Biocompatilibity-related surface characteristics of oxidized NiTi

In the present study, we examined the effect of NiTi oxidation on material surface characteristics related to biocompatibility. Correspondence between electron work function (EWF) and adhesive force predicted by electron theory of adsorption as well as the effect of surface mechanical stress on the adhesive force were studied on the nonoxidized and oxidized at 350, 450, and 600 degrees C NiTi alloy for medical application. The adhesive force generated by the material surface towards the drops of alpha-minimal essential medium (alpha-MEM) was used as a characteristic of NiTi adsorption properties. The study showed that variations in EWF and mechanical stress caused by surface treatment were accompanied by variations in adhesive force. NiTi oxidation at all temperatures used gave rise to decrease in adhesive force and surface stress values in comparison to the nonoxidized state. In contrary, the EWF value revealed increase under the same condition. Variations in surface oxide layer thickness and its phase composition were also followed. The important role of oxide crystallite size in EWF values within the range of crystallite dimensions typical for NiTi surface oxide as an instrument for the fine regulation of NiTi adsorption properties was demonstrated. The comparative oxidation of pure titanium and NiTi showed that the effect of Ni on the EWF value of NiTi surface oxide is negligible.     

Modelling of protein adsorption on polymeric surfaces

A new method of calculation of protein adsorption on polymeric surfaces is presented. The method considers the thermodynamic equilibrium of a three-component system-water, protein, and polymer surface-and describes the protein concentration profile based on the interaction energy parameters for protein-polymer, water-polymer, and protein-water contacts. Calculation of these parameters calls for introduction of the energies arising from the dispersive forces, the hydrophobic effect, and the Drago et al. (1971) electron donor-acceptor interactions. Comparison with experimental results of protein adsorption on various polymeric surfaces gave satisfactory agreement.     

ATR-FTIR analysis of protein adsorption on polymeric surfaces

ATR-FTIR spectroscopy was used to study the adsorption of bovine serum albumin on poly(vinyl chloride), poly(vinylidene fluoride), poly(methyl methacrylate), and poly(ethylene glycol)-grafted poly(methyl methacrylate) surfaces. The results were analyzed in terms of the amount of albumin adsorbed on the various polymeric surfaces when immersed in a 1 g/l protein solution. Experimental results were compared with the Van Straaten and Peppas model (1990) and satisfactory agreement was obtained. It was confirmed that poly(ethylene glycol) surfaces exhibited the lowest albumin adsorption patterns.     

Modelling of protein adsorption on polymeric surfaces.

A new method of calculation of protein adsorption on polymeric surfaces is presented. The method considers the thermodynamic equilibrium of a three-component system--water, protein, and polymer surface--and describes the protein concentration profile based on the interaction energy parameters for protein-polymer, water-polymer, and protein-water contacts. Calculation of these parameters calls for introduction of the energies arising from the dispersive forces, the hydrophobic effect, and the Drago et al. (1971) electron donor-acceptor interactions. Comparison with experimental results of protein adsorption on various polymeric surfaces gave satisfactory agreement.     

ATR-FTIR analysis of protein adsorption on polymeric surfaces

ATR-FTIR spectroscopy was used to study the adsorption of bovine serum albumin on poly(vinyl chloride), poly(vinylidene fluoride), poly(methyl methacrylate), and poly(ethylene glycol)-grafted poly(methyl methacrylate) surfaces. The results were analyzed in terms of the amount of albumin adsorbed on the various polymeric surfaces when immersed in a 1 g/1 protein solution. Experimental results were compared with the Van Straaten and Peppas model (1990) and satisfactory agreement was obtained. It was confirmed that poly(ethylene glycol) surfaces exhibited the lowest albumin adsorption patterns.     

The spatial resolution of protein adsorption on surfaces of heterogeneous metallic biomaterials

This study was designed to examine the heterogeneity of the adsorption of proteins onto metallic materials. The materials studied included pure Ag, Au, and Ti and sintered Ag 10% Ti and Ag 10% Ta. The distribution of the protein adsorption was studied using I-125 labeled albumin detected by microautoradiography. The surface morphology of the specimens was examined in the scanning electron microscope prior to exposure to the protein solution. A heterogeneous distribution in albumin adsorption was observed over the Ag surface. Similar regions were observed over parts of the mixed metal specimens, but superimposed on this pattern were distinct regions of very low protein adsorption which appeared to correlate closely with the regions of Ti or Ta observed in the scanning electron microscope. A uniform distribution of adsorbed albumin was observed on the Au and Ti, with Au giving a much denser microautoradiograph than Ti. This work demonstrates that variations in the protein adsorption to heterogeneous materials can be observed on a microscopic scale.     

Protein adsorption on a copolymer having pendant monosaccharide groups. Relationship between surface free energy and protein adsorption

Protein adsorption on copolymer films having pendant monosaccharide groups was investigated from the viewpoint of surface chemistry. The copolymers were synthesized by the copolymerization of a monomer having a pendant monosaccharide (GEMA) and methyl methacrylate (MMA). The contact angles of methylene iodide and air bubble on the GEMA-MMA copolymer films were measured in water, and the surface free energy of the copolymer films in water was calculated from the contact angles. With increasing GEMA content, the amount of protein adsorbed on the GEMA-MMA copolymer films decreased gradually. The relationship between protein adsorption and the surface free energy of the copolymer films in water is discussed. The differences between surface free energy before and after protein adsorption on the copolymer surface is closely related to the amount of adsorbed protein.     

Molecular simulation of protein adsorption and desorption on hydroxyapatite surfaces

Protein adsorption and desorption on material surfaces play a key role in the biocompatibility of medical implants, biomineralization and protein separation. In this report, the adsorption and desorption behavior of the 10th type III module of fibronectin (FN-III(10)) with different orientations on hydroxyapatite (HAP) (001) surface were systematically studied by molecular dynamics (MD) and steered MD simulations. These studies show that the electrostatic energy plays a dominant role in the interaction between the model protein and the HAP surface. The values of the interaction energy not only relates to the number of adsorbed sites but also the type. The charged -COO(-) and -NH(3)(+) are the strongest groups that interact with the surface, while other groups like charged guanido group, neutral amino and hydroxyl groups have considerable interactions with the surface. The effects of these groups on interaction energy were quantitatively investigated.     

Blood compatibility of surfaces with superlow protein adsorption

In this work, five self-assembled monolayers (SAMs) and three polymeric brushes with very low fibrinogen adsorption were prepared. The five SAMs are oligo(ethylene glycol) (OEG), phosphorylcholine (PC), oligo(phosphorylcholine) (OPC), and two mixed positively and negatively charged SAMs of SO(3)(-)/N(+)(CH(3))(3) (SA/TMA) and COO(-)/N(+)(CH(3))(3) (CA/TMA). Three polymer brushes were prepared on gold surfaces via surface-initiated atom transfer radical polymerization (ATRP) using three monomers, sulfobetaine methacrylate (SBMA), carboxybetaine methacrylate (CBMA), and oligo(ethylene glycol) methyl ether methacrylate (OEGMA). Surface plasmon resonance (SPR) measurements show that although all of these surfaces are "nonfouling" to fibrinogen adsorption from buffer solution, their protein adsorption from undiluted human blood plasma varies widely. Polymer brushes exhibit much lower protein adsorption from plasma than any of the five SAMs tested. However, platelet adhesion measurements on plasma-preadsorbed surfaces show that all of these surfaces have very low platelet adhesion. Clotting time measurements using recalcified platelet poor plasma (PPP) incubation with the eight types of surfaces show that they do not shorten clotting times. Linear polymers of polySBMA and polyCBMA with similar molecular weights were also synthesized and characterized. In the presence of polyCBMA linear polymers, the clotting time of PPP was prolonged and increased with the concentration of the polymer, while no anticoagulant activity was observed for the polySBMA or PEG polymers. The unique anticoagulant activity of polyCBMA, as well as its high plasma protein adsorption resistance, makes polyCBMA a candidate for blood-contacting applications.     

The influence of protein adsorption on nanoparticle association with cultured endothelial cells

As materials are produced at smaller scales, the properties that make them especially useful for biological applications such as drug delivery, imaging or sensing applications also render them potentially harmful. There has been a reasonable amount of work addressing the interactions of biological fluids at material surfaces that demonstrates the high affinity of protein for particle surfaces and some looking at the role of particle surface chemistry in cellular associations, but mechanisms have been too little addressed outside the context of intended, specific interactions. Here, using cultured endothelium as a model for vascular transport, we demonstrate that the capacity of nanoparticle surfaces to adsorb protein is indicative of their tendency to associate with cells. Quantification of adsorbed protein shows that high binding nanoparticles are maximally coated in seconds to minutes, indicating that proteins on particle surfaces can mediate cell association over much longer time scales. We also remove many of the most abundant proteins from culture media which alters the profile of adsorbed proteins on nanoparticles but does not affect the level of cell association. We therefore conclude that cellular association is not dependent on the identity of adsorbed proteins and therefore unlikely to require specific binding to any particular cellular receptors.     

Competitive plasma protein adsorption onto fluorinated polyimide surfaces

A series of fluorinated polyimides cured at different temperatures was prepared, and plasma protein adsorption and platelet adhesion onto the polyimide films were evaluated in vitro using scanning electron microscopy, a micro-bicinchoninic acid protein assay, and a gold-colloid-labeled immunoassay. In particular, we focused on competitive plasma protein adsorption onto polyimide film because elucidation of the competitive adsorption mechanism is needed for a good understanding of in vivo biocompatibility of polyimide. Interestingly, the trend of IgG adsorption onto the polyimide surface measured in human plasma was completely contrary to that observed with IgG dissolved in PBS, and the adsorption increased with an increase in the curing temperature. We propose that the human plasma F(c) region in IgG might selectively adsorb onto polyimide film cured at high temperatures because of competitive plasma protein adsorption to the surface.     

Computer modelling of the adsorption of proteins on solid surfaces under the influence of double layer and van der waals energy

The study of protein interactions with surfaces is important in many branches of biomedical engineering. A computer model has been set up in order to aid the understanding and prediction of the likelihood of protein adsorption at a surface and of coagulation between two proteins. In this model, a protein is represented as a hard sphere, neglecting conformation changes which may occur during the adsorption process. The sphere is assumed to be in a medium whose properties are described by the ionic strength, the pH and the dielectric permittivity. It is considered to interact both with an infinite plane, representing the surface, and with another sphere, representing another protein. The model focuses on the total interaction energy between a protein and a surface and between two proteins. The energy is expressed according to the DLVO theory of colloidal stability, which assumes that the adsorption behaviour of proteins at a surface depends, first, on the van der Waals interactions energy and, second, on the electrostatic double layer interaction energy. The conditions under which adhesion is prevented correspond to the presence of local extremes of the enegy function, whereas the conditions under which adhesion is likely to take place correspond to absence of local extremes.     

Leukocyte adhesion on model surfaces under flow: effects of surface chemistry, protein adsorption, and shear rate

The effect of specific chemical functionalities on the adhesion of polymorphonuclear leukocytes (PMNs) under flow was investigated using a set of well-characterized, chemically functionalized surfaces prepared by self-assembly of alkanethiolate monolayers on gold surfaces. Terminal functionalities included CH(3), CH(2)OH, COOH, and (OCH(2)CH(2))(3)OH groups. A new surface modification was used to incorporate a phosphorylcholine moiety on the hydroxyl-terminated monolayer. Surface modification was verified using contact-angle measurements, ellipsometry, and X-ray photoelectron spectroscopy. Adhesion on the surfaces was studied in the presence and absence of pre-adsorbed fibrinogen. Fibrinogen adsorption on self-assembled monolayers (SAMs) was quantified using radioisotope detection. PMN adhesion was found to be dependent on the monolayer's terminal functionality. Adhesion was higher on the hydrophobic CH(3) surface and the polar COOH monolayer. Leukocyte adhesion was least on the phosphorylcholine-rich surface, followed by the ethylene-oxide-containing monolayer. Cell adhesion also was low on the hydrophilic OH monolayer. Attachment was decreased with increasing shear rate, exhibiting a three-fold decrease between 20 and 100 s(-1). Fibrinogen adsorption was higher on the CH(3) monolayer but comparable for the other four SAMs. Preincubation of the surfaces with fibrinogen decreased adhesion on all SAMs examined.     

The effect of protein adsorption on the anticoagulant activity of surface immobilized heparin

The anticoagulant activity of albumin-heparin conjugates covalently immobilized on carboxylated polystyrene beads was determined before and after exposure to different plasma/PBS dilutions using a thrombin inhibition assay, a FXa inhibition assay, and a modified aPTT assay. Exposure of albumin-heparin modified surfaces (alb-hep surfaces) to plasma dilutions resulted in surfaces with a lower anticoagulant activity than surfaces which were not exposed to plasma dilutions. The reduction of the activity increased up to ±80% when the surfaces were exposed to solutions containing more than 70% plasma. Alb-hep surfaces incubated in plasma which was preexposed to heparin-Sepharose retained 30% of their initial activity. These observations were attributed to non-specific adsorption of plasma proteins onto the surface and to interaction of heparin binding proteins with the immobilized heparin. Both processes result in a decreased accessibility of the immobilized heparin and thus in a reduced anticoagulant activity displayed by the heparinized surface. Identification of adsorbed proteins with SDS gel electrophoresis and immunoblotting revealed that many different proteins were present at the heparinized surface. Only small differences were observed between the gel electrophoresis pattern of adsorbed proteins obtained from heparinized surfaces and from a surface containing immobilized albumin.     

The impact of dendrimer-grafted modifications to model silicon surfaces on protein adsorption and bacterial adhesion

In the oral cavity, omnipresent salivary protein films (pellicle) mediate bacterial adhesion and biofilm formation on natural tissues as well as on artificial implant surfaces, which may cause serious infectious diseases like periimplantitis. The purpose of this in?vitro study was to investigate the adsorption/desorption behaviour of human saliva on model surfaces grafted with polyamidoamine (PAMAM) dendrimer molecules compared to self-assembled monolayers (SAMs) exhibiting the same terminal functions (-NH(2), -COOH) by two complementary analytical methods. Furthermore, the role of saliva conditioning of PAMAM and analogous SAM modifications on the adhesion of Streptococcus gordonii DL1, an early oral colonizer, was investigated. In contrast to SAMs, PAMAM-grafted surfaces showed reduced streptococcal adherence in the absence of pre-adsorbed saliva similar to the level obtained for poly(ethylene glycol) (PEG) coatings. Moreover, coatings of PAMAM-NH(2) maintained their bacteria-repellent behaviour even after saliva-conditioning. As a general outcome, it was found that lower amounts of protein adsorbed on PAMAM coatings than on analogous SAMs. Since this study demonstrates that covalently bound PAMAM dendrimers can modulate the oral bacterial response, this approach has significant potential for the development of anti-adhesive biomaterial surfaces that are conditioned with proteinaceous films.     

Polyurethane surfaces modified by amphiphilic polymers: effects on protein adsorption

Surface modification of polyurethane (PUR) surfaces was carried out by using three different amphiphilic polymers. Two of the polymers were graft copolymers, having backbones consisting of poly(methyl methacrylate-co-ethylhexyl acrylate) and poly(styrene-co-acrylamide), respectively, and poly(ethylene oxide) PEO 2000 grafts. The third polymer was a commercially available poly(ethylene oxide-b-propylene oxide-b-ethylene oxide) block copolymer, Pluronic 9400. The polymers were designated ACRY, STY2, and PE94, respectively. Surface modification was achieved by adsorption of the amphiphilic polymers at PUR surfaces from an aqueous solution, or by blending the amphiphiles into a PUR solution, followed by solution casting of films. The accumulation of the amphiphilic polymers at the PUR surfaces was observed by XPS and contact angle measurements. The ACRY and PE94 polymers were shown to adsorb poorly at the PUR surface, but gave strong surface effects when present in the PUR matrix. Protein adsorption was measured under static as well as under flow conditions. The modified surfaces had generally lower adsorption of blood proteins (HSA, Fg and IgG) than the unmodified PUR surfaces. ACRY blend modified surfaces had the lowest adsorption.     

Calculation of solvation interaction energies for protein adsorption on polymer surfaces.

Of the interactions that govern protein adsorption on polymer surfaces, solvation interactions (repulsive hydration and attractive hydrophobic interactions) are thought to be among the most important. The solvation interactions in protein adsorption, however, have not been dealt with in theoretical calculation of the adsorption energy owing to the difficulties in modelling such interactions. We have evaluated the solvation interaction energies using the fragment constant method of calculating the partition coefficients of amino acids. The fundamental assumption of this approach is that the partition coefficients of amino acids between water and organic solvent phases are related to the free energies of transfer from bulk water to the polymer surface. The X-ray crystallographic protein structures of lysozyme, trypsin, immunoglobulin Fab, and hemoglobin from the Brookhaven Protein Data Bank were used. The model polymer surfaces were polystyrene, polypropylene, polyethylene, poly(hydroxyethyl methacrylate) [poly(HEMA)], and poly(vinyl alcohol). All possible adsorption orientations of the proteins were simulated to study the effect of protein orientation on the solvation interactions. Protein adsorption on either hydrophobic or hydrophilic polymer surfaces was examined by considering the sum of solvation and other interaction energies. The results showed that the contribution of the solvation interaction to the total protein adsorption energy was significant. The average solvation interaction energy ranged from -259.1 to -74.1 kJ/mol for the four proteins on the hydrophobic polymer surfaces, such as polystyrene, polypropylene, and polyethylene. On the other hand, the average solvation interaction energies on hydrophilic surfaces such as poly(HEMA) and poly(vinyl alcohol) were larger than zero. This indicates that repulsive hydration interactions are in effect for protein adsorption on hydrophilic polymer surfaces. The total interaction energies of the proteins with hydrophobic surfaces were always lower than those with more hydrophilic surfaces. This trend is in agreement with the experimental observations in the literature. This study suggests that consideration of the solvation interaction energies is necessary for accurate calculation of the protein adsorption energies.