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Effect of external stresses on calcium phosphate glass investigated by IR spectroscopy.

The effect of external stress on calcium phosphate glasses was investigated because they experience high stress in practical uses such as mastication. In order to understand the structural change of calcium phosphate glasses caused by an applied stress, the IR reflection peak shift of the phosphate structural band of the calcium phosphate glass fiber due to a bending stress was measured. The peak shift represents the change of the P-O-P bond angle. The IR reflection peak of the phosphate structural bands near approximately 1300 and approximately 910 cm(-1) shifted to a higher wavenumber under tensile stress and to a lower wavenumber under compressive stress when calcium phosphate fiber with a uniform structure was employed. This indicates that the P-O-P bond angle increases under tensile stress and decreases under compressive stress. The extents of both peak shifts were larger with a lower [corrected] Ca/P ratio than with a higher [corrected] Ca/P ratio. Therefore, phosphate glass with a higher Ca/P ratio is expected to withstand loads under external stress.

Biocompatible Materials↗

Immobilization of luciferase from a firefly lantern extract on glass strips as an alternative strategy for luminescent detection of ATP.

The bioluminescent reaction catalysed by firefly luciferase has become widely established as an outstanding analytical system for assay of ATP. When used in solution, luciferase is unstable and cannot be re-used, a problem that can be partially circumvented by immobilizing the enzyme on solid substrates. Transparent glass is especially advantageous over alternative immobilizing matrices, since it allows most of the emitted photons to be detected. We report a new method for luciferase immobilization on glass which does not require prior silanization and glutaraldehyde activation, thus saving preparation time and minimizing enzyme inactivation. Our method is based on the co-immobilization by adsorption of luciferase (from a firefly lantern extract) and poly-L-lysine (PL) on non-porous glass strips. Luciferase immobilized in this way exhibits minimal variations in intersample activity, high sensitivity for ATP detection (linear luminescence responses down to 50 nmol/L) and good stability (full activity for at least 60 days when stored at -80 degrees C). PL-mediated immobilization of luciferase on glass strips provides an attractive strategy for the design of specific ATP biosensors, with potential in industry, environmental screening, medicine and biological research.

Adenosine Triphosphate↗

Characterization of bioactive glass-reinforced HAP-polymer composites.

The effect of bioactive glass on the mechanical properties of hydroxyapatite-Ca polyacrylate composites was studied. Powder mixtures of tetracalcium phosphate (TetCP), poly(acrylic-co-itaconic) and bioactive glass (up to 50% by weight) were hot pressed for 30 min at 300 degrees C and 40 kpsi. Tensile strengths, elastic moduli, and microstructures of the composites produced were investigated. Results showed the mechanical properties of these composites were enhanced by the addition of bioactive glass. The highest values of tensile strength and elastic modulus were achieved with the addition of 10% bioactive glass. Composites were immersed in SBF for up to 10 days, then in 1.5 simulated body fluid (SBF) for a week. The changes in the concentrations of Ca, P, and Si ions of these solutions were measured. The microstructures of these composites after SBF immersion were also evaluated. Concentrations of Ca, P, and Si increased with the time of immersion in SBF owing to the formation of an apatite layer on their surfaces as found by SEM with energy-dispersive spectroscopy attachment.

Biocompatible Materials↗

Effect of varying physical properties of porous, surface modified bioactive glass 45S5 on osteoblast proliferation and maturation.

The objective of this study was to determine the effect of porous bioactive glass (45S5) substrate characteristics on the expression and maintenance of the osteoblastic phenotype. We cultured ROS 17/2. 8 cells on substrates with different pore size and porosity for periods up to 14 days and analyzed the characteristics of the cells and extracellular matrix. Results of the study show that the glass substrates supported the proliferation and growth of osteoblast-like cells. Although the morphologies of the cells differed on the various substrates, their shape and the extent of membrane ruffling suggested that they maintained high levels of metabolic activity. Cells on all substrates expressed high levels of alkaline phosphatase activity and produced extracellular matrices that mineralized to form nonstoichiometric, carbonated, calcium-deficient apatites. An important finding was that at a given porosity of 44%, the pore size neither directed nor modulated the in vitro expression of the osteoblastic phenotype. In contrast, porosity did affect cellular function. We noted that at an average pore size of 92 microm, as the porosity increased from 35 to 59%, osteoblast activity was reduced. As designed in this experiment, an increase in the porosity led to a corresponding increase in total surface area of the specimens. With increasing porosity and surface area, glass reactions in the media may persist for longer durations at higher intensities, thereby affecting local media composition. As such, we suggest that extensive conditioning treatments before cell seeding can reduce this effect. Our results also revealed that the expression of the osteoblastic phenotype is enhanced by the ongoing glass dissolution. The reaction pathway at the origin of this effect still needs to be elucidated. Taken together, the findings support the overall hypothesis that in vitro cell activity can be controlled by a careful selection of substrate properties.

Alkaline Phosphatase↗

Low-temperature synthesis, structure, and bioactivity of gel-derived glasses in the binary CaO-SiO2 system.

Glasses in the binary system CaO-SiO2 for which the molar fraction of CaO is 0 < or = x < or = 0.50, were prepared by means of a sol-gel route starting from tetraethylorthosilicate and calcium nitrate. The textural features of the monoliths obtained were characterized using N2 gas adsorption, helium ultrapycnometry, and mercury porosimetry. In vitro bioactivity tests were performed in simulated body fluid (SBF). The ionic concentration of the SBF after glass immersion was analyzed using inductively coupled plasma atomic emission spectroscopy. The surfaces of the specimens were characterized using X-ray diffraction and scanning electron microscopy coupled with energy dispersive X-ray analysis before and after in vitro testing. The textural characterization revealed that the glasses were mesoporous with cylindrical pores with average diameters ranging from 25 to 663 A depending on the molar fraction of CaO. The in vitro studies showed that all binary CaO-SiO2 gel-glass compositions produced were bioactive. These results indicate that the binary gel-derived CaO-SiO2 system exhibits a level of bioactivity over a similar molar range of SiO2 content as the previously studied ternary CaO-P2O5-SiO2 system.

Adsorption↗

Fabrication of a glass-implemented microcapillary electrophoresis device with integrated contactless conductivity detection.

Glass microdevices for capillary electrophoresis (CE) gained a lot of interest in the development of micrototal analysis systems (microTAS). The fabrication of a microTAS requires integration of sampling, chemical separation and detection systems into a microdevice. The integration of a detection system into a microchannel, however, is hampered by the lack of suitable microfabrication technology. Here, a microfabrication method for integration of insulated microelectrodes inside a leakage-free microchannel in glass is presented. A combination of newly developed technological approaches, such as low-temperature glass-to-glass anodic bonding, channel etching, fabrication of buried metal interconnects, and deposition of thin plasma-enhanced chemical vapour deposition (PECVD) silicon carbide layers, enables the fabrication of a CE microdevice with an integrated contactless conductivity detector. The fabrication method of this CE microdevice with integrated contactless conductivity detector is described in detail. Standard CE separations of three inorganic cations in concentrations down to 5 microM show the viability of the new microCE system.

Biosensing Techniques↗

Lead exposure in stained glass workers.

To evaluate lead exposure in stained glass workers, we measured blood lead levels in 12 professional glass workers, in 5 hobbyists, and in 4 workers' family members. Professional workers lead levels (mean 20.7 micrograms/dl) were higher than hobbyists' (11.6 micrograms/dl) (P = 0.02) or family members' (11.3 micrograms/dl). Levels increased with years worked, hours worked per week, and percentage of work involving lead. The mean lead concentration in settled dust samples from a stained glass workshop was 11,000 parts per million. Stained glass workers are at increased risk of lead exposure.

Environmental Exposure↗

Surface transformation of bioactive glass in bioreactors simulating microgravity conditions. Part I: experimental study.

Surface modified bioactive glass with surface properties akin to those of the bone mineral phase is an attractive candidate for use as a microcarrier material for 3-D growth of bone-like tissue in rotating wall vessel bioreactors (RWVs). The critical surface properties of this material are the result of reaction in solution. Because an RWV environment is completely different from conditions previously employed for bioactive glass testing, a detailed study of the surface reactions is warranted. Under properly chosen conditions, RWVs can also provide a simulated microgravity environment for the bioactive glass (BG) particles. In this sense, this study is also a report on the behavior of a bioactive material under microgravity conditions simulated on earth. A high aspect ratio vessel (HARV) and carefully selected experimental conditions enabled the simulation of microgravity in our laboratory. A complimentary numerical study was simultaneously conducted to ascertain the appropriateness of the experimental parameters (particle size, particle density, medium density, medium viscosity, and rotational speed) that ensure simulated microgravity conditions for the glass particles in the HARV. Physiological solutions (pH 7.4) with and without electrolytes, and also with serum proteins, were used to study the change in surface character resulting from simulated microgravity. Control tests at normal gravity, both static and dynamic, were also conducted. Solution and surface analyses revealed major effects of simulated microgravity. The rates of leaching of constituent ions (Si-, Ca-, and P-ions) were greatly increased in all solutions tested. The enhanced dissolution was followed by the enhanced formation of bone-like minerals at the BG surface. This enhancement is expected to affect adsorption of serum proteins and attachment molecules, which, in turn, may favorably affect bone cell adhesion and function. The findings of the study are important for the use of bioactive materials as microcarriers to generate and analyze 3-D bone-like tissue structures in bioreactors under microgravity conditions or otherwise.

Bioreactors↗

Miniaturized amperometric flow immunoassay system using a glass fiber membrane modified with anion.

This paper describes a miniaturized amperometric flow immunoassay system using a glass fiber membrane modified with anion. The glass fiber membrane was functionally modified with gamma-glycidoxypropyltrimethoxysilane and sodium thiosulfate and was used for separation of protein. Anti-human chorionic gonadotrophin (HCG) immunoglobulin G (IgG) antibody conjugated with ferrocenemonocarboxylic acid (Fc), namely, Fc-conjugated IgG (Fc-IgG), was used as a novel analytical reagent. HCG and Fc-IgG complexes were separated from free Fc-IgG based on differences in isoelectric point (pI) using the glass fiber membrane modified with a thiosulfonyl acid functional group. The assay yields a linear relationship between current and HCG concentration in the range of 0-2000 mIU/mL. This simple technique enables the assay of HCG within 2 min. The modified glass fiber membrane was regenerated by occasional elution with malonate buffer (pH 6.0) containing 0.5 M NaCl, to remove free Fc-IgG. Free Fc-IgG recovered in this manner could be reused up to eight times without significant decreases in sensitivity. This miniaturized amperometric flow immunoassay requires only minute quantities of serum and generates highly reproducible results.

Animals↗

Capillary zone electrophoresis of amino acids on a hybrid poly(dimethylsiloxane)-glass chip.

Poly(dimethylsiloxane) (PDMS)-PDMS and hybrid PDMS-glass devices have been characterized and compared in terms of current-voltage linearity, contact angle, electroosmotic velocity, electroosmotic mobility, and electrokinetic potential in dependence on the surface treatment. The hybrid PDMS-glass microfluidic devices have further been tested as on-chip capillary electrophoresis systems for the separation of fluorescently labeled amino acids. It has been demonstrated that different methods of surface pretreatment of the PDMS-glass devices result in significantly different separation performance, with plate numbers varying from 650 to 57 000 in dependence on the surface state and the nature of the amino acids. Electrophoretic separations of amino acids have been achieved within tens of seconds with detection limits of less than 2 microM (approximately 2 x 10(-16) to 2.5 x 10(-16) mol quantities at injection volumes of 110-120 pL). The detected amounts of fluorescein isothiocyante (FITC)-amino acids are at least ten times lower, since the amino acid:FITC ratio is 10:1 mol. The results demonstrate the perspective of such hybrid PDMS-glass microfluidic systems and the methods to modify their surfaces for on-chip separation methods for biomolecules.

Amino Acids↗

Restorative service patterns in Australia: amalgam, composite resin and glass ionomer restorations.

AIM: To examine the provision of amalgam, composite resin and glass ionomer restorations, and to assess whether these main restorative services varied by patient, visit and oral health characteristics. DESIGN: A cross-sectional survey incorporating a log of service items provided on a typical day. SETTING: Australian private general practice. METHODS: Data on services and patients were collected by a mailed survey from a random sample of dentists from each State/Territory in Australia in 1998-99 with a response rate of 71%. MAIN OUTCOME MEASURES: Rates per visit of amalgam, composite resin and glass ionomer restorations among dentate adults who had received a restoration. RESULTS: Analysis showed older patients had lower amalgam rates but higher glass ionomer rates, composite resin rates were lower at emergency visits, capital city patients had higher amalgam rates but lower composite resin rates, patients with decayed teeth had higher amalgam and composite resin rates, and use of restorative materials varied by clinical problem. CONCLUSIONS: Despite widespread use of alternative materials, amalgam rates remained high in circumstances such as replacement restorations and restorations involving more than one surface. Other restorative materials also had specific applications. Both amalgam and composite resins were provided at higher rates to patients with active caries but composite resins were also used at higher rates for aesthetic problems. Glass ionomer restorations were used at higher rates for initial and one-surface restorations, and for conditions such as root caries and dentinal sensitivity.

Adult↗

In vitro and in vivo dissolution behavior of a dysprosium lithium borate glass designed for the radiation synovectomy treatment of rheumatoid arthritis.

Dysprosium lithium borate (DyLB) glass microspheres were investigated for use in the radiation synovectomy treatment of rheumatoid arthritis. In vitro testing focused on weight loss and cation dissolution from glass microspheres immersed in simulated synovial fluid (SSF) at 37 degrees C for up to 64 days. In vivo testing was performed by injecting glass microspheres into the stifle joints of Sprague-Dawley rats and monitoring the biodegradability of the microspheres and the tissue response within the joints. The DyLB microspheres reacted nonuniformly in SSF with the majority of lithium and boron being dissolved, whereas nearly all of the dysprosium (>99.7%) remained in the reacted microspheres. Because the DyLB glasses released negligible amounts of dysprosium while reacting with SSF, they are considered safe for radiation synovectomy from the standpoint of unwanted radiation release from the joint capsule. Furthermore, the DyLB microspheres fragmented, degraded, and reacted with body fluids while in the joints of rats without histologic evidence of joint damage.

Algorithms↗

Materials, clinical and morphological evaluation of custom-made bioreactive-glass-coated canine hip prostheses.

Bioreactive-glass-coated hip prostheses were implanted in dogs for periods of time ranging from 3 to 17 months. The prostheses were tailor-made for each animal through the use of standard radiographs. However, such a procedure does not necessarily yield a close apposition between the glass layer and bone. As a result, the initial stability of the component can be lost before bone bonding is established. Secondary to this phenomenon, the micromotion can lead to a continuous disruption of the top film on the glass despite the evidence of various reaction layers on the glass surface.

Animals↗

Attachment and spreading of human gingival fibroblasts on potentially bioactive glasses in vitro.

The attachment and spreading of cultured fibroblasts on potentially bioactive glasses (bioglasses) of ten different compositions were studied. Human gingival fibroblasts were allowed to attach and spread on bio-glasses for 1-72 h. Unreactive silica glass and cell culture polystyrene served as controls. The attachment and spreading of cells were examined by 3H-thymidine labeling of cells, planimetric analysis, cytological staining, immunocytochemistry, and scanning electron microscopy. The cell attachment to bioglasses and silica glass and the cell spreading on bioglasses were slower and cell morphology more elongated compared to control plastic. In spite of great differences in bioglass compositions no great differences in cell behavior on these surfaces were detected. Thus the initial events in the tissue-implant interface might be independent on the bioglass composition, and furthermore the differences in the organization of the tissue-implant interface in vivo might depend on the nature of the surrounding tissues and subsequent changes of the implant surface and the extracellular environment.

Adult↗

Solutions able to reproduce in vivo surface-structure changes in bioactive glass-ceramic A-W.

High-strength bioactive glass-ceramic A-W was soaked in various acellular aqueous solutions different in ion concentrations and pH. After soaking for 7 and 30 days, surface structural changes of the glass-ceramic were investigated by means of Fourier transform infrared reflection spectroscopy, thin-film x-ray diffraction, and scanning electronmicroscopic observations, in comparison with in vivo surface structural changes. So-called Tris buffer solution, pure water buffered with trishydroxymethyl-aminomethane, which had been used by various workers as a "simulated body fluid," did not reproduce the in vivo surface structural changes, i.e., apatite formation on the surface. A solution, ion concentrations and pH of which are almost equal to those of the human blood plasma--i.e., Na+ 142.0, K+ 5.0, Mg2+ 1.5, Ca2+ 2.5, Cl- 148.8, HCO3- 4.2 and PO4(2-) 1.0 mM and buffered at pH 7.25 with the trishydroxymethyl-aminomethane--most precisely reproduced in vivo surface structure change. This shows that careful selection of simulated body fluid is required for in vitro experiments. The results also support the concept that the apatite phase on the surface of glass-ceramic A-W is formed by a chemical reaction of the glass-ceramic with the Ca2+, HPO4(2-), and OH- ions in the body fluid.

Biocompatible Materials↗

Histological and biochemical evaluation of osteoblasts cultured on bioactive glass, hydroxylapatite, titanium alloy, and stainless steel.

We investigated the behavior of fetal rat osteoblasts cultured on four bone replacing materials: bioactive glass, hydroxylapatite, a titanium alloy, and stainless steel. The cultures were histologically examined for individual cell morphology and osteoblast expression after several periods of time using scanning electron, fluorescence, and normal light microscopy. Other cultures were used for biochemical determinations of alkaline phosphatase activity (APA) and DNA content. Osteoblasts cultured on bioactive glass showed a better osteoblast-like morphology and a higher proliferation rate, leading to confluent cultures with higher cell density and a generally better expression of the osteoblast phenotype in comparison with the other substrates. The confluent bioactive glass cultures also showed significantly higher DNA content and APA as well as the calculated APA/DNA ratio. Based on the evaluation of histological and biochemical parameters we conclude that osteoblasts cultured on bioactive glass show a generally better osteoblast character than on the other materials.

Actins↗

Interactions of plasminogen and fibrinogen with model silica glass surfaces: adsorption from plasma and enzymatic activity studies.

The adsorption of fibrinogen and plasminogen from plasma to silica glass, sulfonated silica glass, and lysine-derivatized silica glass has been investigated. The data indicate that the sulfonated material has a high affinity for both fibrinogen and plasminogen, but that the ratio of plasminogen to fibrinogen is greater on the lysine-derivatized surface. The adsorption data also suggest plasminogen as a possible contributor to the fibrinogen Vroman effect, whereby initially absorbed fibrinogen is displaced from the surface. The plasmin activity of plasminogen adsorbed to the lysine-derivatized silica glass and its sulfonated precursor was assessed by both a chromogenic substrate assay and a radioimmunoassay for the plasmin cleavage product of fibrinogen, the B beta 1-42 peptide. The data indicate that 1) the adsorbed plasminogen is not inherently plasmin-like; 2) the enzymatic activity associated with the bound plasminogen is significantly enhanced on both surfaces in the presence of activator; and 3) in the presence of activator, the plasmin activity per mole of bound plasminogen on the lysinized material is approximately a factor of two greater than on the sulfonated material based on the chromogenic substrate assay, and a factor of four greater based on the B beta 1-42 radioimmunoassay. The lysinized material thus exhibits several properties that are different from its sulfonated precursor. It adsorbs more plasminogen relative to fibrinogen after the Vroman peak, and this adsorbed plasminogen appears to be in a conformation that is more readily activated to plasmin. Once activated, the surface bound plasmin shows enhanced ability to cleave either a low molecular weight chromogenic substrate or a macromolecular substrate.(ABSTRACT TRUNCATED AT 250 WORDS)

Adsorption↗

Inhibition of platelet spreading from plasma onto glass by an adsorbed layer of a novel fluorescent-labeled poly(ethylene oxide)/poly(butylene oxide) block copolymer: characteristics of the exclusion zone probed by means of polystyrene beads and macromolecules.

We have investigated the anti-adhesive properties of a newly synthesized fluorescent triblock copolymer containing poly(ethylene oxide). This adsorbs from aqueous solution onto glass that has been rendered hydrophobic. When the polymer-treated surface was exposed to human platelet-rich plasma (PRP) or whole blood at 37 degrees C, platelet adhesion and spreading were prevented. Avid adhesion and rapid platelet spreading occurred along tracks scraped in the adsorbed polymer coating, as seen by video-enhanced interference reflection microscopy. Leukocytes from whole blood are eventually able to adhere to the polymer-treated surface and were seen to remove labeled polymer from their vicinity and accumulate it at the cell body. Interferometry using polystyrene spheres showed that they do not adhere to polymer-coated glass and are unable to approach closer than 70-95 nm. On scraped tracks, beads make molecular contacts with the glass. Because the fully extended solvated (EO)400 arms may extend up to 100 nm from the glass, this suggests that the polymer forms a monolayer with the hydrophilic arms projecting into the water, whereas the hydrophobic (BO)55 segment binds the molecule to the hydrophobic surface. Another tri-bloc copolymer with shorter hydrophilic arms allows particles to approach more closely.

Adsorption↗