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Exposure to exogenous particles is of increasing concern to human health. Characterizing the concentrations, chemical species, distribution, and involvement of the stimulus with the tissue microanatomy is essential in understanding the associated biological response. However, no single imaging technique can interrogate all these features at once, which confounds and limits correlative analyses. Developments of synchronous imaging strategies, allowing multiple features to be identified simultaneously, are essential to assess spatial relationships between these key features with greater confidence. Here, we present data to first highlight complications of correlative analysis between the tissue microanatomy and elemental composition associated with imaging serial tissue sections. This is achieved by assessing both the cellular and elemental distributions in three-dimensional space using optical microscopy on serial sections and confocal X-ray fluorescence spectroscopy on bulk samples, respectively. We propose a new imaging strategy using lanthanide-tagged antibodies with X-ray fluorescence spectroscopy. Using simulations, a series of lanthanide tags were identified as candidate labels for scenarios where tissue sections are imaged. The feasibility and value of the proposed approach are shown where an exposure of Ti was identified concurrently with CD45 positive cells at sub-cellular resolutions. Significant heterogeneity in the distribution of exogenous particles and cells can be present between immediately adjacent serial sections showing a clear need of synchronous imaging methods. The proposed approach enables elemental compositions to be correlated with the tissue microanatomy in a highly multiplexed and non-destructive manner at high spatial resolutions with the opportunity for subsequent guided analysis.
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Elementos de la Serie de los Lantanoides , Microscopía , HumanosRESUMEN
With the advent of nanotechnology, there has been an extensive interest in the antimicrobial potential of metals. The rapid and widespread development of antimicrobial-resistant and multidrug-resistant bacteria has prompted recent research into developing novel or alternative antimicrobial agents. In this study, the antimicrobial efficacy of metallic copper, cobalt, silver and zinc nanoparticles was assessed againstEscherichia coli(NCTC 10538),S. aureus(ATCC 6538) along with three clinical isolates ofStaphylococcus epidermidis(A37, A57 and A91) and three clinical isolates ofE. coli(Strains 1, 2 and 3) recovered from bone marrow transplant patients and patients with cystitis respectively. Antimicrobial sensitivity assays, including agar diffusion and broth macro-dilution to determine minimum inhibitory and bactericidal concentrations (MIC/MBC) and time-kill/synergy assays, were used to assess the antimicrobial efficacy of the agents. The panel of test microorganisms, including antibiotic-resistant strains, demonstrated a broad range of sensitivity to the metals investigated. MICs of the type culture strains were in the range of 0.625-5.0 mg ml-1. While copper and cobalt exhibited no difference in sensitivity between Gram-positive and Gram-negative microorganisms, silver and zinc showed strain specificity. A significant decrease (p< 0.001) in the bacterial density ofE. coliandS. aureuswas demonstrated by silver, copper and zinc in as little as two hours. Furthermore, combining metal nanoparticles reduced the time required to achieve a complete kill.
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Antiinfecciosos , Nanopartículas del Metal , Humanos , Cobre , Plata/farmacología , Zinc , Cobalto , Staphylococcus aureus , Antiinfecciosos/farmacología , Antibacterianos/farmacología , Pruebas de Sensibilidad MicrobianaRESUMEN
Traditional osteosarcoma therapies tend to focus solely on eradicating residual cancer cells and often fail to promote local bone regeneration and even inhibit it due to lack of precise control over target cells, i.e., the treatment affects both normal and cancer cells. Typically, multistep procedures are required for optimal efficacy. Here, we found that a silica-based bioactive material containing 3 mol % gallium oxide selectively kills human osteosarcoma cells and presents excellent in vivo osteointegration, while showing no local or systemic toxicity. Cell culture media conditioned with the proposed material was able to kill 41% of osteosarcoma cells, and no significant deleterious effect on normal human osteoblasts was observed. In addition, rats treated with the gallium-doped material showed excellent material-bone integration with no sign of local toxicity or implant rejection. Systemic biocompatibility investigation did not indicate any sign of toxicity, with no presence of fibrosis or cellular infiltrate in the histological microstructure of the liver and kidneys after 56 days of observation. Taken together, these results show that synergistic bone regeneration and targeted cancer therapy can be combined, paving the way toward new bone cancer treatment approaches.
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Neoplasias Óseas , Galio , Osteosarcoma , Animales , Neoplasias Óseas/tratamiento farmacológico , Galio/química , Galio/farmacología , Vidrio/química , Humanos , Osteosarcoma/tratamiento farmacológico , Ratas , Dióxido de SilicioRESUMEN
The substitution of calcium with strontium in bioactive materials has been promising but there has been some concern over the material instability and possible toxicity. The aim of this research was the synthesis and characterization of calcium and strontium substituted bioactive materials and assessment of interactions with local tissues and peripheral elemental migration in an animal model. A bioactive glass, hydroxyapatite and hydraulic calcium silicate with 50% or 100% calcium substitution with strontium were developed and the set materials were characterized immediately after setting and after 30 and 180-days in solution. Following subcutaneous implantation, the local (tissue histology, elemental migration) and systemic effects (elemental deposition after organ digestion) were assessed. The strontium-replaced silicate cements resulted in the synthesis of partially substituted phases and strontium leaching at all-time points. The strontium silicate implanted in the animal model could not be retrieved in over half of the specimens showing the high rate of material digestion. Tissue histology showed that all materials caused inflammation after 30 days of implantation however this subsided and angiogenesis occurred after 180 days. Strontium was not detected in the local tissues or the peripheral organs while all calcium containing materials caused calcium deposition in the kidneys. The tricalcium silicate caused elemental migration of calcium and silicon in the local tissues shown by the elemental mapping but no deposition of calcium was identified in the peripheral organs verified by the assessment of the digested tissues. Strontium can substitute calcium in bioactive materials without adverse local or systemic effects.
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Calcio , Estroncio , Compuestos de Calcio , Durapatita , Ensayo de Materiales , Silicatos/farmacología , Silicio , Estroncio/farmacologíaRESUMEN
The emergence of antimicrobial resistant strains bacteria and a decline in the discovery of new antibiotics has led to the idea of combining various antimicrobials to treat resistant strains and/or polymicrobial infections. Metal oxide-doped glasses have been extensively investigated for their antimicrobial potential; however to date, most experiments have focused on single metal species in isolation. The present study investigates the antimicrobial potential of sodium calcium phosphates (P2O5)50(Na2O)20(CaO)30-X(MO)X, where M is cobalt, copper, or zinc as single species. In addition, this work studied the effect of co-doping glasses containing two different metal ions (Co + Cu, Co + Zn, and Cu + Zn). The antimicrobial efficacy of all glasses was tested against Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacterial strains, as well as a fungal strain (Candida albicans). Minimum inhibitory and bactericidal concentrations and time kill/synergy assays were used to assess the antimicrobial activity. An enhanced antimicrobial effect, at 5 mg/mL concentration, was exhibited by cobalt, copper, and zinc oxide glasses alone and in combinations. A synergistic antimicrobial effect was observed by Cu + Co and Cu + Zn against E. coli and Cu + Zn against S. aureus.
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Antiinfecciosos , Óxido de Zinc , Antibacterianos/farmacología , Antiinfecciosos/farmacología , Bacterias , Cobalto/farmacología , Cobre/farmacología , Farmacorresistencia Bacteriana , Escherichia coli , Fosfatos/farmacología , Staphylococcus aureusRESUMEN
Forty-eight spring barley genotypes were evaluated for deoxynivalenol (DON) concentration under natural infection across 5 years at Harrington, Prince Edward Island. These genotypes were also evaluated for Fusarium head blight (FHB) severity and DON concentration under field nurseries with artificial inoculation of Fusarium graminearum by the grain spawn method across 2 years at Ottawa, Ontario, and one year at Hangzhou, China. Additionally, these genotypes were also evaluated for FHB severity under greenhouse conditions with artificial inoculation of F. graminearum by conidial suspension spray method across 3 years at Ottawa, Ontario. The objective of the study was to investigate if reactions of barley genotypes to artificial FHB inoculation correlate with reactions to natural FHB infection. DON concentration under natural infection was positively correlated with DON concentration (r = 0.47, P < 0.01) and FHB incidence (r = 0.56, P < 0.01) in the artificially inoculated nursery with grain spawn method. Therefore, the grain spawn method can be used to effectively screen for low DON. FHB severity, generated from greenhouse spray, however, was not correlated with DON concentration (r = 0.12, P > 0.05) under natural infection and it was not correlated with DON concentration (r = -0.23, P > 0.05) and FHB incidence (r = 0.19, P > 0.05) in the artificially inoculated nursery with grain spawn method. FHB severity, DON concentration, and yield were affected by year, genotype, and the genotype × year interaction. The effectiveness of greenhouse spray inoculation for indirect selection for low DON concentration requires further studies. Nine of the 48 genotypes were found to contain low DON under natural infection. Island barley had low DON and also had high yield.
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KEY MESSAGE: QTL analyses of two bi-parental mapping populations with AC Barrie as a parent revealed numerous FHB-resistance QTL unique to each population and uncovered novel variation near Fhb1. Fusarium head blight (FHB) is a destructive disease of wheat worldwide, leading to severe yield and quality losses. The genetic basis of native FHB resistance was examined in two populations: a recombinant inbred line population from the cross Cutler/AC Barrie and a doubled haploid (DH) population from the cross AC Barrie/Reeder. Numerous QTL were detected among the two mapping populations with many being cross-specific. Photoperiod insensitivity at Ppd-D1 and dwarfing at Rht-B1 and Rht-D1 was associated with increased FHB susceptibility. Anthesis date QTL at or near the Vrn-A1 and Vrn-B1 loci co-located with major FHB-resistance QTL in the AC Barrie/Reeder population. The loci were epistatic for both traits, such that DH lines with both late alleles were considerably later to anthesis and had reduced FHB symptoms (i.e., responsible for the epistatic interaction). Interestingly, AC Barrie contributed FHB resistance near the Fhb1 locus in the Cutler population and susceptibility in the Reeder population. Analyses of the Fhb1 candidate genes PFT and TaHRC confirmed that AC Barrie, Cutler, and Reeder do not carry the Sumai-3 Fhb1 gene. Resistance QTL were also detected at the expected locations of Fhb2 and Fhb5. The native FHB-resistance QTL detected near Fhb1, Fhb2, and Fhb5 do not appear to be as effective as Fhb1, Fhb2, and Fhb5 from Sumai-3. The presence of awns segregated at the B1 awn inhibitor locus in both populations, but was only associated with FHB resistance in the Cutler/AC Barrie population suggesting linkage caused the association rather than pleiotropy.
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Resistencia a la Enfermedad/genética , Enfermedades de las Plantas/genética , Sitios de Carácter Cuantitativo , Triticum/genética , Alelos , Mapeo Cromosómico , Fusarium/patogenicidad , Genes de Plantas , Genotipo , Fenotipo , Enfermedades de las Plantas/microbiología , Triticum/microbiologíaRESUMEN
Photo-activated resin composites are widely used in industry and medicine. Despite extensive chemical characterisation, the micro-scale pattern of resin matrix reactive group conversion between filler particles is not fully understood. Using an advanced synchrotron-based wide-field IR imaging system and state-of-the-art Mie scattering corrections, we observe how the presence of monodispersed silica filler particles in a methacrylate based resin reduces local conversion and chemical bond strain in the polymer phase. Here we show that heterogeneity originates from a lower converted and reduced bond strain boundary layer encapsulating each particle, whilst at larger inter-particulate distances light attenuation and monomer mobility predominantly influence conversion. Increased conversion corresponds to greater bond strain, however, strain generation appears sensitive to differences in conversion rate and implies subtle distinctions in the final polymer structure. We expect these findings to inform current predictive models of mechanical behaviour in polymer-composite materials, particularly at the resin-filler interface.
RESUMEN
OBJECTIVE: The structure of the polymer phase of dental resin-based-composites is highly sensitive to photo-polymerisation variables. The objective of this study was to understand how different polymer structures, generated with different photo-polymerisation protocols, respond to thermal perturbation. METHODS: Experimental resins were prepared from a series of Bis-GMA/TEGDMA blends (40/60, 50/50 and 60/40 wt.%), with either Camphorquinone/DMAEMA or Lucirin TPO as the photo-initiator system. Resins were photo-polymerised, in a disc geometry, at either relatively 'high' (3000 mW cm-2 for 6 s) or 'low' (300 mW cm-2 for 60 s) irradiances ensuring matched radiant exposures (18 J cm-2). Specimens were heated, from 20-160 °C at a rate of 5 °C min-1, whilst simultaneous synchrotron X-ray scattering measurements were taken at 5 °C increments to determine changes in polymer chain segment extension and medium-range order as a function of temperature. For each unique resin composition (n = 3), differential scanning calorimetry was used to measure glass transition temperatures using the same heating protocol. A paired t-test was used to determine significant differences in the glass transition temperature between irradiance protocols and photo-initiator chemistry at É = 0.05. RESULTS: Resins pre-polymerised through the use of TPO and or high irradiances demonstrated a reduced rate of chain extension indicative of lower thermal expansion and a larger decrease in relative order when heated below the glass transition temperature. Above the transition temperature, differences in the rate of chain extension were negligible, but slower converted systems showed greater relative order. There was no significant difference in the glass transition temperature between different photo-initiator systems or irradiance protocols. SIGNIFICANCE: The evolution of chain extension and medium-range order during heating is dependent on the initial polymer structure which is influenced by photo-polymerisation variables. Less ordered systems, generated at faster rates of reactive group conversion displayed reduced chain extension below the glass transition temperature and maintained lower order throughout heating.
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Resinas Compuestas , Resinas Sintéticas , Bisfenol A Glicidil Metacrilato , Ensayo de Materiales , Polietilenglicoles , Polimerizacion , Ácidos Polimetacrílicos , Resinas de Plantas , TemperaturaRESUMEN
In vitro and in vivo experiments were undertaken to evaluate the solubility, apatite-forming ability, cytocompatibility, osteostimulation, and osteoinduction for a series of Nb-containing bioactive glass (BGNb) derived from composition of 45S5 Bioglass. Inductively coupled plasma optical emission spectrometry (ICP-OES) revealed that the rate at which Na, Ca, Si, P, and Nb species are leached from the glass decrease with the increasing concentration of the niobium oxide. The formation of apatite as a function of time in simulated body fluid was monitored by 31P Magic Angle Spinning (MAS) Nuclear magnetic resonance spectroscopy. Results showed that the bioactive glasses: Bioglass 45S5 (BG45S5) and 1 mol%-Nb-containing-bioactive glass (BGSN1) were able to grow apatite layer on their surfaces within 3 h, while glasses with higher concentrations of Nb2 O5 (2.5 and 5 mol%) took at least 12 h. Nb-substituted glasses were shown to be compatible with bone marrow-derived mesenchymal stem cells (BMMSCs). Moreover, the bioactive glass with 1 mol% Nb2 O5 significantly enhanced cell proliferation after 4 days of treatment. Concentrations of 1 and 2.5 mol% Nb2 O5 stimulated osteogenic differentiation of BMMSCs after 21 days of treatment. For the in vivo experiments, trial glass rods were implanted into circular defects in rat tibia in order to evaluate their osteoconductivity and osteostimulation. Two morphometric parameters were analyzed: (a) thickness of new-formed bone layer and (b) area of new-formed subperiostal bone. Results showed that BGNb bioactive glass is osteoconductive and osteostimulative. Therefore, these results indicate that Nb-substituted glass is suitable for biomedical applications.
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Células de la Médula Ósea/metabolismo , Cerámica , Vidrio , Células Madre Mesenquimatosas/metabolismo , Niobio , Osteogénesis/efectos de los fármacos , Tibia , Animales , Cerámica/química , Cerámica/farmacología , Vidrio/química , Niobio/química , Niobio/farmacología , Ratas , Ratas Wistar , Tibia/lesiones , Tibia/metabolismoRESUMEN
Here, we investigated the biocompatibility of a bioactive sodium calcium silicate glass containing 2.6 mol% Nb2 O5 (denoted BGPN2.6) and compare the results with the archetypal 45S5 bioglass. The glass bioactivity was tested using a range of in vitro and in vivo experiments to assess its suitability for bone regeneration applications. in vitro studies consisted of assessing the cytocompatibility of the BGPN2.6 glass with bone-marrow-derived mesenchymal stem cells (BM-MSCs). Systemic biocompatibility was verified by means of the quantification of biochemical markers and histopathology of liver, kidneys, and muscles. The glass genotoxicity was assessed using the micronucleus test. The regeneration of a calvarial defect was assessed using both qualitative and quantitative analysis of three-dimensional microcomputed tomography images. The BGPN2.6 glass was not cytotoxic to BM-MSCs. It is systemically biocompatible causing no signs of damage to high metabolic and excretory organs such as the liver and kidneys. No mutagenic potential was observed in the micronucleus test. MicroCT images showed that BGPN2.6 was able to nearly fully regenerate a critical-sized calvarial defect and was far superior to standard 45S5 Bioglass. Defects filled with BGPN2.6 glass showed over 90% coverage compare to just 66% for 45S5 Bioglass. For one animal the defect was completely filled in 8 weeks. These results clearly show that Nb-containing bioactive glasses are a safe and effective biomaterial for bone replacement.
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Regeneración Ósea/efectos de los fármacos , Sustitutos de Huesos/farmacología , Cerámica/farmacología , Niobio/farmacología , Animales , Línea Celular , Vidrio , Humanos , Osteoblastos/citología , Osteoblastos/efectos de los fármacos , Ratas , Cráneo/efectos de los fármacos , Cráneo/lesionesRESUMEN
Phosphate-based glasses are materials of great interest for the regeneration and repair of damaged hard or soft tissues. They have the desirable property of slowly dissolving in the physiological environment, eventually being totally replaced by regenerated tissue. Being bioresorbable, they can simultaneously induce tissue regeneration and deliver therapeutic agents (e.g. antibacterial ions) in a controlled way. In this work, we have synthesised a series of glasses in the P2O5-CaO-Na2O system doped with Ag2O using the coacervation method. The addition of silver is known to provide the glass with antibacterial properties due to the release of Ag+ ions into the body fluid. The coacervation method is a facile, water-based technique which offers significant advantages over the conventional melt-quench route for preparing phosphate-based glasses which requires melting of metal oxide powders at high temperatures (1000-1200 °C). The properties of the initial colloidal polyphosphate systems (coacervates) as a function of the Ag2O content were characterised using rheology and liquid state 31P NMR. The effect of Ag+ addition on the final dried glasses was investigated using thermal analysis, Raman spectroscopy and X-ray diffraction. The antibacterial activity was assessed against Staphylococcus aureus (S. aureus), a bacterial strain commonly found in post-surgery infections. A dose-dependent antimicrobial effect was seen with an increasing silver content.
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Antibacterianos/química , Fosfatos/química , Plata/química , Coloides , Vidrio/química , Óxidos , Polifosfatos , Plata/farmacología , Compuestos de Plata , Staphylococcus aureus/efectos de los fármacosRESUMEN
In this study, a series of phosphate-based glasses; (P2O5)50(Na2O)20(CaO)30-x (ZnO)x were prepared with increasing concentration of zinc oxide to determine the antimicrobial effect against clinically relevant microorganisms. The addition of 1 and 3â¯mol% zinc oxide decreased glass degradation however a higher dissolution rate was observed for 5 and 10â¯mol% ZnO. The antimicrobial results showed a concentration dependent effect on the viability of microorganisms. When in direct contact zinc doped glasses showed a complete kill, within 24â¯h, against Escherichia coli and a significant (pâ¯<â¯0.01) kill was observed against Staphylococcus aureus however the effect of dissolution products was not seen until 48â¯h. Furthermore, the cytotoxic studies showed no toxic effects on the viability of uroepithelial cells. This study has shown that zinc doped phosphate-based glasses can potentially be used to prevent/treat catheter associated urinary tract infections.
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Antibacterianos/química , Catéteres , Infecciones por Escherichia coli/prevención & control , Escherichia coli/crecimiento & desarrollo , Vidrio/química , Fosfatos/química , Infecciones Estafilocócicas/prevención & control , Staphylococcus aureus/crecimiento & desarrollo , Infecciones Urinarias/prevención & control , Zinc/química , Línea Celular , Humanos , Infecciones Urinarias/etiologíaRESUMEN
Biological exposures to micro- and nano-scale exogenous metal particles generated as a consequence of in-service degradation of orthopaedic prosthetics can result in severe adverse tissues reactions. However, individual reactions are highly variable and are not easily predicted, due to in part a lack of understanding of the speciation of the metal-stimuli which dictates cellular interactions and toxicity. Investigating the chemistry of implant derived metallic particles in biological tissue samples is complicated by small feature sizes, low concentrations and often a heterogeneous speciation and distribution. These challenges were addressed by developing a multi-scale two-dimensional X-ray absorption spectroscopic (XAS) mapping approach to discriminate sub-micron changes in particulate chemistry within ex-vivo tissues associated with failed CoCrMo total hip replacements (THRs). As a result, in the context of THRs, we demonstrate much greater variation in Cr chemistry within tissues compared with previous reports. Cr compounds including phosphate, hydroxide, oxide, metal and organic complexes were observed and correlated with Co and Mo distributions. This variability may help explain the lack of agreement between biological responses observed in experimental exposure models and clinical outcomes. The multi-scale 2D XAS mapping approach presents an essential tool in discriminating the chemistry in dilute biological systems where speciation heterogeneity is expected. SIGNIFICANCE: Metal implants are routinely used in healthcare but may fail following degradation in the body. Although specific implants can be identified as 'high-risk', our analysis of failures is limited by a lack of understanding of the chemistry of implant metals within the peri-prosthetic milieu. A new approach to identify the speciation and variability in speciation at sub-micron resolution, of dilute exogenous metals within biological tissues is reported; applied to understanding the failure of metallic (CoCrMo) total-hip-replacements widely used in orthopedic surgery. Much greater variation in Cr chemistry was observed compared with previous reports and included phosphate, hydroxide, oxide, metal and organic complexes. This variability may explain lack of agreement between biological responses observed in experimental exposure models and clinical outcomes.
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Prótesis de Cadera/efectos adversos , Metales/efectos adversos , Falla de Prótesis , Humanos , Nanopartículas/química , Espectrometría por Rayos X , Espectroscopía de Absorción de Rayos XRESUMEN
Bioactive phosphate glasses are of considerable interest for a range of soft and hard tissue engineering applications. The glasses are degradable and can release biologically important ions in a controlled manner. The glasses can also potentially be used as an antimicrobial delivery system. In the given study, novel cobalt-doped phosphate-based glasses, (P2O5)50(Na2O)20(CaO)30-x(CoO)x where 0 ≤ x (mol %) ≤ 10, were manufactured and characterized. As the cobalt oxide concentration increased, the rate of dissolution was observed to decrease. The antimicrobial potential of the glasses was studied using direct and indirect contact methods against both Escherichia coli (NCTC 10538) Staphylococcus aureus (ATCC 6538) and Candida albicans (ATCC 76615). The results showed strong, time dependent, and strain specific antimicrobial activity of the glasses against microorganisms when in direct contact. Antimicrobial activity (R) ≥ 2 was observed within 2 h against Escherichia coli, whereas a similar effect was achieved in 6 h against Staphylococcus aureus and Candida albicans. However, when in indirect contact, the dissolution products from the bioactive glasses failed to show an antimicrobial effect. Following direct exposure to the glasses for 7 days, osteoblast-like SAOS-2 cells showed a 5-fold increase in VEGF mRNA while THP-1 monocytic cells showed a 4-fold increase in VEGF mRNA expression when exposed to 10% CoO-doped glass compared with the cobalt free control glass. Endothelial cells stimulated with conditioned medium taken from cell cultures of THP-1 monocytes exposed to 10% CoO doped glass showed clear tubelike structure (blood vessel) formation after 4 h.
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Synchrotron radiation X-ray fluorescence microscopy is frequently used to investigate the spatial distribution of elements within a wide range of samples. Interrogation of heterogeneous samples that contain large concentration ranges has the potential to produce image artefacts due to the profile of the X-ray beam. The presence of these artefacts and the distribution of flux within the beam profile can significantly affect qualitative and quantitative analyses. Two distinct correction methods have been generated by referencing the beam profile itself or by employing an adaptive-thresholding procedure. Both methods significantly improve qualitative imaging by removing the artefacts without compromising the low-intensity features. The beam-profile correction method improves quantitative results but requires accurate two-dimensional characterization of the X-ray beam profile.
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OBJECTIVES: To measure the spatial distribution of crystallographic strain in tooth enamel induced by the photo-polymerisation of a dimethacrylate resin based composite cavity restoration. METHODS: Six sound first premolar teeth, allocated into two groups (n=3), were prepared with mesio-occlusal distal cavities. The enamel was machined at the point of maximum convexity on the outer tooth to create a vertical fin of thickness 100µm and 0.5mm depth to allow for synchrotron X-ray diffraction measurements. 2D diffraction patterns were used to determine crystallite orientation and quantify changes in the hydroxyapatite crystal lattice parameters, before and after photo-polymerisation of a composite material placed in the cavity, to calculate strain in the respective axis. The composite was photo-polymerised with either relatively high (1200mWcm-2, group 1) or low (480mWcm-2, group 2) irradiances using LED or quartz halogen light sources, respectively. A paired t-test was used to determine significant differences in strain between irradiance protocols at É=0.001. RESULTS: Photo-polymerisation of the composite in the adjacent cavity induced significant changes in both the crystallographic c and a axes of the enamel measurement area. However the magnitude of strain was low with â¼0.1% difference before and after composite photo-polymerisation. Strain in enamel was not uniformly distributed and varied spatially as a function of crystallite orientation. Increased alignment of crystallites perpendicular to the cavity wall was associated with higher c axis strain. Additionally, strain was significantly greater in the c (p<0.001) and a axis (p<0.001) when using a high irradiance photo-polymerisation protocol. SIGNIFICANCE: Although cuspal deflection is routinely measured to indirectly assess the 'global' effect of composite shrinkage on the tooth-restoration complex, here we show that absolute strains generated in enamel are low, indicating strain relief mechanisms may be operative. The use of low irradiance protocols for photo-polymerisation resulted in reduced strain.
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Resinas Compuestas/química , Preparación de la Cavidad Dental/métodos , Esmalte Dental/química , Durapatita/química , Sincrotrones , Diente Premolar , Resinas Compuestas/efectos de la radiación , Cristalografía por Rayos X , Esmalte Dental/efectos de la radiación , Filtración Dental , Durapatita/efectos de la radiación , Humanos , Procesamiento de Imagen Asistido por Computador , Técnicas In Vitro , Polimerizacion , Estrés Mecánico , Difracción de Rayos XRESUMEN
We use a combination of classical molecular dynamics simulation and neutron diffraction to identify the atomic structure of five different Mg-Zn-Ca bulk metallic glasses, covering a range of compositions with substantially different behaviour when implanted in vitro. There is very good agreement between the structures obtained from computer simulation and those found experimentally. Bond lengths and the total correlation function do not change significantly with composition. The zinc and calcium bonding shows differences between composition: the distribution of Zn-Ca bond lengths becomes narrower with increasing Zn content, and the preference for Zn and Ca to avoid bonding to themselves or each other becomes less strong, and, for Zn-Ca, transforms into a positive preference to bond to each other. This transition occurs at about the same Zn content at which the behaviour on implantation changes, hinting at a possible structural connection. A very broad distribution of Voronoi polyhedra are also found, and this distribution broadens with increasing Zn content. The efficient cluster packing model, which is often used to describe the structure of bulk metallic glasses, was found not to describe these systems well.
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Defining the mechanisms underlying the programming of early life growth is fundamental for improving adult health and wellbeing. While the association between maternal diet, offspring growth and adult disease risk is well-established, the effect of father's diet on offspring development is largely unknown. Therefore, we fed male mice an imbalanced low protein diet (LPD) to determine the impact on post-fertilisation development and fetal growth. We observed that in preimplantation embryos derived from LPD fed males, expression of multiple genes within the central metabolic AMPK pathway was reduced. In late gestation, paternal LPD programmed increased fetal weight, however, placental weight was reduced, resulting in an elevated fetal:placental weight ratio. Analysis of gene expression patterns revealed increased levels of transporters for calcium, amino acids and glucose within LPD placentas. Furthermore, placental expression of the epigenetic regulators Dnmt1 and Dnmt3L were increased also, coinciding with altered patterns of maternal and paternal imprinted genes. More strikingly, we observed fetal skeletal development was perturbed in response to paternal LPD. Here, while offspring of LPD fed males possessed larger skeletons, their bones comprised lower volumes of high mineral density in combination with reduced maturity of bone apatite. These data offer new insight in the underlying programming mechanisms linking poor paternal diet at the time of conception with the development and growth of his offspring.
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Blastocisto/metabolismo , Proteínas en la Dieta , Epigénesis Genética , Desarrollo Fetal , Regulación del Desarrollo de la Expresión Génica , Desarrollo Musculoesquelético , Complicaciones del Embarazo/metabolismo , Efectos Tardíos de la Exposición Prenatal/metabolismo , Deficiencia de Proteína/metabolismo , Animales , Blastocisto/patología , Femenino , Masculino , Ratones , Embarazo , Complicaciones del Embarazo/patología , Efectos Tardíos de la Exposición Prenatal/patología , Deficiencia de Proteína/patologíaRESUMEN
In this study, we have developed a series of novel gallium oxide doped bioactive glasses to specifically target osteosarcoma cells while aiding new bone formation. The results show that osteosarcoma (Saos-2) cell death is induced through the addition of gallium oxide. Relative to the gallium-free control glass (0% Ga) glasses containing 1, 2, and 3% Ga decreased Saos-2 cell viability in a dose dependent manner. After 72 h in media preconditioned with 3% Ga Saos-2 cell viability was reduced by over 50%. Corresponding studies undertaken on primary normal human osteoblast cells (NHOst) demonstrated no adverse effects to the gallium containing glasses. Hydroxyapatite formation was observed for all glasses when exposed to simulated body fluid.