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Modified capillary electrophoresis system for peptide, protein and double-stranded DNA analysis.

The results of high-performance capillary electrophoresis (HPCE) studies of peptide, protein and double-stranded DNA separations on a laboratory-made HPCE system are presented. Parameters of the HPCE system are given. The new method of capillary surface modification by grafting poly(glycidyl methacrylate) is described. The problems of HPCE biopolymer analysis connected with the sample-wall interactions are discussed.

DNA↗

Molecularly imprinted polymer membranes for substance-selective solid-phase extraction from water by surface photo-grafting polymerization.

Hydrophilized polyvinylidene fluoride microfiltration membranes were surface-modified in the presence of a template (terbumeton) in methanol with a graft copolymer of a functional monomer (2-acrylamido-2-methyl-1-propane sulfonic acid, AMPS, methacrylic acid, MAA, or acrylic acid, AA) and a cross-linker (N,N'-methylene-bis-acrylamide) using UV irradiation and benzophenone as photoinitiator. As result, membranes covered with a thin layer of imprinted polymer selective to terbumeton were obtained. Blank membranes were prepared with the same monomer composition, but in the absence of the template. The membranes' capacity to adsorb terbumetone from aqueous solution was evaluated yielding information regarding the effect of polymer synthesis (type and concentration of functional monomer, concentration of cross-linker) on the resulting membranes' recognition properties. UV spectroscopic studies of the interactions with terbumetone revealed that AMPS forms a stronger complex than MAA and AA. In agreement with that finding, imprinting with AMPS gave higher affinities than with MAA and AA. The terbumeton-imprinted membranes showed significantly higher sorption capability to this herbicide than to similar compounds (atrazine, desmetryn, metribuzine). With the novel surface modification technology, the low non-specific binding properties of the hydrophilized microfiltration membrane could successfully be combined with the receptor properties of molecular imprints, yielding substance-specific molecularly imprinted polymer composite membranes. The high affinity of these synthetic affinity membranes to triazine herbicides together with their straightforward and inexpensive preparation provides a good basis for the development of applications of imprinted polymers in separation processes such as solid-phase extraction.

Membranes, Artificial↗

High-performance liquid chromatography of sulfur-containing amino acids and related compounds with amperometric detection at a modified electrode.

Organic disulfides generally are not oxidized at bare electrodes under conditions that are suited to routine amperometric detection, and thiols are typically oxidized in a manner that leads to partial blockage of the surface. Modification of a carbon electrode with a film of Ru(III,IV) oxide stabilized with cyanocross-links permits the amperometric detection of cystine, cysteine, glutathione, methionine, and glutathione disulfide under conditions compatible with their chromatographic separation on a strong cation-exchange column. Detection limits of 0.2-0.6 microM and linear dynamic ranges of at least 1-50 microM were obtained. The electrode was stable for at least 11 days with a pH 1 citrate, phosphate mobile phase.

Amino Acids, Sulfur↗

Controlling hydrolysis and dispersion of AlN powders in aqueous media.

Aqueous suspensions of aluminum nitride (AlN) powders have been prepared in the presence of different surface-active agents, namely, H3PO4 and an anionic surfactant, to avoid the hydrolysis of AlN powders and to enhance dispersion. The most determinant parameters to the hydrolysis process (DeltapH and time of contact) and the stabilization of AlN particles in water (surface crystallinity, surface chemical modification, and surface ionic charge) were seen to be strongly dependent on the acidic agent. The H3PO4 treatment was effective against hydrolysis of AlN due to the formation of a phosphate-based protection layer on the particles' surface, and, although it keeps the pH of the suspension below 4, it does not guarantee a good dispersion. The individual adsorption of the anionic surfactant at the surface of AlN particles suspensions did not completely suppress the hydrolysis but it did enhance the degree of dispersion. A proper combination of the two types of surface-active agents enabled the preparation of AlN aqueous suspensions of relatively low viscosity and high AlN concentration, which can be a good starting point for aqueous-based colloidal shaping techniques or for freeze granulation or spray drying to obtain suitable granulate powder characteristics for dry-pressing technologies. An adsorption mechanism of the surface-active agents onto the particles' surface is proposed and supported by NMR and FT-IR analyses.

Journal Article↗

Optical properties of nanocrystalline Y2O3:Eu depending on its odd structure.

The structure of nanocrystalline Y2O3:Eu prepared by a combustion reaction was analyzed by XRD and high-resolution electron microscopy. Compared with a large-scale particles, 5-nm Y2O3:Eu particles presented as distorted crystallite and rough surfaces. Luminescent and absorption properties of nano-Y2O3:Eu showed remarkably particle size effects. At Y2O3:Eu particle sizes smaller than 10 nm some new results were observed: (a) a red shift of the charge-transfer-state absorption; (b) new emission bands of Eu3+ in the 5D0 --> 7F2 region; (c) luminescent decay of energy level 5D0 of Eu3+ turning to a two-step exponential; and (d) a pronounced increase in quenching concentration and much lower phonon density compared with those of the bulk material. All these phenomena can be attributed to the effect of the softened lattice and surface state of the nanomaterial. The latter was confirmed by stronger excitation by the host absorption after the surface modification.

Journal Article↗

Micropatterning of Ni particles on a BaTiO3 green sheet using a self-assembled monolayer.

We studied surface modification of ceramic BaTiO(3) green sheets including polyvinylbutyral (PVB) as an organic binder and successfully fabricated a patterned self-assembled monolayer (SAM) which has a region of amino groups and a region of silanol groups on the green sheet. The patterned SAM was then immersed in water containing Ni particles that show negative zeta potential in water. Ni particles were attracted to the amino groups of the patterned SAM that show positive zeta potential by attractive electrostatic interaction. Consequently, a micropattern of Ni particles was achieved using a SAM at room temperature in water. This process can be used to fabricate a thin uniform internal Ni electrode for miniaturized high-performance multilayer ceramic capacitors (MLCC).

Journal Article↗

Direct coating of gold nanoparticles with silica by a seeded polymerization technique.

Gold nanoparticles prepared through a conventional citrate-reduction method were directly coated with silica by means of a seeded polymerization technique based on the Stöber method. The method required no surface modification. The addition of tetraethylorthosilicate and water prior to ammonia was found to be critical to obtain a proper coating. The silica shell thickness was varied from 30 to 90 nm for TEOS concentrations of 0.0005-0.02 M at 10.9 M of water and 0.4 M of ammonia. The optical spectra of the core-shell gold-silica composite particles agreed with predictions of Mie theory.

Journal Article↗

Immobilization of heavy metals from aqueous solutions using polyacrylamide grafted hydrous tin (IV) oxide gel having carboxylate functional groups.

A new adsorbent containing a carboxylate group has been prepared by the surface modification of a polyacrylamide grafted hydrous tin (IV) oxide gel. The product exhibits a very high adsorption potential for Pb(II), Hg(II) and Cd(II). The effect of initial metal ion concentration, adsorbent dose, pH, concentration of light metal ions, and temperature on metal removal has been studied. The process follows a first-order rate kinetics. The intraparticle diffusion of metal ions through pores in the adsorbent was shown to be the main rate limiting step. The equilibrium data fit well with the Langmuir adsorption isotherm. The selectivity order of the adsorbent is Pb(II) > Hg(II) > Cd(II). Adsorption rate constants and thermodynamic parameters were also presented to predict the nature of adsorption. The method was applied on synthetic wastewaters. Acid regeneration has been tried for several cycles with a view to recover the adsorbed metal ions and also to restore the sorbent to its original state.

Adsorption↗

Molecular mechanisms of improved adhesion and growth of an endothelial cell line cultured on polystyrene implanted with fluorine ions.

Endothelial cells derived from the bovine pulmonary artery (line CPAE, CCL 209, American Tissue Culture Collection, Rockville, MD, USA) were cultured on pristine or fluorine ion-irradiated polystyrene (5 x 10(12) or 5 x 10(14) F ions/cm2, 150 keV). At 24-h post-seeding interval, the number of cells which adhered to the ion-modified polystyrene was significantly higher than on the unmodified material (+20 and +58% in cultures with the polystyrene irradiated by lower and higher ion doses, respectively). On day 7, the populations cultured on the irradiated substrates grew to higher densities, exceeding the controls at the lower and higher ion doses by 69 and 180%, respectively. The cells on ion-implanted samples were also larger (+70-95% and +90-99% at the lower and higher ion doses, respectively) and contained more protein (+16% at both ion doses). As was shown by ELISA, the polystyrene irradiated by the higher ion dose enhanced the expression of a cytoskeletal protein, vimentin (+65%) and protein of focal adhesion plaques, talin (+15%). The content of integrin alpha5beta1 (VLA-5), receptor for fibronectin, was increased at both lower and higher ion doses (+22 and +57%). In contrast to this, the content of ICAM-1 and vinculin was similar in cells grown on both pristine and ion-irradiated growth substrates. Moreover, the expression of VCAM-1 and ELAM-1 was lower by 11-14% in both ion dose groups. The present study has shown that ion implantation of polymers improves the adhesion and growth of endothelial cells without elevating the expression of immunoglobulin and selectin types of adhesion molecules. This surface modification should promote colonization of an artificial vascular prosthesis by endothelial cells and make it less vulnerable by immune system cells of the recipient.

Animals↗

Behavior of osteoblasts on a type I atelocollagen grafted ozone oxidized poly L-lactic acid membrane.

With oxidizing poly-L-lactic acid (PLLA) surface by ozone, peroxide groups are easily generated on the surface. Those peroxides are broken down by redox-coupling reaction, and provide active species that initiate grafting by reaction with the collagen molecules. The surface density of generated peroxide on a PLLA surface was determined by an iodide method. The maximum concentration of peroxide was about 2.87 x 10(-8) mol/cm2 when ozone oxidation was performed at 60 V for 60 min. After the surface oxidation, type I atelocollagen was grafted onto PLLA surface. All physical measurements on the collagen-grafted surface indicated that the PLLA surface was effectively grafted with type I atelocollagen. Behavior of rat calvaria osteoblasts on type I atelocollagen grafted PLLA (PLLA + COL) surface was observed. Initial attachment of osteoblasts on the surface was significantly enhanced, and it is assumed that the atelocollagen matrix supported the initial attachment and growth of cells. Collagenous protein synthesis of osteoblasts was maintained at relatively low level in the early stage of proliferation due to the primarily existing grafted type I atelocollagen, and then increased in 7 days as the osteoblast differentiated. After 7 days, collagenous protein synthesis in osteoblasts was activated. Alkaline phosphatase (ALPase) activity and mineralization by osteoblasts were promoted on PLLA + COL surface. In comparison with PLLA + COL, non-treated PLLA and tissue culture plate (TCPS) did not show any feature expressed in osteoblasts' maturation up to 9 days in this experiment. The grafted type I atelocollagen provided a favorable matrix for cell migration in relation with collagenase expression. Ozone oxidation might be a favorable method for surface modification of PLLA membranes by collagen grafting, and cell behavior could be modulated by the grafted collagen.

Animals↗

Electrochemical studies on the stability and corrosion resistance of titanium-based implant materials.

The corrosion susceptibility of Ti, Ti-6A1-4V and Ti-45Ni was studied in a buffered saline solution using anodic polarisation and electrochemical impedance measurements. Pitting potentials as low as + 250 mV(SCE) were recorded for Ti-45Ni and once initiated pits continued to propagate at potentials as low as -150 mV(SCE). It was possible to increase the pitting potential of Ti-45Ni to values greater than +800 mV(SCE) using a H2O2 surface treatment procedure; however, this surface modification process had no beneficial effect on the rate of pit repassivation. Impedance spectra, recorded under open-circuit conditions, were modelled using a dual oxide film model; a porous outer layer and an inner barrier oxide layer. The nature of this porous outer layer was found to depend on the nature of the electrode material and the presence of phosphate anions in the saline-buffered solution. The porous layers formed on Ti-45Ni and Ti-6Al-4V in the presence of phosphate anions had low resistances typically between 10 and 70 ohm cm2. Much higher porous layer resistances were recorded for Ti and also for Ti-45Ni and Ti-6Al-4V in the absence of the phosphate anions.

Alloys↗

Comparison of corneal epithelial cellular growth on synthetic cornea materials.

The application of artificial corneas for severely wounded ocular surfaces has always encountered the problem of biocompatibility with corneal epithelial cells (CECs). For the eye to stay healthy, it must continually have a complete sheet of CECs across the artificial corneal surface. Various surface modifications of different polymeric materials have been examined to determine which have the best cellular growth rates. A mathematical model of corneal cell growth profiles on synthetic materials was formulated based upon a linear mitotic growth rate. Experimental data reported for the CEC growth on modified poly(vinyl alcohol), silicone rubber, polystyrene, and polycarbonate was analyzed using the model to estimate the linear mitotic rate constant (k). The model proved to be useful in comparing data from different investigators. Plasma-induced graft copolymerized poly(hydroxyethyl methacrylate) (pHEMA) on silicone rubber provided the best growth rate from this particular set of data.

Animals↗

Physical properties and blood compatibility of surface-modified segmented polyurethane by semi-interpenetrating polymer networks with a phospholipid polymer.

Segmented polyurethanes, (SPU)s, are widely used in the biomedical fields because of their excellent mechanical property. However, when blood is in contact with the SPU, non-specific biofouling on the SPU occurs which reduces its mechanical property. To obtain novel blood compatible elastomers, the surface of the SPU was modified with 2-methacryloyloxyethyl phosphorylcholine (MPC) by forming a semi-interpenetrating polymer network (semi-IPN). The SPU film modified by MPC polymer with the semi-IPN (MS-IPN film) was prepared by visible light irradiation of the SPU film in which the monomers were diffused. X-ray photoelectron spectroscopy confirmed that the MPC units were exposed on the MS-IPN film surface. The mechanical properties of the MS-IPN film characterized by tensile testing were similar to those of the SPU film. Platelet adhesion on MS-IPN films was also investigated before and after stress loading to determine the effects of the surface modification on the blood compatibility. Many platelets did adhere on the SPU film before and after stress loading. On the other hand, the MS-IPN film prevented platelet adhesion even after repeated stress loading.

Blood Platelets↗

Binding of D-mannose to poly(2-hydroxyethyl methacrylate) hydrogels by azo coupling.

Crosslinked hydrogels based on poly(2-hydroxyethyl methacrylate) poly(HEMA) were prepared and modified with tyramine [4-(2-aminoethyl)phenol] to enable azo coupling with mannosyl derivatives, copolymerization of HEMA with polymerizable activated esters and surface modification reaction with 4-nitrophenyl chlorocarbonate for activation of the hydrogels were used.

Biocompatible Materials↗

Towards biomimetic scaffolds: anhydrous scaffold fabrication from biodegradable amine-reactive diblock copolymers.

The development of biomimetic materials and their processing into three-dimensional cell carrying scaffolds is one promising tissue engineering strategy to improve cell adhesion, growth and differentiation on polymeric constructs developing mature and viable tissue. This study was concerned with the fabrication of scaffolds made from amine-reactive diblock copolymers, N-succinimidyl tartrate monoamine poly(ethylene glycol)-block-poly(D,L-lactic acid), which are able to suppress unspecific protein adsorption and to covalently bind proteins or peptides. An appropriate technique for their processing had to be both anhydrous, to avoid hydrolysis of the active ester, and suitable for the generation of interconnected porous structures. Attempts to fabricate scaffolds utilizing hard paraffin microparticles as hexane-extractable porogens failed. Consequently, a technique was developed involving lipid microparticles, which served as biocompatible porogens on which the scaffold forming polymer was precipitated in the porogen extraction media (n-hexane). Porogen melting during the extraction and polymer precipitation step led to an interconnected network of pores. Suitable lipid mixtures and their melting points, extraction conditions (temperature and time) and a low-toxic polymer solvent system were determined for their use in processing diblock copolymers of different molecular weights (22 and 42 kDa) into highly porous off-the-shelf cell carriers ready for easy surface modification towards biomimetic scaffolds. Insulin was employed to demonstrate the principal of instant protein coupling to a prefabricated scaffold.

Biocompatible Materials↗

Tissue engineering of the meniscus.

Meniscus lesions are among the most frequent injuries in orthopaedic practice and they will inevitably lead to degeneration of the knee articular cartilage. The fibro-cartilage-like tissue of the meniscus is notorious for its limited regenerative capacity. Tissue engineering could offer new treatment modalities for repair of meniscus tears and eventually will enable the replacement of a whole meniscus by a tissue-engineered construct. Many questions remain to be answered before the final goal, a tissue-engineered meniscus is available for clinical implementation. These questions are related to the selection of an optimal cell type, the source of the cells, the need to use growth factor(s) and the type of scaffold that can be used for stimulation of differentiation of cells into tissues with optimal phenotypes. Particularly in a loaded, highly complex environment of the knee, optimal mechanical properties of such a scaffold seem to be of utmost importance. With respect to cells, autologous meniscus cells seems the optimal cell source for tissue engineering of meniscus tissue, but their availability is limited. Therefore research should be stimulated to investigate the suitability of other cell sources for the creation of meniscus tissue. Bone marrow stroma cells could be useful since it is well known that they can differentiate into bone and cartilage cells. With respect to growth factors, TGF-beta could be a suitable growth factor to stimulate cells into a fibroblastic phenotype but the problems of TGF-beta introduced into a joint environment should then be solved. Polyurethane scaffolds with optimal mechanical properties and with optimal interconnective macro-porosity have been shown to facilitate ingrowth and differentiation of tissue into fibro-cartilage. However, even these materials cannot prevent cartilage degeneration in animal models. Surface modification and/or seeding of cells into the scaffolds before implantation may offer a solution for this problem in the future.This review focuses on a number of specific questions; what is the status of the development of procedures for lesion healing and how far are we from replacing the entire meniscus by a (tissue-engineered) prosthesis. Subquestions related to the type of scaffold used are: is the degree of tissue ingrowth and differentiation related to the initial mechanical properties and if so, what is the influence of those properties on the subsequent remodelling of the tissue into fibro-cartilage; what is the ideal pore geometry and what is the optimal degradation period to allow biological remodelling of the tissue in the scaffold. Finally, we will finish with our latest results of the effect of tear reconstruction and the insertion of prostheses on articular cartilage degradation.

Animals↗

Covalent immobilization of hirudin improves the haemocompatibility of polylactide-polyglycolide in vitro.

A biodegradable polymer, poly(D,L-lactide-co-glycolide) RESOMER RG756, was modified by surface immobilization of recombinant hirudin (r-Hir) with glutaraldehyde as coupling reagent to improve the blood contacting properties of the polymer. The activity of immobilized hirudin on the polymer was estimated by a chromogenic assay to about 2.5 ATU r-Hir cm-2. The improvement of the haemocompatibility of the modified RG756 was evaluated in terms of platelet adhesion/activation, whole blood clotting times and clot formation rate. Fluorescence microscopy revealed that surface modification with r-Hir resulted in decreased platelet adhesion and activation. An ELISA for P-selectin, a marker of platelet activation, was used to confirm this result. Clotting time experiments demonstrated significantly prolonged non-activated partial thromboplastin times, and a decreased clot formation rate of whole blood in contact with r-Hir modified RG756 compared with the plain polymer. Comparison of immobilized r-Hir with bound heparin yielded equivalent improvement of blood-contacting properties of the investigated polymers. These in vitro investigations indicate that the immobilization of r-Hir on RG756 is a useful method to improve the blood contacting properties of polylactides/polyglycolides and other polymers as well.

Antithrombins↗

Understanding and controlling the bone-implant interface.

A goal of current implantology research is to design devices that induce controlled, guided, and rapid healing. In addition to acceleration of normal wound healing phenomena, endosseous implants should result in formation of a characteristic interfacial layer and bone matrix with adequate biomechanical properties. To achieve these goals, however, a better understanding of events at the interface and of the effects biomaterials have on bone and bone cells is needed. Such knowledge is essential for developing strategies to optimally control osseointegration. This paper reviews current knowledge of the bone-biomaterial interface and methods being investigated for controlling it. Morphological studies have revealed the heterogeneity of the bone-implant interface. One feature often reported, regardless of implant material, is an afibrillar interfacial zone, comparable to cement lines and laminae limitantes at natural bone interfaces. These electron-dense interfacial layers are rich in noncollagenous proteins, such as osteopontin and bone sialoprotein. Several approaches, involving alteration of surface physicochemical, morphological, and/or biochemical properties, are being investigated in an effort to obtain a desirable bone-implant interface. Of particular interest are biochemical methods of surface modification, which immobilize molecules on biomaterials for the purpose of inducing specific cell and tissue responses or, in other words, to control the tissue-implant interface with biomolecules delivered directly to the interface. Although still in its infancy, early studies indicate the value of this methodology for controlling cell and matrix events at the bone-implant interface.

Adsorption↗