Low-density Lipoprotein Adsorption Research Articles

Simultaneous Rosiglitazone Release and Low-Density Lipoprotein Removal by Chondroitin Sodium Sulfate/Cyclodextrin/Poly(acrylic acid) Composite Adsorbents for Atherosclerosis Therapy.

Atherosclerosis (AS) is characterized by the accumulation of substantial low-density lipoprotein (LDL) and inflammatory response. Hemoperfusion is commonly employed for the selective removal of LDL from the body. However, conventional hemoperfusion merely focuses on LDL removal and does not address the symptom of plaque associated with AS. Based on the LDL binding properties of acrylated chondroitin sodium sulfate (CSA), acrylated beta-cyclodextrin (CD) and acrylic acid (AA), along with the anti-inflammatory property of rosiglitazone (R), the fabricated AA-CSA-CD-R microspheres could simultaneously release R and facilitate LDL removal for hemoperfusion. The AA and CSA offer electrostatic adsorption sites for LDL, while the CD provides hydrophobic adsorption sites for LDL and weak binding sites for R. According to the Sips model, the maximum static LDL adsorption capacity of AA-CSA-CD-R is determined to be 614.73 mg/g. In dynamic simulated perfusion experiments, AA-CSA-CD-R exhibits an initial cycle LDL adsorption capacity of 150.97 mg/g. The study suggests that the weakened inflammatory response favors plaque stabilization. The anti-inflammatory property of the microspheres is verified through an inflammation model, wherein the microsphere extracts are cocultured with mouse macrophages. Both qualitative analysis of iNOS\TNF-α and quantitative analysis of IL-6\TNF-α collectively demonstrate the remarkable anti-inflammatory effect of the microspheres. Therefore, the current study presents a novel blood purification treatment of eliminating pathogenic factors and introducing therapeutic factors to stabilize AS plaque.

Adsorption of Low Density Lipoproteins on an Azobenzene-Containing Polymer Brush: Modelling by Coarse-Grained Molecular Dynamics

One of the strategies to reduce the contents of low density lipoproteins (LDLs) in blood is a hemoperfusion, when they are selectively retracted from plasma by an adsorber located outside the patient’s body. Recently, a photo-controllable smart surface was developed experimentally, that is characterized by high selectivity and reusability. It comprises a nanocarrier functionalised by a brush of azobenzene-containing polymer chains. We present a mesoscopic model that mimics principal features of this setup and focus our study on the role played by the length and flexibility of the chains and grafting density of a brush. The model for LDL comprises a spherical core covered by a shell of model phospholipids. The model is studied via coarse-grained molecular dynamics simulation, performed with the aid of the GMBOLDD package modified for the case of soft-core beads. We examined the dependencies of the binding energy on both the length of polymer chains and the grafting density of a brush and established optimal conditions for adsorption. These are explained by competition between the concentration of azobenzenes and phospholipids in the same spatial region, flexibility of polymer chains, and excluded volume effects.

Fabrication and Study of Dextran/Sulfonated Polysulfone Blend Membranes for Low-Density Lipoprotein Adsorption.

The abnormal increase in low-density lipoprotein (LDL) in human blood is a main independent risk factor for the pathogenesis of atherosclerosis, whereas a reduced LDL level effectively lowers morbidity. It is important to develop LDL adsorption materials with high efficiency and selectivity, as well as to simplify their fabrication processes. In this paper, polysulfone (PSF), sulfonated polysulfone (SPSF), and sulfonated polysulfone/dextran (SPSF/GLU) membranes were successfully fabricated for LDL adsorption using a solution casting technique. Attenuated total reflectance Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy measurements confirmed the success of the preparation. The water contact angle decreased from 89.7 ± 3.4° (PSF) to 76.4 ± 3.2° (SPSF) and to 71.2 ± 1.9° (SPSF/GLU), respectively. BSA adsorption testing showed that the SPSF/GLU with surface enrichment of sulfonate groups and glycosyl groups possessed higher resistance to protein solution. The adsorption and desorption behaviors of the studied samples in single-protein or binary-protein solutions were systematically investigated by enzyme-linked immunosorbent assay (ELISA), The results showed that SPSF/GLU, which had excellent resistance to protein adsorption, possessed a similar adsorption capacity to that of PSF. SPSF membrane exhibited excellent selective affinity for LDL in single and binary protein solutions, suggesting potential applications in LDL removal.

Simple emulsion template method towards self-anticoagulant and high-efficiency carboxymethyl chitosan-based adsorbent for low-density lipoprotein from whole blood

In this study, carboxymethyl chitosan (CMCS)-based double cross-linked network spheres, are constructed by an emulsion template method. The first crosslinking network comes from the chelation between the CMCS and zinc ions, while the second one is based on the covalent crosslinking of 2-acrylamido-2-methyl-1-propanesulfonicacid (AMPS); and the as-prepared spheres are termed as ECMCS@AMPSs. The emulsion template method endows the ECMCS@AMPSs with large pore area (58.3 m2/g) and ultra-high porosity (92.3 %). Thanks to the heparin-mimicking functional groups of –OH, –COO- and -SO3-, the ECMCS@AMPSs show excellent anticoagulant properties without the activation of the complement system, contact system and platelets. Compared with the spheres without emulsification, the porous structure of the ECMCS@AMPSs results in a 5.3-fold increase in low-density lipoprotein (LDL) adsorption capacity in hypercholesterolemia plasma. Most importantly, in vitro simulated hemoperfusion experiment shows that the cumulative adsorption capacity of LDL by the ECMCS@AMPSs is as high as 41.66 mg/g in 1 h, which is approximately 4.2 times that of high-density lipoprotein (HDL). In conclusion, the ECMCS@AMPSs with self-anticoagulant and high-efficiency adsorption of LDL fabricated via simple emulsion template method have great clinical application prospects in the field of lipoprotein apheresis.

Photorenewable Azobenzene Polymer Brush-Modified Nanoadsorbent for Selective Adsorption of LDL in Serum

The elevated concentration of low-density lipoprotein (LDL) is recognized as a leading factor of hyperlipidemia (HLP), and selective adsorption of serum LDL is regarded as a practical therapy. Based on the superior structure-function characteristics of stimuli-responsive materials, a photorenewable nanoadsorbent (SiO2@Azo@Gly) with high selectivity and reusability was developed using azobenzene as the functional ligand. Its principle was certified by the preparation of silicon nanoparticles with atom transfer radical polymerization (ATRP)-initiating groups via a sol-gel reaction and their subsequent grafting of azobenzene polymer brushes by surface-initiated ATRP, followed by modification with glycine. Immobilization of carboxylated azobenzene polymer brushes onto the nanoparticles endowed SiO2@Azo@Gly with high adsorption selectivity and reusability. The advanced nanoadsorbent exhibited excellent LDL adsorption capacity at about 27 mg/g and could be regenerated by illumination with high efficiency (circulations ≥ 5); this was further verified by transmission electron microscopy (TEM) and Fourier-transform infrared (FTIR) analysis. SiO2@Azo@Gly also demonstrated superior adsorption efficiency and selectivity in serum from HLP patients, the respective adsorption capacities of LDL, triglyceride, and total cholesterol were about 15.65, 24.48, and 28.36 mg/g, and the adsorption to high-density lipoprotein (cardioprotective effect) was only about 3.66 mg/g. Green regeneration of the nanoadsorbent could be achieved completely through a simple photoregeneration process, and the recovery rate was still 97.9% after five regeneration experiments.

Biomembrane-mimetic hemoperfusion adsorbent for efficient removal of low-density lipoprotein from hyperlipemia blood.

Lowering of low-density lipoprotein (LDL) levels in blood of patients with hyperlipidaemia can effectively prevent the progression of atherosclerosis and coronary heart disease. The present study demonstrated a facile synthesis strategy to prepare biomembrane-mimetic LDL adsorbent (PVA@COOH-PE) via directional immobilization of phospholipid onto macro-porous cross-linked poly(vinyl alcohol) spheres. The binding between the prepared adsorbent and LDL particles simulates the cytosolic lipid droplets to form a lipid-packing structure. The adsorbent possesses satisfactory removal efficiency for LDL and total cholesterol (TCH) in hyperlipemia serum, while remains high-density lipoprotein (HDL) concentration within the normal range. The adsorption capacities for LDL and TCH are about 1.13 and 1.74 mg/ml respectively, which are nearly three and four times higher than that of HDL (0.42 mg/ml). The adsorbent also possesses satisfactory anticoagulant properties, causes negligible effect on blood cells and produces low hemolysis ratios. The excellent blood compatibility plus LDL removal efficiency of PVA@COOH-PE indicates its good application prospect as hemoperfusion adsorbent in the treatment of hyperlipidaemia.

Familial Hypercholesterolemia and Lipoprotein Apheresis.

Lipoprotein apheresis has been developed as the treatment for refractory familial hypercholesterolemia (FH) to remove low-density lipoprotein (LDL), which is the main pathogenic factor. Currently, three procedures are available in Japan, including the plasma exchange, double-membrane filtration, and selective LDL adsorption. Selective LDL adsorption, which was developed in Japan, has been one of the most common treatment methods in the world. Lipoprotein apheresis enabled the prevention of atherosclerosis progression even in homozygous FH (HoFH) patients. However, in our observational study, HoFH patients who started lipoprotein apheresis in adulthood had a poorer prognosis than those who started in childhood. Therefore, HoFH patients need to start lipoprotein apheresis as early as possible. Although the indication for lipoprotein apheresis in heterozygous FH (HeFH) patients has been decreasing with the advent of strong statins, our observational study showed that HeFH patients who discontinued lipoprotein apheresis had a poorer prognosis than patients who continued apheresis therapy. These results suggest that it is beneficial for very-high-risk HeFH patients to be treated by lipoprotein apheresis even if their LDL cholesterol is controlled well by lipid-lowering agents. Since launching a new class of lipid-lowering agents, proprotein convertase subtilisin/kexin type 9 (PCSK9) antibody and microsome triglyceride transfer protein inhibitors, the indication for lipoprotein apheresis in FH has been changing. However, despite the development of these drugs, lipoprotein apheresis is still an option with a high therapeutic effect for FH patients with severe atherosclerotic cardiovascular disease.

Low density lipoprotein adsorption on a titanium surface and its effect on osteoblast behaviors.

Objective: This study aims to investigate the adsorption of low density lipoprotein (LDL) on a titanium surface and to explore its effect on osteoblast behaviors. Materials and methods: LDL adsorption on a titanium surface was analyzed using LDL assay and X-ray photoelectron spectroscopy (XPS). Physical properties, including topography, surface roughness and wettability of a control smooth titanium surface and a LDL pre-adsorbed titanium surface, were assessed. Subsequently, the adhesion, proliferation and differentiation abilities of MC3T3-E1 cells (an osteoblast-like cell line) on the surfaces of control titanium and LDL pre-adsorbed titanium were investigated. Results: LDL assay and XPS confirmed LDL adsorption on the titanium surface. The maximum adsorption of LDL on the titanium surfaces was observed after 150 minutes of incubation. In comparison with the control smooth titanium surface, the roughness and hydrophilicity of the LDL pre-adsorbed titanium surface were significantly altered. Furthermore, in vitro studies demonstrated that LDL adsorption obviously attenuated the adhesion, proliferation and differentiation of MC3T3-E1 cells on the titanium surface. Conclusion: LDL could adsorb on a titanium surface. Meanwhile, LDL adsorption changed the characteristics of the titanium surface, which, in turn, negatively regulated osteoblast behaviors.

A mussel-inspired approach towards heparin-immobilized cellulose gel beads for selective removal of low density lipoprotein from whole blood

In this study, we report a mussel-inspired approach to fabricate heparin-immobilized cellulose (HeTaCe) gel beads with self-anticoagulative and biocompatible properties which can selectively remove low density lipoprotein (LDL) from whole blood directly. First, a phase inversion technique was applied to prepare cellulose gel beads. Then the as-prepared gel beads were dipped into a mixed solution of heparin and tannic acid in phosphate buffered saline (PBS, pH 8.5) to obtain HeTaCe gel beads. Blood compatibility experiments indicated that the HeTaCe gel beads could suppress complement activation as well as contact activation and prolong the clotting times to the upper detect limits (activated partial thromboplastin time >600 s and thrombin time >180 s) of the automated blood coagulation analyzer. An ideal adsorption capacity of LDL in vitro was achieved by the HeTaCe gel beads with an amount of 79.1 mg/g. Besides, dynamic column adsorption test further demonstrated a selective adsorption of LDL without a significant reduction of high density lipoprotein (HDL) in a simulative hemoperfusion system. It is believed that the HeTaCe gel beads will be quite appealing to future clinical practice aiming at lowering LDL and improving the outcomes of patients with high cardiovascular risk.

Selective and Regenerable Surface Based on β-Cyclodextrin for Low-Density Lipoprotein Adsorption.

Cyclodextrins (CDs) are a family of cyclic oligosaccharides, whose unique hydrophilic outer surface and lipophilic central cavity facilitate the formation of inclusion complexes with various biomolecules, such as cholesterol and phospholipids, via multi-interactions. Low-density lipoprotein (LDL) is the main carrier of cholesterol in bloodstream and is associated with the progression of atherosclerosis. The surface of LDL is composed of a shell of phospholipids monolayer containing most of the free unesterified cholesterol as well as the single copy of apolipoprotein B-100. To date, various LDL adsorbents have been fabricated to interact with the biomolecules on LDL surface. Owing to its elegant structure, CD is considered to be a promising choice for preparation of more economical and effective LDL-adsorbing materials. Therefore, in this study, interaction between β-CD and LDL in solution was investigated by dynamic light scattering, circular dichroism, and ultraviolet spectroscopy. Further, a supramolecular surface based on β-CD was simply prepared by self-assembled monolayer on gold surface. The effect of hydrogen bond and the cavity of β-CD on the interaction between β-CD and LDL was particularly explored by surface plasmon resonance (SPR) analysis. The SPR results showed that such β-CD-modified surface exhibited good selectivity and could be largely regenerated by sodium dodecyl sulfate wash. This study may extend the understanding of the interaction between LDL and LDL adsorbent or the design and development of more efficient and lower-cost LDL adsorbents in the future.

Anionic glycosylated polysulfone membranes for the affinity adsorption of low-density lipoprotein via click reactions.

Low-density lipoprotein (LDL) adsorbents normally contain negative charged ligand to induce electrostatic interaction with the positively charged regions of LDL. Furthermore, saccharide is another common component which share in most of the LDL-adsorbents and the LDL-receptor (LDLR). Such structural similarity impels us to investigate the synergistic effect of anionic and saccharide on LDL recognition. For this purpose, an anionic glycosylated membrane of which surface composition can be controlled by click reaction with mutable glycosyl/sulfonyl ratios was prepared. The obtained membrane showed better LDL adsorption/desorption property and the adsorption amount for LDL at an optimum feed ratio. This finding highlights the role of synergistic effect of anionic and saccharide, which offer a new strategy for designing LDL adsorbent with high efficiency.

Facile preparation of heparinized polysulfone membrane assisted by polydopamine/polyethyleneimine co-deposition for simultaneous LDL selectivity and biocompatibility

Low-density lipoprotein (LDL) gains worldwide attention for decades as the key risk factor to atherosclerosis that progressively deteriorating into cardiovascular diseases. Until recent years, LDL-apheresis comes to be extensively used as a direct and efficient LDL removal method, with LDL adsorption materials particularly important. In this paper, a new strategy based on the co-deposition of polydopamine (PDA) with polyethylenimine (PEI) onto polysulfone (PSf) membranes, then subsequent heparinization by amino-carbonyl reactions, to achieve LDL selectivity and simultaneous biocompatibility, is proposed. Surface properties of modified PSf membranes are characterized by ATR-FTIR, XPS, FESEM, Zeta potential and WCA measurements. LDL adsorption ability is investigated by ELISA, while blood biocompatibility is evaluated by platelet adhesion experiments. Results suggest that heparin-modified PSf membranes show high selectivity for LDL removal and fine biocompatibility in contact with plasma, as excellent potential materials for LDL-apheresis.

Fe3O4@ZnO core-shell nanocomposites for efficient and repetitive removal of low density lipoprotein in plasma and on blood vessel

Low density lipoprotein (LDL)-apheresis therapy, which directly removes LDL from plasma by LDL-adsorbents in vitro is found to be clinically effective and safe to lower the LDL content in blood to prevent cardiovascular disease. Thus, developing excellent LDL adsorbents are becoming more and more attractive. Herein, functional Fe3O4@ZnO core–shell nanocomposites have been synthesized by a facile and eco-friendly two-step method. Not only do they possess high LDL adsorption (in PBS/plasma as well as on blood vessels) and favorable magnetic targeting ability but they can also be reused conveniently, which offer the Fe3O4@ZnO core–shell nanocomposites significant potential in the removal of LDL in vitro and in vivo.

Surface glycosylation of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) membrane for selective adsorption of low-density lipoprotein

A novel method of constructing a glycosylated surface on poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] membrane surface for the selective adsorption of low-density lipoprotein (LDL) was developed, which involved the photoinduced graft polymerization of acrylic acid followed by the chemical binding of carboxyl groups with glucosamine in the presence of 1-ethyl-3-(dimethyl-aminopropyl) carbodiimide hydrochloride and N-hydroxy-succinimide. The chemical structures of the fabricated membranes were characterized by attenuated total reflectance Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. Zeta potential and water contact angle measurements were performed to investigate the surface charge and wettability of the membranes, respectively. An enzyme linked immunosorbent assay was used to measure the LDL adsorption on the plain and modified membrane surfaces. It was found that the surface glycosylation of P(3HB-co-4HB) membrane greatly enhanced the affinity interactions with LDL and the absorbed LDL could be easily desorbed with eluents, indicating a specific and reversible binding of LDL to the surface. Furthermore, the hemocompatibility of glycosylated membrane was improved as examined by platelet adhesion. The results suggest that the glycosylated P(3HB-co-4HB) membrane is promising for application in LDL apheresis therapy.

Immobilization of sodium alginate sulfates on polysulfone ultrafiltration membranes for selective adsorption of low-density lipoprotein

A novel method for the immobilization of sodium alginate sulfates (SAS) on polysulfone (PSu) ultrafiltration membranes to achieve selective adsorption of low-density lipoprotein (LDL) was developed, which involved the photoinduced graft polymerization of acrylamide on the membrane and the Hofmann rearrangement reaction of grafted acrylamide followed by chemical binding of SAS with glutaraldehyde. The surface modification processes were confirmed by attenuated total reflectance Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy characterization. Zeta potential and water contact angle measurements were performed to investigate the surface charge and wettability of the membranes. An enzyme-linked immunosorbent assay was used to measure the binding of LDL on plain and modified PSu membranes. It was found that the PSu membrane immobilized with sodium alginate sulfates (PSu-SAS) greatly enhanced the selective adsorption of LDL from protein solutions and the absorbed LDL could be easily eluted with sodium chloride solution, indicating a specific and reversible binding of LDL to SAS, mainly driven by electrostatic forces. Furthermore, the PSu-SAS membrane showed good blood compatibility as examined by platelet adhesion. The results suggest that the PSu-SAS membranes are promising for application in simultaneous hemodialysis and LDL apheresis therapy.

Bioinspired Multiple-Interaction Model Revealed in Adsorption of Low-Density Lipoprotein to Surface Containing Saccharide and Alkanesulfonate

A new "multiple-interaction model" for low-density lipoprotein (LDL) adsorption to a specific surface containing saccharide and alkanesulfonate ligands is proposed. The model suggests that there are interactions of the saccharide component beyond electrostatic interactions of the alkanesulfonate component that both influence the LDL adsorption process. This concept of multiple interactions between saccharide and LDL was inspired by the similarity in structures of LDL receptors (LDLR), heparin, and heparans used in LDL-apheresis. The model was confirmed by SPR analysis by the adsorption maxima on SAM surfaces with different compositions of saccharide and alkanesulfonate and additionally by CD detection of the conformation of LDL when in contact with saccharide.

Successful treatment of monomorphic primary central nervous system post‐transplantation lymphoproliferative disorder 5 years after kidney transplantation

A 31-year-old man underwent living-related kidney transplantation in 2004 as a consequence of primary focal segmental glomerulosclerosis (FSGS). Four years after the transplantation, we confirmed nephrotic syndrome caused by recurrent FSGS. We performed plasmapheresis and low-density lipoprotein adsorption. We also combined steroid therapy with a reduction in the dose of tacrolimus and an increased dose of mycophenolate mofetil. The nephrotic syndrome improved dramatically with this combined therapeutic approach. However, 10months after these treatments, he revisited our hospital because of altered consciousness. We detected multiple tumor masses in his brain that were ring enhanced on contrast magnetic resonance imaging. Consequently, we suspected primary central nervous system post-transplantation lymphoproliferative disorder (CNS-PTLD). We performed a craniotomy to biopsy the brain tumors. The biopsy specimen showed Epstein-Barr virus-associated diffuse large B-cell lymphoma. There is no definitive treatment for CNS-PTLD. Therefore, we treated the primary CNS-PTLD successfully with whole-brain radiation and discontinuation of immunosuppression therapy.

In vitro experimental study on removing low density lipoprotein with HB-H-8 resin adsorption

Objective Currently,adsorption materials applied to remove low density lipoprotein (LDL) still remain defects.HB-H-8 resin has not been put in use in removing LDL.This study focused on primary evaluation of adsorption capability of HB-H-8 resin.Methods HB-H-8 resin was used in the adsorption of LDL of patient that was higher than the normal range of blood sample in dynamic test and static test in vitro.Results The time of adsorption saturation was two hours,and the optimum temperature was 37℃.The results of adsorption in vitro showed that the average adsorption rates of LDL and high density lipoprotein (HDL) were 63.2％ and 1.9％,respectively.Conclusion HB-H-8 resin has LDL higher adsorption capability,thus made it to be a potential blood purification material for the removal of LDL. Key words: Low density lipoprotein; High density lipoprotein; Hemoperfusion; Adsorption

LDL-apheresis: technical and clinical aspects.

The prognosis of patients suffering from severe hyperlipidemia, sometimes combined with elevated lipoprotein (a) levels, and coronary heart disease refractory to diet and lipid-lowering drugs is poor. For such patients, regular treatment with low-density lipoprotein (LDL) apheresis is the therapeutic option. Today, there are five different LDL-apheresis systems available: cascade filtration or lipid filtration, immunoadsorption, heparin-induced LDL precipitation, dextran sulfate LDL adsorption, and the LDL hemoperfusion. There is a strong correlation between hyperlipidemia and atherosclerosis. Besides the elimination of other risk factors, in severe hyperlipidemia therapeutic strategies should focus on a drastic reduction of serum lipoproteins. Despite maximum conventional therapy with a combination of different kinds of lipid-lowering drugs, sometimes the goal of therapy cannot be reached. Hence, in such patients, treatment with LDL-apheresis is indicated. Technical and clinical aspects of these five different LDL-apheresis methods are shown here. There were no significant differences with respect to or concerning all cholesterols, or triglycerides observed. With respect to elevated lipoprotein (a) levels, however, the immunoadsorption method seems to be most effective. The different published data clearly demonstrate that treatment with LDL-apheresis in patients suffering from severe hyperlipidemia refractory to maximum conservative therapy is effective and safe in long-term application.

Preparation of sulfonated porous carbon nanotubes/activated carbon composite beads and their adsorption of low density lipoprotein

The high level of low density lipoprotein (LDL) in plasma is the main cause of atherosclerosis. Hemoperfusion is an ideal therapy to lower the level of LDL in human blood system while therapeutic effect is determined by the adsorbent. The adsorbent must have suitable pore structure and specific functional groups. Carbon nanotubes (CNTs) could be a new adsorbent material because CNTs have high specific surface area and they can be modified by a variety of functional groups. Porous carbon composite beads with the CNTs and phenolic resin mixture were synthesized by suspension polymerization, following with carbonization and steam-activation. Then the porous composite beads were sulfonated with a sulfanilic acid anhydrous by diazotization and coupling reaction. The potential application of the sulfonated porous composite beads in adsorbing low density lipoprotein (LDL) from human serum was investigated. The results showed that the sulfonic acid groups on the composite beads could lower LDL levels greatly by electrostatic interaction with electropositive LDL. The higher 20-100nm pore volume the composite beads had, the more LDL they could adsorb. The 20-100nm pore volume was enhanced by adding more CNTs and improving CNTs dispersion (ultrasonic crushing). The sulfonated composite beads containing 45wt% CNTs presented the highest adsorption capacity to LDL 10.46mg/g, showing a good prospect as LDL adsorbent in hemoperfusion.