Interaction Of Blood Proteins Research Articles

Surface modification strategies for improved hemocompatibility of polymeric materials: a comprehensive review.

Polymeric biomaterials are a widely used class of materials due to their versatile properties. However, as with all other types of materials used for biomaterials, polymers also have to interact with blood. When blood comes into contact with any foreign body, it initiates a cascade which leads to platelet activation and blood coagulation. The implant surface also has to encounter a thromboinflammatory response which makes the implant integrity vulnerable, this leads to blood coagulation on the implant and obstructs it from performing its function. Hence, the surface plays a pivotal role in the design and application of biomaterials. In particular, the surface properties of biomaterials are responsible for biocompatibility with biological systems and hemocompatibility. This review provides a report on recent advances in the field of surface modification approaches for improved hemocompatibility. We focus on the surface properties of polysaccharides, proteins, and synthetic polymers. The blood coagulation cascade has been discussed and blood - material surface interactions have also been explained. The interactions of blood proteins and cells with polymeric material surfaces have been discussed. Moreover, the benefits as well as drawbacks of blood coagulation on the implant surface for wound healing purposes have also been studied. Surface modifications implemented by other researchers to enhance as well as prevent blood coagulation have also been analyzed.

Impact of spontaneous liposome modification with phospholipid polymer-lipid conjugates on protein interactions

ABSTRACT Liposome surface coating has been studied to avoid the immunological responses caused by the complement system, and alternative materials to poly(ethylene glycol) (PEG) have been explored recently since the production of anti-PEG IgM antibodies has been found in humans. We previously reported a liposome coating with poly(2-methacryloyloxyethyl phosphorylcholine) (poly(MPC))-conjugated lipids (PMPC-lipids) and demonstrated its protective effect on blood protein interactions. Here, we attempted to modify the liposome surface by exogenously adding PMPC-lipids, which were spontaneously incorporated into the outer membrane via hydrophobic interactions. The polymerization degree of the PMPC segment was regulated from 10 to 100. The incorporated ratio of PMPC-lipid increased with a decrease in the degree of PMPC polymerization. Due to surface modification with PMPC-lipids, increase in the length of the PMPC-chain increased the size of the liposomes. The modified liposomes were kept stable for 14 d in terms of their size, polydispersity, and surface properties, where approximately 70% of PMPC-lipids were incorporated into the liposome surface. We demonstrated that liposome surface modification with PMPC-lipids can inhibit protein adsorption when exposed to serum, regardless of the degree of polymerization of PMPC. In addition, the PMPC-lipid modified surface was not recognized by the anti-PEG IgM antibody, whereas PEG-lipid was recognized by the antibody. Thus, we successfully fabricated an inert liposome surface via spontaneous modification with PMPC-lipids, where only the outer bilayer surface was modified. This technique can be available for full loading of water-soluble active pharmaceutical ingredient inside the modified liposome.

Hemostasis in the Lab, Approaching to the Correct Diagnosis in the Coagulopathies

Coagulation is the result of a coordinated interaction of blood proteins, circulating cells, endothelium cells and extracellular matrix proteins in the vessel wall. This process works in conjunction with other mechanisms that have to keep the blood in an effective flow, without hemorrity when damage occurs or without a thrombus when clotting begins. Most of the time, there is not so much information that a clotting test gives us, however, in the face of a real clinical scenario that concurrent with a personal haemorrhage, this is essential to take action. In addition, there is a thrombophilic scenario that has to be investigated. In all of them, it is important to note that doctors have to take into account that the pre-analytical phase plays a fundamental role, so it is very important to look around any situation that may impact the patient's results. There is a wide range of laboratory tests to evaluate different mechanisms of hemostasis, according to the history and characteristic of the clinical patient. With this in mind, there is a right time to ask for the right test, to get a better result for diagnosis and treatment. Analytical quality is the set of actions that are carried out in the laboratory with the aim of obtaining an accurate result, reflecting the real situation of the patient. Grossly, platelets, clotting factors and the fibrinolytic system could be studied, not all patients need all tests, they could be solved with a simple test, but may require a deep focus; that is why the patient with a history of mucocutaneous bleeding has to be studied for thrombocytopaties or VWD; a patient with muscle or joint bleeding has to be a factor deficiency test. We propose an initial approach to detecting the pathological situations essential to make decisions. Each person has different symptoms and different approaches, which is why identification and the right tests give us the greatest opportunity to get the right diagnosis.

Controlling protein interactions in blood for effective liver immunosuppressive therapy by silica nanocapsules.

Immunosuppression with glucocorticoids is a common treatment for autoimmune liver diseases and after liver transplant, which is however associated with severe side-effects. Targeted delivery of glucocorticoids to inflammatory cells, e.g. liver macrophages and Kupffer cells, is a promising approach for minimizing side effects. Herein, we prepare core-shell silica nanocapsules (SiO2 NCs) via a sol-gel process confined in nanodroplets for targeted delivery of dexamethasone (DXM) for liver immunosuppressive therapy. DXM with concentrations up to 100 mg mL-1 in olive oil are encapsulated while encapsulation efficiency remains over 95% after 15 days. Internalization of NCs by non-parenchymal murine liver cells significantly reduces the release of inflammatory cytokines, indicating an effective suppression of inflammatory response of liver macrophages. Fluorescent and magnetic labeling of the NCs allows for monitoring their intracellular trafficking and biodegradation. Controlled interaction with blood proteins and good colloidal stability in blood plasma are achieved via PEGylation of the NCs. Specific proteins responsible for stealth effect, such as apolipoprotein A-I, apolipoprotein A-IV, and clusterin, are present in large amounts on the PEGylated NCs. In vivo biodistribution investigations prove an efficient accumulation of NCs in the liver, underlining the suitability of the SiO2 NCs as a dexamethasone carrier for treating inflammatory liver diseases.

Superhydrophobic blood-compatible surfaces: state of the art

Various strategies have been developed to improve blood compatibility of biomaterials, mostly through surface modification techniques. Superhydrophobic surfaces are among the promising approaches controlling the interaction of proteins and cells with external surfaces in medical applications. This review summarizes the recent studies on hemocompatibility of superhydrophobic surfaces. The review begins with principle theories on wettability of surfaces, and then evaluates the interaction of blood proteins and platelets with superhydrophobic surfaces and discusses the conflicting results. Finally, future challenges in utilizing superhydrophobic surfaces for blood contacting applications are also addressed.

Nitric oxide releasing vascular catheters for eradicating bacterial infection.

The interaction of blood proteins with an implant surface is not only a fundamental phenomenon but is also key to several important medical complications. Plasma proteins binding on the surface of intravascular catheters can promote bacterial adhesion leading to the risk of local and systemic complications such as catheter-related blood infections (CRBIs). The incidences of CRBIs in the United States amount to more than 250,000 cases/year with an attributable mortality of up to 35% and an annual healthcare expenditure of $2.3 billion approximately. This demands the development of truly nonthrombogenic and antimicrobial catheters. In the present study, catheters were fabricated by incorporating a nitric oxide (NO) donor molecule, S-nitroso-N-acetyl-penicillamine (SNAP) in a hydrophobic medical grade polymer, Elasteon-E2As. NO offers antithrombotic and antibacterial attributes without promoting drug resistance and cytotoxicity. E2As-SNAP catheters were first coated with fibrinogen, a blood plasma protein plays a key role in clot formation and eventual bacterial adhesion to the implant surface. The suitability of the catheters for biomedical applications was tested in vitro for contact angle, NO release kinetics, inhibition of bacteria, and absence of cytotoxicity toward mammalian cells. The highly hydrophobic catheters released NO in the physiological range that inhibited >99% bacterial viability on fibrinogen-coated catheters in a 24 h study. No toxic response of E2As-SNAP catheters leachate was observed using a standard cytotoxicity assay with mouse fibroblast cells. Overall, the results showed that the E2As-SNAP catheters can inhibit viable bacteria even in the presence of blood proteins without causing a cytotoxic response. The fundamentals of this study are applicable to other blood-contacting medical devices as well. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2849-2857, 2018.

Understanding the interactions of human follicle stimulating hormone with single-walled carbon nanotubes by molecular dynamics simulation and free energy analysis.

Interactions of carbon nanotubes (CNTs) and blood proteins are of interest for nanotoxicology and nanomedicine. It is believed that the interactions of blood proteins and glycoproteins with CNTs may have important biological effects. In spite of many experimental studies of single-walled carbon nanotubes (SWCNT) and glycoproteins with different methods, little is known about the atomistic details of their association process or of structural alterations occurring in adsorbed glycoproteins. In this study, we have applied molecular dynamics simulation to investigate the interaction of follicle stimulating hormone (hFSH) with SWCNT. The aim of this work is to investigate possible mechanisms of nanotoxicity at a molecular level. We present details of the molecular dynamics, structure, and free energy of binding of hFSH on the surface of SWCNT. We find that hFSH in aqueous solution strongly adsorbs onto SWCNT via their concave surface as evidenced by high binding free energies for residues in both protein subunits. It was found that hydrophobic, π-cation, and π-π stacking interactions are the main driving forces for the adsorption of the protein at the nanotube surface.

The interaction of blood proteins with brucine

The features of brucine (BC) binding to two blood proteins, bovine hemoglobin (BHb), and bovine serum albumin (BSA), were investigated via fluorescence, circular dichroism and UV/Vis absorption spectroscopy. The results revealed that BC caused the fluorescence quenching of blood proteins by the formation of BC-protein complex. The corresponding thermodynamic parameters were measured at different temperatures. The process of binding BC molecule on protein was a spontaneous molecular interaction procedure in which entropy increased and Gibbs free energy decreased. Hydrophobic and electrostatic interactions play a major role in stabilizing the complex. The molecular docking has been employed to explore the binding site of the BC in BHb and BSA on the Autodock 4.2. The distances r between BC and protein were calculated to be 4.93 and 5.08nm for BHb, and BSA, respectively. The effect of BC on the conformation of blood proteins was analyzed using CD, synchronous fluorescence and three-dimensional fluorescence spectra.

Preparation and characterization of poly(hydroxyethyl methacrylate-co -poly(ethyleneglycol-methacrylate)/hydroxypropyl-chitosan) hydrogel films: Adhesion of rat mesenchymal stem cells

This study examined the effects of the surface properties of the materials, such as the hydroxyl, methyl and amino groups, on rat bone marrow derived Mesenchymal Stem Cell (MSC) seeding. A series of hydrogels were prepared in film form using 2-hydroxyethyl methacrylate (pHEMA), poly(ethyleneglycol) methacrylate (PEG-MA), and/or hydroxypropyl-chitosan (HPC). The physicochemical properties of these hydrogel films, such as water content, functional groups, contact angle, surface energy and thermal properties were affected by the composition of the materials. The ability of the MSCs to form colonies, as well as their viability on these materials were also analyzed. The water content of the hydrogel films increased with increasing PEG-MA and HPC ratio in the hydrogel. Contact angle measurements of the surface of the hydrogel films demonstrated that all the materials gave rise to a significantly hydrophilic surface compared to pure pHEMA. The blood protein interactions and platelet adhesion were reduced significantly on the surface of the materials upon the incorporation of PEG-MA compared to the control pure pHEMA and vice versa for HPC. The ability of the MSCs to adhere and form colonies on these materials was also analyzed. The results showed that these materials are suitable candidates to isolate and expand MSCs. Open image in new window

Analysis of blood proteins on protein arrays with a spectral surface plasmon resonance biosensor

Abstract We investigated whether blood proteins can be analyzed on protein arrays with a spectral surface plasmon resonance (SPR) biosensor. We modified gold arrays with a mixture of mercaptoundecanoic acid and mercaptohexanol, and immobilized blood proteins-hemoglobin (Hb) and haptoglobin (Hp), or their antibodies. We analyzed interactions of Hb with Hp on protein arrays by measuring the shift of resonance wavelength, and a significant interaction was observed when Hp was used as a ligand. Then, we fabricated antibody arrays with antibodies against Hb and Hp, and analyzed binding of blood proteins onto the arrays. Anti-Hb showed a specific interaction with Hb on the antibody arrays. Interaction of anti-Hp with Hp was specific when Hp was used as capture molecules. The shift of resonance wavelength caused by the interaction of blood proteins was explained by changes of refractive index on the gold surfaces of protein arrays. The present work suggests that spectral SPR biosensors can be used as a useful tool for the analysis of blood proteins on protein arrays.

Interactions of blood proteins with poly(isobutylcyanoacrylate) nanoparticles decorated with a polysaccharidic brush

The aim of this work was to examine the in vitro interactions of core-shell poly(isobutylcyanoacrylate)-polysaccharide nanoparticles (NP) with blood proteins. The particles were prepared by initiating the emulsion polymerization of isobutylcyanoacrylate (IBCA) in the presence of dextran 71 or 15 kDa, heparin, a blend of dextran 71 and heparin, or dextran sulphate in aqueous medium at pH 1. The mechanisms of polymerisation were redox radical (Rad) or anionic (An), resulting in differences in the spatial arrangement of the polysaccharide chains at the NP surface, i.e. “loops” and “trains” by anionic polymerization, “brush” by radical polymerization. Surface composition of NPs was determined by X-ray photo-electron spectroscopy (XPS) and surface charge by zeta potential measurements. In the presence of citrated blood plasma, efficacy of the steric repulsive effect of the NP dextran shell towards protein adsorption decreased in the order: Dex71-Rad>Dex15-Rad>Dex71-An⪢Dex15-An. Dextran-coated NPs adsorbed ApoA-I and fibrinogen from plasma. Concerning activation of complement in serum, the effect was sharp: Dex71-Rad was a very low activator whereas Dex15-An, Dex15-Rad and Dex71-An were strong activators. In citrated plasma, the steric repulsive effects of Hep-Rad and Dex-Hep-Rad NPs were similar to Dex71-An, and Dex-Sulph-Rad NPs adsorbed twice more proteins than Hep-Rad. Hep-Rad, Dex-Hep-Rad and Dex-Sulph-Rad NPs adsorbed IgG and fibrinogen. Complement was not activated in serum in the presence of Hep-Rad and Dex-Hep-Rad and a slight adsorption of C3 was noted. C3 was completely adsorbed on Dex-Sulph-Rad. The exquisite sensitivity of blood proteins to differences in the nature and outermost structure of the polysaccharides-coated NPs is highlighted by the present results.

Investigation of in vitro interactions between different polymeric surfaces and blood proteins via phagocytosis phenomena

The effect of various polymeric materials on blood components and their in vitro phagocytosis was the object of the present research. Polystyrene- (PS) and polymethylmetacrylate- (PMMA) based microspheres were produced by phase-inversion polymerization and chemically modified to obtain different surface hydrophilicities. The interactions between blood proteins and chemically- and biologically-modified surfaces were investigated and compared to plain microspheres. Adsorption properties of albumin, fibrinogen and total immunoglobulines on microspheres were tested. Hydrophilic surfaces have high ability for human serum albumin (HSA) adsorption, which also leads to less phagocytosis of microspheres in vitro. In the case of activated PMMAPVA microspheres, both protein adsorption and phagocytosis were significant. Interaction of blood proteins with microspheres did not cause any change in phagocytosis by leukocytes and monocytes. BSA adsorption on microspheres with different hydrophilicities showed the same blood protein adsorption results and phagocytosis was not detected. On the other hand, the highest level of phagocytosis was found with fibronectin-modified microspheres. The changes occurring in intrinsic and extrinsic coagulation mechanisms were determined by measuring the activated partial tromboplastin time (APTT) and the prothrombin time (PT). PT values of blood samples did not increase by treatment with microspheres, except for PS/HEMA, while chemical modification caused important prolongation in APTT.

Opsono-recognition of liposomes by tissue macrophages

On exposure to blood, liposomes of different morphology and lipid composition attract different arrays of proteins. The mode of interaction of blood proteins with liposomes vary in quantity and conformation. These differences may explain different observed patterns in blood clearance and tissue distribution of liposomes. In this paper, an attempt is made to assess the role of serum and its components in regulating liposome recognition by isolated phagocytes of rat liver and spleen.

Blood protein interactions with chromium surfaces

Protein adsorption, contact activation, and complement activation were studied on thin evaporated films of chromium (Cr) in vitro. The surfaces were, prior to the experiments, cleaned in either ethanol and water, or in a basic peroxide solution (RCA standard clean 1, SC-1 Surface spectroscopic studies of the outermost oxides showed a significant reduction of carbon contaminants after washing in SC-1 but also suggested an increase in the oxidation state as compared with the ethanol-washed surfaces. In situ ellipsometry combined with antibody techniques was used to determine protein deposition and antibody binding onto surfaces after incubations in heparin plasma or in normal serum. Incubation times from I to 10 min in serum showed increased depositions of serum and antibodies to complement factor 3c (C3c) and was larger on ethanol-washed surfaces than on surfaces washed in SC-1. ELISA methods indicated increased amounts of iC3b in serum for both surfaces, but no presence of C3 convertases (C4d or Bb fractions). A low or transient complement activation via the classical pathway was indicated on ethanol washed Cr, since deposition of secondary antibodies to complement factor Iq (Clq) was observed only after short incubation times in serum. No procoagulant activity of Cr was indicated, since only low amounts of antibodies to factor XII (F XII), prekallikrein (PKK), and high molecular weight kiniogen (HMWK) bound to the surfaces after incubations in heparin plasma. These results were confirmed using a colorimetric assay where the relative amounts of free plasma kallikrein was assessed using a chromogenic substrate, H-D-Pro-Phe-Arg-pNA (S-2302).

Blood protein interactions with titanium surfaces

Protein adsorption and complement activation were studied on thin evaporated films of titanium (Ti). The surfaces were cleaned in either a radio frequency (RF) plasma unit, or washed sequentially in trichloroethylene, acetone, ethanol, and water. Both methods resulted in hydrophilic surfaces with low carbon contamination on the outermost oxide ( ~ 11-13 at%). In situ ellipsometry suggested that Ti is an intrinsic coagulation activator in vitro, since significant amounts of factor XII (F XII) and high molecular weight kininogen (HMWK) were found on the surfaces after 1 min incubation in heparin plasma. Ellipsometry, performed after serum incubations ranging from 15 s to 30 min showed that the total amount of serum proteins and the deposition of antibodies to complement factor 3c (C3c) increased with serum incubation time. ELISA methods showed increased levels of free iC3b in serum after 10 min incubation of the surfaces, but no detectable amounts of C3 convertase fractions C4d or Bb. Ellipsometric results indicated, however, an increased deposition of antibodies to C I q and IgG on Ti after short serum incubation times. The combined results indicate that Ti-surfaces initially activate complement through the classical pathway. The activation then continues via a positive amplification loop where increased amounts of C3 are deposited on the surfaces via the alternative pathway.

Protein adsorption on low temperature isotropic carbon: V. How is it related to its blood compatibility?

Based on our research on blood protein interactions with low temperature isotropic carbon (LTIC) and data from the literature, we propose that the carbon surface has strong interactions with adsorbed proteins. In this paper we focus on how a relatively blood-compatible material interacts with plasma proteins. We present our results on the structure and properties of the LTIC surface utilizing SEM, STM, XPS, and contact angle measurements. We briefly review protein adsorption on LTIC using DSC, impedance, radioisotopes, and two-dimensional gel electrophoresis. LTIC is characterized by a microporous, oxidized, hydrophobic, and domain mosaic structure. Surface polishing smoothens the roughness and removes the porosity, while largely destroying the ordered atomic texture, making the surface more random and more amorphous. The LTIC surface denatures all adsorbed proteins studied. The rate of protein adsorption is high and the surface concentration is large. The LTIC surface adsorbs all proteins without preference. The surface also tenaciously retains proteins such that they cannot be displaced by buffer or exchanged by proteins in solution. We conclude that LTIC accomplishes its blood compatibility through a passivating film of strongly adsorbed bland proteins, which do not interact with platelets nor participate in blood coagulation. We also suggest mechanisms for the production of such a film by the LTIC surface.

Use of an Optical-Fiber-Based Biosensor to Study the Interaction of Blood Proteins with Solid Surfaces

An optical fiber-based laser biosensor has been used to investigate adsorption of blood lipoproteins and immunoglobulin G to the glass optical-fiber surface. While lipoproteins readily interact (adsorb) with unmodified optical-fiber surface, immunoglobulin G does not bind to the glass surface at a concentration of 5 μg/mL. However, upon glass surface modification with silane, immunoglobulin G binds significantly to the optical fiber. High-density lipoproteins and low-density lipoproteins show a differential interaction with the glass surface, and their interaction was altered upon modification of the glass surface with cholesterol. With these results in mind, the utility of the biosensor for the study of protein interaction with biorelevant surfaces has been outlined.

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.

Protein interactions with solid surfaces following contact with plasma and blood

AbstractTwo aspects of blood protein interactions with solid surfaces are discussed. The first is competitive adsorption among the many proteins in the blood. The general factors which determine the composition of the adsorbed layer in any multicomponent protein system are considered and then related to current knowledge of competitive adsorption in blood. The latter pertains largely to the so‐called Vroman effect whereby proteins are adsorbed and displaced from the surface in an ordered sequence. Secondly the possible transformation of adsorbed enzyme precursors into active enzymes by the agency of the surface is discussed. The example of the plasminogen‐plasmin system is used to illustrate this discussion.