Efficacy and stability of a novel silica supplement for improving bone development in broilers.

1. The essentiality of silicon for skeletal development has been established, but the adequacy of bioavailable silicon supply in broiler diets has not been considered for 30 years, despite average daily weight gain of birds increasing by almost a third over that time. Therefore, two studies were undertaken to investigate whether modern strains of broiler chicken benefit from diet supplementation with bioavailable silica. 2. Trial 1 was a 2x2x2 factorial study where six replicate pens of seven chicks were fed one of the eight freshly prepared diets from hatch to 21 days of age, with bodyweight gain and feed intake recorded weekly. Diets combined the following factors: silicon supplement fed at 0 ppm or 1000 ppm, phytase levels of either 0 FTU/kg or 1500 FTU/kg and either 0.6% or 0.7% Ca. Tibia were analysed for bone breaking strength, extent of tibial dyschondroplasia and feet measured for bone ash and pododermatitis score. 3. Trial 2 used a 0.7% Ca with 1500 FTU phytase diet as the control and compared this with the same diet containing either 1000 ppm silicon (MONO-Si) freshly added each week or 1000 ppm silicon added in a single, advance-prepared batch per feeding phase. Each diet was fed to nine pens of seven birds from 0 to 35 d with feed consumption and weight recorded weekly. Two birds per pen were euthanised on d 14, 21 and 35 and tibias collected for measurement of bone breaking strength, ash and mineral content. Serum was collected for Si content. 4. Univariate analysis of means from each trial showed that silica supplementation improved bird weight gain over the starter phase, though there was no effect on feed conversion. 5. Bone strength improved with added silica in both studies, without affecting bone mineral content; indicating that modern strains of broiler may require dietary supplementation with bioavailable silicon.

Bioavailability of a novel form of silicon supplement.

In this study, we assessed uptake and potential efficacy of a novel, pH neutral form of silicon supplement in vitro and using broiler chickens as a model species. In vitro bioavailability of this supplement was significantly higher than other commercial supplements tested, all of which claim available silica content. To confirm bioavailability of the new supplement in vivo, a broiler chick feeding trial reported blood uptake that was significantly higher than a Bamboo-derived silicon supplement. We assessed dose response of the novel supplement in a further study with increased dose related levels of silicon being detected in the blood and tibia. We found tibia and foot ash residue as a percentage of dry mass was higher with inclusion of the novel supplement in the diet, particularly in young birds and that this was followed by significant increase in tibia breaking strength. This novel supplement may therefore have applications in the improvement of bone integrity, with implications for the reduction of lameness in broilers. These results indicate the novel silica supplement is readily absorbed in chicks, and transported in the blood supply to sites such as the skeleton due to it being present in a non-condensed, monomeric form. There is potential for wider application of this silica supplement in other species where bone breakages are a problem, including high performance sport.

HPV-associated cervical cancer cells targeted by triblock copolymer gold nanoparticle curcumin combination.

OBJECTIVE: Curcumin (diferuloylmethane) has promising anti-cervical cancer properties but requires a stabilizing complex such as the Pluronic triblock copolymer gold nanoparticle (GNP). The objectives were to study cytotoxicity of curcumnin and to determine the effect of copolymer GNPs curcumnin complex on cancer cell necrosis. MATERIALS AND METHODS: The HeLa cells were maintained in Eagle Minimal Essential Medium, fetal bovine serum, and antibiotics, and passaged until 60% confluency was reached. The cells were exposed to either: (1) control medium, (2) 50 µM curcumin, (3) 100 µM curcumin, (4) 50 µM curcumnin with copolymer GNPs complex, or (5) 100 µM curcumnin with copolymer GNPs complex. The treated cells were incubated at 37°C with 5% CO(2) in air for 24 hours, and analyzed for viability, apoptosis or necrosis using the dual stains fluorescence procedure. RESULTS: A dose-dependent increase in the HeLa necrosis was observed with increasing curcumnin concentrations. Cytotoxic effect was decreased by five- to ten-fold when the curcumin was complexed with copolymer GNPs. There were more apoptotic HeLa cells at the higher concentration of curcurnin but combination with copolymer GNPs resulted in decreased apoptosis. Cell viability was higher in curcumnin with copolymer GNPs (74.4 ± 4.8 versus 2.3 ± 2.2% live, mean ± SEM, with and without copolymer GNPs, respectively). CONCLUSION: Curcumin increased HeLa cancer cell necrosis but its cytotoxicity was decreased by copolymer GNPs. The results suggested that this specific copolymer GNP did not enhance the curcumnin bioavailability in cultured cells possibly due to formation of copolymer GNP aggregates.

Enhancing in vitro biocompatibility and corrosion protection of organic-inorganic hybrid sol-gel films with nanocrystalline hydroxyapatite.

Application of novel organic-inorganic hybrid sol-gel coatings containing dispersed hydroxyapatite (HAp) particles improves the biocompatibility, normal human osteoblast (NHOst) response in terms of osteoblast viability and adhesion of a Ti6Al4V alloy routinely used in medical implants. The incorporation of HAp particles additionally results in more effective barrier proprieties and improved corrosion protection of the Ti6Al4V alloy through higher degree of cross-linking in the organopolysiloxane matrix and enhanced film thickness.

Triethylphosphite as a network forming agent enhances in vitro biocompatibility and corrosion protection of hybrid organic-inorganic sol-gel coatings for Ti6Al4V alloys.

The biocompatibility and life of metallic implants can be enhanced through improving the biocompatibility and corrosion protection characteristics of the coatings used with these materials. In this study, triethylphosphite (TEP) was used to introduce phosphorus into organic-inorganic hybrid silica based sol-gel coatings prepared using γ-methacryloxypropyltrimethoxysilane and tetramethylorthosilicate. Addition of TEP dramatically increased the rate of intermolecular condensation and resulted in materials showing greater cross-linking. Protein (fibrinogen) uptake, osteoblast in vitro biocompatibility and corrosion resistance was enhanced in coatings containing TEP. Although higher concentrations of phosphorus supported the greatest improvement in biocompatibility, a compromise in the phosphorus concentration used would be required if corrosion resistance was most desirable parameter for optimisation. Films prepared by dip coating on Ti6Al4V alloys from these sols offer a promising alternative to wholly metallic prostheses.

Construction of silica-enhanced S-layer protein cages.

The work presented here shows for the first time that it is possible to silicify S-layer coated liposomes and to obtain stable functionalized hollow nano-containers. For this purpose, the S-layer protein of Geobacillus stearothermophilus PV72/p2 was recombinantly expressed and used for coating positively charged liposomes composed of dipalmitoylphosphatidylcholine, cholesterol and hexadecylamine in a molar ratio of 10:5:4. Subsequently, plain (uncoated) liposomes and S-layer coated liposomes were silicified. Determination of the charge of the constructs during silicification allowed the deposition process to be followed. After the particles had been silicified, lipids were dissolved by treatment with Triton X-100 with the release of previously entrapped fluorescent dyes being determined by fluorimetry. Both, ζ-potential and release experiments showed differences between silicified plain liposomes and silicified S-layer coated liposomes. The results of the individual preparation steps were examined by embedding the respective assemblies in resin, ultrathin sectioning and inspection by bright-field transmission electron microscopy (TEM). Energy filtered TEM confirmed the successful construction of S-layer based silica cages. It is anticipated that this approach will provide a key to enabling technology for the fabrication of nanoporous protein cages for applications ranging from nano medicine to materials science.

A multi-technique study of the effect of aqueous aluminium speciation on hydrolytic gelation of aluminium (oxy)hydroxide.

The sol-gel transformation of aqueous solutions of aluminium ions into aluminium (oxy)hydroxides induced by the addition of a 'soft base'-'Tris-buffer' (pK(a)=8.2) has been investigated using monotonous single-batch titrations and a combination of four complimentary techniques for monitoring pH, conductivity, viscosity and ultrasound parameters (velocity and attenuation). The multi-probe monitoring of the formation of aluminium (oxy)hydroxides enabled important stages of the sol-gel transformation process including: the structural conversion of aluminium Keggin-like polynuclear clusters into nanoparticles of aluminium (oxy)hydroxide; the aggregation of primary nuclei of aluminium (oxy)hydroxide into larger particles, and the 'arrested growth' of the aggregates with the formation of the three-dimensional gel network to be followed. The effect of aluminium ion molecular speciation on the sol-gel transformation stages is discussed. The data presented show that ultrasonic spectrometry, although a novel tool for sol-gel studies, is beneficial for the non-invasive monitoring of the latter stages of aluminium (oxy)hydroxide formation and its eventual breakdown in the presence of excess base.

Electron stimulated reactions of methyl iodide coadsorbed with amorphous solid water.

The electron stimulated reactions of methyl iodide (MeI) adsorbed on and suspended within amorphous solid water (ice) were studied using a combination of postirradiation temperature programmed desorption and reflection absorption infrared spectroscopy. For MeI adsorbed on top of amorphous solid water (ice), electron beam irradiation is responsible for both structural and chemical transformations within the overlayer. Electron stimulated reactions of MeI result principally in the formation of methyl radicals and solvated iodide anions. The cross section for electron stimulated decomposition of MeI is comparable to the gas phase value and is only weakly dependent upon the local environment. For both adsorbed MeI and suspended MeI, reactions of methyl radicals within MeI clusters lead to the formation of ethane, ethyl iodide, and diiodomethane. In contrast, reactions between the products of methyl iodide and water dissociation are responsible for the formation of methanol and carbon dioxide. Methane, formed as a result of reactions between methyl radicals and either parent MeI molecules or hydrogen atoms, is also observed. The product distribution is found to depend on the film's initial chemical composition as well as the electron fluence. Results from this study highlight the similarities in the carbon-containing products formed when monohalomethanes coadsorbed with amorphous solid water are irradiated by either electrons or photons.

Wetting and wetting transitions on copper-based super-hydrophobic surfaces.

Rough and patterned copper surfaces were produced using etching and, separately, using electrodeposition. In both of these approaches the roughness can be varied in a controlled manner and, when hydrophobized, these surfaces show contact angles that increase with increasing roughness to above 160 degrees . We show transitions from a Wenzel mode, whereby the liquid follows the contours of the copper surface, to a Cassie-Baxter mode, whereby the liquid bridges between features on the surface. Measured contact angles on etched samples could be modeled quantitatively to within a few degrees by the Wenzel and Cassie-Baxter equations. The contact angle hysteresis on these surfaces initially increased and then decreased as the contact angle increased. The maximum occurred at a surface area where the equilibrium contact angle would suggest that a substantial proportion of the surface area was bridged.

Kinetics of electron-induced decomposition of CF2Cl2 coadsorbed with water (ice): a comparison with CCl4.

The kinetics of decomposition and subsequent chemistry of adsorbed CF(2)Cl(2), activated by low-energy electron irradiation, have been examined and compared with CCl(4). These molecules have been adsorbed alone and coadsorbed with water ice films of different thicknesses on metal surfaces (Ru; Au) at low temperatures (25 K; 100 K). The studies have been performed with temperature programmed desorption (TPD), reflection absorption infrared spectroscopy (RAIRS), and x-ray photoelectron spectroscopy (XPS). TPD data reveal the efficient decomposition of both halocarbon molecules under electron bombardment, which proceeds via dissociative electron attachment (DEA) of low-energy secondary electrons. The rates of CF(2)Cl(2) and CCl(4) dissociation increase in an H(2)O (D(2)O) environment (2-3x), but the increase is smaller than that reported in recent literature. The highest initial cross sections for halocarbon decomposition coadsorbed with H(2)O, using 180 eV incident electrons, are measured (using TPD) to be 1.0+/-0.2 x 10(-15) cm(2) for CF(2)Cl(2) and 2.5+/-0.2 x 10(-15) cm(2) for CCl(4). RAIRS and XPS studies confirm the decomposition of halocarbon molecules codeposited with water molecules, and provide insights into the irradiation products. Electron-induced generation of Cl(-) and F(-) anions in the halocarbon/water films and production of H(3)O(+), CO(2), and intermediate compounds COF(2) (for CF(2)Cl(2)) and COCl(2), C(2)Cl(4) (for CCl(4)) under electron irradiation have been detected using XPS, TPD, and RAIRS. The products and the decomposition kinetics are similar to those observed in our recent experiments involving x-ray photons as the source of ionizing irradiation.

Topography driven spreading.

Roughening a hydrophobic surface enhances its nonwetting properties into superhydrophobicity. For liquids other than water, roughness can induce a complete rollup of a droplet. However, topographic effects can also enhance partial wetting by a given liquid into complete wetting to create superwetting. In this work, a model system of spreading droplets of a nonvolatile liquid on surfaces having lithographically produced pillars is used to show that superwetting also modifies the dynamics of spreading. The edge speed-dynamic contact angle relation is shown to obey a simple power law, and such power laws are shown to apply to naturally occurring surfaces.

The systematic study of aluminium speciation in medium concentrated aqueous solutions.

Industrial applications and environmental problems involving the aqueous chemistry of aluminium require an understanding of the speciation of this metal ion at a wide range of concentrations. The formation of polynuclear species is of special interest due to the complexity of the hydrolysis mechanisms and the diversity of the hydrolysis products. Kinetic aspects of speciation are also important considering the different stability ranges of polycationic species formed during the hydrolysis process. In the present paper we report results of systematic studies on the formation of aluminium polycations at room temperature. Automated potentiometric titrations have been used to study the hydrolysis of aluminium-ions in solutions (0.01-0.2 M) on a short time scale (2 min between titrant additions). (27)Al NMR spectroscopy and dynamic light scattering have been used for investigations on a longer time scale (24 h). The effects of alkali strength (KOH, NH(4)OH and KHCO(3)) and concentration (0.45-2.0 M), counterion identity (Cl(-), NO(3)(-), SO(4)(2-)) and ionic strength have been investigated. Optimum conditions for the generation of Al(13)-mer are proposed on short and long time scales. On a short time scale, aluminium chloride and nitrate should be used as starting materials, KOH and KHCO(3) should be used for hydrolysis and experiments conducted at low ionic strength. For solutions that have been left to age, there is a considerable hydrolysis window that can be used to generate significant quantities of the Al(13)-mer that vary little with the alkali used. Al(13)-mer species are not generated from alum as the precursor. The presence of sulphate ions alters the pathway of aluminium polymerisation to form polymeric and solid materials. On the basis of the potentiometric titration data, dynamic light scattering and (27)Al NMR measurements evidence is provided for the detrimental role of sulphate-ions in the formation of Al(13)-mer and an alternative mechanism of aluminium ion polycondensation is proposed, based on the increased stability of monomeric and oligomeric species (dimer and trimer) in the presence of sulphate-ions.

Biosilicification: the role of the organic matrix in structure control.

Silicon (although never in the elemental form) is present in all living organisms and is required for the production of structural materials in single-celled organisms through to higher plants and animals. Hydrated amorphous silica is a mineral of infinite functionality and yet it is formed into structures with microscopic and macroscopic form. Research into the mechanisms controlling the process have highlighted proteins and proteoglycans as possible control molecules. Such molecules are suggested to play a critical role in the catalysis of silica polycondensation reactions and in structure direction. This article reviews information on silica form and function, silica condensation chemistry, the role of macromolecules in structure control and in vitro studies of silica formation using biomolecules extracted from biological silicas. An understanding of the mechanisms by which biological organisms regulate mineral formation will assist in our understanding of the essentiality of silicon to life processes and in the generation of new materials with specific form and function for industrial application in the 21st century.

Model studies of the precipitation of silica in the presence of aluminium; implications for biology and industry.

The unique chemical affinity between the oxides of silicon and aluminium has been cited as a potential route for the amelioration of the detrimental effects of aluminium in the environment and in biological systems. A greater understanding of silicon-aluminium interactions may assist in this endeavour and also provide a means of overcoming silica fouling problems encountered by industry which are exacerbated by the presence of aluminium. It is also conceivable that this increased knowledge may demonstrate a positive use for aluminium in the processing of the silicon dioxide phase. In this study we report the effect of aluminium ions, derived from aluminium chloride, on silicic acid species obtained from potassium catecholato complexes of silicon at circumneutral pH at the molar ratios 1000Si:Al, 100Si:Al and 50Si:Al. Silica and low levels of aluminium-rich silica materials were formed with Si:Al ratios of about 3.5:1 comparable with the element ratios detected in senile plaques and aluminium-rich scale. A kinetic study showed that aluminium in the reaction medium slowed down the rate of formation of one of the silica species formed early in the condensation process, e.g. trimers, but increased the rate at which silicic acid was removed from sub 1 nm diameter particles. The materials precipitated in the presence of aluminium were composed of smaller particles and aggregates with smaller pores (Si100:Al and Si50:Al systems) or larger pores (Si1000:Al) compared to the control. The nature of the interactions responsible for these differences is discussed. The effects described here demonstrate the ability of silica and aluminium to interact under conditions such as those found in biological systems. That silica reacts with aluminium in the presence of catechol supports the protective role assigned to silicon.

Aspects of the bioinorganic chemistry of silicon in conjunction with the biometals calcium, iron and aluminium.

Silicon, in its various forms (silicic acid (Si(OH)4) through to hydrated amorphous silica (SiOn(OH)4-2n, n = 2-4) is important in geological and biological processes. Silicon is considered to be an 'essential' element for some plants and higher animals and when present as silica imparts structural, defensive and photosynthetic advantages to many plants. For the majority of essential elements specific molecular binding sites have been identified, but this is not the case for silicon. This has lead to the proposal that silicon may not act directly on biological systems but may exert its influence via interactions with biometals. Metals for which this may be a viable option include aluminium, iron and calcium, all of which can be found co-located with silicon in both minerals and living organisms. This article briefly reviews the pertinent solution and solid state chemistry of silicon in relation to aluminium, iron and calcium. Examples where silicon and an additional metal are found in solid state phases are described. The biological systems where these deposits are found are extremely complex and preliminary results from a model system designed to study silicon-biometal interactions are described.

Evaluation of the osteoblast response to a silica gel in vitro.

Many bioactive glasses and glass ceramics contain silica, yet the effect of silica on the osteoblast is not well understood. The osteoblast cell response to a silica surface, without the interference of the other ions present in glasses and glass ceramics has been investigated. A silica sol-gel was prepared which gave a molar ratio of 1:4:4 tetraethyl orthosilicate (TEOS): ethanol:acidified water 0.2 M HCl) and spin cast on to thermanox discs. The gel was characterized in terms of bioactivity and release of silicic acid. Primary human osteoblasts (HOBs) were seeded on the surface of upright or inverted silica discs. Cell activity (alamar blue reduction), number (DNA content) and differentiation (alkaline phosphatase activity, nodule formation and mineralization) were measured. There was no apparant difference in cell number, activity or alkaline phosphatase activity between silica discs and controls. Nodules formed much earlier on the silica surfaces and these eventually mineralized. Nodule formation was reproducibly enhanced on the silica surface and less markedly on the inverted discs. It is likely that both the surface characteristics of the silica gel and silicic acid release from the disc affect osteoblast behaviour.

A physico-chemical approach to morphogenesis: the roles of inorganic ions and crystals.

We consider morphogenesis with special references to the development of mineral frameworks, organic filamentous structures and the location of enzymes, including ion-pumps, in membranes. Starting from a description of the morphology of inorganic crystals we analyse so-called equilibrium growth, i.e. growth at constant shape, both outside and inside biological systems. It is shown that an initial small spherical cell in which linear, ordered, inorganic or organic features are built will become distorted. The distortion is due to stresses which affect membrane curvature and consequently rearrange enzymes in membranes. The cell system can rapidly attain a steady-state of development, ('equilibrium') growth, of fixed morphology. After a considerable growth period the cell may cease to grow or the steady state may be broken and a transition can then occur to a quite new morphology. Examples are taken mostly from unicellular organisms but the ideas apply to multi-cellular systems.

Structural aspects of biogenic silica.

The objectives of this paper are to discuss the characterization of biogenic silica in terms of structural properties, and to elucidate the mechanisms of structural organization within biological systems. The scale of organization is a critical factor in the characterization of biosilicification processes, and order at the nanometre, micrometre and macroscopic levels is described. Molecular order is discussed in the light of high-resolution transmission electron microscopy and solid-state NMR results obtained from samples of biogenic silica. Microscopic organization is expressed in a range of structural motifs, e.g. gels, sheets, fibres, tubes and globular assemblies, and reflects the infinitely adaptive morphology of biogenic silica. Macroscopic structures such as curved rods, spicules, perforated plates, teeth and reticular frameworks can be assembled from these microscopic motifs. The mechanisms of structural organization involve spatial (scalar and vectorial) constraints, ordered particle aggregation and chemical regulation. The possible importance of organic surfaces is discussed.