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Methods for labeling quantum dots to biomolecules.

Semiconductor quantum dots (QDs) are thought to be the most promising fluorescent probe for biological and biomedical applications because of their unique size-dependent optical and electronic properties. QD bioconjugates provide important advantages over conventional organic dyes. This review compares the optical properties of nanoparticles and dyes and describes the surface modification of the QDs and the methods for biomolecules conjugating to QDs. In addition, the effect of QDs on normal physiology in real biological applications is discussed.

Fluorescent Dyes↗

[Development of the vascular prosthesis research].

The search for a nonthrombogenic material with the potential for use in small diameter vascular graft applications continues to be a field of extensive investigation. This article describes the choice of biomaterials used as vascular prosthesis, the innovation of construction of tissue-engineered blood vessels, the indispensability, methods and the effect produced by surface modification of vascular prosthesis. The article also points out that research achievements of vascular prosthesis must be made with the exploitation of new nonthrombogenic biomaterial and the development of tissue engineering.

Biocompatible Materials↗

Bioelectrocatalytic signaling from immunosensors with back-filling immobilization of glucose oxidase on biorecognition surfaces.

We report a novel method of electrochemical signaling from antigen-antibody interactions at immunoelectrodes with bioelectrocatalyzed enzymatic signal amplification. For the immunosensing surface construction, a poly(amidoamine) G4-dendrimer was employed not only as a building block for the electrode surface modification but also as a matrix for ligand functionalization. As a model biorecognition reaction, the dinitrophenyl (DNP) antigen-functionalized electrode was fabricated and an anti-DNP antibody was used. Glucose oxidase (GOX) was chosen to amplify electrochemical signal by enzymatic catalysis. The signal amplification strategy introduced in this study is based on the back-filling immobilization of biocatalytic enzyme to the immunosensor surface, circumventing the use of an enzyme-labeled antibody. The non-labeled native antibody was biospecifically bound to the immobilized ligand, and the activated enzyme (periodate-treated GOX) reacted and "back-filled" the remaining surface amine groups on the dendrimer layer by an imine formation reaction. From the bioelectrocatalyzed signal registration with the immobilized GOX, the surface density of biospecifically bound antibody could be estimated. The DNP functionalization reaction was optimized to facilitate the antibody recognition and signaling reactions, and approximately 6% displacement of surface amine to DNP was found to be an optimum. From quartz crystal microbalance measurement, immunosensing reaction timing and the surface inertness to the nonspecific biomolecular binding were tested. By changing the surface functionalization level of DNP in the calibration experiments, immunosensors exhibited different dynamic detection ranges and limits of detection, supporting the capability of parameters modulation for the immunosensors. For the anti-DNP antibody assay, the fabricated immunosensor having 65% functionalization ratio exhibited the linear detection range of 10(-4) to 0.1 g/L protein and a limit of detection around 2 x 10(-5) g/L.

2,4-Dinitrophenol↗

Preliminary beta-tricalcium phosphate coating prepared by discharging in a modified body fluid enhances collagen immobilization onto titanium.

Because of its excellent scaffold properties toward bone cells, collagen has been recognized as a promising extracellular matrix protein for surface modification of titanium implants. Hydroxyapatite (HA) coatings have been investigated as a preliminary coating for collagen immobilization on titanium implants. However, the composition of HA-collagen is recognized as being difficult, and while many studies have suggested that biodegradable beta-tricalcium phosphate (beta-TCP) has better osteoconductivity than HA, the efficiency of preliminary beta-TCP coating for collagen immobilization on titanium surfaces has yet to be evaluated. This investigation aimed to evaluate the applicability of HA and beta-TCP coatings, prepared by discharging in modified body fluids, as preliminary collagen coatings. To increase collagen induction on preliminary HA and beta-TCP coatings, we used a new cathodic polarization method. X-ray photoelectron spectroscopy revealed that the bonding strength between the collagen NH(+) amino groups of collagen and phosphate (PO(4) (3-)) was greater on the beta-TCP coating than the HA coating. The preliminary beta-TCP coating was tightly crosslinked with RCOO(-) carboxyl groups of the collagen molecules and showed high cellular responses, even in the early stage of cell cultivation. Thus, this coating was found to be more effective than HA as a preliminary coating for collagen immobilization on titanium implants.

Animals↗

Surface characterization of recycled tire rubber to be used in cement paste matrix.

The surface modification of tire rubber after treatment with saturated NaOH aqueous solution was investigated by HATR infrared analysis, potentiometric titration, and contact angle measurements. Infrared analysis of the powdered treated rubber showed a decrease in absorption at 1540, 1450, and 1395 cm(-1). This decrease is attributed to the removal of zinc stearate, an additive present in tire formulations that often migrates and diffuses to the surface, resulting in poor adhesion between the rubber and other materials. The potentiometric titration of the suspension of powdered rubber in 0.1 M NaCl showed that more hydrochloric acid was consumed by the untreated rubber, most likely a result of the hyrdrolysis of the zinc stearate to the organic acid. Contact angles of flat tire pieces showed an homogeneity enhancement of the treated rubber surface. The decrease of the zinc stearate on the treated rubber surface explains the improvement in the adhesion of this material to the cement matrix, observed in a previous research. The promising results of this study are a starting point for future research on incorporating rubber particles into cementitious materials as a means of successfully utilizing the vast amounts of tire waste currently in landfills.

Journal Article↗

Phosphate removal from wastewaters using a weak anion exchanger prepared from a lignocellulosic residue.

Surface modifications of lignocellulosic residues has become increasingly important for improving their applications as adsorbents. In this study a new adsorbent system (BS-DMAHP) containing dimethylaminohydroxypropyl (DMAHP) weak base groups was prepared by the reaction of banana stem (BS), a lignocellulosic residue with epichlorohydrin and dimethylamine followed by treatment of hydrochloric acid. The original BS and BS-DMAHP were characterized with the help of surface area analyzer, infrared spectroscopy (IR) and scanning electron microscopy (SEM). Surface charge density of the samples as a function of pH was investigated using potentiometric titrations. Adsorbent exhibits very high adsorption potential for phosphate and more than 99.0% removal was achieved in the pH range of 5.0-7.0. Adsorption has been found to be concentration dependent and endothermic and follows a reversible second-order kinetics. The Langmuir and Freundlich isotherm models were applied to describe the equilibrium data. Equilibrium data agreed very well with the Langmuir model. Adsorption experiments were conducted using a commercial chloride form Duolite A-7, a weak base anion exchanger. The removal efficiency was tested using fertilizer industry wastewater. Adsorbed phosphate on BS-DMAHP can be recovered by treating with 0.1 M NaOH solution. A stability test operated for four cycles indicate a capacity loss of < 12.0%.

Adsorption↗

Protein-like molecular architecture: biomaterial applications for inducing cellular receptor binding and signal transduction.

The development of biomaterials with desirable biocompatibility has presented a difficult challenge for tissue engineering researchers. First and foremost, materials themselves tend to be hydrophobic and/or thrombogenic in nature, and face compatibility problems upon implantation. To mediate this problem, researchers have attempted to graft protein fragments onto biomaterial surfaces to promote endothelial cell attachment and minimize thrombosis. We envisioned a novel approach, based on the capability of biomolecules to self-assemble into well-defined and intricate structures, for creating biomimetic biomaterials that promote cell adhesion and proliferation. One of the most intriguing self-assembly processes is the folding of peptide chains into native protein structures. We have developed a method for building protein-like structural motifs that incorporate sequences of biological interest. A lipophilic moiety is attached onto a N alpha-amino group of peptide chain, resulting in a "peptide-amphiphile." The alignment of amphiphilic compounds at the lipid-solvent interface is used to facilitate peptide alignment and structure initiation and propagation, while the lipophilic region absorbs to hydrophobic surfaces. Peptide-amphiphiles containing potentially triple-helical or alpha-helical structural motifs have been synthesized. The resultant head group structures have been characterized by CD spectroscopy and found to be thermally stable over physiological temperature ranges. Triple-helical peptide-amphiphiles have been applied to studies of surface modification and cell receptor binding. Cell adhesion and spreading was promoted by triple-helical peptide-amphiphiles. Cellular interaction with the type IV collagen sequence alpha 1(IV) 1263-1277 increased signal transduction, with both the time and level of induction dependent upon triple-helical conformation. Collectively, these results suggest that peptide-amphiphiles may be used to form stable molecular structure on biomaterial surfaces that promote cellular activities and improve biocompatibility.

Amino Acid Sequence↗

Fabrication of thromboresistant multilayer thin film on plasma treated poly (vinyl chloride) surface.

Layer-by-layer deposited anticoagulant multilayer films were prepared on ammonia plasma treated poly (vinyl chloride) (PVC). Fourier transform infrared spectroscopy-attenuated total reflectance (FTIR-ATR) and contact angle results revealed the presence of -NH2 on the ammonia plasma treated PVC surfaces and the layer-by-layer self-assembly process. The stability of multilayer film was studied with the radio labeled method. The remainder bovine serum albumin (BSA) in cross-linked 5(heparin/BSA) multilayer films dipped in phosphate buffered saline (PBS, pH 7.4) was more than 90% in 40 days. The static platelet adhesion result indicated the anticoagulant multilayer films deposited on the plasma treated PVC reduced platelet adhesion drastically and no thrombus forming. The plasma recalcification time revealed that the multilayer modified surfaces greatly prolonged the plasma recalcification time. Such an easy processing and shape-independent method may have good potential for surface modification of cardiovascular devices.

Blood Platelets↗

Supported planar bilayer formation by vesicle fusion: the interaction of phospholipid vesicles with surfaces and the effect of gramicidin on bilayer properties using atomic force microscopy.

We have used magnetic alternating current mode atomic force microscopy (MAC-AFM) to investigate the formation of supported phospholipid bilayers (SPB) by the method of vesicle fusion. The systems studied were dioleoylphosphatidylcholine (DOPC) on mica and mica modified with 3-aminopropyl-triethoxy-silane (APTES), and DOPC vesicles with gramicidin incorporated on mica and APTES-modified mica. The AFM images reveal three stages of bilayer formation: localized disklike features that are single bilayer footprints of the vesicles, partial continuous coverage, and finally complete bilayer formation. The mechanism of supported phospholipid bilayers formation is the fusion of proximal vesicles, rather than surface disk migration. This mechanism does not appear to be affected by incorporation of gramicidin or by surface modification. Once formed, the bilayer develops circular defects one bilayer deep. These defects grow in size and number until a dynamic equilibrium is reached.

Anti-Bacterial Agents↗

Electrochemical evaluation of the interfacial capacitance upon phosphorylation of amino acid analogue molecular films.

An approach based on electrochemistry to differentiate between phosphorylated and nonphosphorylated amino acid analogues adsorbed on gold is presented. Analogues of serine, threonine, and tyrosine, containing thiohexadecyl headgroups, were synthesized and assembled on gold, and the surface capacitance was evaluated using electrochemical impedance spectroscopy. A procedure for deprotection of tert-butyl phosphate protecting groups, on the monolayer, is also described. Characterizations of the assembled analogues by cyclic voltammetry, infrared spectroscopy, and ellipsometry are used to confirm the insulating properties of the monolayers and the outcome of surface modifications. The results from cyclic voltammetry show good insulating properties for the monolayers even after phosphate deprotection. The infrared measurements reveal well-ordered monolayers, and the thickness from ellipsometry is in good agreement with expectations from molecular modeling. The impedance experiments show a capacitance increase up to 0.6 microF/cm2 as phosphate groups are introduced. The results in this study indicate the possibility of using a surface chemical and impedance spectroscopy approach to detect the kinase/phosphatase activity and kinetics involved in phosphorylation reactions.

Amino Acids↗

Nanoscavenger based dispersion preconcentration; sub-micron particulate extractants for analyte collection and enrichment.

A new approach has been developed for the preconcentration of analytes from solution using nanoscavengers; monodisperse functionalised particles of sub-micron dimensions, that can be suspended as a quasi-stable sol in an aqueous solution, and quantitatively recovered with the analyte by conventional filtration. No external agitation of the sample is required as the particles move naturally through the sample by Brownian motion, convection and sedimentation. By careful choice and control of their particle size and surface chemistries, nanoscavengers can be designed to suit a number of different analytical problems. Surface modification of these nanometre-sized particles, through the grafting of complexing or partitioning functional groups, can produce nanoscavengers having affinities for specific analytes and operating through a wide range of mechanisms from covalent bonding to hydrophobic interaction. The approach is illustrated by the development of an extraction-based preconcentration of metals from solution that employs sub-micron Stöber-type silica spheres, the surfaces of which have been modified using chelating diamine and dithiocarbamate groups. The concept has potentially widespread applicability as it is neither limited to metal extractions, nor to the use of silica-based nanoscavengers. Minimal involvement of organic solvents make nanoscavengers a potentially environmentally benign ("green") alternative to many conventional solvent extraction techniques.

Journal Article↗

Photolithographic generation of protein micropatterns for neuron culture applications.

Standard positive photoresist techniques were adapted to generate micropatterns of proteins on glass and oxide surfaces. Both lift-off and plasma-etching techniques were used to transfer the photoresist pattern into a layer of covalently immobilised protein. The surface properties of the areas adjacent to the patterns were altered by chemical surface modification. Using a combination of the lift-off and the etching process complementary patterns of two different proteins were generated. The biochemical and biological functionality of the protein patterns were assessed by immunostaining and by investigating the outgrowth of neurites from neurons plated on the patterned substrates. The investigated patterning processes are compatible with microstructuring and thin film processes, and may be used to generate functional surfaces for sensor and neuron culture applications.

Animals↗

Proteoglycans at the bone-implant interface.

The widespread success of clinical implantology stems from bone's ability to form rigid, load-bearing connections to titanium and certain bioactive coatings. Adhesive biomolecules in the extracellular matrix are presumably responsible for much of the strength and stability of these junctures. Histochemical and spectroscopic analyses of retrievals have been supplemented by studies of osteoblastic cells cultured on implant materials and of the adsorption of biomolecules to titanium powder. These data have often been interpreted to suggest that proteoglycans permeate a thin, collagen-free zone at the most intimate contact points with implant surfaces. This conclusion has important implications for the development of surface modifications to enhance osseointegration. The evidence for proteoglycans at the interface, however, is somewhat less than compelling due to the lack of specificity of certain histochemical techniques and to possible sectioning artifacts. With this caveat in mind, we have devised a working model to explain certain observations of implant interfaces in light of the known physical and biological properties of bone proteoglycans. This model proposes that titanium surfaces accelerate osseointegration by causing the rapid degradation of a hyaluronan meshwork formed as part of the wound-healing response. It further suggests that the adhesive strength of the thin, collagen-free zone is provided by a bilayer of decorin proteoglycans held in tight association by their overlapping glycosaminoglycan chains.

Animals↗

Osteoblast response and calcium deposition on phospholipid modified surfaces.

In this study, the effect of calcium phosphate complexed phospholipid (Ca-PL-PO4) coatings on solid surfaces on the in vitro calcium (Ca) deposition and on the osteoblast responses was evaluated. Commercially available phospholipids were converted to their Ca-PL-PO4, and were coated on glass Petri dishes. The coated dishes were immersed in the simulated body fluid for up to 14 days under sterilized conditions at 37 degrees C, and the amount of calcium (Ca) deposited was quantified. Similarly, by measuring the alkaline phosphatase specific activity, the differentiation of osteoblast precursor cells were evaluated after seeding the cells on Ca-PL-PO4 coated cell culture plastics. It was observed that all Ca-PL-PO4 enhanced Ca deposition on coated surfaces. The, polar head group of phospholipids in coated surfaces was observed to have an influence on the Ca deposition as well as the osteoblast differentiation. Among the phospholipids evaluated, phosphatidylserine (Ca-PS-PO4) exhibited the strongest calcium deposition and more enhanced alkaline phosphatase specific activity. It was therefore concluded from this study that Ca-PS-PO4 surface modification may be an alternative method for enhancing bone-implant interactions.

Body Fluids↗

Liposome-mediated drug targeting in topical and regional therapies.

Liposome-mediated drug targeting is reviewed in four major categories of topical and regional therapies: wounds and burns, ocular, intraperitoneal, and pulmonary. A survey of the data in the field is preceded by definitions of carrier-mediated drug targeting, in particular for topical and regional treatments. The ability of liposomes to meet essential requirements for task performance and liposome surface-modification as the major approach to endow liposomes with targeting abilities are reviewed. Analysis of current findings in the field shows that (1) most studies explored regular liposomes that were unable to meet the essential requirements for targeting and (2) in vivo drug targeting in topical and regional therapies has been achieved rarely and seldom attempted, yet there are encouraging indications from a few studies that using surface-modified liposomes such targeting is feasible. Both established and novel liposomal systems attest to this feasibility and point out future directions. The former can be found by revisiting immunoliposomes that were initially designed for systemic administration but might well fit topical and regional cases. The latter is exemplified by bioadhesive liposomes, designed specifically for topical/regional therapies. It is concluded that careful implementation of such approaches could be successful for the achievement of liposome-mediated drug targeting in topical and regional therapies.

Administration, Topical↗

Shear bond strength of veneering composite resin to titanium nitride coating alloy deposited by radiofrequency sputtering.

OBJECTIVE: The success of dental restorations utilizing composite resin over cast alloy has been variable and affected by micro-leakage. The purpose of this study was to investigate the shear bond strength between a composite resin and cast dental alloy coated with a layer of titanium nitride (TiN). METHODS: Cast disk specimens made of Au-Pd-Ag alloy were randomly divided into four groups. The first group was left uncoated as a control while one group was coated with a metal conditioner, and the other two were coated with TiN at either 200 or 500 W with a radiofrequency magnetron sputtering system. Each group was then bonded to one of two types of light-activated composite resin material. Shear bond strengths were tested and compared for each specimen. The bond strength of each group was analyzed by 2-way ANOVA and by post hoc multiple comparison tests (Fisher's PLSD). RESULTS: As the interaction among groups was not significant (p=0.27) by 2-way ANOVA, TiN coated groups were found to have improved shear bond strength compared with non-coated groups without metal conditioner for both resin composite materials by multiple comparison tests. Furthermore, TiN coating is significantly stronger than non-coated groups with metal conditioner for one of the two resin composite materials examined (p<0.01). CONCLUSIONS: The results suggested that metal surface modification by coated TiN represents a technique that may significantly improve the bond between the Au-Pd-Ag alloy and veneering resin composite.

Acrylic Resins↗

Immobilization of gold nanorods onto acid-terminated self-assembled monolayers via electrostatic interactions.

We report the immobilization of gold nanorods onto self-assembled monolayers (SAMs) of 16-mercaptohexadecanoic acid (16-MHA). The simple two step protocol involves formation of a SAM of 16-MHA molecules onto gold-coated glass slides and subsequent immersion of these slides into the gold nanorod solution. The nanorods, formed by a seed-mediated, surfactant-assisted synthesis protocol, are stabilized in solution due to surface modification by the surfactant cetyltrimethylammonium bromide (CTAB). Attractive electrostatic interactions between the carboxylic acid group on the SAM and the positively charged CTAB molecules are likely responsible for the nanorod immobilization. UV-vis spectroscopy has been used to follow the kinetics of the nanorod immobilization. The nature of interaction between the gold nanorods and the 16-MHA SAM has been probed by Fourier transform infrared spectroscopy (FTIR). The surface morphology of the immobilized rods is studied by scanning electron microscopy (SEM) and atomic force microscopy (AFM) measurements. SEM was also used to determine the density of the immobilized nanorods as a function of the pH of immobilization. Control over the surface coverage of the immobilized gold nanorods has been demonstrated by simple pH variation. Such well-dispersed immobilized gold nanorods with control over the surface coverage could be interesting substrates for applications such as surface-enhanced Raman spectroscopy (SERS).

Journal Article↗

Cytotoxicity of colloidal CdSe and CdSe/ZnS nanoparticles.

Cytotoxicity of CdSe and CdSe/ZnS nanoparticles has been investigated for different surface modifications such as coating with mercaptopropionic acid, silanization, and polymer coating. For all cases, quantitative values for the onset of cytotoxic effects in serum-free culture media are given. These values are correlated with microscope images in which the uptake of the particles by the cells has been investigated. Our data suggest that in addition to the release of toxic Cd(2+) ions from the particles also their surface chemistry, in particular their stability toward aggregation, plays an important role for cytotoxic effects. Additional patch clamp experiments investigate effects of the particles on currents through ion channels.

Animals↗