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PubMed1997

Retention of enzymatic activity immobilized on silanized Co-Cr-Mo and Ti-6Al-4V.

Biochemical surface modification of biomaterials utilizes immobilized biomolecules to induce preferred tissue responses. Operational stability, or retention of biological activity, of biochemically modified biomaterials is a fundamental determinant of their usefulness. The present study investigated retention of enzymatic activity immobilized on silanized Co-Cr-Mo and Ti-6Al-4V. A model enzyme, trypsin, was immobilized on monolayers and multilayers of silane deposited from aqueous or organic solutions of gamma-aminopropyltriethoxysilane (APS). Trypsin-conjugated biomaterials were incubated in cell culture medium at 37 degrees C for up to 96 h, and the residual immobilized activity was measured. Retention of bioactivity in physiological saline was dependent on the type of material and on the method of silanization. Activity of enzyme adsorbed on the metals was lost within 24-48 h. Both mono- and multilayers of APS deposited on Co-Cr-Mo by aqueous silanization effectively retained enzymatic activity throughout the 96 h incubation period. The monolayer retained approximately 23% of the activity initially present, and the multilayers retained approximately 50% of the initial activity. Organic silanization of Co-Cr-Mo was marginally effective as it initially slowed the loss of activity. However, all activity was lost by 48-72 h of incubation. Neither organic nor aqueous silanization enhanced retention of enzymatic activity on Ti-6Al-4V.

Alloys↗

PubMed2000

Blood compatibility of particulate carriers investigated by cell electrophoresis and particle electrophoresis: a preliminary report.

Polyethylene glycol (PEG) and dextran were covalently coupled, or only adsorbed, to the surface of three kinds of inorganic particles in order to shield their surface and reduce their nonspecific binding to red blood cells. Surface modification as well as interaction of particles with red blood cells was followed up by particle electrophoresis. This allowed a quick evaluation of the efficiency of polymer coupling. Moreover, the nonspecific binding of particles to red blood cells was easily investigated with cell electrophoresis, showing the inhibitory effect of immobilized PEG-5000 or dextran. The electrophoretic mobility analysis presented here may be used for screening blood compatibility of particulate drug carriers and could be helpful in formulating long-living circulating particles.

Animals↗

PubMed2000

Development, characterization, and anti-microbial efficacy of hydroxyapatite-chlorhexidine coatings produced by surface-induced mineralization.

The surface-induced mineralization (SIM) technique was used to produce hydroxyapatite (HAP) coatings on external fixation pins with the antimicrobial agent, chlorhexidine, incorporated within the coating. The SIM process involved surface modification of the substrate with organic functional groups followed by immersion in aqueous supersaturated calcium phosphate solutions. X-ray diffraction spectra confirmed that hydroxyapatite coatings were formed. Chlorhexidine was incorporated into the coating by placing the substrate into various chlorhexidine solutions in between mineralization cycles. Total uptake was measured by dissolution of the coating into a 0.1 M nitric acid solution and measuring the chlorhexidine concentration using UV spectroscopy at 251 nm. Release rates were measured by submersion of coated substrates into saline solutions and measuring chlorhexidine UV absorbency at 231 nm as a function of time. Results show an initial rapid release followed by a period of slower sustained release. The anti-microbial efficacy of the HAP-chlorhexidine coatings was evaluated in vitro using a Staphylococcus aureus cell culture. Initial results show a large "inhibition zone" formed around the chlorhexidine/HAP coating vs. coatings with HAP only. This preliminary work clearly demonstrates that SIM HAP coatings have great potential to locally deliver antimicrobial agents such as chlorhexidine at implantation sites, which may greatly reduce the incidence of pin tract infection that occurs in external fixation.

Anti-Infective Agents, Local↗

PubMed2001

Preservation of platelet function on 2-methacryloyloxyethyl phosphorylcholine-graft polymer as compared to various water-soluble graft polymers.

The chemical structures of water-soluble polymers grafted onto PE surfaces affect platelet function when the platelets contact the polymer surfaces. To improve our understanding of this effect, this study sought to control the blood/materials interaction on the surfaces of polyethylene (PE) by grafting with various water-soluble polymers. Such polymers as poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC), poly(acrylamide) (PAAm), poly(N-vinylpyrrolidone) (PVPy), and poly[monomethacryloyl poly(ethylene glycol)] (PMPEG) were grafted on low-density PE sheets by photoinduced graft polymerization. Both the PE bags modified with water-soluble polymers and those nonmodified were prepared by heat processing. Activation of platelets after storage in the PE bags was evaluated by measuring the cytoplasmic free calcium ion concentration ([Ca(2+)]i). The concentration of [Ca(2+)]i of platelets in contact with the PE surface grafted with PMPC was the same as that of native platelets and significantly less than that in contact with other PE surfaces grafted with water-soluble polymers. The number of adherent platelets was effectively decreased on PE surfaces grafted with PMPC and PMPEG, as compared with nontreated PE. The aggregation ability of platelets was also measured after storage of platelet-rich plasma in the PE bags. The PE surface grafted with PMPC effectively maintained aggregation ability as compared with both the nontreated PE and with PE grafted with PAAm, PVPy, and PMPEG. It was concluded that for preserving platelet function, PMPC was the most effective of these water-soluble polymers used for surface modification.

Animals↗

PubMed1981

The ultrastructure of the thin limbs of Henle in Kidneys of the desert heteromyid (Perognathus penicillatus).

The thin limbs of both long- and short-looped nephrons in Perognathus kidneys were studied with transmission and scanning electron microscopy. The superficial nephrons have a short thin limb located in the vascular bundles of the outer medulla and are characterized by a simple, low-lying epithelium (0.4 +/- 0.1 mu thickness). In contrast, the first descending part of the thin limb of the majority of midcortical and juxtamedullary nephrons has a relatively thick epithelium (1.7 +/- 0.6 mu in thickness) with marked lateral and basal interdigitation and a dense surface covering of microvilli. The remaining part of the long descending thin limb is relatively simple with a low-lying epithelium (0.6 +/- 0.1 mu in thickness), decorated on its surface by sparse microplicae. The bend of the loop and the ascending limb are covered by a very simple low-lying epithelium (0.6 +/- 0.2 mu in thickness) with relatively little surface modification. The extreme urine-concentrating ability of Perognathus does not appear to be due to the development of a unique thin loop epithelium but rather to the extensive length of the inner and outer medulla.

Animals↗

PubMed1975

Transmission and scanning electron microscope studies of calcified cartilage resorption.

The authors' previous report (Savostin-Asling and Asling, '73) demonstrated that Meckel's carilage is a favorable site for study of calcified cartilage resorption. In the present study the ultrastructural features at this resorption front have been examined by transmission and scanning electron microscopes (19-day rat retus). Multinucleated giant cells chondroclasts) dominated the erosion front. The many features which they showed in common with osteoclasts included abundant mitochondria, vacuolation, lysonsomes, sparsity of rough-sufaced endoplasmic reticulum, and deep infoldings at loci of contact with calcified matrix. Crumbling of matrix (with mineral crystals penetrating between these foldings) and fragmentation of collagen fibrils were also seen. The propensity of chondroclasts for spanning several opened lacunae provided special opportunity to demonstrate cell surface modifications in presence or absence of matrix contact. Amebiod processes extending into lacunae were seen by both transmission and scanning procedures; they were sometimes tipped with a veil of filamentous processes as small as 0.3 mum in diameter. Most hypertrophic chondrocytes. when released from lacunae, appeared to be disintegrating. However, in accord with previous evidence of their possible merger with chondroclasts (in light microscopic studies) there was also evidence for breakdown of cell walls between a chondroclast and a chondrocyte in intimate contact, with possibility of cytoplasmic continuity.

Animals↗

PubMed1976

A scanning electron microscope study of the interstitial tissue of the canine testis.

Scanning electron microscopy (SEM) is a potent tool that is especially valuable in interpreting the three-dimensional relationships of cells within tissues. This type of information is obtainable from thin sections in transmission electron microscopy (TEM) only by reconstructions of serial sections. The arrangement of the interstitial cells of the testis in relation to the capillaries and lymphatic channels, in particular, is easier to visualize in SEM than in TEM. Cytoplasmic constituents, as well as cell surface modifications, are demonstrable by this technique. The presence of droplets, presumably lipid droplets, both within and on the Leydig cells and the lymphatic endothelial cells, is quite evident. Other cytoplasmic structures are also apparent. For example, the possible functional significance of "openings" that are seen by SEM on the septa that surround lipid droplets is discussed relative to the appearance of the same area as seen in thin sections or in freeze-fracture replicas. SEM should become a very useful method for studying cytological and morphological alterations that occur in testicular tissue that is subjected to physical or chemical manipulation.

Animals↗

PubMed1980

A new organic solvent for use in the clearing of tissues. I. Soft tissue histology.

Histosol is a non-flammable solvent mixture of synthetic aromatic hydrocarbons with a flash point of 124 degrees F (T.C.C.). It has a lower vapor pressure and evaporation rate than other organic solvents, such as xylene, routinely used as clearing and deparaffinizing agents. Although both xylene and Histosol clear and deparaffinize soft organ tissues effectively in the preparation of permanently mounted stained slides, Histosol appears, in many instances, to be the choice solvent: tissues are easier to section; cell borders and cell surface modifications are most distinct; cytoplasmic eosinophilia is more vivid; and nuclear detail is improved. Of prime importance, Histosol is a safer and more efficient solvent for use in histological and pathological laboratories.

Animals↗

PubMed2006

Electrospun scaffold tailored for tissue-specific extracellular matrix.

The natural extracellular matrix (ECM) is a complex structure that is built to meet the specific requirements of the tissue and organ. Primarily consisting of nanometer diameter fibrils, ECM may contain other vital substances such as proteoglycans, glycosaminoglycan and various minerals. Current research in tissue engineering involves trying to replicate the ECM such that it provides the environment for tissue regeneration. Electrospinning is a versatile process that results in nanofibers by applying a high voltage to electrically charge a liquid. A variety of polymers and other substances have been incorporated into the artificial nanofibrous scaffold. Surface modification and cross-linking of the nanofibers are some ways to improve the biocompatibility and stability of the scaffold. Electrospun scaffolds with oriented nanofibers and other assemblies can be constructed by modifying the electrospinning setup. Using electrospinning, researchers are able to specifically tailor the electrospun scaffold to meet the requirements of the tissue that they seek to regenerate. In vitro and in vivo experiments demonstrate that electrospun scaffolds hold great potential for tissue engineering applications.

Animals↗

PubMed2002

Capillary electrophoresis of Cr(VI) reducer Arthrobacter oxydans.

Rapid and effective separation of bacteria Arthrobacter oxydans was performed using capillary electrophoresis. For optimal separation of bacteria the influence of buffer concentration, pH and applied voltage were studied. It was found that the most appropriate conditions for electrophoretic mobility measurements are as follows: applied voltage 6-14 kV; buffer concentration 5-10 mM pH 6-8. At the stationary phase of growth there are always two main heterogeneous peaks. They are connected with the morphology of bacteria as well as with cell aggregation. The heterogeneity of samples may be explained by surface modifications of bacterial cells.

Arthrobacter↗

PubMed1996

Influence of fibronectin on HIV-1 infection and capability of binding to platelets.

We have previously demonstrated that fibronectin (FN) can bind HIV-1 envelope proteins, in particular gp 120. The aim of the present study was to determine some biological effects of this phenomenon. Pretreating HIV-1 with human FN increased the infectivity of HIV-1, when a low concentration of the virus was used. In contrast, an RGD-containing pentapeptide (Gly-Arg-Gly-Asp-Ser), which is a fundamental binding site of FN, reduced the infectivity of a suspension of HIV-1 at high concentrations of the virus. It is likely that FN bridges the cell surface and the virions, while the RGD-containing pentapeptide may saturate the HIV-1 binding sites for cell surface receptors. Moreover, gp 120 was bound to the FN present on the surface of platelets. The specificity of this binding was confirmed by the inhibition obtained by pretreating platelets with anti-FN antibodies. The consequence of the surface modifications of the platelets could explain the thrombocytopenia that frequently occurs in patients infected with HIV and suggests also the possibility that platelets could be a vehicle for the virus in the circulation.

Blood Platelets↗

PubMed2003

A molecular approach to self-assembly of trimethylsilylacetylene derivatives on gold.

We recently discovered that a linear multifunctional trimethylsilylacetylene (TMSA) compound forms long-range and highly stable self-assembled monolayers (SAMs) on reconstructed Au(111). To better understand the interactions governing self-assembly in this new system, we synthesized a series of homologue organosilanes and performed scanning tunneling microscopy (STM) measurements at the Au(111)/n-tetradecane interface. The four TMSA-terminated linear silanes that we synthesized self-assemble in similar ways on gold, with the molecules standing upright on the surface. In contrast, compounds with a slightly modified terminal group but the same polyunsaturated linear chain above the TMSA head do not self-assemble. In particular, substituting a methyl group of TMSA with a more bulky one prevents self-assembly. Removing the C triple bond C triple bond of TMSA or substituting the Si atom by a C atom also hinders self-assembly. Finally, substituting one methyl group of TMSA by a hydrogen atom induces self-assembly but in a different geometry, with the molecules lying flat on the gold surface in a quasi-epitaxy mode. Our molecular approach demonstrates the key role played by the TMSA head in self-assembly, its origin being twofold: 1) the TMSA layers are commensurate to the Au(111) adlattice along the <112> direction, and 2) the C triple bond C triple bond of TMSA activates the Si atom and induces the creation of a surface Si-Au chemical bond. The highly stable TMSA-based SAMs appear then as promising materials for applications in surface modification.

Journal Article↗

PubMed2006

Use of carbonaceous polysaccharide microspheres as templates for fabricating metal oxide hollow spheres.

A general method for the synthesis of metal oxide hollow spheres has been developed by using carbonaceous polysaccharide microspheres prepared from saccharide solution as templates. Hollow spheres of a series of metal oxides (SnO2, Al2O3, Ga2O3, CoO, NiO, Mn3O4, Cr2O3, La2O3, Y2O3, Lu2O3, CeO2, TiO2, and ZrO2) have been prepared in this way. The method involves the initial absorption of metal ions from solution into the functional surface layer of carbonaceous saccharide microspheres; these are then densified and cross-linked in a subsequent calcination and oxidation procedure to form metal oxide hollow spheres. Metal salts are used as starting materials, which widens the accessible field of metal oxide hollow spheres. The carbonaceous colloids used as templates have integral and uniform surface functional layers, which makes surface modification unnecessary and ensures homogeneity of the shell. Macroporous films or cheese-like nanostructures of oxides can also be prepared by slightly modified procedures. XRD, TEM, HRTEM, and SAED have been used to characterize the structures. In a preliminary study on the gas sensitivity of SnO2 hollow spheres, considerably reduced "recovery times" were noted, exemplifying the distinct properties imparted by the hollow structure. These hollow or porous nanostructures have the potential for diverse applications, such as in gas sensitivity or catalysis, or as advanced ceramic materials.

Crystallography, X-Ray↗

PubMed2006

Silica-coated Ln3+-Doped LaF3 nanoparticles as robust down- and upconverting biolabels.

The preparation of nearly monodisperse (40 nm), silica-coated LaF(3):Ln(3+) nanoparticles and their bioconjugation to FITC-avidin (FITC=fluorescein isothiocyanate) is described in this report. Doping of the LaF(3) core with selected luminescent Ln(3+) ions allows the particles to display a range of emission lines from the visible to the near-infrared region (lambda=450-1650 nm). First, the use of Tb(3+) and Eu(3+) ions resulted in green (lambda=541 nm) and red (lambda=591 and 612 nm) emissions, respectively, by energy downconversion processes. Second, the use of Nd(3+) gave emission lines at lambda=870, 1070 and 1350 nm and Er(3+) gave an emission line at lambda=1540 nm by energy downconversion processes. Additionally, the Er(3+) ions gave green and red emissions and Tm(3+) ions gave an emission at lambda=800 nm by upconversion processes when codoped with Yb(3+) (lambda(ex)=980 nm). Bioconjugation of avidin, which has a bound fluorophore (FITC) as the reporter, was carried out by means of surface modification of the silica particles with 3-aminopropyltrimethoxysilane, followed by reaction with the biotin-N-hydroxysuccinimide activated ester to form an amide bond, imparting biological activity to the particles. A 25-fold or better increase in the FITC signal relative to the non-biotinylated silica particles indicated that there is minimal nonspecific binding of FITC-avidin to the silica particles.

Avidin↗

PubMed2005

Si-C linked organic monolayers on crystalline silicon surfaces as alternative gate insulators.

Herein, the influence of silicon surface modification via Si-C(n)H(2n+1) (n=10,12,16,22) monolayer-based devices on p-type 100 and n-type 100 silicon is studied by forming MIS (metal-insulator-semiconductor) diodes using a mercury probe. From current density-voltage (J-V) and capacitance-voltage (C-V) measurements, the relevant parameters describing the electrical behavior of these diodes are derived, such as the diode ideality factor, the effective barrier height, the flatband voltage, the barrier height, the monolayer dielectric constant, the tunneling attenuation factor, and the fixed charge density (Nf). It is shown that the J-V behavior of our MIS structures could be precisely tuned via the monolayer thickness. The use of n-type silicon resulted in lower diode ideality factors as compared to p-type silicon. A similar flatband voltage, independent of monolayer thickness, was found, indicating similar properties for all silicon-monolayer interfaces. An exception was the C10-based monolayer device on p-type silicon. Furthermore, low values of N(f) were found for monolayers on p-type silicon (approximately 6 x 10(11) cm(-2)). These results suggest that Si--C linked monolayers on flat silicon may be a viable material for future electronic devices.

Journal Article↗

PubMed2006

Effects of polymer architecture and composition on the adhesion of poly(tetrafluoroethylene).

Poly(glycidyl methacrylate), PGMA, chains in linear and arborescent structures were incorporated onto surfaces of poly(tetrafluoroethylene), PTFE, films by hydrogen plasma and ozone treatment and atom transfer radical polymerization. The epoxide groups of the PGMA chains were further reacted with acetic acid (AAc), oxalic acid (XAc), allyl amine (AA), and ethylenediamine (EDN) to introduce hydroxyl and amine groups to the surfaces of the PTFE films. Surface characterizations performed by Fourier Transform infrared attenuated total reflectance (FTIR-ATR) spectroscopy and X-ray photoelectron spectroscopy (XPS) confirmed the surface modification and the chemical structure. The PGMA chains in arborescent structures show a high effectiveness for the enhancement of the adhesion of PTFE films. The adhesion of PTFE films was also significantly enhanced by ring-opening reactions of the PGMA epoxide groups with acetic acid and amine compounds. A high value of 9.5 N cm(-1) in the optimum 180 degrees peel strength test was observed with PTFE/copper assemblies.

Journal Article↗

PubMed2003

Fabrication of porous polymer monoliths covalently attached to the walls of channels in plastic microdevices.

UV-initiated grafting of plastic tubes and microfluidic chips with ethylene diacrylate followed by the preparation of porous polymer monoliths has been studied. The first step affords a thin grafted layer of polymer with a multiplicity of pendent double bonds that are then used in the second step for covalent attachment of the monolith to the wall. As clearly seen on scanning electron micrographs, this procedure prevents the formation of voids at the monolith-channel interface a problem that has always plagued approaches involving bulk polymerization in nontreated channels due to the shrinkage of the monolith during the polymerization process and its lack of compatibility with the material of the device. Irradiation with UV light through a photomask allows precise patterning specifying both the area subjected to surface modification and the location of the monolith within specific areas of the device.

Microfluidics↗

PubMed2003

Sol-gel column technology for capillary isoelectric focusing of microorganisms and biopolymers with UV or fluorometric detection.

The sol-gel surface modification is used for capillary isoelectric focusing of microorganisms and biopolymers. The coating strongly decreases the electroosmotic flow so that it enables the use of the short capillaries down to 100 mm in the separation length. The examples of capillary isoelectric focusing of the low-molecular-mass pI markers and mixed cultures of microbial populations of Escherichia coli, Candida albicans, Staphylococcus epidermidis, and Enteroccocus faecalis with UV detection are shown. It is possible to quantify bacterial cells according to their peak areas; the minimum detectable number of microbial cells was 5 x 10(2)-1 x 10(3). The compatibility of sol-gel capillaries with sensitive fluorometric detection of fluorescent pI markers together with fluorescein labeled proteins is demonstrated.

Bacteria↗