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PubMed2001

Application of oxygen ion implantation to titanium surfaces: effects on surface characteristics, corrosion resistance, and bone response.

BACKGROUND: The surface oxide layer of titanium plays a decisive role in determining biocompatibility. However, there are some reports demonstrating that the natural oxide film may not be sufficiently protective in the aggressive biologic environment. PURPOSE: The goal of this study was to examine the effectiveness of a thick oxide layer on corrosion resistance in vitro and the bone formation around titanium implants in vivo. MATERIALS AND METHODS: A plasma source ion implantation (PSII) method was used to increase the thickness of the surface oxide layer. Several instruments were employed to confirm the surface properties before and after the surface modification. Potentiodynamic polarization measurements in a phosphate-buffered saline (PBS) solution were carried out to investigate corrosion resistance in vitro. Bone formation around this surface-modified specimen was examined in a rabbit model and assessed in histomorphometry. RESULTS: Improved corrosion resistance was demonstrated by the potentiodynamic polarization measurements. Light microscopic histomorphometry showed that all implants were in contact with bone and had some proportion of bone within the threads at 4 weeks; however, there were no significant differences compared with as-machined controls. CONCLUSIONS: The results indicate that in spite of improved corrosion resistance in vitro, a thick oxide layer fabricated with the PSII method does not influence early bone formation around titanium implants in vivo.

Animals↗

PubMed2003

Platelet adhesion onto segmented polyurethane surfaces modified by carboxybetaine.

Polyurethanes are widely used as blood-contacting biomaterials due to their good biocompatibility and mechanical properties. Nevertheless, their blood compatibility is still not adequate for more demanding applications. Surface modification is an effective way to improve the hemocompatibility for biomaterials. The purpose of present study was to synthesize a novel nonthrombogenic biomaterial by modifying the surface of polyurethane with Zwitterions of carboxybetaine monomer. The films of polyurethane were grafted with two kinds of carboxybetaine by a three-step procedure. In the first step, the film surfaces were treated with hexamethylene diisocyanate (HDI) in toluene at 50 degrees C in the presence of di-n-butyl tin dilaurate (DBTDL) as a catalyst. The extent of the reaction was measured by ATR-FT-IR spectra: a maximum number of free NCO group was obtained after a reaction time of 90 min. In the second step, the hydroxyl group of N,N-dimethylethylethanolamine (DMEA) or 4-dimethylamino-1-butanol (DMBA) was allowed to react in toluene with isocyanate groups bound on surface. In the third step, carboxybetaines were formed in the surface through the ring-opening reaction between tertiary amine of DMEA or DMBA and beta-propiolactone (PL). It was characterized by ATR-FT-IR and XPS that the grafted surfaces were composed of carboxybetaine. The results of the contact angle measurements showed that they were strongly hydrophilic. Platelet adhesion tests showed that films grafted carboxybetaine have good blood compatibility, as featured by the low platelet adhesion.

Betaine↗

PubMed1992

ESCA study on dental alloy surfaces modified by Ga-Sn alloy.

A new, simple surface modification method for adherend metals has been developed. It gives high bond strength and superior water durability to dental precious-metal alloys bonded with 4-META/MMA-TBB resin. However, there was no effect on the bonding of Ag-In-Zn alloy and base-metal alloys. In the present study, the alloy surfaces modified by the new method were analyzed by ESCA and SEM for determination of details of the modification effect. A new alloying layer containing Ga and Sn was formed on the precious-metal alloys. The main factor for excellent adhesion to be achieved was the formation of a very thin layer of Ga2O3 and SnO2, less than 1-2nm thick, on the alloy surface. A thicker modified layer, as formed on the Ag-In-Zn and Ni-Cr alloys, led to low bonding ability.

Acrylic Resins↗

PubMed2005

[Modification of MSWI fly ash by using anionic chelating surfactant].

This paper aims at the reutilization and stabilization of municipal solid waste incineration (MSWI) fly ash to produce a high value added product. The effects of anionic chelating surfactants on the surface modification and fixing capacity of MSWI fly ash were primarily explored. Based on the indexes of active ratio and analysis of IR-spectrometer, the active ratio of ACS1 modified fly ash can be found higher than 95% under the condition of surfactant dosage at 7.0mL/100g fly ash, modification time at 15min and temperature at 75 degrees C . Moreover, Anionic chelating surfactant shows a strong fixing capacity for heavy metals by the United States Environmental Protection Agency's Toxicity Characteristic Leaching Procedure (TCLP).

Cadmium↗

PubMed2003

Surface engineering of living myoblasts via selective periodate oxidation.

Cell surface molecules are vital for normal cell activity. To study the functions of these molecules or manipulate cell behavior, the ability to decorate cell surfaces with bioactive molecules of our choosing is a potentially powerful technique. Here, we describe the molecular engineering of living L6 myoblast monolayers via selective periodate oxidation of sialic acid residues and the application of this surface modification in the artificial aggregation of cells. The aldehyde groups generated by this reaction were used to selectively ligate a model molecule, biotin hydrazide, to the cell surfaces. Flow cytometry analysis after staining with fluorescently conjugated avidin revealed a concentration-dependent increase in fluorescence compared to untreated cells with a maximal shift of 345.1 +/- 27.4-fold and an EC(50) of 17.4 +/- 1.1 microM. This mild oxidation reaction did not affect cell number, viability, or morphology. We then compared this chemical technique with the metabolic incorporation of reactive cell surface ketone groups using N-levulinoylmannosamine (ManLev). In this cell line, only a 22.3-fold fluorescence shift was observed compared to untreated cells when myoblasts were incubated with a high concentration of ManLev for 48 hours. Periodate oxidation was then used to modify myoblast surfaces to induce cell aggregation. Crosslinking biotinylated myoblasts, which do not spontaneously aggregate in culture, with avidin resulted in the rapid formation of millimeter-sized, multicellular structures. These data indicate that sodium periodate treatment is an effective, noncytotoxic method for the in vitro molecular engineering of living cell surfaces with the potential for cell biology and tissue engineering applications.

Aldehydes↗

PubMed2000

SiO(x) Monolayer Overcoating Effect on the TiO(2) Photocatalytic Oxidation of Cetylpyridinium Bromide.

The effect of SiO(x) monolayer coverage on the rate of TiO(2) photocatalytic oxidation of cetylpyridinium bromide (CPB) in aqueous solutions has been studied. The rate of CPB removal from the solution (5<pH<7) increases with the surface modification at concentrations below 4.5x10(-4) M and its promoting effect is enhanced with decreasing concentration. A modified Langmuir-Hinshelwood model, where the rate of surface reaction is assumed to be proportional to the coverage of CPB at the photostationary state is proposed for dilute reaction systems. As a result of the kinetic analyses, the acceleration of the reaction with the SiO(x) monolayer coverage can be attributed to the increase in the rate of adsorption due to the electrostatic attraction of cetylpyridinium ion. The suppression of Br(-) adsorption is also suggested as a minor contribution. Copyright 2000 Academic Press.

Journal Article↗

PubMed1994

A cell surface ELISA for the screening of monoclonal antibodies to antigens on viable cells in suspension.

To simplify the screening of monoclonal antibodies to different human T cell surface molecules a live cell enzyme-linked immunosorbent assay (cell ELISA) has been established and optimized. The assay was performed in 96-well plates. By using living human T lymphocytes in suspension surface modification by fixation or insolubilization of the cells was avoided. Several parameters influencing sensitivity and specificity were studied. About 150 ng/ml of mouse monoclonal antibodies to cell surface antigens could be detected when using 5 x 10(4) cells per well and a 1/1000 dilution of the anti-mouse IgG-alkaline phosphatase conjugate. This sensitivity permitted the primary screening of cell specific antibodies from hybridoma supernatants. The same detection limit was obtained in flow cytometric analysis. If required, the sensitivity of the cell ELISA could be increased using higher cell numbers and conjugate concentration. When analysing different cell lines with selected antibodies the cell ELISA was found to be as sensitive and specific as the fluorescence assay. The assay was applied to the screening of supernatants from hybridomas developed against human T helper cell clones and the detection of V beta specificities of T cell clones.

Antibodies, Monoclonal↗

PubMed2006

Electrogenerated indium tin oxide-coated glass surface with photosensitive interfaces: surface analysis.

We present herein a photo-immobilization technique for the localized and specific conjugation of biochip platforms with different proteinaceous bioreceptors, such as antigen or antibodies. This methodology based on a photoactivable electrogenerated polymer film, pyrrole-benzophenone, allows the covalent immobilization of biomolecules through light mediation. The surface-conductive glass platform electropolymerized with poly(pyrrole-benzophenone) thin film may then be used to affinity-coat the chip with molecular recognition probes. This glass chip electroconductive surface modification is done by the deposition of a thin layer of indium tin oxide (ITO). Thereafter, pyrrole-benzophenone monomers are electropolymerized onto the conductive metal oxide surface and then exposed to an antigen Staphylococcal Enterotoxin B (SEB)) solution and illuminated with UV light (wavelength approximately 345 nm) through a mask. As a result of the photochemical reaction, a pattern thin layer of the antigen was covalently bound to the benzophenone-modified surface. Then the sample to be analyzed, along with its specific target antibody (anti-SEB antibodies), is introduced onto the glass surface and left to react with the previously photo-immobilized antigen. When the immuno-reaction is completed, the specifically attached immunoglobulin analytes are detected by using secondary antibodies conjugated with Fluorescein isothiocyanate (FITC). The fluorescence signal emanating from the biochip surface is then quantified by two methods, using a filtered intensified charge-coupled device (CCD) camera and a grating spectrometer.

Antibodies↗

PubMed2005

Photo-chemically patterned polymer surfaces for controlled PC-12 adhesion and neurite guidance.

The in vitro assembling of cellular networks offering control over cell positions and connectivities by patterned culture substrates is a valuable tool for neuroscience research and other applications in cell biology. We developed a versatile technique based on polymer surface modification which allows the patterning of different cell lines for advanced tissue engineering, among them are Pheochromocytoma cells (PC-12). In contrast to other techniques applied for surface patterning, the presented photo patterning by deep UV irradiation is applicable to the widely used cell culture substrate material polystyrene (PS) and should be easily performed in most laboratories. Irradiation of polystyrene with UV radiation of lambda = 185 nm yields mainly carboxyl groups at the polymer surface which can be used to control the spontaneous competitive protein adsorption from serum containing culture media [Welle A, Gottwald E. UV-based patterning of polymeric substrates for cell culture applications. Biomed. Microdev. 2002;4:33-41] or to serve as defined coupling sites for controlled protein/peptide immobilization. Extending our previous studies on patterning hepatoma cells and fibroblasts via spatially defined plasma protein adsorption, we here describe an advanced application to produce patterns of cell repellent albumin domains and cell attractive laminin regions for the patterning of Pheochromocytoma cells.

Animals↗

PubMed1999

Prolonged circulation time of doxorubicin-loaded liposomes coated with a modified polyvinyl alcohol after intravenous injection in rats.

The purpose of this study was to evaluate the functions of a modified polyvinyl alcohol (PVA-R), which has a hydrophobic moiety, as a coating material for liposomes to be loaded with the anticancer drug, doxorubicin. The size controlled liposomes (egg phosphatidylcholine: cholesterol=1:1 molar ratio) were prepared by the hydration method followed by extrusion. Drug encapsulation and surface modification with polymers (PVA and PVA-R) were carried out simultaneously using a modified pH gradient method. The existence of a thick polymer layer on the surface of the liposomes was confirmed by an increase in particle size and the amount of polymer on the liposomal surface, especially for the PVA-R-coated liposomes. The effects of polymer coating on the behavior of the liposomes in vivo were evaluated by measuring the circulation time and biodistribution of the drug after i.v. administration of the liposomal drug in rats. The PVA-R-coated liposomes showed a more prolonged circulating time for the drug with less uptake by the reticuloendothelial system after i.v. administration in rats, compared with non-coated liposomes. These results confirm that polymer possessing a hydrophobic anchor at its end, like PVA-R, is a suitable material for modifying the surface of doxorubicin-loaded liposomes to improve their stability in the circulating blood.

Animals↗

PubMedDate not supplied

Folate-conjugated iron oxide nanoparticles for solid tumor targeting as potential specific magnetic hyperthermia mediators: synthesis, physicochemical characterization, and in vitro experiments.

New folate-conjugated superparamagnetic maghemite nanoparticles have been synthesized for the intracellular hyperthermia treatment of solid tumors. These ultradispersed nanosystems have been characterized for their physicochemical properties and tumor cell targeting ability, facilitated by surface modification with folic acid. Preliminary experiments of nanoparticles heating under the influence of an alternating magnetic field at 108 kHz have been also performed. The nanoparticle size, surface charge, and colloidal stability have been assessed in various conditions of ionic strength and pH. The ability of these folate "decorated" maghemite nanoparticles to recognize the folate receptor has been investigated both by surface plasmon resonance and in folate receptor expressing cell lines, using radiolabeled folic acid in competitive binding experiments. The specificity of nanoparticle cellular uptake has been further investigated by transmission electron microscopy after incubation of these nanoparticles in the presence of three cell lines with differing folate receptor expression levels. Qualitative and quantitative determinations of both folate nanoparticles and nontargeted control nanoparticles demonstrated a specific cell internalization of the folate superparamagnetic nanoparticles.

Cell Line, Tumor↗

PubMed2005

Surface grafted antibodies: controlled architecture permits enhanced antigen detection.

The attachment of antibodies to substrate surfaces is useful for achieving specific detection of antigens and toxins associated with clinical and field diagnostics. Here, acrylated whole antibodies were produced through conjugation chemistry, with the goal of covalently photografting these proteins from surfaces in a controlled fashion, to facilitate rapid and sensitive antigenic detection. A living radical photopolymerization chemistry was used to graft the acrylated whole antibodies on polymer surfaces at controlled densities and spatial locations by controlling the exposure time and area, respectively. Copolymer grafts containing these antibodies were synthesized to demonstrate two principles. First, PEG functionalities were introduced to prevent nonspecific protein interactions and improve the reaction kinetics by increasing solvation and mobility of the antibody-containing chains. Both of these properties lead to sensitive (pM) and rapid (<20 min) detection of antigens with this surface modification technique. Second, graft composition was tailored to include multiple antibodies on the same grafted chains, establishing a means for simultaneously detecting multiple antigens on one grafted surface area. Finally, the addition of PEG spacers between the acrylate functionality and the pendant detection antibodies was tuned to enhance the detection of a short-half-life molecule, glucagon, in a complex biological environment, plasma.

Acrylates↗

PubMed1981

Ageing of rabbit red cells in vitro: membrane modifications and their possible role in red cell survival in vivo.

In vitro incubation induces, in rabbit red cell membranes, significant modifications consisting mainly in a decrease of sialic acid and galactose. In vivo the life span of incubated erythrocytes seems to be correlated to the degree of surface alterations and ATP depletion: larger surface modifications and energy charge reduction induce shorter survival time. It can therefore be postulated that incubation of red cell in vitro can cause an ageing process similar to that occurring physiologically in vivo.

Adenosine Triphosphate↗

PubMed2002

Grafting sulfobetaine monomer onto the segmented poly(ether-urethane) surface to improve hemocompatibility.

Polyurethanes are widely used as blood-contacting biomaterials, due to their good biocompatibility and mechanical properties. Nevertheless, their blood compatibility is still not adequate for more demanding applications. Surface modification is an effective way to improve the hemocompatibility for biomaterials. The purpose of the present study was to synthesize a novel nonthrombogenic biomaterial by modifying the surface of polyurethane. Ozonization was used to introduce active peroxide groups onto the segmented poly(ether-urethane) (SPEU) film surface and graft polymerization of N,N'-dimethyl (methacryloyloxyethyl) ammonium propanesulfonate (DMAPS), a sulfobetaine structure, onto the ozone-activated SPEU surface was conducted. The SPEU-g-PDMAPS film was characterized by ATR-FTIR, XPS, and contact angle measurements. ATR-FTIR and XPS confirmed the graft polymerization. The grafted film possessed a relatively hydrophilic surface, as revealed by contact angle measurement. The blood compatibility of the grafted films was evaluated by a platelet-rich plasma (PRP) adhesion study and scanning electron microscopy, using SPEU film as the reference. No platelet adhesion was observed for the grafted films incubated with PRP at 37 degrees C for 60 and 180 min. This new sulfobetaine structure grafted biomaterial might have potential for biomedical applications.

Betaine↗

PubMed2006

Prolonged bleeding-free period following prophylactic infusion of recombinant factor VIII reconstituted with pegylated liposomes.

Prophylactic treatment for hemophilia A involves infusion of factor VIII (FVIII) concentrates every 2 to 3 days. Liposomes can be efficacious vehicles for medicines, and surface modification by PEGylation can prolong liposome circulation time. When reconstituted with PEGylated liposomes (PEGLip's), recombinant FVIII binds noncovalently but with high affinity to the external liposome surface. This preparation showed prolongation of FVIII half-life and increased protection from bleeding in preclinical models. Here we report a blinded, controlled, crossover, multicenter clinical study that evaluated the time free from bleeding episodes in patients with hemophilia A during prophylaxis with standard rFVIII (no liposomes) or PEGLip rFVIII (PEGLip reconstituted) at 25 and 35 IU/kg doses. Of 24 enrolled patients, 23 were eligible for efficacy analysis. Mean number of days without bleeds was 7.2 +/- 1.7 with standard rFVIII compared with 13.3 +/- 4.8 with PEGLip rFVIII at the 35 IU/kg dose and 5.9 +/- 1.7 with standard rFVIII versus 10.9 +/- 2.9 with PEGLip rFVIII at the 25 IU/kg dose (P < .05 between treatment groups for each dose). PEGLip rFVIII was well tolerated. These data suggest that reconstitution of rFVIII with PEGLip's may reduce the frequency of treatment during prophylaxis.

Adolescent↗

PubMed2001

Specific inhibition of C3 to facilitate general complement inhibition on endotoxin affinity sorbents for apheresis applications.

Complement activation, as a result of human blood exposure to biomaterial surfaces, continues to be a concern in medical applications. The purpose of this study was to identify sorbent(s) and surface modification(s) that allow specific removal of endotoxin, while minimizing complement activation. Maleic anhydride (MA) modification of Sepharose CL-4B, cellulose, and Toyopearl HW-65F resulted in reduced generation of C3a, a marker of complement activation, by two orders of magnitude over unmodified surfaces. Surfaces modified with both MA and polymyxin B (PMB), utilized for binding endotoxin, reduced complement activation in a similar manner. Western Blot analysis of the larger C3 cleavage product C3dg showed a similar reduction, for all MA-modified sorbents, as observed for C3a by ELISA. C3alpha43 levels (constituent of iC3b and C3c) were also reduced, although only MA-Sepharose CL-4B levels were similar to C3a. Activation of C5, measured as the SC5b-9 complex, was also reduced by two orders of magnitude after MA modification of Sepharose CL-4B; the decrease was similar to all chemical modifications tested. PMB immobilized via CNBr on MA-modified cellulose maintained its endotoxin-binding capacity, while the latter was eliminated when PMB was immobilized via CNBr to MA-modified Sepharose CL-4B and Toyopearl HW-65F.

Adsorption↗

PubMed2003

In vivo studies of polypyrrole/peptide coated neural probes.

Neural probes are micromachined multichannel electrode arrays that facilitate the functional stimulation and recording of neurons in the peripheral and central nervous system. For long-term implantations, surface modification is necessary for maintaining the stable connection between electrodes and neurons. The conductive polymer polypyrrole (PPy) and synthetic peptide DCDPGYIGSR were co-deposited on the electrode surface by electrochemical polymerization. The stability of PPy/DCDPGYIGSR coatings was tested in soaking experiments. It was found that the peptide was entrapped in the PPy film and did not diffuse away within 7 weeks of soaking in DI water. Coated probes were implanted in guinea pig brain for periods of 1, 2 and 3 weeks. Recording tests were performed and the impedance was monitored. The explanted probes and tissue were examined by immunocytochemical studies. Significantly more neurofilament positive staining was found on the coated electrode which indicated that the coatings had established strong connections with the neuronal structure in vivo. Good recordings were obtained from the coated sites that had neurons attached. First week tissue sections had no significant gliosis. In week 2, a layer of non-neuronal tissue consisting of mostly meningeal fibroblasts and ECM protein including at least fibronectin was formed around the probe tracks of both coated and uncoated probes. Astrocytes started to form a loosely organized layer by the end of the third week.

Animals↗

PubMed2003

3D protein microarrays: performing multiplex immunoassays on a single chip.

The enzyme-linked immunosorbent assay (ELISA) is typically applied in the format of microtiter plates. To increase throughput and reduce consumption of precious samples, efforts have been made to transfer ELISA to the microchip format using conventional microarrays, microfluidic systems, and chips bearing microwells. However, all three formats lack the possibility to screen several analytes on several immobilized binders at a time or require complicated liquid handling, surface modifications, and additional equipment. Here, we describe an immunoassay performed on a standard microscope slide without the requirement for wells or tubes to separate the samples using standard surfaces and machinery already available for microarray technology. The new multiple spotting technique (MIST) comprises immobilization of a binder onto a surface and subsequent spotting of the second compound on the same spot, on top of the immobilized binder. We show that the analytes bind their ligands immediately within the confined space of separate droplets on the chip surface, thereby eliminating the need for extra incubation time. We illustrate the feasibility of the new technique by spotting dilution rows of proteins or monoclonal and polyclonal antibodies on top of their immobilized binders. Moreover, we demonstrate specificity by applying a mixture of antibodies in a multiplex format and demonstrate that the technique is compatible with conventional microarray protocols, such as total incubation. Finally, we indicate that the technique is capable of quantifying as little as 400 zmol (240,000 molecules) of analyte.

Antibodies↗