Ultrasmall silica nanoparticles directly ligate the T cell receptor complex.

The impact of ultrasmall nanoparticles (<10-nm diameter) on the immune system is poorly understood. Recently, ultrasmall silica nanoparticles (USSN), which have gained increasing attention for therapeutic applications, were shown to stimulate T lymphocytes directly and at relatively low-exposure doses. Delineating underlying mechanisms and associated cell signaling will hasten therapeutic translation and is reported herein. Using competitive binding assays and molecular modeling, we established that the T cell receptor (TCR):CD3 complex is required for USSN-induced T cell activation, and that direct receptor complex-particle interactions are permitted both sterically and electrostatically. Activation is not limited to αß TCR-bearing T cells since those with γδ TCR showed similar responses, implying that USSN mediate their effect by binding to extracellular domains of the flanking CD3 regions of the TCR complex. We confirmed that USSN initiated the signaling pathway immediately downstream of the TCR with rapid phosphorylation of both ζ-chain-associated protein 70 and linker for activation of T cells protein. However, T cell proliferation or IL-2 secretion were only triggered by USSN when costimulatory anti-CD28 or phorbate esters were present, demonstrating that the specific impact of USSN is in initiation of the primary, nuclear factor of activated T cells-pathway signaling from the TCR complex. Hence, we have established that USSN are partial agonists for the TCR complex because of induction of the primary T cell activation signal. Their ability to bind the TCR complex rapidly, and then to dissolve into benign orthosilicic acid, makes them an appealing option for therapies targeted at transient TCR:CD3 receptor binding.

Silicon Forms a Rich Diversity of Aliphatic Polyol Complexes in Aqueous Solution.

A detailed examination of aqueous Si complexation by alditols and aldonic acids was conducted using high-sensitivity 29Si NMR spectroscopy of isotopically enriched solutions combined with theoretical modeling. Contrary to previous thinking, we have established that aliphatic polyols do not require a threo pair of hydroxy groups to form hypercoordinated Si complexes, although formation constants may be orders of magnitude higher if they are present. Thirteen distinctly different molecular assemblages containing 4-, 5-, or 6-coordinate Si centers have been identified, with significant concentrations of 5-coordinate Si bis-ligand complex being detected even under biologically relevant solution conditions.

Identification of a mammalian silicon transporter.

Silicon (Si) has long been known to play a major physiological and structural role in certain organisms, including diatoms, sponges, and many higher plants, leading to the recent identification of multiple proteins responsible for Si transport in a range of algal and plant species. In mammals, despite several convincing studies suggesting that silicon is an important factor in bone development and connective tissue health, there is a critical lack of understanding about the biochemical pathways that enable Si homeostasis. Here we report the identification of a mammalian efflux Si transporter, namely Slc34a2 (also termed NaPiIIb), a known sodium-phosphate cotransporter, which was upregulated in rat kidney following chronic dietary Si deprivation. Normal rat renal epithelium demonstrated punctate expression of Slc34a2, and when the protein was heterologously expressed in Xenopus laevis oocytes, Si efflux activity (i.e., movement of Si out of cells) was induced and was quantitatively similar to that induced by the known plant Si transporter OsLsi2 in the same expression system. Interestingly, Si efflux appeared saturable over time, but it did not vary as a function of extracellular [Formula: see text] or Na+ concentration, suggesting that Slc34a2 harbors a functionally independent transport site for Si operating in the reverse direction to the site for phosphate. Indeed, in rats with dietary Si depletion-induced upregulation of transporter expression, there was increased urinary phosphate excretion. This is the first evidence of an active Si transport protein in mammals and points towards an important role for Si in vertebrates and explains interactions between dietary phosphate and silicon.

Effects of carbonation on the leachability and compressive strength of cement-solidified and geopolymer-solidified synthetic metal wastes.

The effects of accelerated carbonation on the compressive strength and leachability of fly ash-based geopolymer and ordinary portland cement (OPC) doped with Cd(II), Cr(III), Cr(VI), Cu(II), Pb(II) or Zn(II) salts were investigated. Cement was effective at immobilizing Cd, Cr(III), Cu, Pb and Zn under both the Synthetic Precipitation Leaching Procedure (SPLP) and the Toxicity Characteristic Leaching Procedure (TCLP), but ineffective for retaining Cr(VI). Carbonated cement maintained its ability to immobilize Cd, Cr(III), Pb and Zn, but, under acidic TCLP conditions, was much worse at retaining Cu. Geopolymer was effective at immobilizing Cr(III) and Cu, and, to a lesser degree, Cd, Pb and Zn in SPLP leaching tests. Only Cr(III) was immobilized under comparatively acidic TCLP testing conditions. Carbonation did not change the metal retention capacity of the geopolymer matrix. Metal doping caused compressive strengths of both geopolymer and cement to decrease. Carbonation increased the compressive strength of cement, but decreased that of the geopolymer. Geochemical equilibrium modeling provided insight on the mechanisms of metal immobilization.

Aqueous alkali-metal silicate anions containing fully condensed four-coordinate sites.

Conspicuous absence: (29)SiNMR studies show that highly condensed, four-coordinate Si-containing anions are common in concentrated alkali-metal silicate solutions, but reveal no evidence for the existence of specialized zeolite building units. The results add to the mounting evidence that silicate polymerization simply proceeds through stepwise condensation of monosilicate tetrahedra.

Divergent effects of orthosilicic acid and dimethylsilanediol on cell survival and adhesion in human osteoblast-like cells.

Although dietary silicon (Si) is recognized to be an important factor for the growth and development of bone and connective tissue, its biochemical role has yet to be identified. The predominant Si-containing species in blood and other biofluids is orthosilicic acid, Si(OH)(4). Dimethylsilanediol, (CH(3))(2)Si(OH)(2), is an environmental contaminant that results from decomposition of silicone compounds used in personal hygiene, health care and industrial products. We examined the in vitro effects of both Si species on the survival (colony forming efficiency), proliferation (DNA content), differentiation (alkaline phosphatase activity) and adhesion (relative protein content) of the human osteoblast-like cell lines Saos-2 and hFOB 1.19. Orthosilicic acid yielded a small, dose-dependent decrease in Saos-2 cell survivability up to its 1,700 micromol/L solubility limit, by which point survival was 20% less than that of untreated cells. This negative association, although small, correlated with a reduction in the proliferation and adhesion of Saos-2 cells as well as of hFOB 1.19 and osteoclast-like GCT cells. By contrast, dimethylsilanediol treatment had no discernable influence on Saos-2 survivability at concentrations up to 50 micromol/L, and yet significantly enhanced cell survival at higher doses. Moreover, dimethylsilanediol did not affect proliferation or adhesion of any cell line. The findings show that orthosilicic acid and dimethylsilanediol affect osteoblast-like cells very differently, providing insight into the mechanism by which silicon influences bone health, although the specific site of Si activity remains unknown. There was no evidence to suggest that dimethylsilanediol is cytotoxic at environmental/physiological concentrations.

Effects of sucrose and sorbitol on cement-based stabilization/solidification of toxic metal waste.

The effects of sucrose or sorbitol addition on the hydration, unconfined compressive strength and leachability of Portland cement pastes containing 1% Pb and 1% Zn were studied as a function of time. Whereas Pb and Zn were found to shorten the time to achieve maximum hydration of Portland cement, the combination of these metals with 0.15 wt% sucrose or 0.40 wt% sorbitol retarded the setting of cement by at least 7 and 28 days, respectively, without affecting the strength at 56 days. The leachability of Pb and Zn evaluated by the TCLP 1311 protocol at 56 and 71 days was slightly reduced or unchanged by the addition of sucrose or sorbitol. SEM-EDS and XRD analyses revealed that ettringite precipitation was favored whereas the formation of CSH gel, which accounts for most of the strength of hydrated cement, was delayed in cement pastes containing both metals and sucrose or sorbitol. These results indicate that controlled additions of sucrose or sorbitol can add flexibility to the handling of cement-treated metal waste, particularly when it needs to be transported by truck or pipeline between the treatment plant and the disposal site, without affecting its long-term performance.

Non-Functionalized Ultrasmall Silica Nanoparticles Directly and Size-Selectively Activate T Cells.

Sub-micron-sized silica nanoparticles, even as small as 10-20 nm in diameter, are well-known for their activation of mononuclear phagocytes. In contrast, the cellular impact of those <10 nm [ i.e., ultrasmall silica nanoparticles (USSN)] is not well-established for any cell type despite anticipated human exposure. Here, we synthesized discrete populations of USSN with volume median diameters between 1.8 to 16 nm and investigated their impact on the mixed cell population of human primary peripheral mononuclear cells. USSN 1.8-7.6 nm in diameter, optimally 3.6-5.1 nm in diameter, induced dose-dependent CD4 and CD8 T-cell activation in terms of cell surface CD25 and CD69 up-regulation at concentrations above 150 µM Sitotal (∼500 nM particles). Induced activation with only ∼2.4 µM particles was (a) equivalent to that observed with typical positive control levels of Staphylococcal enterotoxin B (SEB) and (b) evident in antigen presenting cell-deplete cultures as well as in a pure T-cell line (Jurkat) culture. In the primary mixed-cell population, USSN induced IFN-γ secretion but failed to induce T-cell proliferation or the secretion of IL-2, IL-10, or IL-4. Collectively, these data indicate that USSN initiate activation, with Th1 polarization, of T cells via direct particle-cell interaction. Finally, similarly sized iron hydroxide particles did not induce the expression of T-cell activation markers, indicating some selectivity of the ultrasmall particle type. Given that humans may be exposed to ultrasmall particles and that these materials have emerging bioclinical applications, their off-target immunomodulatory effects via direct T-cell activation should be carefully considered.

Adsorption of Amorphous Silica Nanoparticles onto Hydroxyapatite Surfaces Differentially Alters Surfaces Properties and Adhesion of Human Osteoblast Cells.

Silicon (Si) is suggested to be an important/essential nutrient for bone and connective tissue health. Silicon-substituted hydroxyapatite (Si-HA) has silicate ions incorporated into its lattice structure and was developed to improve attachment to bone and increase new bone formation. Here we investigated the direct adsorption of silicate species onto an HA coated surface as a cost effective method of incorporating silicon on to HA surfaces for improved implant osseointegration, and determined changes in surface characteristics and osteoblast cell adhesion. Plasma-sprayed HA-coated stainless steel discs were incubated in silica dispersions of different concentrations (0-42 mM Si), at neutral pH for 12 h. Adsorbed Si was confirmed by XPS analysis and quantified by ICP-OES analysis following release from the HA surface. Changes in surface characteristics were determined by AFM and measurement of surface wettability. Osteoblast cell adhesion was determined by vinculin plaque staining. Maximum Si adsorption to the HA coated disc occurred after incubation in the 6 mM silica dispersion and decreased progressively with higher silica concentrations, while no adsorption was observed with dispersions below 6 mM Si. Comparison of the Si dispersions that produced the highest and lowest Si adsorption to the HA surface, by TEM-based analysis, revealed an abundance of small amorphous nanosilica species (NSP) of ~1.5 nm in diameter in the 6 mM Si dispersion, with much fewer and larger NSP in the 42 mM Si dispersions. 29Si-NMR confirmed that the NSPs in the 6 mM silica dispersion were polymeric and similar in composition to the larger NSPs in the 42 mM Si dispersion, suggesting that the latter were aggregates of the former. Amorphous NSP adsorbed from the 6 mM dispersion on to a HA-coated disc surface increased the surface's water contact angle by 53°, whereas that adsorbed from the 42 mM dispersion decreased the contact angle by 18°, indicating increased and decreased hydrophobicity, respectively. AFM showed an increase in surface roughness of the 6 mM Si treated surface, which correlated well with an increase in number of vinculin plaques. These findings suggest that NSP of the right size (relative to charge) adsorb readily to the HA surface, changing the surface characteristics and, thus, improving osteoblast cell adhesion. This treatment provides a simple way to modify plasma-coated HA surfaces that may enable improved osseointegration of bone implants.

Silicon-29 NMR Studies of Tetraalkylammonium Silicate Solutions. 1. Equilibria, (29)Si Chemical Shifts, and (29)Si Relaxation.

The addition of tetraalkylammonium cations to aqueous silicate solutions enhances the abundance of symmetric, cagelike, polysilicate anions including the cubic octamer, Si(8)O(20)(8)(-). The equilibrium ratio of tetramethylammonium (TMA) cations to the octameric silicate anion is 8:1 for solutions with a concentration ratio [OH(-)]:[Si] >/= 1:1. Evidence indicates that organocations directly associate with cagelike polyanions to form a protective shell of hydrophobic hydration that impedes hydrolysis of the central anion.

Silicon-29 NMR Studies of Tetraalkylammonium Silicate Solutions. 2. Polymerization Kinetics.

The kinetics of formation of the silicate cubic octamer, Q(3)(8), in aqueous tetramethylammonium (TMA) silicate solutions was investigated by (29)Si NMR. The rate equation for solutions at pH 13.2-13.6 is d[Q(3)(8)]/dt = k(f) [H(+)](1.6)(+/-)(0.1)[TMA(+)](0.36)(+/-)(0.08)[Si](0.8)(+/-)(0.3) where k(f) = (2.2 +/- 0.8) x 10(16) mol(-)(1.8) kg(1.8) s(-)(1) at 296 K. The findings prove unequivocally that alkylammonium cations participate directly in the formation and subsequent stabilization of cagelike polysilicate anions. This implies a radically different mechanistic role than "templating" for alkylammonium cations in the synthesis of molecular sieves.

The silicon supplement 'Monomethylsilanetriol' is safe and increases the body pool of silicon in healthy Pre-menopausal women.

BACKGROUND: Monomethylsilanetriol (MMST) has been used for decades as an oral silicon supplement for bone and connective tissue health, although there are no formal data on its in vivo utilisation or safety following sustained dosing. METHODS: To investigate whether MMST contributes to the body pool of silicon and, secondly, to establish its safety following 4 weeks' supplementation in humans, twenty-two healthy pre-menopausal women (22-38 years) were recruited and supplemented with MMST at the maximum daily recommended dose (10.5 mg Si/day) for 4 weeks in a double-blind, randomised, placebo-controlled, cross-over design (i.e. 8 weeks in total). Fasting serum and urine samples were collected at baseline and at the end of the 4-week supplementation/placebo periods for analysis of total silicon by inductively coupled plasma optical emission spectrometry, MMST by proton nuclear magnetic resonance spectroscopy and full serum biochemistry. Participants also reported on, by questionnaire, their health, well-being and quality of life at 0, 4 and 8 weeks. RESULTS: Overall, 4-weeks supplementation with MMST significantly increased total fasting Si concentrations in serum and urine (P ≤ 0.003; paired t-test). MMST was semi-quantifiable in serum and quantifiable in urine, but only accounted for ca. 50% and 10%, respectively, of the increased total-Si concentration. There were no reported adverse effects (i.e. changes to health and well-being) or serum biochemical changes with MMST versus placebo. CONCLUSIONS: Our data indicate that oral MMST is safe, is absorbed and undergoes sufficient metabolism in vivo to raise fasting serum silicon levels, consistent with other well absorbed forms of dietary silicon (e.g. orthosilicic acid). It thus appears to be a suitable silicon supplement.

Do zeolite precursor species really exist in aqueous synthesis media?

The authors of a recent report in this journal (Houssin, et al., Phys. Chem. Chem. Phys., 2003, 5, 3518, ) claim that a tetrapropylammonium (TPA) silicate mixture with molar composition 4.41 TPAOH : 10 SiO(2) : 117 H(2)O contains high concentrations of two silicate oligomers, the prismatic double five-ring and a novel pentacyclic dodecamer. The latter species is derivative of the framework structure of silicalite-1, a MFI-type zeolite that spontaneously crystallizes from this system, and, indeed, the authors declare it to be a "nanoprecursor" in the TPA-mediated growth of silicalite-1. Using two-dimensional (29)Si COSY NMR spectroscopy to examine an equivalent mixture enriched in the (29)Si isotope, we show that the published peak assignments are erroneous. The signal assigned to the double five-ring corresponds, in reality, to the well-characterized pentacyclic octamer, while the three signals assigned to the pentacyclic dodecamer arise from three completely separate species. We find no evidence, therefore, to support the suggestion that silicalite-1 is formed by sequential condensation of precursor species any more complex than the simple orthosilicate anion.

The structure of aqueous pentaoxo silicon complexes with cis-1,2-dihydroxycyclopentane and furanoidic vicinal cis-diols.

Addition of cis-1,2-dihydroxycyclopentane to aqueous alkaline silicate solutions results in the spontaneous formation of three organosilicate species, each with a 2:1 ligand to Si ratio and a pentacoordinated silicon centre. By using a mixture of both cis-1,2-dihydroxycyclopentane and 1,4-anhydroerythritol we show unambiguously that all three species are diastereomers of the monomeric bis(diolato)-hydroxo complex, [(L=)(2)SiOH](-)(where L represents the cis-diol ligand), thus clarifying the general assignment of (29)Si NMR spectra reported for silicate solutions containing furanoidic sugars with vicinal cis-diol functionality, such as ribose.