Design of functional hollow fiber membranes modified with phospholipid polymers for application in total hemopurification system

In this study, we prepared cellulose acetate (CA) hollow fiber membranes (HFMs) modified with poly (2-methacryloyloxyethyl phosphorylcholine (MPC)-co-n-butyl methacrylate)(PMB30 and PMB80) by the dry-jet wet spinning process. The physical and chemical structures of the HFMs were controlled in order to design highly functional HFMs that had suitable performance to each targeting HFM device used in a total hemopurification system. The CA HFMs modified with the MPC polymer, such as CA/PMB30, CA/PMB80, and CA/PMB30-80 HFMs, were successfully prepared by controlling the spinning conditions. The modified HFMs showed an improved performance in solute and water permeability, due to the modification by the hydrophilic MPC polymers. The CA/PMB30 and CA/PMB80 showed a high potential in an application for a high performance hemocompatible plasmapheresis and hemofilter device. Furthermore, CA/PMB30-80 HFM, modified asymmetrically with PMB30 and PMB80, showed a potential for application in an advanced total hemopurification system as a highly functional scaffold for a biohybrid renal tubule, or a liver assist bioreactor device, because of their enhanced permeability, hemocompatibility, and cytocompatibility.     

Cellulose acetate hollow fiber membranes blended with phospholipid polymer and their performance for hemopurification

Commercially available hollow fiber membranes (HFMs) made from synthetic polymers, including cellulose acetate (CA) HFMs, used as hemopurification membranes, need to improve in hemocompatibility, by suppressing protein adsorption and clot formation. In this study, CA HFMs blended with 2-methacryloyloxyethyl phosphorylcholine (MPC) copolymer (PMB30 composed of MPC and n-butyl methacrylate (BMA)) were prepared by a dry-jet wet spinning process. Their performances were evaluated by characterizing their properties such as structure, permeability and protein adsorption. CA/PMB30-blend HFMs showed structure changes such as increase of porosity, development of large pores and decreasing of the thickness of the active layer. And the structure and permeability of CA/PMB30-blend HFMs were controllable by changing preparation conditions. Also, the CA/PMB30-blend HFMs had good permeability, low protein adsorption and low fouling property during the permeability experiment in comparison with CA HFMs, because the hydrophilic and hemocompatible MPC copolymer (PMB30) existed on the surface of the HFM.     

High functional hollow fiber membrane modified with phospholipid polymers for a liver assist bioreactor

For practical application of a liver assist system with a tissue-conjugated hollow fiber membrane (HFM) bioreactor used in an extracorporeal therapy, it would require a highly sophisticated HFM which has both hemocompatibility on one side and cytocompatibility on the other side. In this study, we present a cellulose acetate (CA) HFM modified with 2-methacryloyloxyethyl phosphorylcholine (MPC) copolymers (PMB30 (MPC-co-n-butyl methacrylate) and PMA30 (MPC-co-methacrylic acid) for preparing a novel liver assist HFM bioreactor. A CA/PMB-PMA30 HFM modified asymmetrically on the inner and outer surface with the PMB30 and PMA30 was prepared successfully. Analysis with an X-ray photoelectron spectroscope showed that the intensity of the phosphorus atom attributed to the MPC units on the outer surface of the modified HFM was stronger than that of the inner surface. The PMA30 was immobilized on the outer surface of the CA/PMB30 blend HFM by a chemical condensation reaction. The CA/PMB-PMA30 HFM showed good water and solute permeability in comparison with the CA HFM. The morphologies of the adherent hepatocytes were round in shape in comparison with the cells that adhered on CA HFM. Furthermore, hepatocytes cultured on the inner surface of the CA/PMB-PMA30 HFM showed higher functional expression in terms of urea synthesis and albumin synthesis than that of the CA HFM.     

Facilitated transport of alkali metal picrates through acetylcellulose blend membranes containing synthetic macrolides

Permeation of alkali metal picrates through acetylcellulose membranes containing synthetic macrolides ( 4 and 5 ) was examined in tris-buffer solutions (pH 7,8) to evaluate the effectiveness of the macrolides as ion carriers. The macrolides 4 and 5 are 20- and 25-membered macrocyclic oligoesters, respectively, consisting of alternating tetrahydropyran ring and ester moieties, which were prepared by cationic ring-opening polymerization of 6,8-dioxabicyclo[3.2.1]octan-7-one ( 1 ). Ion permeability through the blend membranes increased with increasing macrolide content. The macrolide 5 facilitated ion transport more effectively than the macrolide 4 . Dependence of ion permeability on solute concentration in a source phase was analyzed by using Lineweaver-Burk's equation, and it was revealed that the ion transport through the blend membranes obeyed kinetics of a Michaelis-Menten type. Ion selectivity sequence for the permeation of alkali metal picrates through the membrane containing 5 was, although not remarkable, Na⁺ > K⁺ > Cs⁺, in sharp contrast to the previously reported ion selectivity sequence for the transport mediated by the macrolides through liquid organic membranes.     

Enhanced biocompatibility and antibacterial property of polyurethane materials modified with citric acid and chitosan

Citric acid (CA) and chitosan (CS) were covalently immobilized on polyurethane (PU) materials to improve the biocompatibility and antibacterial property. The polyurethane pre-polymer with isocyanate group was synthesized by one pot method, and then grafted with citric acid, followed by blending with polyethersulfone (PES) to prepare the blend membrane by phase-inversion method so that chitosan can be grafted from the membrane via esterification and acylation reactions eventually. The native and modified membranes were characterized by attenuated total reflectance-Fourier transform infrared spectroscope, X-ray photoelectron spectroscopy, scanning electron microscopy, water contact angle measurement, and tensile strength test. Protein adsorption, platelet adhesion, hemolysis assay, activated partial thromboplastin time, prothrombin time, thrombin time, and adsorption of Ca²⁺ were executed to evaluate the blood compatibility of the membranes decorated by CA and CS. Particularly, the antibacterial activities on the modified membranes were evaluated based on a vitro antibacterial test. It could be concluded that the modified membrane had good anticoagulant property and antibacterial property.     

Permeability of PEUU membranes: their modification towards blood compatibility

An attempt was made to develop haemodialysis membranes using polyether urethane urea synthesised in our laboratory. It was observed that the processing parameters such as precipitation medium, precipitation temperature etc. can influence the porosity of the membrane and subsequently the permeability property. It was also noted that the permeability of the dried membrane was negligible even though it was kept in distilled water overnight before use. The effect of pH on permeability through the membrane was studied by dialysis experiment using mixtures of various components such as urea, creatinine, uric acid, inulin, albumin, NaCl and KCl at various pH. Standard cellulose acetate (CA) membrane was used for comparison. Membranes were also prepared using biomer solution by precipitating in distilled water at room temperature and the monomer, 2-hydroxy ethyl methacrylate (HEMA) was grafted onto it by glow discharge technique. It was found that the permeability was increased by HEMA grafting with some loss of tensile strength and strain. A comparative study of fibrinogen adsorption during dialysis and adsorption by direct exposure of samples to a mixture containing urea, uric acid, creatinine, dextran, fibrinogen and electrolytes like sodium and potassium ions was also done with 125I labelled fibrinogen. Platelet adhesion studies indicated that the number of adhered platelets was less on the HEMA grafted samples which may enhance blood compatibility. Finally, the membranes were subjected to different sterilization processes possible under wet conditions such as glutaraldehyde treatment and autoclaving. The contact angle, permeability, mechanical property and platelet adhesion studies indicated that the sterilization method can affect the performance of the membrane.     

A novel approach for blood purification: mixed-matrix membranes combining diffusion and adsorption in one step

Hemodialysis is a commonly used blood purification technique in patients requiring kidney replacement therapy. Sorbents could increase uremic retention solute removal efficiency but, because of poor biocompatibility, their use is often limited to the treatment of patients with acute poisoning. This paper proposes a novel membrane concept for combining diffusion and adsorption of uremic retention solutes in one step: the so-called mixed-matrix membrane (MMM). In this concept, adsorptive particles are incorporated in a macro-porous membrane layer whereas an extra particle-free membrane layer is introduced on the blood-contacting side of the membrane to improve hemocompatibility and prevent particle release. These dual-layer mixed-matrix membranes have high clean-water permeance and high creatinine adsorption from creatinine model solutions. In human plasma, the removal of creatinine and of the protein-bound solute para-aminohippuric acid (PAH) by single and dual-layer membranes is in agreement with the removal achieved by the activated carbon particles alone, showing that under these experimental conditions the accessibility of the particles in the MMM is excellent. This study proves that the combination of diffusion and adsorption in a single step is possible and paves the way for the development of more efficient blood purification devices, excellently combining the advantages of both techniques.     

Tissue response to poly(L-lactic acid)-based blend with phospholipid polymer for biodegradable cardiovascular stents

A temporary cardiovascular stent device by bioabsorbable materials might reduce late stent thrombosis. A water-soluble amphiphilic phospholipid polymer bearing phosphorylcholine groups (PMB30W) was blended with a high-molecular-weight poly(l-lactic acid) (PLLA) to reduce unfavorable tissue responses at the surface. The PLLA implants and the polymer blend (PLLA/PMB30W) implants were inserted into subcutaneous tissues of rats, the infrarenal aorta of rats, and the internal carotid arteries of rabbits. After 6 months subcutaneous implantation, the PLLA/PMB30W maintained high density of phosphorylcholine groups on the surface without a significant bioabsorption. After intravascular implantation, the cross-sectional areas of polymer tubing with diameters less than 1.6?mm were histomorphometrically measured. Compared to the PLLA tubing, the PLLA/PMB30W tubing significantly reduced the thrombus formation during 30?d of implantation. Human peripheral blood mononuclear cells were cultured on the PLLA and the PLLA/PMB30W to compare inflammatory reactions. Enzyme-linked immunosorbent assay quantified substantially decreased proinflammatory cytokines in the case of the PLLA/PMB30W. They were almost the same level as the negative controls. Thus, we conclude that the phosphorylcholine groups could reduce tissue responses significantly both in vivo and in vitro, and the PLLA/PMB30W is a promising material for preparing temporary cardiovascular stent devices.     

Transport of water vapour and gases in modified cellulose acetate matrices. Influence of the nature of the penetrant on diffusion and relaxation kinetics

Membranes developed by modification of a cellulose acetate (CA) matrix were studied by means of the flow method CA matrix is modified by poly(4-vinylpyridine) (PVP), or an interpenetrating network (IPN) agent. The obtained diffusion coefficients, permeability coefficients and relaxation kinetic parameters for oxygen, carbon dioxide and water vapour enabled us to consider not only the transport properties of the membranes but also the influence of the relaxation processes on the membrane permeability and stability. The results show that the interactions between water molecules and polar groups on the polymer chains cause the membrane material to swell. This swelling process provokes an increase in the permeation flux of ca. 20–30%. Oxygen causes a consolidation of unstable membranes of the cellulose acetate matrix modified by IPN but not of blends with PVP. The consolidation process reduces the membrane permeability by ca. 38%, while carbon dioxide has no effects on the membrane properties. Membranes based on a modified cellulose acetate matrix have higher permeability than pure CA membranes.     

Protein adsorption and separation on amphoteric chitosan/carboxymethylcellulose membranes

This article reported the preparation of an amphoteric natural polymeric membrane-macroporous chitosan (CS)/carboxymethylcellulose (CMC) blend membrane and the utilization of such a membrane on the membrane chromatography for bioseparation. The membranes were prepared by solution blending of CS and CMC solution, and using silica particles as porogen. Both glutaraldehyde and epichlorohydrin were used as crosslinking agent to increase its chemical stability in aqueous solution. Such a natural polymeric membrane can be served as an amphoteric membrane because of the amino group on CS and the carboxymethyl group on CMC, in which the surface charge can be changed with the environmental pH. Ovalbumin (pI = 4.6) and lysozyme (pI = 11) were selected as model proteins. These two proteins adsorption on different CS/CMC blend membranes with different initial protein concentrations at different pH values were investigated in batch systems. The results indicated that the maximum adsorption for lysozyme and ovalbumin was at pH 9.2 and 4.8 respectively, and the adsorption capacity on the membrane both increased with the increase of initial protein concentration. Though the adsorption mechanism of lysozyme and ovalbumin was found not the same, the maximum adsorption capacity of two proteins on the membranes was quite similar (about 250 mg/g). Moreover, the desorption ratio of both proteins was found to be more than 90% that implied CS/CMC blend membrane could separate proteins by adsorption-desorption process. Finally, both lysozyme and ovalbumin were successfully separated from their binary mixture only by adjusting the pH of the feed and the desorption solution.     

Immunopurification of the blood group RhD protein from human erythrocyte membranes

Rh proteins are membrane proteins encoded by genes at the blood group RH locus. They are of paramount importance in transfusion medicine, but their function is still unknown. Biochemical and biophysical studies of these proteins are scarce since only minute amounts of the very hydrophobic Rh proteins, can be purified from human erythrocytes. Recently, a human monoclonal antibody (LOR-15C9) was described as having the unique property to recognize the Rh30 protein carrying the major blood group D specificity (RhD protein), either in a membrane detergent extract or when blotted on a membrane. In this report, we describe one-step purification of the RhD protein from detergent extracts of red cell membranes, based on immunoaffinity chromatography carried out with immobilized LOR-15C9 IgG. The technique yielded RhD protein with high purity which was devoid of other associated proteins (RhAG, CD47, LW and GPB) that comprise the Rh complex in the erythrocyte membrane. By contrast immunoprecipitation performed with the same antibody led to co-isolation of both RhD and RhAG.     

Clinical characterization of a new polymeric membrane for use in renal replacement therapy

Renal replacement therapy makes extensive use of semi-permeable membranes, ideal requirements for such membranes are good solute transport characteristics and a low reactivity with blood. Membranes manufactured from synthetic polymers fulfil these requirements. Such membranes have asymmetric and anisotropic structures characterized by a dense layer with which the blood is in contact supported by a thicker solid structure with containing interlinked voids, providing support. The nature of the structures are critically dependent upon the polymer blend and the control of parameters during manufacture such as the temperature or additive concentrations. In this prospective study, we have evaluated the clinical performance of a new membrane manufactured from a blend of polyamide, polyarylethersulfone and polyvinylpyrrolidone (Polyflux, Gambro GmbH, Hechingen, Germany), and compared it with that of polysulfone blended with polyvinylpyrrolidone (Fresenius Polysulfone, Fresenius Medical Care, Bad Homburg, Germany), a material widely acknowledged as providing an optimal biocompatibility in terms of solute removal and complement activation. The clearance of small molecules (urea, creatinine, phosphate) for both membranes was comparable. Both membranes removed beta2 microglobulin during treatment (50.2% reduction with Polyflux and 54.5% reduction with polysulfone. This removal due to the non-selectivity of the membranes was associated with protein loss during therapy which was similar for both the membranes (7.7 g). The biocompatibility profiles of the membranes indicated slight neutropenia and platelet adhesion and minimal C3a, C5a and SC5b-9 generation which were independent of the membrane material. These findings indicate that despite the differences in microstructure of the membranes, their functional performance in the clinical setting is comparable.     

Modification of polysulfone with phospholipid polymer for improvement of the blood compatibility. Part 1. Surface characterization.

To improve the surface blood compatibility of polysulfone (PSf) membranes, we prepared novel polymeric additives which have suitable blood compatibility. They were polymers with a phosphorylcholine group, a 2-methacryloyloxyethyl phosphorylcholine (MPC) unit. The MPC polymer could be blended with polysulfone by a solvent evaporation method during membrane processing, and a transparent membrane could be obtained. The mechanical properties of the blend membrane were similar to that of the original PSf membrane. Surface analysis of the blend membrane by X-ray photoelectron spectroscopy and dynamic contact angle measurement revealed that the MPC unit in the polymeric additive was concentrated on the surface of the membrane. The blend membrane significantly reduced plasma protein adsorption compared with that of the PSf membrane.     

壳聚糖与硫酸软骨素共混膜性质的研究

以壳聚糖和硫酸软骨素按一定比例制备出共混膜，研究了膜片的透光性、含水量、渗透性、力学性质、表面结构、生物降解性、生物相容性等性质。结果表明该共混膜具有较好的透光性、通透性、生物降解性和生物相容性，膜表面较粗糙。以此共混膜为载体培养兔角膜基质细胞，发现细胞在此共混膜上生长良好。制备膜片随着加入CaSO4量的增加，膜的通透性也随之增加。     

Diffusion transport mechanisms in synthetic polymer membranes for haemodialysis

New synthetic polymer membranes have been developed as replacements for the cellulose films currently used in haemodialysis. Cationic, anionic and chemically modified membranes have been prepared. Permeabilities of the synthetic membranes to blood components have been measured and compared with standard 150 PT and 300 PT cellulose films. Results obtained with cellulose films confirm the previous conclusion that permeability is principally in inverse proportion to the molecular volume of the diffusing solute. Cationic and anionic membranes possess selectivity to certain blood poisons. Cationic membranes chemically modified with ethylene oxide permit mass transfer rates for specific metarials which are dependent on the degree and conditions of reaction achieved. A new mechanism of solute transport for perm-selective membranes is proposed, since the improved film permeability of the new materials is not explicable solely on the basis of pore density or dimensions.     

聚砜中空纤维血液透析膜的制备与表征

为满足医学治疗的需要,提出一种以期用于血液透析的聚砜中空纤维膜制备方法。以聚砜(PSF)为膜材料,一缩二乙二醇(DEG)为致孔剂,聚乙烯吡咯烷酮(PVP-K30)为改性剂,N,N-二甲基乙酰胺(DMAC)为溶剂,应用非溶剂致相分离法(NIPS)制备中空纤维血液透析膜。采用扫描电镜、透析实验、接触角和孔隙率测试、拉伸测试等方法,表征不同质量分数的聚砜(17%、19%、21%、23%)对膜的微观结构、渗透性能、亲水性和机械性能的影响。利用肌酐、磷酸氢二钠和维生素B₁₂配制成模拟液,进行模拟血液透析实验,测试不同聚砜质量分数的透析膜对溶质的清除效果。对不同分子量(1 000、2 000、4 000、6 000、10 000、20 000)聚乙二醇(PEG)的水溶液进行溶质截留测试,计算得到透析膜的平均孔径和切割分子量。结果表明:当铸膜液中PSF的质量分数为21%,添加剂DEG和PVP的质量分数均为10%时,能够得到性能最优的透析膜;所得透析膜超滤系数达185.1 mL/m²·h·mmHg,对中小型分子的清除率达50%,断裂强度达5.22 MPa,膜表面孔径为5.00 nm,切割分子量为14 200。基于其良好的渗透性能和机械性能,该聚砜中空纤维膜有成为理想的血液透析膜材料的潜在可能。     

Red cell membrane CO2 permeability in normal human blood and in blood deficient in various blood groups, and effect of DIDS.

The red cell membrane has an exceptionally high permeability for CO2, PCO2 approximately 0.15 cm/s, which is two to three orders of magnitude greater than that of some epithelial membranes and similarly greater than the permeability of the red cell membrane for HCO3-. As shown previously, this high PCO2 can be drastically inhibited by 10 microM 4,4'-diisothiocyanato-2,2'-stilbenedisulfonate (DIDS), indicating that membrane proteins may be involved in this high gas permeability. Here, we have studied the possible contribution of several blood group proteins to CO2 permeation across the red cell membrane by comparing PCO2 of red cells deficient in specific blood group proteins with that of normal red cells. While PCO2 of normal red cells is approximately 0.15 cm/s and that of Fy(null) and Jk(null) red cells is similar, PCO2's of Colton null (deficient in aquaporin-1) and Rh(null) cells (deficient in Rh/RhAG) are both reduced to about 0.07 cm/s, i.e. to about one half. In addition, the inhibitory effect of DIDS is about half as great in Rh(null) and in Colton null red cells as it is in normal red cells. We conclude that aquaporin-1 and Rh/RhAG proteins contribute substantially to the high permeability of the human red cell membrane for CO2. Together these proteins are responsible for 50% or more of the CO2 permeability of red cell membranes. The CO2 pathways of both proteins can be partly inhibited by DIDS, which is why this compound very effectively reduces membrane CO2 permeability.     

Fabrication of zein/hyaluronic acid fibrous membranes by electrospinning

Electrospinning of biopolymers, such as proteins and polysaccharides, has recently attracted much attention for the fabrication of scaffolds for tissue engineering. In this report, zein/hyaluronic acid (HA) blend fibrous membranes were electrospun and characterized. To facilitate the compatibility of zein and HA, poly(vinyl pyrrolidone) (PVP) was introduced into aqueous ethanol solutions of the blend. A series of zein/HA/PVP blend fibrous membranes with different volume ratios were successfully electrospun. The effect of blend composition on the morphology of electrospun fibrous membranes was investigated by scanning electron microscopy. The average diameter of blend fibers increased with increasing the content of zein component. The electrospun zein/HA/PVP fibrous membranes were then cross-linked by methylene diphenyl diisocyanate (MDI). The morphology of the cross-linked zein/HA/PVP fibrous membranes changed slightly compared with un-cross-linked membranes. Tensile tests demonstrated that the mechanical properties of the zein/HA/PVP fibrous membranes were improved by cross-linking.     

Endotoxin removal from albumin and saline solutions

Microporous membranes have been developed which can remove endotoxins selectively from electrolyte and albumin solutions by regioselective adsorption in the membrane matrix and outside surface of the membrane. The membranes were prepared in the form of hollow fibre membranes in a continuous process. By varying the membrane preparation parameters, different pore sizes and adsorption capacities could be realized, thus broadening applications for biological purification. Dynamic adsorption capacities for endotoxin from albumin and saline solution were determined and were found to be in the range of 0.2 and 0.1 microg endotoxin/g membrane, respectively, suggesting different adsorption mechanisms.     

Fabrication of zein/hyaluronic acid fibrous membranes by electrospinning

Electrospinning of biopolymers, such as proteins and polysaccharides, has recently attracted much attention for the fabrication of scaffolds for tissue engineering. In this report, zein/hyaluronic acid (HA) blend fibrous membranes were electrospun and characterized. To facilitate the compatibility of zein and HA, poly(vinyl pyrrolidone) (PVP) was introduced into aqueous ethanol solutions of the blend. A series of zein/HA/PVP blend fibrous membranes with different volume ratios were successfully electrospun. The effect of blend composition on the morphology of electrospun fibrous membranes was investigated by scanning electron microscopy. The average diameter of blend fibers increased with increasing the content of zein component. The electrospun zein/HA/PVP fibrous membranes were then cross-linked by methylene diphenyl diisocyanate (MDI). The morphology of the cross-linked zein/HA/PVP fibrous membranes changed slightly compared with un-cross-linked membranes. Tensile tests demonstrated that the mechanical properties of the zein/HA/PVP fibrous membranes were improved by cross-linking.