Biomedical subjects

N D Spencer

Publications and source records attributed to N D Spencer.

15 recordsLinked to original sources

PubMed2005

Protein-mediated boundary lubrication in arthroplasty.

Wear of articulated surfaces can be a major lifetime-limiting factor in arthroplasty. In the natural joint, lubrication is effected by the body's natural synovial fluid. Following arthroplasty, and the subsequent reformation of the synovial membrane, a fluid of similar composition surrounds the artificial joint. Synovial fluid contains, among many other constituents, a substantial concentration of the readily adsorbing protein albumin. The ability of human serum albumin to act as a boundary lubricant in joint prostheses has been investigated using a pin-on-disc tribometer. Circular dichroism spectroscopy was employed to follow the temperature- and time-dependent conformational changes of human serum albumin in the model lubricant solution. Effects of protein conformation and polymer surface hydrophilicity on protein adsorption and the resulting friction in the boundary lubrication regime have been investigated. Unfolded proteins preferentially adsorb onto hydrophobic polymer surfaces, where they form a compact, passivating layer and increase sliding friction-an effect that can be largely suppressed by rendering the substrate more hydrophilic. A molecular model for protein-mediated boundary friction is proposed to consolidate the observations. The relevance of the results for in vivo performance and ex vivo hip-joint testing are discussed.

Adsorption↗

PubMed2004

Interaction forces and morphology of a protein-resistant poly(ethylene glycol) layer.

The molecular interactions on a protein-resistant surface coated with low-molecular-weight poly(ethylene glycol) (PEG) copolymer brushes are investigated using the extended surface forces apparatus. The observed interaction force is predominantly repulsive and nearly elastic. The chains are extended with respect to the Flory radius, which is in agreement with qualitative predictions of scaling theory. Comparison with theory allows the determination of relevant quantities such as brush length and adsorbed mass. Based on these results, we propose a molecular model for the adsorbed copolymer morphology. Surface-force isotherms measured at high resolution allow distinctive structural forces to be detected, suggesting the existence of a weak equilibrium network between poly(ethylene glycol) and water--a finding in accordance with the remarkable solution properties of PEG. The occurrence of a fine structure is interpreted as a water-induced restriction of the polymer's conformational space. This restriction is highly relevant for the phenomenon of PEG protein resistance. Protein adsorption requires conformational transitions, both in the protein as well as in the PEG layer, which are energetically and kinetically unfavorable.

Adsorption↗

PubMed2003

Surface modification of PLGA microspheres.

Microspheres made of poly(lactic-co-glycolic acid) (PLGA) are biocompatible and biodegradable, rendering them a promising tool in the context of drug delivery. However, nonspecific adsorption of plasma proteins on PLGA micro- and nanospheres is a main limitation of drug targeting. Poly(L-lysine)-g-poly(ethylene glycol) (PLL-g-PEG), physisorbed on flat metal oxide surfaces, has previously been shown to suppress protein adsorption drastically. The goal of our work was to characterize the efficiency of the protein repellent character of PLL-g-PEG on PLGA microspheres and to show the feasibility of introducing functional groups on the PLGA microspheres via functionalized PLL-g-PEG. To quantify the adsorbed amount of protein, a semiquantitative method that uses confocal laser scanning microscopy (CLSM) was applied. The first part of the experiment confirms the feasibility of introducing specific functional groups on PLL-g-PEG-coated PLGA microspheres. In the second part of the experiment, PLL-g-PEG-coated PLGA microspheres show a drastic decrease of adsorbed proteins by two orders of magnitude in comparison to uncoated PLGA microspheres. Low protein-binding, functionalizable microspheres provide a fundamental basis for the design of drug delivery and biosensor systems.

Biosensing Techniques↗

PubMed2003

Chemically patterned, metal oxide based surfaces produced by photolithographic techniques for studying protein- and cell-surface interactions I: Microfabrication and surface characterization.

Chemical patterns on smooth wafer substrates comprising areas with two different metals have been produced by vacuum metal deposition and photolithographic techniques. The combination of metals has been chosen from the series titanium (Ti), aluminium (Al), vanadium (V) and niobium (Nb), producing patterns (dots and stripes with dimensions of 50, 100 and 150 micrometer) with one of the metals as the background and with the second metal (foreground pattern) deposited on the background metal. The structure and chemical composition of the patterned surfaces were evaluated by scanning electron microscopy, X-ray photoelectron spectroscopy and imaging time-of-flight secondary-ion mass spectrometry. The surfaces proved to be geometrically well defined with the expected surface-chemical composition, i.e. a surface oxide (passive) film essentially composed of TiO(2),Al(2)O(3),V(2)O(5), or Nb(2)O(5). Ti/Ti patterned surfaces were produced as controls and found to show no chemical composition contrast. The surface roughness of the pattern was greater than that of the background by a factor of 2-3, but was still extremely smooth with Ra<2nm. The patterns serve as model surfaces for studying in vitro the behaviour of cells as well as the adsorption of serum proteins on different metal oxides, which will be reported in a companion paper. These surfaces can be used to compare and contrast the response of osteoblasts to Ti and other alloy components, such as Al, V, or Nb, which are used in load-bearing medical implants.

Aluminum↗

PubMed2002

Optical grating coupler biosensors.

By incorporating a grating in a planar optical waveguide one creates a device with which the spectrum of guided lightmodes can he measured. When the surface of the waveguide is exposed to different solutions, the peaks in the spectrum shift due to molecular interactions with the surface. Optical waveguide lightmode spectroscopy (OWLS) is a highly sensitive technique that is capable of real-time monitoring of these interactions. Since this integrated optical method is based on the measurement of the polarizability density (i.e., refractive index) in the vicinity of the waveguide surface, radioactive, fluorescent or other kinds of labeling are not required. In addition, measurement of at least two guided modes enables the absolute mass of adsorbed molecules to be determined. In this article, the technique will be described in some detail, and applications from different areas will be discussed. Selected examples will be presented to demonstrate how monitoring the modification of different metal oxides with polymers and the response of the coated oxides to biofluids help in the design of novel biomaterials; how OWLS is useful for accurate bioaffinity sensing, which is a key issue in the development of new drugs; and how the quantitative study of protein-DNA/RNA and cell surface interactions can enhance the understanding of processes in molecular and cellular biology.

Adsorption↗

PubMed2002

Comparative investigation of the surface properties of commercial titanium dental implants. Part I: chemical composition.

The surfaces of five commercially available titanium implants (Brånemark Nobel Biocare, 3i ICE, 3i OSSEOTITE, ITI-TPS, and ITI-SLA) were compared by scanning electron microscopy, X-ray photoelectron spectroscopy, time-of-flight secondary ion mass spectroscopy. All five implant types were screw-shaped and fabricated from commercially pure (cp) titanium, but their surface properties differed both as regards surface morphology and surface chemical composition. The macro- and microstructure of the implant surfaces were investigated by scanning electron microscopy. The surfaces chemical composition was determined using the surface-sensitive analytical techniques of X-ray photoelectron spectroscopy and time-of-flight secondary ion spectrometry. Surface topographies were found to reflect the type of mechanical/chemical fabrication procedures applied by the manufacturers. The titanium oxide (passive) layer thickness was similar (5-6 nm) and typical for oxide films grown at or near room temperature. A variety of elements and chemical compounds not related to the metal composition were found on some implant types. They ranged from inorganic material such as sodium chloride to specific organic compounds believed to be due to contamination during fabrication or storage. The experimental findings are believed to make a contribution to a better understanding of the interplay between industrial fabrication procedure and physico-chemical implant surface properties.

Journal Article↗

PubMed2001

Density fluctuations under confinement: when is a fluid not a fluid?

Knowing the behavior of a fluid in small volumes is essential for the understanding of a vast array of common problems in science, such as biological interactions, fracture propagation, and molecular tribology and adhesion, as well as pressure solvation and other geophysical processes. When a fluid is confined, its phase behavior is altered and excluded-volume effects become apparent. Pioneering measurements performed with the surface forces apparatus have revealed so-called structural or oscillatory solvation forces as well as the occurrence of a finite shear stress, which was interpreted as a solidification transition. Here, we report measurements obtained with an extended surface forces apparatus, which makes use of fast spectral correlation to gain insight into the behavior of a thin film of cyclohexane confined within attoliter volumes, with simultaneous measurement of film thickness and refractive index. With decreasing pore width, cyclohexane is found to undergo a drastic transition from a three-dimensional bulk fluid to a two-dimensional adsorbate with strikingly different properties. Long-range density fluctuations of unexpected magnitude are observed.

Journal Article↗

PubMed2000

Feasibility study of an online toxicological sensor based on the optical waveguide technique.

Morphological properties of the cells often change as an early response to the presence of a pharmacologically acting toxic substance [Etcheverry, S.B., Crans, D.C., Keramidas, A.D., Cortizo, A.M., Arch. Biochem. Biophys. 338 (1997) 7-14]. Recently it has been shown that living animal cell adhesion and spreading can be monitored online and quantitatively via the interaction of the cells with the evanescent electromagnetic field present at the surface of an optical waveguide [Ramsden, J.J., Li, S.Y., Heinzle, E., Prinosil, J.E. Cytometry 19 (1995) 97-102]. In the present study, optical waveguide lightmode spectroscopy (OWLS) and confocal laser scanning microscopy (CLSM), which provides information about the shape of the cells at the surface, were compared under identical experimental conditions. This allowed for the correlation between the cell-shape information from CLSM and the cell-surface interaction measurements from OWLS. The proposed design of the microsystem sensor involves the establishment of a cell layer on the surface of the waveguide and the subsequent online measurement of the morphological response of the cells to various toxic substances. In the present study, the setup was evaluated using cells from an osteoblastic MC 3T3-E1 cell line, and sodium hypochlorite was used as model toxic substance. Comparing the OWLS signal to the morphological response measured by CLSM reveals that OWLS is effective in monitoring not only cell attachment and spreading but also the cellular response to toxic compounds (i.e. by means of change in cell morphology). For the model toxin, the OWLS measurements indicate that, at concentrations above 0.01%, the cells exhibit a clearly discernable morphological effect (i.e. a decrease in average cell contact area). Thus, the potential of an on-line sensor based on OWLS to applications in toxicology, pharmacy and biocompatibility was demonstrated.

3T3 Cells↗

PubMed1999

Surface characterization of implant materials c.p. Ti, Ti-6Al-7Nb and Ti-6Al-4V with different pretreatments.

The biocompatibility of commercially pure titanium and its alloys is closely related to their surface properties, with both the composition of the protecting oxide film and the surface topography playing an important role. Surfaces of commercially pure titanium and of the two alloys Ti-6Al-7Nb and Ti-6Al-4V (wt %) have been investigated following three different pretreatments: polishing, nitric acid passivation and pickling in nitric acid-hydrogen fluoride. Nitric acid treatment is found to substantially reduce the concentration of surface contaminants present after polishing. The natural 4-6 nm thick oxide layer on commercially pure titanium is composed of titanium oxide in different oxidation states (TiO2, Ti2O3 and TiO), while for the alloys, aluminium and niobium or vanadium are additionally present in oxidized form (Al2O3, Nb2O5 or V-oxides). The concentrations of the alloying elements at the surface are shown to be strongly dependent on the pretreatment process. While pickling increases the surface roughness of both commercially pure titanium and the alloys, different mechanisms appear to be involved. In the case of commercially pure titanium, the dissolution rate depends on grain orientation, whereas in the case of the two alloys, selective alpha-phase dissolution and enrichment of the beta-phase appears to occur.

Journal Article↗

PubMed1999

Microstructured bioreactive surfaces: covalent immobilization of proteins on Au(1 1 1)/silicon via aminoreactive alkanethiolate self-assembled monolayers.

Micrometer-scale patterns of a defined surface chemistry and structure were produced on both ultraflat Au(1 1 1) and on gold-coated monocrystalline silicon surfaces by a method combining microcontact printing, wet chemical etching and the replacement of etch-resist self-assembled monolayers (SAMs) by functionalized or reactive SAMs. Key steps in this methodology were characterized by X-ray photoelectron spectroscopy (XPS), ellipsometry and contact angle measurements. The covalent immobilization of (functional) biological systems on these surfaces was tested using an N-hydroxysuccinimide ester omega-functionalized disulphide (DSU), which covalently binds primary amines without the need for further activation steps. Atomic force microscope images of native collagen V single molecules immobilized on these patterned surfaces revealed both high spatial resolution and strong attachment to the monolayer/gold surface. Microcontact printing of DSU is shown to be feasible on specially prepared, ultraflat Au(1 1 1) surfaces providing a valuable tool for scanning probe experiments with biomolecules. The retention of enzymatic activity upon immobilization of protein was demonstrated for the case of horseradish peroxidase. The described approach can thus be used to confine biological activity to predetermined sites on microstructured gold/silicon devices - an important capability in biomedical and biomolecular research.

Journal Article↗

PubMed1999

Characterization of anodic spark-converted titanium surfaces for biomedical applications.

The aim of the present study was to characterize the surface morphology, microstructure and the chemical composition of anodic spark-converted titanium surfaces. The coatings were prepared in an electrochemical cell by the anodic spark deposition technique in an aqueous solution of Ca(H2PO4)2. The coatings were characterized by X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), electron probe microanalysis (EPMA) and X-ray diffraction (XRD). The properties of the coatings are described in terms of morphology.

Journal Article↗

PubMed1999

The implant material, Ti6Al7Nb: surface microstructure, composition and properties.

The excellent biocompatibility of titanium and its alloys is intimately related with the properties of the surface in contact with the biological environment, and therefore it is closely connected with the stable, passivating oxide layer that forms on its surface. In the present paper, the oxide layer on the alloy Ti6Al7Nb has been characterized using X-ray photoelectron spectroscopy, scanning Auger microscopy and pH-dependent lateral force microscopy. The alloying elements Al and Nb are incorporated in the oxide layer and detected in their most stable oxidized form, as Al2O3 and Nb2O5. Their distribution in the oxide reflects the underlying alpha-beta microstructure, with enrichment of Al in the alpha- and of Nb in the beta-phase (determined by electron microprobe). Friction measurements (lateral force microscopy) indicate slightly different, pH-dependent, lateral forces above the alpha- and beta-phase structures that point to small local variations in surface charges.

Journal Article↗

PubMed1997

Immobilization of the cell-adhesive peptide Arg-Gly-Asp-Cys (RGDC) on titanium surfaces by covalent chemical attachment.

Surface modification of acid-pretreated titanium with 3-aminopropyltriethoxylsilane (APTES) in dry toluene resulted in covalently bonded siloxane films with surface coverage that was relatively controllable by regulating the reaction conditions. A hetero-bifunctional cross-linker, N-succinimidyl-3-maleimidopropionate (SMP), reacted with the terminal amino groups, forming the exposed maleimide groups. Finally, a model cell-binding peptide, Arg-Gly-Asp-Cys (RGDC), was immobilized on the surface through covalent addition of the cysteine thiol groups to the maleimide groups. X-ray photoelectron spectroscopy, radiolabelling techniques, and ellipsometry were used to quantify and characterize the modified surfaces.

Journal Article↗

PubMed1997

Surface activation of polyetheretherketone (PEEK) and formation of calcium phosphate coatings by precipitation.

Plasma activation of polyetheretherketone (PEEK) surfaces and the influence on coating formation in a supersaturated calcium phosphate solution was investigated in this study. It was observed that plasma treatment in a N2/O2 plasma had a significant effect on the wettability of the PEEK surface. The contact angle decreased from 85 degrees to 25 degrees after plasma treatment. Cell culture testing with osteoblastic cell lines showed plasma activation not to be disadvantageous to cell viability. X-ray photoelectron spectroscopy (XPS) analysis was performed to characterize the chemical composition of the PEEK surfaces. It was observed that the O1s intensity increased with plasma activation time. At the C1s peak the appearance of a shoulder at higher binding energies was observed. Coating of PEEK was performed in a supersaturated calcium phosphate solution. Coating thicknesses of up to 50 microm were achieved after 24 days of immersion. Plasma activation followed by nucleation in a highly saturated hydroxyapatite solution had a positive effect on the growth rate of the layer on PEEK. Chemical analysis revealed that the coating consists of a carbonate-containing calcium phosphate.

Journal Article↗

PubMedDate not supplied

Wavelength-dependent roughness: a quantitative approach to characterizing the topography of rough titanium surfaces.

Topographies of grit-blasted, etched, grit-blasted and etched, and microfabricated and etched surfaces of commercially pure titanium have been investigated. Such surface topographies vary across the scale range of interest for dental implants, extending from nanometers to millimeters. The complete characterization of topography requires the use of complementary methods. This study compared the topographic characterization methods of non-contact laser profilometry, interference microscopy, stereo-scanning electron microscopy (stereo-SEM), and atomic force microscopy. Non-contact laser profilometry was shown to be a useful method to characterize topographic features in the micron to millimeter range, whereas interference microscopy and stereo-SEM can be employed down to the submicron range. Stereo-SEM is particularly useful for quantifying topographies with complex, strongly corrugated ("sharp"), and high-aspect-ratio features and was shown to be complementary to non-contact laser profilometry and interference microscopy. Because of tip-related envelope problems, atomic force microscopy was not found to be suitable for the type of surfaces investigated in this study. Independent of the method used, the commonly used "integral" amplitude roughness parameters, such as Ra, Rq, or Rt, were often of limited value in the description of actual implant surfaces. The application of the wavelength-dependent roughness approach was shown to be an effective method for the description of surface topographies in the complete range of characteristic roughness and is also a useful means of examining the effects of surface treatment processes.

Analysis of Variance↗