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PubMed1989

Evaluation of blood compatibility of PEO grafted and heparin immobilized polyurethanes.

To develop better blood compatible polymer for long-term biomedical applications, poly(ethylene oxide) (PEO) grafted and/or heparin immobilized polyurethanes (PUs) were made by novel surface modification. Their blood compatibilities were investigated using in vitro platelet adhesion test, APTT, PT, immobilized heparin bioactivity measurement, and ex vivo rabbit A-A shunt test. In platelet adhesion tests, PEO grafted PU surfaces, compared to PU control, displayed very little platelet adhesion and activation, and this effect was more significant as the molecular weight of PEO increased from 200 to 2000. Also, the degree of platelet adhesion was lower in the heparinized PU surfaces than that of PU control. The immobilized heparin showed a greater effect on intrinsic blood coagulation factors than on extrinsic ones, whereas the PU-PEO surface was independent of blood coagulation factors. Lowering both in vitro platelet adhesion and activation led to a prolongation in the ex vivo occlusion time. In particular, the heparinized PU-PEO surfaces displayed enhanced blood compatibility due to the synergistic effects of PEO and heparin.

Animals↗

PubMed1989

Non-enzymic post-translational modification of proteins in aging. A review.

Various non-enzymic post-translational changes to proteins occur in vivo and some of these progress with aging. These changes are reviewed and linked to a number of age-related diseases, and to alterations in the charge distribution on protein surfaces. Modification by cyanate and by glucose 6-phosphate causes a partial unfolding of proteins, with loss of tertiary structure but retention of secondary structure. These products are reminiscent of the intermediate state observed during folding and unfolding of some proteins.

Aged↗

PubMed2005

Chitosan based surfactant polymers designed to improve blood compatibility on biomaterials.

We developed chitosan based surfactant polymers that could be used to modify the surface of existing biomaterials in order to improve their blood compatibility. These polymers consist of a chitosan backbone, PEG side chains to repel non-specific protein adsorption, and hexanal side chains to facilitate adsorption and proper orientation onto a hydrophobic substrate via hydrophobic interactions. Since chitosan is a polycationic polymer, and it is thrombogenic, the surface charge was altered to determine the role of this charge in the hemocompatibility of chitosan. Charge had a notable effect on platelet adhesion. The platelet adhesion was greatest on the positively charged surface, and decreased by almost 50% with the neutralization of this charge. A chitosan surface containing the negatively charged SO(3)(-) exhibited the fewest number of adherent platelets of all surfaces tested. Coagulation activation was not altered by the neutralization of the positive charge, but a marked increase of approximately 5-6 min in the plasma recalcification time (PRT) was displayed with the addition of the negatively charged species. Polyethylene (PE) surfaces were modified with the chitosan surfactant resulting in a significant improvement in blood compatibility, which correlated to the increasing PEG content within the polymer. Adsorption of the chitosan surfactants onto PE resulted in approximately an 85-96% decrease in the number of adherent platelets. The surfactant polymers also reduced surface induced coagulation activation, which was indicated by the PEG density dependent increase in PRTs. These results indicate that surface modification with our chitosan based surfactant polymers successfully improves blood compatibility. Moreover, the inclusion of either negatively charged SO(3)(-) groups or a high density of large water-soluble PEG side chains produces a surface that may be suitable for cardiovascular applications.

Adsorption↗

PubMed2006

The ruffled border and attachment regions of the apposing membrane of resorbing osteoclasts as visualized from the cytoplasmic face of the membrane.

The aim of our present research was to visualize how the plasma membrane is modified and how the cytoskeleton interacts with the attachment and ruffled border regions of resorbing osteoclasts. In order to view the surface modification of membranes and associated cytoskeleton, we employed the method of cell-shearing combined with quick-freezing and rotary replication to expose and replicate an extensive area of the cytoplasmic face of the surface membrane of osteoclasts in contact with synthetic apatite as a substratum. The membrane apposed to the apatite was composed of three different domains: the attachment zone, ruffled border and the remainder. In the attachment zone, a highly organized actin filament network formed dot-shaped, F-actin rich adhesion sites, so-called podosomes, and the actin ring. The cytoskeletal filament of podosomes and actin ring appeared to be in direct contact with the cytoplasmic surface of the underlying membrane. Within the actin ring, individually recognizable podosomes were well preserved, which indicates that the actin ring was probably derived from the fusion of podosomes. After shearing at the ruffled border region, the ruffled border projections and membrane regions among the projections were left behind. These ruffled border projections contained the cytoskeletal network. These actin networks also appeared to be in direct contact with the inner side of the ruffled border membrane or in contact with it via membrane-associated particles. At the basal portion of the ruffled border, numerous clathrin-coated patches or pits were well preserved. Deeper clathrin-coated pits and vesicles were also found, which indicates an active site for receptor-mediated endocytotic events. Clathrin sheets were also observed in the cell periphery outside of the actin ring. This type of clathrin sheets adhered to the apatite substrate, but was not anchored to the actin microfilaments. Our study thus clearly visualized the interaction between the cytoskeletal filaments and the underlying membrane at the ruffled border, attachment zone and podosome in osteoclasts cultured on apatitepellets.

Animals↗

PubMed2002

Thermal and chemical modification of dentin by 9-11-microm CO2 laser pulses of 5-100-micros duration.

BACKGROUND AND OBJECTIVES: Previous studies have shown that dentin can be thermally modified by pulsed CO(2) laser irradiation to form a more highly mineralized tissue. The implications are important for the potential laser modification or removal of dentinal and root caries and the transformation of dentin to a more acid resistant mineralized tissue. STUDY DESIGN/MATERIALS AND METHODS: Time resolved radiometry measurements with TEA CO(2) laser pulses were used to determine the magnitude of the absorption coefficients of dentin at the highly absorbed CO(2) laser wavelengths and to measure the temperature excursions during lambda = 9.3, 9.6, 10.3, and 10.6 microm laser irradiation at irradiation intensities of 0.1-8 J/cm(2) per pulse. In addition, photoacoustic and transient reflectance measurements were used to monitor the loss of water and organics and to detect the thresholds for surface modification and tissue ablation. RESULTS: The absorption coefficients were measured to be 5,000; 6,500; 1,200; and 800 cm(-1) at lambda = 9.3, 9.6, 10.3, and 10.6 microm, respectively. The surface temperatures of dentin were markedly higher than those measured on enamel for similar irradiation intensities due to the lower reflectance losses of dentin and the lower thermal diffusivity of dentin at the respective wavelengths. Hence, lower fluences are required for the thermal decomposition of dentin. Ablation typically occurred with the first few laser-pulses during multiple pulse irradiation and eventually ceased after modification of dentin to a more highly mineralized enamel-like tissue. The debris ejected during the initial laser pulses shielded the surface by as much as 60% at the low fluences employed in this study. Optical and electron microscopy and IR spectroscopy indicated that incident laser pulses with incident fluence as low as 0.5 J/cm(2) at 9.3 and 9.6 microm wavelengths with a duration of 5-8-micros were sufficient to induce chemical and morphological changes in dentin. CONCLUSIONS: In this study, the laser parameters for the efficient thermal modification of dentin with minimum heat deposition at CO(2) laser wavelengths were firmly established.

Dental Caries↗

PubMed1997

Ultrastructure of the interface between cultured osteoblasts and surface-modified polymer substrates.

Osteoblasts derived from rat bone marrow cells were cultured on surface-modified poly(ethylene terephthalate) films in the presence of ascorbic acid, beta-glycerophosphate, and dexamethasone. The surfaces employed for cell culture included the untreated hydrophobic surface and three modified surfaces possessing immobilized phosphate polymer chains, collagen molecules, and a thin hydroxyapatite-deposited layer. They all were produced by photo-induced graft polymerization with subsequent surface modifications of the graft chains. The ultrastructural morphology of the substrate/cell interfaces formed in in vitro osteoblast culture on these substrates was studied by transmission electron microscopy. The osteoblasts cultured for 1 week on the modified surfaces showed rough endoplasmic reticula rich in intracellular space and early matrix production in the extracellular space, irrespective of the surface chemistry. After 2 weeks of culture, osteoblasts exhibited active elaboration of extracellular matrix proteins, mostly composed of collagen, on all the surfaces. A remarkable result observed at this stage was direct deposition of an electron-dense, afibrillar layer of 180 nm thickness onto the surface having phosphate polymer chains. This layer became much more electron dense after 3 weeks of culture. Energy dispersive X-ray microanalysis revealed the presence of calcium phosphate in this layer. It was further found that the predeposited hydroxyapatite layer on the phosphate polymer-grafted surface promoted mineral deposition in the extracellular matrix that surrounded cuboid, osteocyte-like cells.

Animals↗

PubMedDate not supplied

Magnetic acoustic resonance immunoassay (MARIA): a multifrequency acoustic approach for the non-labelled detection of biomolecular interactions.

A unique sensing platform, comprising an electromagnetic field detector and an acoustic resonator, has been used as a wireless system for remote sensing of biorecognition events. The MARS (Magnetic Acoustic Resonator Sensor) technique has proven useful for detecting the formation of protein multilayers derived from specific binding phenomena. The technique enables multifrequency analysis, without the need of electrodes attached to the sensing element, and also facilitates the in situ surface modification of the substrate for antibody attachment. The MARS sensor was utilized as the platform on which a standard immunoassay was carried out. Two different conditions for the attachment of the first antibody to the quartz surface were tested: (i) Adsorption of the antibody onto the surface of a bare quartz disc; (ii) covalent immobilization of the antibody to a chemically modified quartz surface. Both methods can be successfully utilized for the 'label-less' detection of the biorecognition event between goat IgG and anti-goat IgG by analysis of the multifrequency spectrum. Covalent attachment of the primary antibody results in a more efficient immobilization, with higher surface density, and a consistently enhanced response for the binding of the secondary antibody. This approach will be of interest to life scientists and biochemists that require high performance assay methodologies that do not use chemical labels.

Acoustics↗

PubMed2003

Impact of surface chemistry and blocking strategies on DNA microarrays.

The surfaces and immobilization chemistries of DNA microarrays are the foundation for high quality gene expression data. Four surface modification chemistries, poly-L-lysine (PLL), 3-glycidoxypropyltrimethoxysilane (GPS), DAB-AM-poly(propyleminime hexadecaamine) dendrimer (DAB) and 3-aminopropyltrimethoxysilane (APS), were evaluated using cDNA and oligonucleotide sub-arrays. Two un-silanized glass surfaces, RCA-cleaned and immersed in Tris-EDTA buffer were also studied. DNA on amine-modified surfaces was fixed by UV (90 mJ/cm(2)), while DNA on GPS-modified surfaces was immobilized by covalent coupling. Arrays were blocked with either succinic anhydride (SA), bovine serum albumin (BSA) or left unblocked prior to hybridization with labeled PCR product. Quality factors evaluated were surface affinity for cDNA versus oligonucleotides, spot and background intensity, spotting concentration and blocking chemistry. Contact angle measurements and atomic force microscopy were preformed to characterize surface wettability and morphology. The GPS surface exhibited the lowest background intensity regardless of blocking method. Blocking the arrays did not affect raw spot intensity, but affected background intensity on amine surfaces, BSA blocking being the lowest. Oligonucleotides and cDNA on unblocked GPS-modified slides gave the best signal (spot-to-background intensity ratio). Under the conditions evaluated, the unblocked GPS surface along with amine covalent coupling was the most appropriate for both cDNA and oligonucleotide microarrays.

DNA, Complementary↗

PubMedDate not supplied

Biomembrane mimicry provides improved thromboresistance for total artificial hearts.

Thromboembolic events remain a significant issue in mechanical circulatory support. The aim of this study was to evaluate the potential benefit of surface modification in total artificial hearts (TAHs) using polymeric phospholipids (biomembrane mimicry). For this purpose, pneumatic TAHs (vacuum formed pellethane housing, hard double flap hinged inflow valves, soft trileaflet polyurethane outflow valves) had their blood-exposed surfaces either modified with polymeric phospholipids or unmodified before evaluation in bovine experiments. Orthotopic implantation of the TAHs was performed with cardiopulmonary bypass (CPB) using tip-to-tip heparin surface coated perfusion equipment and very low systemic heparinization (50 IU/kg bodyweight). After weaning from CPB and stabilizing hemodynamics, circulating heparin was neutralized with protamine (1:1). All animals were totally supported for 24 hours before elective sacrifice. No heparin was added at any time during support. Mean activated coagulation time (ACT) was 167+/-24 s at baseline before heparinization for CPB, 330+/-45 s at the end of CPB, 181+/-25 s after 1 hour of support, 180+/-31 s after 6 hours, and 185+/-28 s after 18 hours. After explantation, the TAHs perfused without anticoagulation were carefully analyzed. Atrial cuff coverage with red clot was 30+/-21% for artificial surfaces modified by biomembrane mimicry versus 100+/-0% for standard control surfaces (p<0.01). The number of macroscopic deposits found on the inflow valves was 1.33+/-0.47 for surfaces modified by biomembrane mimicry versus 3.83+/-1.86 for standard control surfaces (p<0.05). Likewise, on the outflow valves the number of macroscopic deposits was 0.00+/-0.00 for surfaces modified by biomembrane mimicry versus 1.00+/-0.81 for standard control surfaces (p<0.05). We conclude that presence and distribution of red clots and other macroscopic deposits are significantly different for artificial surfaces with biomembrane mimicry versus standard control surfaces. Application of the biomembrane mimicry concept has the potential to provide improved TAHs.

Animals↗

PubMed2001

Fabricating and imaging carbon-fiber immobilized enzyme ultramicroelectrodes with scanning electrochemical microscopy.

The scanning electrochemical microscope (SECM) is used to image the activity of enzymes immobilized on the surfaces of disk-shaped carbon-fiber electrodes. SECM was used to map the concentration of enzymatically produced hydroquinone or hydrogen peroxide at the surface of a 33-microm diameter disk-shaped carbon-fiber electrode modified by an immobilized glucose-oxidase layer. Sub-monolayer coverage of the enzyme at the electrode surface could be detected with micrometer resolution. The SECM was also employed as a surface modification tool to produce microscopic regions of enzyme activity by using a variety of methods. One method is a gold-masking process in which microscopic gold patterns act as mask for producing patterns of chemical modification. The gold masks allow operation in both a positive or negative process for patterning enzyme activity. A second method uses the direct mode of the SECM to produce covalently attached amine groups on the carbon surface. The amine groups are anchors for attachment of glucose oxidase by use of a biotin/avidin process. The effect of non-uniform enzyme activity was investigated by using the SECM tip to temporarily damage an immobilized enzyme surface. SECM imaging can observe the spatial extent and time-course of the enzyme recovery process.

Avidin↗

PubMed1998

Modification of materials formed from poly(L-lactic acid) to enable covalent binding of biopolymers: application to high-density three-dimensional cell culture in foams with attached collagen.

We describe a method for increasing the hydrophilicity of materials formed from biodegradable polymers and introducing chemical functional groups on their surfaces. Poly(L-lactic acid) was blended with poly(epsilon-CBZ-L-lysine) at an 80:20 ratio. Films of the mixture were prepared and foams were made by solvent casting and salt leaching. Amino groups on the surface of the polymer mixture were deprotected by acid hydrolysis. As an example of the applicability of the technique for attachment of biomolecules, we covalently linked collagen to the deprotected amino groups, creating a surface capable of high density growth of a differentiated cell type (bovine adrenocortical cells). The method should be generally useful for surface modification of biodegradable polymer materials used in tissue engineering.

Adrenal Cortex↗

PubMed2003

Evidence that homogenization of BSA-stabilized hexadecane-in-water emulsions induces structure modification of the nonadsorbed protein.

The structural modification of globular proteins (bovine serum albumin, BSA) in the aqueous phase of emulsions produced by homogenization was studied using front-face fluorescence spectroscopy (FFFS). A series of hydrocarbon oil-in-water emulsions (30 wt % n-hexadecane, 0.35 wt % BSA, pH 7.0) were homogenized to differing degrees with a high-speed blender and a high-pressure valve homogenizer. The wavelength of the maximum in the tryptophan emission spectrum (lambda(max)) of serum phases collected from the emulsions by centrifugation was measured and compared to lambda(max) values of BSA solutions subjected to the same homogenization conditions. There was no significant (p < 0.05) change in lambda(max) with homogenization conditions for BSA solutions. In contrast, lambda(max) of serum phases from emulsions blended for 2 min in a high-speed blender was significantly smaller (p < 0.05) than nontreated BSA solutions (Deltalambda(max) = 2 nm). In addition, there was a further significant decrease in lambda(max) of the serum phases with an increasing number of passes of the emulsion through the high-pressure valve homogenizer (e.g., Deltalambda(max) = 4 nm for 12 passes). This study shows that globular proteins present in the aqueous phase of a hexadecane-in-water emulsion after homogenization could be altered, which is probably caused by surface modification of the protein structure during temporary adsorption to emulsion droplet surfaces during homogenization.

Adsorption↗

PubMed2002

Effect of surface treatment of NiTi alloy on its corrosion behavior in Hanks' solution.

The effect of surface modification of NiTi shape memory alloy on its corrosion behavior in Hanks' solution was determined. The near-equi-atomic super elastic NiTi (Ni 55.8 wt %) alloy used for this study was provided by Memry USA. The surfaces of heat-treated samples were modified by mechanical polishing (MP), electropolishing (EP), and electropolishing followed by chemical passivation (CP). As-heat-treated samples with straw-colored oxide finishes (SCO) and blue-colored oxide finishes (BO) also were included in the study. Surface analysis was performed using auger electron spectroscopy (AES), atomic force microscopy (AFM), and contact angle measurements (CAM). It was shown that surface roughness increased in the order CP < EP < SCO < BO < MP. The nickel release within the five groups of NiTi samples, as determined by atomic absorption spectrophotometry, reduced in time over the measured period. The level of Ni ions released over a 25-day immersion period was highest in the SCO sample (0.002 microg/day). This Ni level is negligible compared with the daily intake of Ni in an ordinary diet. The auger electron spectroscopy (AES) analyses indicated that before immersion in Hanks' physiologic solution, the main surface composition of all the samples was titanium and nickel, with a small amount of oxygen, carbon, and sulphur as contaminants. And the surface oxide thickness of the different samples increased in the order CP < EP < MP < BO < SCO. On the other hand, for the electrodes treated under the same conditions, the mean breakdown potential value decreased in the order BO > MP > CP > EP > SCO while the corrosion current density and rate increased in the order CP < SCO < EP < BO < MP.

Alloys↗

PubMed2001

Enzymatic Treatments on Tencel in Water and Microemulsion.

Tencel is a relatively new fabric, obtained from wood pulp, that looks like natural cotton. In order to be suitable for commercial purposes, Tencel must be processed to improve its qualities. In this paper we report our studies on the enzymatic defibrillation of Tencel, in which we checked the different behavior of the same set of enzymes dispersed in pure water and in a microemulsion system. Surface properties, such as scanning electron microscopy, contact angle, porosimetry, breaking load, and thickness, were determined, in order to monitor the surface modification of the fabric upon enzymatic defibrillation, and indicate that the process is more efficient and less damaging when carried out in the microemulsion medium. Furthermore, we chemically modified Tencel by attaching fluorinated chains to the fabric surface. Surface properties show that fluorination of Tencel leads to a high degree of water- and oil-repellency in the fabric. Copyright 2001 Academic Press.

Journal Article↗

PubMed2005

Neurite outgrowth on well-characterized surfaces: preparation and characterization of chemically and spatially controlled fibronectin and RGD substrates with good bioactivity.

Study of axonal growth and ligand-receptor interactions requires specificity and careful characterization of the biomaterial substrates to which the neurons bind. It would be impossible to predict the effects of important variables such as composition, surface density, spatial distribution, and conformation of the ligands on axonal growth of a neuron without highly specific surface characterization. Here, we compare two methods of surface modification (hereafter referred to as "Heterobifunctional Crosslinker" and "Pluronics" methods) used for immobilization of fibronectin (FN) and FN-derived, RGD-containing peptides to the substrates. We also characterized their performance in neurite outgrowth experiments. Various surface analytical techniques such as contact angle measurement, XPS, and time-of-flight secondary ion mass spectrometry (TOF-SIMS) were used for the analysis of the substrates at each step of the two different chemistries involved. FN-patterned surfaces were created by micro-contact printing methods and confirmed by imaging TOF-SIMS, and AFM techniques. After immobilization of FN and/or FN-derived RGD-containing peptide, including the formation of micron-scale patterns of FN, the modified surfaces were plated with neurons from postnatal rat dorsal root ganglia (DRG) and incubated in serum-free medium. Both the peptide- and/or protein-modified substrates supported significantly greater neurite outgrowth than controls, and outgrowth on both substrate chemistries was inhibited by the addition of soluble RGD peptide. Patterned FN surfaces were successful in spatially controlling the neuron attachment and outgrowth.

Adsorption↗

PubMed2002

Soft tissue response to microtextured silicone and poly-L-lactic acid implants: fibronectin pre-coating vs. radio-frequency glow discharge treatment.

From in vitro studies it is known that a plasma-treatment can enhance cell spreading. Similar effects can be observed after pretreatment of the surface with a protein coating, to mediate cell adhesion. The aim of the current study was to evaluate the in vivo effects of these surface modifications, in a three-month experiment in a goat model. We made silicone and poly-L-lactic acid implants with double-sided parallel micro-grooves (depth 1.0 microm, width 10.0 microm), a random surface roughness, or a smooth surface. Implants either received a radio-frequency glow discharge (RFGD) treatment, a fibronectin (Fn) pre-coating, or no pre-treatment. Subsequently, they were inserted into subcutaneous pockets created on the flanks of goats for 1, 3 or 12 weeks. Histological analysis showed that a fibrous tissue capsule had formed around all implants. Histomorphometrical analysis was performed on capsule thickness, capsule quality and the implant-tissue interface quality. Fn-treated surfaces showed a considerable early inflammatory reaction. Besides this, RFGD treatment or Fn pre-coating did not further influence any of the measured parameters. In conclusion, pre-treatment of polymer implant surfaces with Fn or RFGD treatment did not significantly influence tissue reaction around implants with micro-grooved, roughened or smooth surfaces.

Animals↗

PubMed2006

Biocompatible surface preparation using amino-functionalized amylose.

Aminopropyl amyloses with various degrees of substitution (DS) were prepared and investigated with respect to their surface modification properties. Poly(acrylic acid) was grafted to plasma-activated PVDF films, and the functional amylose was bound via amide linkage formation. Layer formation was confirmed by X-ray photoelectron spectroscopy. Contact angle measurements and surface MALDI-TOF mass spectrometry indicated a hydrophilic surface and minimization of protein adsorption.

Amines↗

PubMed2000

Surface modified methotrexate loaded erythrocytes for enhanced macrophage uptake.

The unique property of macrophages to distinguish between 'mature self' and 'senescent self' was applied for the enhanced macrophage uptake upon loading drugs into erythrocytes and provoking environment within which it gets translated into senescent cells. Two different approaches were applied one by depleting the sialic acid from the surface of cells and another by inducing hemichrome formation in the cells. Erythrocytes encapsulation of methotrexate was carried out by preswell dilution technique. Surface modification of erythrocytes was performed using desialation and hemichrome induction by treating with trypsin (Tt) and phenylhydrazine (PhT) respectively. Desialation technique was optimized in terms of concentration of trypsin, incubation temperature and time period. Both of these surface modified erythrocytes were characterized for in vitro macrophage uptake. In vivo organ localization was assessed by recording amount of drug present in different organs. The macrophage uptake of both surface modified erythrocytes was enhanced by almost 3-5 and 5-6 times with Tt and PhT cells, respectively. These new surface modified carriers are found to be excellent candidate for enhanced macrophage uptake and anticipated to be useful for any RES tumors.

Animals↗