Adsorption Of Blood Plasma Proteins Research Articles

Boosting Titanium Surfaces with Positive Charges: Newly Developed Cationic Coating Combines Anticorrosive and Bactericidal Properties for Implant Application.

Along with poor implant-bone integration, peri-implant diseases are the major causes of implant failure. Although such diseases are primarily triggered by biofilm accumulation, a complex inflammatory process in response to corrosive-related metallic ions/debris has also been recognized as a risk factor. In this regard, by boosting the titanium (Ti) surface with silane-based positive charges, cationic coatings have gained increasing attention due to their ability to kill pathogens and may be favorable for corrosion resistance. Nevertheless, the development of a cationic coating that combines such properties in addition to having a favorable topography for implant osseointegration is lacking. Because introducing hydroxyl (-OH) groups to Ti is essential to increase chemical bonds with silane, Ti pretreatment is of utmost importance to achieve such polarization. In this study, plasma electrolytic oxidation (PEO) was investigated as a new route to pretreat Ti with OH groups while providing favorable properties for implant application compared with traditional hydrothermal treatment (HT). To produce bactericidal and corrosion-resistant cationic coatings, after pretreatment with PEO or HT (Step 1), surface silanization was subsequently performed via immersion-based functionalization with 3-aminopropyltriethoxysilane (APTES) (Step 2). In the end, five groups were assessed: untreated Ti (Ti), HT, PEO, HT+APTES, and PEO+APTES. PEO created a porous surface with increased roughness and better mechanical and tribological properties compared with HT and Ti. The introduction of -OH groups by HT and PEO was confirmed by Fourier transform infrared spectroscopy and the increase in wettability producing superhydrophilic surfaces. After silanization, the surfaces were polarized to hydrophobic ones, and an increase in the amine functional group was observed by X-ray photoelectron spectroscopy, demonstrating a considerable amount of positive ions. Such protonation may explain the enhanced corrosion resistance and dead bacteria (Streptococcus aureus and Escherichia coli) found for PEO+APTES. All groups presented noncytotoxic properties with similar blood plasma protein adsorption capacity vs the Ti control. Our findings provide new insights into developing next-generation cationic coatings by suggesting that a tailorable porous and oxide coating produced by PEO has promise in designing enhanced cationic surfaces targeting biomedical and dental implant applications.

Silk fibroin and recombinant elastin blend nano-coatings for implantable medical devices

Silicone breast implants are conventionally used to reconstruct the shape and size of the breast in breast cancer patients. However, rate of failure of implants due to formation of capsular contracture is significantly high. Recent studies have shown that coating the surface of the implant with a biocompatible polymer can mitigate this risk. Here, we have modified the surface of implant using a biocompatible natural biopolymer silk fibroin (SF). Further, we have developed formulations of SF with a recombinantly produced elastin-like-peptide (ELP) and compared their performance with a known anti-fouling hydrophilic polymer - polyethylene oxide (PEO). Microscopic and spectroscopic characterization confirm the formation of uniform coatings. These coatings have been also characterized for their ability to resists crack formation. Further, the coatings have been evaluated for their biological performance. Our studies show that addition of 25 wt% of ELP to SF significantly enhances the crack resistance for the coatings. In addition, SF/ELP coatings reduce the adsorption of blood plasma proteins by more than 80%. Preliminary analysis shows that the SF/ELP blend coatings are non-cytotoxic and support adhesion, growth and proliferation of fibroblast cells. The study therefore demonstrates that SF/ELP coatings have the potential to mitigate the risk of breast implant failure.

Effect of N-vinylimidazole comonomer on blood plasma protein and endogenous toxin adsorption on mesoporous copolymer beads

N-vinylimidazole-divinylbenzene (VI-DVB) copolymer beads with a N-vinylimidazole (VI) weight content ranging from 7% to 15% were studied as possible adsorbents for hemoperfusion and compared with the cross-linked polystyrene beads from commercial devices. All the beads had a large volume fraction of mesopores with diameters of 2–50 nm, quantified by nitrogen adsorption, and BET areas from 500 to 1000 m²/g. Metal-chelate adsorption of copper (II) ions on VI-DVB was high in 0.15 M NaCl (up to 40 μmol/mL), and low in blood plasma (< 1 μmol/mL). Reversible adsorption of blood plasma proteins was high (> 100 mg/mL beads) on all the bead types. Irreversibly adsorbed proteins eluted by 2% SDS included albumin, fibrinogen, fibronectin, and transferrin. Metal chelating contributed to the irreversible adsorption. Unlike the beads from commercial devices, VI-DVB beads adsorbed the LPS endotoxin from plasma. Adsorption of bilirubin and IL-6 was high on all the beads tested, but VI-DVB beads were better adsorbents of bile acids. VI-DVB are promising as components of multifunctional devices for blood purification applicable to treatment of diseases where conventional devices have lower or negligible adsorption capacity.

Improved hemocompatibility and reduced bacterial adhesion on superhydrophobic titania nanoflower surfaces

Thrombosis formation and bacterial infection are key challenges for blood-contacting medical devices. When blood components encounter a device's surface, proteins are adsorbed, followed by the adhesion and activation of platelets as well as an immune response. This culminates in clot formation via the trapping of red blood cells in a fibrin matrix, which can block the device's function and cause severe complications for the patient. In addition, bacteria may adhere to a device's surface. This can lead to the formation of a biofilm, a protective layer for bacteria that significantly increases resistance to antibiotics. Despite years of research, no long-term solutions have been discovered to combat these issues. To impede thrombosis, patients often take antiplatelet drugs for the life of their device, which can cause excess bleeding and other complications. Patients can take antibiotics to fight bacterial infection, but these are often ineffective if biofilms are formed. Superhydrophobic surfaces show promise in reducing both thrombosis and bacterial infection on devices by impeding contact between biological components and the biomaterial. In this study, superhydrophobic titania nanoflower surfaces were successfully fabricated on a titanium alloy Ti-6Al-4V substrate with hydrothermal synthesis and vapor-phase silanization. The surface topography, surface wettability, surface chemistry, and surface crystallography of the surfaces was subsequently characterized. Surface hemocompatibility was investigated through lactate dehydrogenase (LDH) cytotoxicity analysis, blood-plasma protein adsorption, platelet and leukocyte adhesion and activation, and whole blood clotting analysis. Surface bacterial infection was characterized through Gram-positive and Gram-negative bacterial adhesion and biofilm morphology. The results indicated a reduction of protein adsorption, platelet and leukocyte adhesion and activation, bacterial adhesion, and biofilm formation as well as improved contact angle stability compared to control surfaces.

Synthesis of bioactive glass-based coating by plasma electrolytic oxidation: Untangling a new deposition pathway toward titanium implant surfaces

HypothesisAlthough bioactive glass (BG) particle coatings were previously developed by different methods, poor particle adhesion to surfaces and reduced biological effects because of glass crystallization have limited their biomedical applications. To overcome this problem, we have untangled, for the first time, plasma electrolytic oxidation (PEO) as a new pathway for the synthesis of bioactive glass-based coating (PEO-BG) on titanium (Ti) materials. ExperimentsElectrolyte solution with bioactive elements (Na2SiO3-5H2O, C4H6O4Ca, NaNO3, and C3H7Na2O6P) was used as a precursor source to obtain a 45S5 bioglass-like composition on a Ti surface by PEO. Subsequently, the PEO-BG coating was investigated with respect to its surface, mechanical, tribological, electrochemical, microbiological, and biological properties, compared with those of machined and sandblasted/acid-etched control surfaces. FindingsPEO treatment produced a coating with complex surface topography, Ti crystalline phases, superhydrophilic status, chemical composition, and oxide layer similar to that of 45S5-BG (~45.0Si, 24.5 Ca, 24.5Na, 6.0P w/v%). PEO-BG enhanced Ti mechanical and tribological properties with higher corrosion resistance. Furthermore, PEO-BG had a positive influence in polymicrobial biofilms, by reducing pathogenic bacterial associated with biofilm-related infections. PEO-BG also showed higher adsorption of blood plasma proteins without cytotoxic effects on human cells, and thus may be considered a promising biocompatible approach for biomedical implants.

In Vitro Investigation of Hemocompatibility of Hydrothermally Treated Titanium and Titanium Alloy Surfaces.

For decades, titanium and its alloys have been established as a biocompatible material for cardiovascular medical devices such as heart valves, stents, vascular grafts, catheters, etc. However, thrombosis is one of the reasons for implant failure, where blood clot forms on the implant surface, thus obstructing the flow of the blood and that leads to some serious complications. Various surface modification techniques such as heparin modification, albumin coating, surface anodization, plasma etching, and hydrothermal treatments have been explored to improve the hemocompatibility of titanium-based materials. However, there are several limitations related to the robustness of the surfaces and long-term efficacy in vivo. In this study, titanium and its alloy Ti–6Al–4V were hydrothermally treated to form nanostructured surfaces with the aim to enhance their hemocompatibility. These modified surfaces were characterized for their wettability, surface morphology, surface chemistry, and crystallinity. The hemocompatibility of these surfaces was characterized by evaluating blood plasma protein adsorption, platelet adhesion and activation, platelet–leukocyte complex formation, and whole blood clotting. The results indicate lower fibrinogen adsorption, cell adhesion, platelet activation, and whole blood clotting on hydrothermally treated surfaces. Thus, these surfaces may be a promising approach to prevent thrombosis for several titanium blood-contacting medical devices.

Novel chitosan/agarose/hydroxyapatite nanocomposite scaffold for bone tissue engineering applications: comprehensive evaluation of biocompatibility and osteoinductivity with the use of osteoblasts and mesenchymal stem cells.

BackgroundNanocomposites produced by reinforcement of polysaccharide matrix with nanoparticles are widely used in engineering of biomaterials. However, clinical applications of developed novel biomaterials are often limited due to their poor biocompatibility.PurposeThe aim of this work was to comprehensively assess biocompatibility of highly macroporous chitosan/agarose/nanohydroxyapatite bone scaffolds produced by a novel method combining freeze-drying with a foaming agent. Within these studies, blood plasma protein adsorption, osteoblast (MC3T3-E1 Subclone 4 and hFOB 1.19) adhesion and proliferation, and osteogenic differentiation of mesenchymal stem cells derived from bone marrow and adipose tissue were determined. The obtained results were also correlated with materials' surface chemistry and wettability to explain the observed protein and cellular response.ResultsObtained results clearly showed that the developed nanocomposite scaffolds were characterized by high biocompatibility and osteoconductivity. Importantly, the scaffolds also revealed osteoinductive properties since they have the ability to induce osteogenic differentiation (Runx2 synthesis) in undifferentiated mesenchymal stem cells. The surface of biomaterials is extremely hydrophilic, prone to protein adsorption with the highest affinity toward fibronectin binding, which allows for good osteoblast adhesion, spreading, and proliferation.ConclusionProduced by a novel method, macroporous nanocomposite biomaterials have great potential to be used in regenerative medicine for acceleration of the bone healing process.

Biological Response of and Blood Plasma Protein Adsorption on Silver-Doped Hydroxyapatite.

Hydroxyapatite (HAP) is an extensively used orthopedic biomaterial because of its high biocompatibility and osteoconductivity. Implant-related infection is a major cause of orthopedic device failure. Previous research showed that silver-doped hydroxyapatite nanoparticles (Ag-HAP NPs) have prominent antimicrobial activity, but their biocompatibility and plasma protein response remained unexplored. Here we investigated the effects of synthesis conditions on Ag-HAP NP antimicrobial (E. coli and S. epidermidis) activity, biocompatibility, and the adsorption of two blood plasma proteins, human serum albumin (HSA) and fibrinogen (Fib). It was found that synthesis pH affected the Ag content of Ag-HAP NPs and subsequent Ag+ release from the NPs in solution. This, in turn, affected antimicrobial efficiency and cytotoxicity to murine preosteoblast cells (MC3T3-E1). More HSA than Fib was adsorbed on a molar basis. The conformation of HSA changed drastically from predominantly α-helix and minor β-sheet content in solution to greater β-sheet than α-helix content when adsorbed. Correspondingly, the melting temperature Tm of HSA changed significantly from 76 °C in solution to ∼65-66 °C when adsorbed. Fib exhibited a modest decrease in α-helix content while theβ-sheet content increased modestly upon adsorption and its Tm remained unchanged at ∼60 °C. These differences in behavior of HSA and Fib are ascribed to the much smaller size of HSA, which allows a greater molecular packing density on the surface, which induces greater conformational changes. The protein adsorption behavior on Ag-HAP was similar to that on pure HAP. Thus, we show that Ag-HAP NPs have antimicrobial activity without deleterious effects on biocompatibility and blood plasma protein adsorption.

One‐Step Noncovalent Surface Functionalization of PDMS with Chitosan‐Based Bioparticles and Their Protein‐Repellent Properties

AbstractStable hydrophilic and protein‐repellent surfaces from nontoxic polysaccharides are highly desirable in several biomedical applications such as implants and prosthetics. Herein, a water‐based one‐step strategy to functionalize polydimethylsiloxane (PDMS) surfaces with chitosan‐based bioparticles is described to suppress the adsorption of blood plasma proteins, which causes biofilm formation and implant‐associated infection. Micro‐ and nanosized chitosan (Chi) and carboxymethyl chitosan (CMChi) particles, featuring high stability, hydrophilicity, and zwitterionic feature, are synthesized in an ecofriendly solvent water and their interaction with PDMS surfaces is investigated in‐real time over a wide range of pH (3–10) using a quartz crystal microbalance with dissipation (QCM‐D). The results show that both Chi and CMChi form stable colloidal particles at pH close to their zero‐point charge, and thus exhibit maximum adsorption and irreversibly bound layers on PDMS surfaces. Coating from CMChi shows high stability, improved hydrophilicity, and subsequently no adsorption of bovine serum albumin and enhanced resistance to fibrinogen, compared to Chi and native PDMS, as revealed by water contact angle and QCM‐D measurements. The results demonstrate the possibility to develop novel and biobased coating materials to address unspecific protein adsorption not only on silicone‐based implants but also on other biomaterials in general.

Mechanical properties, structure, bioadhesion, and biocompatibility of pectin hydrogels.

The surface structure, biocompatibility, textural, and adhesive properties of calcium hydrogels derived from 1, 2, and 4% solutions of apple pectin were examined in this study. An increase in the pectin concentration in hydrogels was shown to improve their stability toward elastic and plastic deformation. The elasticity of pectin hydrogels, measured as Young's modulus, ranged from 6 to 100 kPa. The mechanical properties of the pectin hydrogels were shown to correspond to those of soft tissues. The characterization of surface roughness in terms of the roughness profile (Ra) and the root-mean-square deviation of the roughness profile (Rq) indicated an increased roughness profile for hydrogels depending on their pectin concentration. The adhesion of AU2% and AU4% hydrogels to the serosa abdominal wall, liver, and colon was higher than that of the AU1% hydrogel. The adhesion of macrophages and the non-specific adsorption of blood plasma proteins were found to increase as the pectin concentration in the hydrogels increased. The rate of degradation of all hydrogels was higher in phosphate buffered saline (PBS) than that in DMEM and a fibroblast cell monolayer. The pectin hydrogel was also found to have a low cytotoxicity. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2572-2581, 2017.

Coarse-grained model of adsorption of blood plasma proteins onto nanoparticles.

We present a coarse-grained model for evaluation of interactions of globular proteins with nanoparticles (NPs). The protein molecules are represented by one bead per aminoacid and the nanoparticle by a homogeneous sphere that interacts with the aminoacids via a central force that depends on the nanoparticle size. The proposed methodology is used to predict the adsorption energies for six common human blood plasma proteins on hydrophobic charged or neutral nanoparticles of different sizes as well as the preferred orientation of the molecules upon adsorption. Our approach allows one to rank the proteins by their binding affinity to the nanoparticle, which can be used for predicting the composition of the NP-protein corona. The predicted ranking is in good agreement with known experimental data for protein adsorption on surfaces.

Adsorption of Free Hemoglobin from Aqueous Solutions and from Blood Plasma on Electrochemically Modified Activated Carbons

Acute blood loss is one of the main contributing factors that determine the severity of a trauma patient’s condition. It is well known, however, that the transfusion of more than 1 L of donor blood plasma may lead to complications. Therefore, the reinfusion of the patient’s own blood lost in the course of trauma to serous cavities undoubtedly has important advantages. One of the “technological” drawbacks of reinfusion is the mechanical destruction of some of the erythrocytes and the resultant contamination of the obtained blood plasma with free hemoglobin [1]. The development of a method of free hemoglobin removal from blood plasma is thus a very important problem.To achieve the above goal, activated carbons modified by chemical and affinity methods to selectively adsorb free hemoglobin were used. It is well known that “soft” ligands (such as iodide) are capable of complexing with the ferrous ion in hemoglobin, while cupric ions bond with the protein subunits of hemoglobin.SKT-6A brand activated carbon was modified by means of electropolymerization of pyrrole (Py) on the surface of carbon in 0.10 potassium iodide/0.10 M pyrrole electrolyte, where iodide ions (I–) serve as a dopant for the conductive polypyrrole (PPy) layer that is formed. Cu2+ ions were immobilized on the surface of SKT-6A at potentials more positive than +480 mV, according to [2]. Spectrophotometry was used to measure the concentration of free hemoglobin in aqueous solutions before and after bringing a solution of free hemoglobin in contact with the carbon sorbent [3], while the adsorbent potential was measured against the Ag/AgCl reference electrode.The results of experiments with the PPy/I– composite showed that the adsorption activity of the composite was increased greatly, as compared to the activity of carbon samples with I– immobilized on their surface in the absence of PPy. The additional immobilization of Cu2+ ions on the SKT-6A carbon surface led to a 16-fold increase in the rate of free hemoglobin adsorption, as compared to unmodified carbon samples, and a nearly fivefold increase compared to carbon modified with PPy doped with I– in the absence of Cu2+.Thus, it was shown that the modification of activated carbon by introducing various ligands led to a significant increase in the adsorption rate of free hemoglobin from aqueous solutions, with the carbon modified with cupric ions [SKT-6A/Cu2+] showing the greatest free hemoglobin removal efficiency from aqueous solutions.When free hemoglobin is being removed from blood plasma, the rate of adsorption on the modified SКТ-6А/Cu2+ carbon is reduced 11-fold compared to adsorption from aqueous solutions. Also, the rate of adsorption of blood plasma proteins of the same carbon is comparable to their adsorption rate on the unmodified SKT-6A (measuring 4-5 percent). However, the presence of free hemoglobin in plasma leads to a significant increase in overall protein adsorption (ca. 20 percent). This observation could be rationalized by considering the possible role of adsorbed hemoglobin on the surface of carbon in initiating the subsequent adsorption of proteins. To test this hypothesis, albumin adsorption experiments were carried out in physiological saline solution in the presence and absence of hemoglobin, showing a 0.6% adsorption of albumin in the presence of hemoglobin and zero adsorption in its absence.As shown by a comparison of the adsorption of hemoglobin and blood plasma proteins on the two modified carbons, the adsorption of proteins is more than doubled on SКТ-6А/Cu2+ compared to the adsorption of proteins on SКТ-6А/PPy/I–. It was therefore suggested that the process of hemoglobin interaction with blood plasma proteins occurs simultaneously with the interaction of proteins with cupric ions immobilized on the modified SКТ-6А/Cu2+ carbon, leading to an increase in adsorption over against SКТ-6А/PPy/I–. The interaction of free hemoglobin in plasma with haptoglobin, a plasma protein that is selective with respect to hemoglobin, must also be taken into account. The observed effect can be rationalized if blood plasma proteins bind with a hemoglobin/haptoglobin complex when the latter is adsorbed on the surface of carbon sorbent.Based on the above hypotheses, the following model of adsorption of blood plasma proteins in the presence of hemoglobin on the surface of modified SКТ-6А/Cu2+ and SКТ-6А/PPy/I– carbons was proposed (Fig. 1).[1] Hamburger J. Presse méd, 1960, 68, 279-281[2] Khubutia M. Sh., Goldin M.M., Kurilkin Yu. A., Goldin Mikh. M., Grafov B. M., Davydov A. D., Kolesnikov V.A. Khimicheskaya promyshlennost’ segodnya, 2011, 7, 51-55.[3] Harboe M. Scandinavian Journal of Clinical and Laboratory Investigation, 1959, 11, 66-70.=========Figure 1 caption: Fig.1. Interaction of free hemoglobin and blood plasma proteins with modified activated carbons: (a) SKT-6A/Cu2+, (b) SKT-6A/PPy/I–.

Hemocompatibility of Chitosan/poly(acrylic acid) Grafted Polyurethane Tubing.

The activation and adhesion of platelets or whole blood exposed to chitosan (CH) grafted surfaces is used to evaluate the hemocompatibility of biomaterials. The biomaterial surfaces are polyurethane (PU) tubes grafted with an inner poly(acrylic acid) (PAA) and an outer CH or quaternary ammonium modified CH (CH-Q) brush. The CH, CH-Q and PAA grafted layers were characterized by ellipsometry and fluorescence microscopy. Material wear tests demonstrate that CH (CH-Q) is stably grafted onto PU tubes upon exposure to saline solution for 7 days. Using quartz-crystal microbalances with dissipation (QCM-D), in-situ adsorption of blood plasma proteins on CH and CH-Q compared to a silicon oxide control was measured. The QCM-D results showed that the physically adsorbed plasma protein layer on CH-Q and CH surfaces is softer and more viscous than the protein layer on the SiO2 surface. The CH-Q layer thus has the weakest interaction with plasma proteins. Whole blood and platelet adhesion was reduced by ~92% on CH-Q, which showed the weakest interaction with plasma protein but more viscous adsorbed plasma protein layer, compared to SiO2. Last, to examine the biologic response of platelets and neutrophils to biomaterial surfaces, CH (CH-Q)/PAA, PAA and PU tubes were tested using a Chandler Loop apparatus as an ex vivo model and flow cytometry. The blood adhesion and biologic response results showed that CH and CH-Q reduced adhesion and activation of platelets and neutrophils and improved hemocompatibility relative to other surfaces (PU and PAA). Our studies demonstrated that the properties of physically adsorbed plasma protein layer on biomaterial surfaces correlates with blood coagulation on biomaterial surfaces.

Dynamic in vitro hemocompatibility testing of poly(ether imide) membranes functionalized with linear, methylated oligoglycerol and oligo(ethylene glycol)

Linear, side-chain methylated oligoglycerols (OGMe) were recently reported as potential surface passivating molecules for improving the protein resistance of cardiovascular application relevant poly(ether imide) (PEI) membranes. A previously reported in vitro screening under static test conditions allowed an end-point evaluation of the adhesion and activation of adherent thrombocytes performed on the material surfaces and revealed similar levels of thrombogenicity on PEI membranes, functionalized with OGMe and oligo(ethylene glycol) (OEG) of similar molecular weight (Mn = 1,300 g·mol-1 - 1,800 g·mol-1). In the present study, we investigated the hemocompatibility of these materials in a dynamic closed loop system, in order to study time-dependent thrombocyte material interactions also of the circulating thrombocytes by mimicking in vivo relevant flow conditions in a dynamic test system with multiple material contacts. Activation and aggregation of circulating thrombocytes as well as complement activation and plasmatic coagulation were evaluated after 40 circulations of thrombocyte rich plasma in the closed loop system. The results of the dynamic tests revealed no differences between the OGMe and OEG functionalized PEI membranes. Furthermore, no differences were observed between the latter and a PEI membrane treated under the conditions of functionalization at pH 11 (PEI-pH11) without an oligoether being present. Blood plasma protein adsorption, as well as activation, and adherence of circulating thrombocytes occurred in a comparable, but minor manner on all investigated PEI membranes. From this we conclude that the OGMe and OEG surface functionalization did not lead to an improvement of the already good hemocompatibility of the PEI-pH11 membrane.

Blood plasma protein adsorption capacity of perfluorocarbon emulsion stabilized by proxanol 268 (in vitro and in vivo Studies)

The adsorption abilities of the perfluorocarbon emulsion stabilized by Proxanol 268 were investigated in vitro and in vivo. In vitro, the saturation point for the blood plasma proteins was nearly reached after five minutes of incubation of the emulsion with human/rabbit blood plasma and was stable for all incubation periods studied. The decrease in volume ratio (emulsion/plasma) was accompanied by the increase in the adsorptive capacity of the emulsion with maximal values at 1/10 (3.2 and 1.5 mg of proteins per 1 ml of the emulsion, for human and rabbit blood plasma, respectively) that was unchanged at lower ratios. In vivo, in rabbits, intravenously injected with the emulsion, the proteins with molecular masses of 12, 25, 32, 44, 55, 70, and 200 kDa were adsorbed by the emulsion (as in vitro) if it was used 6 hours or less before testing. More delayed testing (6 h) revealed elimination of proteins with molecular masses of 25 and 44 kDa and an additional pool of adsorpted new ones of 27, 50, and 150 kDa. Specific adsorptive capacity of the emulsion enhanced gradually after emulsion injection and reached its maximum (3.5-5 mg of proteins per 1 ml of the emulsion) after 24 hours.

Laser-optical investigation of the effect of diamond nanoparticles on the structure and functional properties of proteins

Adsorption of such blood plasma proteins as albumin and g-globulin on diamond nanoparticles of size around 5 nm and around 100 nm is observed and studied using laser-optical methods. The adsorption of blood plasma proteins at physiological pH 7.4 is found weaker than that of enzyme protein lysozyme. The observed variations in the Fourier Transform Infrared (FTIR) spectra of proteins may be due to structural transformations of the adsorbed protein. Using the lysozyme as a test protein we show that the protein adsorption leading to observable changes in the FTIR spectrum (the band of Amide I) also induces a significant decrease in the protein functional activity. It is also found that the influence of ∼5-nm diamond nanoparticles on the protein structure and functions is more significant than that of ∼100-nm nanodiamonds.

Oxidation of proteins adsorbed on hemodialysis membranes and model materials

The cleaning of cellulosic hemodialysis membrane Cuprophan and model materials (glass; polystyrene and polypropylene, as such and surface-oxidized), conditioned by adsorption of blood plasma proteins (HSA, fibrinogen, IgG) was investigated in vitro. Sodium hypochlorite (NaClO) and Renalin, a product containing hydrogen peroxide and peroxyacetic acid, were used as cleaning reagents. X-ray photoelectron spectroscopy and the use of radiolabeled fibrinogen demonstrated the presence of varying amounts of a polypeptidic residue, with sulfur brought to a high oxidation stage (sulfonate-like). The trends were the same for the three proteins regarding the effectiveness of the oxidizer and the influence of the surface properties. NaClO was much more effective than Renalin to remove the adsorbed proteins. The proteins adsorbed on Cuprophan were more sensitive to the oxidizers, when compared with proteins adsorbed on other materials. This may be due to both the lower protein-surface affinity, as indicated by radiochemical measurements, and the sensitivity of the material itself to the oxidizer, as revealed by weight loss measurements. These results support the attribution of hemocompatibility improvement after regeneration of Cuprophan with Renalin to the masking of the activating surface by a residue from previously adsorbed proteins.

A Simple Method for Biocompatible Polymer Based Spatially Controlled Adsorption of Blood Plasma Proteins to a Surface

Using the soft lithographic technique, microcontact printing, we demonstrate an example of how a biocompatible polymer can be easily patterned upon another polymer substrate, creating a surface with two spatially different properties. A poly(dimethylsiloxane) mold allowed the patterning of a negative replica of an E-PROM microchip, forming a spatially defined pattern with a period of approximately 1 μm. The amphiphilic biocompatible polymer, poly(lactic acid)−poly(ethylene glycol) (PLA−PEG), was used as the “ink” in order to block off areas of a hydrophobic, polystyrene (PS) substrate. This created a surface with two properties: 2−3 μm strips of PLA−PEG polymer, which resists protein adsorption, divided by micrometer strips of PS. The ability of the patterning technique to provide a true heterogeneous surface was analyzed using atomic force microscopy, while fluorescence microscopy provided a high-contrast method by which to trace the position of specific molecules. We believe this technique to be an elegant demonstration of how the properties of polymers can be exploited in order to arrange molecules at a surface, avoiding the more difficult use of gold, self-assembly, or self-assembled monolayers.

Blood plasma proteins on polyurethane and alkylsiloxane plasma-treated polyurethane surfaces. Dynamic approach by stimulus-response technique

In this study, interactions of blood proteins (i.e. albumin and fibrinogen) with polyurethane biomaterial surfaces were investigated in an in vitro bead column test circuit using a stimulus-response technique. The dynamic sorption process of radiolabelled proteins on the surfaces was followed by detecting the radioactivity at the exit stream of the column, which was the response of a pulse stimulus at the inlet. The mathematical model was described and solved using 'parameter estimation by cybernetic moment technique', and the adsorption rate constants of plasma proteins on different biomaterial surfaces were calculated. By evaluation of the response curves with standard and cybernetic moment techniques, the following results were obtained. Albumin and fibrinogen adsorption is competitive, and the competition is strongly dependent upon the surface characteristics of the biomaterial. Preadsorption or preferential adsorption of albumin decreases the fibrinogen adsorption, and therefore increases the biocompatibility of material surface. Adsorption of blood plasma proteins are irreversible. The moment technique can also be used for the evaluation of stimulus-response data of biological systems, to determine the process parameters.

Investigation of the adsorption of blood plasma proteins by activated carbon fiber material

The authors study the adsorption of fibrinogen, albumin, and gamma globulin by carbon fibrous materials by physical immobilization of protein ligands on their surface. The adsorption of proteins from model solutions under standard conditions was studied by an indirect method according to the decrease in the concentration of the adsorbate in solution, determining the protein content. The adsorption of the same proteins from the plasma and their desorption from activated carbon fibrous materials were estimated by a direct radiometric method using /sup 125/I-labeled proteins.