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Gas chromatographic-mass spectrometric determination of tiopronin in human blood using acrylic acid esters as a derivatization reagent for the thiol group.

A gas chromatographic-mass spectrometric method for determining tiopronin, which has a thiol group, in human blood has been described. To prevent the oxidative degradation of tiopronin in the blood, its thiol group was immediately protected by treatment with isobutyl acrylate, which reacted readily with tiopronin in a 0.1 M Na2HPO4 solution (pH 9.1). The reaction was quantitative within 30 min. The blood sample was deproteinized and purified by a combination of liquid-liquid extraction and solid-phase extraction. Finally, the carboxyl moiety of the ester adduct was derivatized to the pentafluorobenzyl ester. The derivatives of tiopronin and the internal standard were analysed with gas chromatography-mass spectrometry. The precision of the method was satisfactory, and the calibration curve had good linearity in the concentration range investigated. The limit of determination of tiopronin in blood was estimated to be ca. 1 ng/ml.

Acrylates↗

Clinical management of the acrylic splint Herbst appliance.

This article describes one variation in Herbst appliance design--the acrylic splint Herbst. Topics discussed include early fixed appliance treatment before Herbst therapy, impression taking, bite registration, and evaluation of appliance fabrication. The specifics of appliance delivery, including bonding of the maxillary appliance when indicated, are also discussed as are the techniques of appliance advancement and removal.

Acrylic Resins↗

Improved technique for immunoelectron microscopy. How to prepare epoxy resin to obtain approximately the same immunogold labeling for epoxy sections as for acrylic sections without any etching.

The purpose of this study was to improve the immunogold labeling of epoxy sections and to increase our knowledge of the mechanism for how antigens become immunolabeled on resin sections. Tissues from pancreas, thyroid and fibrin clots were embedded in an epoxy resin and LR-White. The epoxy mixture was composed and treated in different ways, especially with respect to altered amounts of accelerator (DMP-30). Immunogold labeling was performed with anti-glucagon, anti-thyroglobulin and anti-fibrinogen respectively. By increasing the amount of DMP-30 in the infiltration steps and/or embedding step, we observed a significant rise in the immunogold labeling. For the largest proteins the labeling was up to 8 times more intense than the labeling achieve with epoxy sections produced by 'normal' amount of accelerator in the embedding mixture and without accelerator in the infiltration mixture. For the smallest protein, glucagon, the differences were almost absent. The labeling of thyroglobulin and fibrinogen on the high accelerator epoxy sections was up to 70% of the labeling of LR-White sections, while conventional epoxy sections showed a labeling of 5-10% of that obtained with acrylic labeling. The cutting qualities of the high-accelerator blocks were similar to that of conventional epoxy embedding. The ultrastructure of the sections from the high-accelerator epoxy blocks were good, and the contrast was improved when tannic acid was used as enhancer. Our theory to explain the improved labeling is that the antigens are less tightly incorporated in the polymer network when the concentration of the accelerator is increased. The method outlined significantly improves the detectability of antigens on epoxy sections, which is the embedding resin routinely used in many laboratories.

Acrylic Resins↗

The theoretical relationship of immunogold labeling on acrylic sections and epoxy sections.

The purpose of this study was to predict the ratio of immunogold labeling of LR-White sections and epoxy sections using theoretical methods. Tissues used in the experiments were pancreas, pituitary, kidney, thyroid and fibrin. Antigens used as test proteins were glucagon, somatostatin, thyroglobulin, chromogranin A, ACTH (adrenocorticotropt hormone), amyloid A and fibrinogen. These are proteins of different sizes. The quotient labelingLR-White/labelingepoxy was deduced theoretically and compared to calculations based on practical immunogold experiments. The theoretically deduced formula showed acceptable correlation to these calculations. This study gives a theory--expressed mathematically--for what is happening on the molecular level at the surface of resin sections in immunoelectron microscopy. The theory explains why acrylic resins normally are better suited for immunoelectron microscopy than epoxy sections, and indicates increased usefulness of epoxy sections when the diameter of the protein carrying the epitope decreases.

Acrylic Resins↗

New aspects of bacterial ultrastructure as revealed by modern acrylics for electron microscopy.

Modern acrylics can be used over a wide temperature range (+60 degrees C to -80 degrees C) for infiltration, embedding, and polymerization. They can be used in procedures involving chemical fixation or rapid freezing. This flexibility allows for studies to be carried out upon the effects that different parameters involved in preparing biological tissue for microscopy have upon structure and retention of immunoreactivity. With most preparative methods contributions have been made to our knowledge on bacterial structure in gram-negative and gram-positive cells. The future should lie in integrating the advantages of the various methods for the purpose of advancing our understanding of bacterial structure/function.

Acrylic Resins↗

Human osteosarcoma cell adhesion behaviour on hydroxyapatite integrated chitosan-poly(acrylic acid) polyelectrolyte complex.

A novel degradable composite system has been prepared by integrating hydroxyapatite, Ca(10)(PO(4))(6)(OH)(2), (HAP) in a polyelectrolyte complex matrix of chitosan (CHI) and poly(acrylic acid) (PAA). The composite was formulated by integrating 80 wt.% HAP in the polyelectrolyte complex matrix of CHI and PAA in the ratio 40/60 (designated as CPH). The composite could be easily fabricated into clinically significant shapes by a simple moulding procedure intended for bone graft applications. The adhesion behaviour of human osteosarcoma (HOS) cells on this degradable composite system was studied by selecting the polyelectrolyte complex, CHI/PAA 40/60 (designated as CP) as control sample. Light microscopic observations show that cells around CPH retained the typical morphology of HOS cells while cells around the polyelectrolyte complex showed a cytotoxic effect. The adhesion behaviour as well as morphological responses of the seeded cells was further investigated by scanning electron microscopy. The scanning electron micrographs of the polyelectrolyte complex, CP, showed the presence of rounded cells with raised nuclear regions, indicating delayed spreading; cells adhered on CPH were flattened with filopodia and showed good attachment and spreading, indicating better adhesion onto the HAP integrated composite. Comparing the MTT assay for quantitative evaluation of cell viability, CPH showed a higher percentage of metabolically active cells compared to CP.

Acrylic Resins↗

Pigment release and secondary glaucoma after implantation of single-piece acrylic intraocular lenses in the ciliary sulcus.

PURPOSE: To report the association of sulcus-fixated, single-piece hydrophobic acrylic intraocular lenses (HAIOL) with pigment release and secondary glaucoma. DESIGN: Interventional case series. METHODS: HAIOL was implanted in the ciliary sulcus of 20 eyes that developed posterior capsule rupture during phacoemulsification. We analyzed postoperative best-corrected visual acuity, manifest refraction, frequency of intraocular lens malpositioning, and postoperative complications. RESULTS: Postoperative best-corrected visual acuity was 20/40 or better in all eyes. The mean postoperative sphere was -0.5 +/- 0.7 diopters (range +1.25 to -2.00); the mean postoperative cylinder was -1.2 +/- 0.8 diopters (range 0 to -2.50). None of the HAIOLs was decentered or dislocated, and none required repositioning. Seven eyes (35%) developed pigment release; three eyes (15%) developed secondary glaucoma. CONCLUSIONS: Implantation of single-piece HAIOLs in the ciliary sulcus is associated with pigment release and secondary glaucoma. Alternative methods of aphakic correction should be considered.

Acrylic Resins↗

Inhibition of HIV-1 TAR RNA-Tat peptide complexation using poly(acrylic acid).

HIV-1 is regulated at the transcriptional level by the interaction of Tat protein with the transactivation responsive region (TAR) RNA, a 59-base stem-loop structure located at the 5'-end of all nascent HIV-1 transcripts. Here, by targeting the Tat peptide, we found that negatively charged poly(acrylic acid) (PAA) had high affinity with Tat peptide and could inhibit the interaction of TAR with Tat. Therefore, PAA could block HIV replication by binding to Tat not to TAR RNA, providing a new thinking for the design of novel anti-HIV drugs.

Acrylic Resins↗

Surface modification of ultra thin poly (epsilon-caprolactone) films using acrylic acid and collagen.

Poly (epsilon-caprolactone) (PCL) has been used as a bioresorbable polymer in numerous medical devices as well as for tissue engineering applications. Its main advantage is its biocompatibility and slow degradation rate. PCL surface, however, is hydrophobic and cell-biomaterial interaction is not the best. We attempt for the first time to modify an ultra thin PCL surface with collagen. The PCL film was prepared using solvent casting and biaxial stretching technique developed in our laboratory. This biaxial stretching produced an ultra thin PCL 3-7 microm thick, ideal for membrane tissue engineering applications. The PCL film was pretreated using Argon plasma, and then UV polymerized with acrylic acid (AAc). Collagen immobilization was then carried out. The modified film surface was characterized by Fourier Transform Infrared (FT-IR) and X-ray Photoelectron Spectroscopy (XPS). Water contact angles were also measured to evaluate the hydrophilicity of the modified surface. Results showed that the hydrophilicity of the surface has improved significantly after surface modification. The water contact angle dropped from 66 degrees to 32 degrees. Atomic Force Microscopy (AFM) showed an increase in roughness of the film. A change from 46 to 60 nm in the surface morphology was also observed. The effect of cells attachment on the PCL film was studied. Human dermal fibroblasts and myoblasts attachment and proliferation were improved remarkably on the modified surface. The films showed excellent cell attachment and proliferation rate.

Acrylates↗

In situ polymerizable polyethyleneglycol containing polyesterpolyol acrylates for tissue sealant applications.

Polyesterpolyol macromers were prepared with succinic acid and polyethylene glycols (PEG) of different molecular weights. The resulting polyols were acrylated to render them photo-cross-linkable. They could be very rapidly cross-linked into non-tacky films with long-wavelength UV radiation. The resulting products were characterized by gel content, water equilibrium swell, cross-link density, Tg , tensile strength, degradation and in vitro burst strengths. Though all of them formed transparent contact lens like films, increasing the PEG molecular weight has resulted in polymers with higher hydrophilicity resulting in higher swelling, faster degradation, higher tensile strength, elongation at break and burst strength. Addition of vinyl pyrrolidinone as a reactive diluent has increased the mechanical as well as burst strength of the polymer. In vitro release of sulfamethoxazole entrapped in these cross-linked matrices was also studied.

Acrylates↗

Stable plasma-deposited acrylic acid surfaces for cell culture applications.

Continuous and modulated glow discharges were used to deposit thin films from acrylic acid vapors. Different deposition regimes were investigated, and their effect on chemical composition, morphology and homogeneity of the coatings, as well as on their stability in water and resistance to sterilization. Stable films were utilized in cell adhesion experiments with human fibroblasts.

Acrylates↗

Acrylic bone cements modified with beta-TCP particles encapsulated with poly(ethylene glycol).

Beta-tricalcium phosphate (beta-TCP) has been encapsulated with poly(ethylene glycol) (PEG) to improve the filler/cement interface, and it was later incorporated to a poly(methyl methacrylate) bone cement in order to obtain cements with improved stability in the long term. Size and size distribution of the agglomerates forming the initial powder was drastically changed after its dispersion in a PEG aqueous solution. Whereas the initial beta-TCP particles had a 584 microm average diameter, the treated particles (TCP-PEG) presented more than 60% of the particles in a range of 2-6 microm. The effect of adding the treated particles to an acrylic cement was evaluated in terms of curing parameters, in vitro behaviour and mechanical performance. The presence of the TCP-PEG particles did not affect either peak temperature or setting time, indicating a good homogeneity of polymerising mass in contrast to the effect observed with the plain beta-TCP particles, which gave rise to higher setting times. In vitro behaviour studies revealed hydration degree values of the modified cements comparable to that of PMMA cements. Early stages of water uptake was Fickian in nature for all the experimental formulations indicating that the water absorption followed a diffusion controlled mechanism. After 3 months of storage in SBF the experimental formulations presented values of compressive strength in the range 76-78 MPa, higher than the minimum required by ISO 5833 (70 MPa) and those of tensile strength in the range 42-48 MPa, higher than the minimum reported for commercial formulations (30 MPa), but no significant differences in the strengths and elastic modulus were observed with the treatment of the filler particles. This observation was confirmed by ESEM analysis of the tensile fracture surfaces, which revealed a rather good cohesion between the bioceramic particles with some gaps around them, independently of the type of particles. The themogravimetric analysis of dry and wet specimens showed a higher dissolution rate of the plain beta-TCP particles in comparison to the encapsulated ones, indicating that the PEG adsorbed on the surface of the TCP particles could be a way to control the resorbability of the bioceramic component.

Absorption↗

Glucose-responsive UV polymerised dextran-concanavalin A acrylic derivatised mixtures for closed-loop insulin delivery.

A novel UV polymerised glucose-responsive mixture containing concanavalin A (con A) and dextran was synthesised and characterised as a "smart" biomaterial to form the basis of a closed-loop delivery device. Dextran and con A precursors were modified with acrylic side groups and then UV polymerised to produce covalently bonded mixtures which were examined by FTIR. The viscoelastic properties of these polymerised mixtures containing glucose concentrations between 0% and 5% w/w were also examined using oscillatory rheometry within the linear viscoelastic range across a frequency range of 0.01-50 Hz. As the formulation glucose concentration was raised, a graded decrease in storage modulus, loss modulus and complex viscosity when compared at 1 Hz was observed. Increasing the mixture irradiation time produced viscosity profiles at higher values throughout the glucose concentration range. The subsequent testing of such formulations in in vitro diffusion experiments revealed that the leaching of the mixture components is formulation dependent and is restricted significantly in the covalently bonded mixtures. Insulin delivery in response to glucose in the physiologically relevant glucose concentration range was demonstrated using the novel polymerised mixture at 37 degrees C. The performance of this covalently cross-linked glucose-responsive biomaterial has been improved in terms of increased mixture stability with reduced component leaching. This could, therefore be used as the basis of the design of a closed-loop drug delivery device for therapeutic agents used for the management of diabetes mellitus.

Acrylic Resins↗

The effect of degree of acrylic derivatisation on dextran and concanavalin A glucose-responsive materials for closed-loop insulin delivery.

Formulations of dextran methacrylate (dex-MA) and concanavalin A methacrylamide (con A-MA) were photo-polymerized to produce covalently cross-linked glucose-responsive materials for the basis of a closed-loop insulin delivery device. The viscoelastic properties of these polymerised materials were tested rheologically in the non-destructive oscillatory mode within the linear viscoelastic range at glucose concentrations between 0% and 5% w/w. The degree of acrylic substitution was varied for the dex-MA and con A-MA, and as the formulation glucose concentration was raised, a graded decrease in storage modulus, loss modulus and complex viscosity when compared at 1 Hz was observed for each cross-linked material. Increasing the degree of substitution (DS) of the derivatised dextran produced viscosity profiles at higher values throughout the glucose concentration range. A comparison with non-polymerised mixtures shows similar rheological properties but at much lower values across the chosen glucose concentration range. High-pressure liquid chromatography analyses and in vitro diffusion experiments showed that there were optimum degrees of derivatisation to minimise dex-MA and con A-MA component leach from the material. The in vitro diffusion experiments also showed that differential delivery of insulin in response to glucose was possible with candidate polymerised glucose-responsive formulations, thus highlighting the potential of such a novel glucose-sensitive material to be used as part of implantable closed-loop insulin delivery device.

Acrylates↗

Drug transport mechanisms and release kinetics from molecularly designed poly(acrylic acid-g-ethylene glycol) hydrogels.

Controlled drug release devices of pH-sensitive, complexing poly(acrylic acid-g-ethylene glycol) (P(AA-g-EG)) hydrogels were prepared by free radical solution UV polymerization. The effects of hydrogel composition, polymerization conditions and surrounding environment on theophylline release kinetics and drug transport mechanisms were evaluated in these P(AA-g-EG) polymer networks. Release studies indicated a dependence of the theophylline release kinetics and diffusion coefficients on the hydrogel structure, polymerization conditions and pH of the environment. The theophylline transport mechanism was studied by fitting experimental data to five different model equations and calculating the corresponding parameters. The Akaike information criterion was also considered to elucidate the best-fit equation. Results indicated that in most release cases, the drug release mechanism was anomalous (non-Fickian). This indicates that such systems may, under certain conditions, provide release characteristics approaching zero-order release. The pH of the dissolution medium appeared to have a strong effect on the drug transport mechanism. At more basic pH values, Case II transport was observed, indicating a drug release mechanism highly influenced by macromolecular chain relaxation. The results obtained in this research work lead us to the conclusion that P(AA-g-EG) hydrogels can be successfully used as drug delivery systems. Their versatility to be designed with specifically tuned release properties renders these biomaterials promising pharmaceutical carriers for therapeutic agents.

Acrylic Resins↗

A novel method to construct a third-generation biosensor: self-assembling gold nanoparticles on thiol-functionalized poly(styrene-co-acrylic acid) nanospheres.

A novel method for fabrication of horseradish peroxidase (HRP) biosensor has been developed by self-assembling gold nanoparticles on thiol-functionalized poly(styrene-co-acrylic acid) (St-co-AA) nanospheres. At first, a cleaned gold electrode was immersed in thiol-functionalized poly(St-co-AA) nanosphere latex prepared by emulsifier-free emulsion polymerization of St with AA and function with dithioglycol to assemble the nanospheres, then gold nanoparticles were chemisorbed onto the thiol groups. Finally, horseradish peroxidase was immobilized on the surface of the gold nanoparticles. The sensor displayed an excellent electrocatalytical response to reduction of H2O2 without the aid of an electron mediator. The sensor was highly sensitive to hydrogen peroxide with a detection limit of 4.0 micromoll(-1), and the linear range was from 10.0 micromoll(-1) to 7.0 mmoll(-1). The biosensor retained more than 97.8% of its original activity after 60 days of use. Moreover, the studied biosensor exhibited good current repeatability and good fabrication reproducibility.

Acrylic Resins↗

Biosensors based on acrylic microgels: a comparative study of immobilized glucose oxidase and tyrosinase.

Acrylic microgels are proposed as enzyme immobilizing support in amperometric biosensors. Two enzymes, glucose oxidase and tyrosinase, were entrapped in this matrix and their behaviour is compared. The optimum cross-linking of the polymeric matrix required to retain the enzyme, and to allow the diffusion of the substrate is different for each enzyme, 3.2% for glucose oxidase and 4.5% for tyrosinase. The effect of pH and temperature on the biosensor responses has been studied by experimental design methodology and predictions have been compared with independently performed experimental measurements. A quadratic effect of the variables studied (pH and T) on the biosensor response and the small or null interaction between them was confirmed. The pH results obtained with both methods are coincident revealing an reversible effect on the enzyme. However, the temperature optimum value obtained by experimental design was 10 degrees C lower as a result of an activity decay due to irreversible thermal denaturation of both enzymes.

Acrylic Resins↗

Effect of solution pH and ionic strength on the stability of poly(acrylic acid)-encapsulated multiwalled carbon nanotubes aqueous dispersion and its application for NADH sensor.

The optimal conditions to prepare water-soluble, stable poly(acrylic acid)-wrapped multiwalled carbon nanotubes (PAA-MWNTs) complex is presented. PAA-MWNTs shows high stability within weak acid to weak basic pH condition. The complex also shows good endurance to moderate ionic strengths in the buffer solution. The PAA-MWNTs complex film-covered electrode demonstrates stable, excellent electrocatalytic activity to oxidize NADH, which makes it possible to prepare NADH sensor at a low potential of approximately 0.13 V (versus Ag/AgCl) with the linear range of 4-100 microM by differential pulse voltammetry.

Acrylic Resins↗