-extruded composites with a column network microstructure agrees well with the prediction from Halpin-Tsai equation.     

Growth of a-b-axis orientation ZnO films with zinc vacancies by SSCVD

Zinc oxide (ZnO) films were grown on silicon (1 0 0) substrates by single-source chemical vapor deposition (SSCVD). X-ray diffraction (XRD) showed that ZnO thin films have a polycrystalline hexagonal wurtzite structure with (1 0 0) and (1 0 1) orientation, i.e., a-b-axis orientation. Atomic force microscopy (AFM) and scanning electronic microscopy (SEM) showed the films to be of relatively high density with a smooth surface. X-ray photoelectron spectroscopy (XPS) showed that the deposited films were very close to stoichiometry but contained a small number of zinc instead of O vacancies as normally found with ZnO films produced by other methods. These results were also confirmed by photoluminescence (PL) measurements.     

Effects of LP-MOCVD prepared TiO2 thin films on the in vitro behavior of gingival fibroblasts

We report on the in vitro response of human gingival fibroblasts (HGF-1 cell line) to various thin films of titanium dioxide (TiO₂) deposited on titanium (Ti) substrates by low pressure metal-organic chemical vapor deposition (LP-MOCVD). The aim was to study the influence of film structural parameters on the cell behavior comparatively with a native-oxide covered titanium specimen, this objective being topical and interesting for materials applications in implantology. HGF-1 cells were cultured on three LP-MOCVD prepared thin films of TiO₂ differentiated by their thickness, roughness, transversal morphology, allotropic composition and wettability, and on a native-oxide covered Ti substrate. Besides traditional tests of cell viability and morphology, the biocompatibility of these materials was evaluated by fibronectin immunostaining, assessment of cell proliferation status and the zymographic evaluation of gelatinolytic activities specific to matrix metalloproteinases secreted by cells grown in contact with studied specimens. The analyzed surfaces proved to influence fibronectin fibril assembly, cell proliferation and capacity to degrade extracellular matrix without considerably affecting cell viability and morphology. The MOCVD of TiO₂ proved effective in positively modifying titanium surface for medical applications. Surface properties playing a crucial role for cell behavior were the wettability and, secondarily, the roughness, HGF-1 cells preferring a moderately rough and wettable TiO₂ coating.     

In vitro bioactivity of chitosan/poly (d,l-lactide-co-glycolide) composites

Porous composites scaffolds of chitosan/poly(d,l-lactide-co-glycolide) were fabricated for tissue engineering applications by thermally induced phase separation and lyophilization techniques. The in vitro bioactivity evaluation of the scaffolds was carried out by analyzing the apatite layers produced on them using SBF as incubation medium. The apatite formation was analyzed using FTIR spectroscopy and Field emission scanning electron microscopy coupled to energy-dispersive electron X-ray spectroscopy. The cumulative results obtained from the IR spectra and SEM-EDS suggest that the developed composites might have potential applications in tissue engineering.     

钛酸钡/硅酸钙复合生物活性压电陶瓷的制备及性能研究

压电材料产生的电信号能够促进成骨细胞增殖分化, 但不具有良好的诱导矿化能力; 生物活性材料在生理环境下能够诱导类骨羟基磷灰石的沉积, 但又不能产生电信号促进成骨。因此, 开发出一种既能产生电信号, 又能诱导矿化沉积的复合生物活性压电材料, 具有重要意义。本研究以钛酸钡为压电组分, 以硅酸钙为生物活性组分, 采用固相烧结法制备了钛酸钡/硅酸钙复合生物活性压电陶瓷, 测试了压电性能, 并用体外矿化实验评价了诱导矿化能力。硅酸钙复合含量达到30%时, 复合陶瓷仍具有一定的压电性能(d₃₃=4 pC·N^-1), 并且能够在模拟体液中诱导磷酸钙沉积。钛酸钡与硅酸钙的复合能够同时具有压电性和生物活性, 为骨修复材料提供了新的选择。     

Cytocompatibility property evaluation of gas pressure sintered SiAlON-SiC composites with L929 fibroblast cells and Saos-2 osteoblast-like cells

Although the oxide ceramics have widely been investigated for their biocompatibility, non-oxide ceramics, such as SiAlON and SiC are yet to be explored in detail. Lack of understanding of the biocompatibility restricts the use of these ceramics in clinical trials. It is hence, essential to carry out proper and thorough study to assess cell adhesion, cytocompatibility and cell viability on the non-oxide ceramics for the potential applications. In this perspective, the present research work reports the cytocompatibility of gas pressure sintered SiAlON monolith and SiAlON-SiC composites with varying amount of SiC, using connective tissue cells (L929) and bone cells (Saos-2). The quantification of cell viability using MTT assay reveals the non-cytotoxic response. The cell viability has been found to be cell type dependent. An attempt has been made to discuss the cytocompatibility of the developed composites in the light of SiC content and type of sinter additives.     

Characterization of electron irradiated GaN n-p diode

Electron-beam irradiated GaN n⁺-p diodes were characterized by deep level transient spectroscopy (DLTS) and optical responsivity measurements. The GaN n⁺-p diode structures were grown by metal organic chemical vapor deposition technique, and the electron irradiation was done by the energies of 1 MeV and 2 MeV with dose of 1 × 10¹⁶ cm^− 2. In DLTS measurement, the defect states of E_c − 0.36 eV and E_c − 0.44 eV in the electron irradiated diodes appeared newly. The optical responsivity of GaN n⁺-p diode was characterized in ultra-violet region, and then the maximum optical responsivity at 350 nm was decreased after electron-beam irradiation.