Preparation of blood-compatible hollow fibers from a polymer alloy composed of polysulfone and 2-methacryloyloxyethyl phosphorylcholine polymer

Blood-compatible hollow fibers were successfully prepared from a polymer alloy composed of polysulfone (PSf) and the 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer. To improve the hydrophilicity, fouling-resistance characteristics, and blood compatibility of the PSf hollow fiber in a hemodialyzer, an MPC polymer that can be blended with PSf was synthesized in order to prepare the polymer alloy (PSf/MPC polymer). The contents of the MPC polymer blended in the PSf were 7 and 15 wt%. The PSf/MPC polymer hollow fiber could be prepared by both wet and dry-wet processing methods. The hollow fiber took an asymmetric structure, that is, the hollow-fiber membrane had a dense skin layer on the porous sponge-like structure. The mechanical strength was higher than that of conventional PSf hollow fibers for hemodialysis. The surface characterization of the PSf/MPC polymer hollow fiber by x-ray photoelectron spectroscopy revealed that the MPC units were concentrated at the surface. The permeability for solutes through the PSf/MPC polymer hollow fibers was measured for 4 h. The permeabilities of both a low-molecular-weight compound and protein were greater than those of the PSf hollow fibers. The amount of adsorbed protein was lower on the PSf/MPC polymer hollow fiber when compared to that of the PSf hollow fiber. Moreover, platelet adhesion was also effectively inhibited on the PSf/MPC polymer hollow fiber. Based on these results, the addition of the MPC polymer to the PSf is a very useful method to improve the functions and blood compatibility of the hollow fiber.     

Preparation and performance of protein-adsorption-resistant asymmetric porous membrane composed of polysulfone/phospholipid polymer blend

To obtain protein-adsorption-resistant membrane for hemodialysis, we prepared a polymer blend composed of polysulfone and 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer (PSf/MPC polymer). The content of the MPC polymer in the PSf was 7 and 15 wt%. The asymmetric porous membrane was obtained by the dry/wet membrane processing method. The surface characterization of the PSf/MPC polymer membrane by X-ray photoelectron spectroscopy revealed that the MPC polymer located at the surface. The mechanical strength of the PSf/MPC polymer membrane did not change compared with that of the PSf membrane. On the other hand, the permeability of solute below a molecular weight (Mw) of 2.0 x 10(4) through the PSf membrane increased with the addition of the MPC polymer, which is considered to be an effect of the hydrophilic character of the MPC polymer. The amount of protein adsorbed on the PSf membrane from plasma was reduced by the addition of the MPC polymer. The permeability of low-molecular-weight protein (Mw = 1.2 x 10(4)) did not change even after the PSf/MPC polymer membrane was contacted with plasma protein solution for 4 h, whereas it decreased dramatically in the case of the PSf membrane. Platelet adhesion was also effectively suppressed on the PSf/MPC polymer membrane. Based on these results, the MPC polymer could serve as a doubly functional polymeric additive, that is, to generate a protein-adsorption-resistant characteristic and to render the membrane hydrophilic.     

A new method to evaluate the local clearance at different annular rings inside hemodialyzers

Recent research indicated that the dialysate flow distribution inside a hemodialyzer was not uniform ("channeling" of the dialysate flows). However, effect of the channeling on the solute clearance has not been directly and quantitatively examined. In this report, a novel experiment approach is presented to test the hypothesis that hollow fibers in different regions within a given hemodialyzer may contribute differently to the solute clearance. Water solution with urea (molecular weight 60) and creatinine (molecular weight 113) were made as "blood," and pure water was used as dialysate. Two high flux dialyzers, dialyzer A (cellulose triacetate) and dialyzer B (polyethersulfone), were used in this study. The hollow fiber potting area at the blood inlet of a dialyzer was divided into equal area concentric rings. In each experiment, only one of the rings was open for blood flow, and the other rings were blocked by epoxy. The "blood" was pumped at 120 ml/min while the dialysate flow rate (Qd) varied at 500, 800, and 1,000 ml/min, respectively. The solute clearance with a specific ring open (local clearance) was determined by measuring solute (urea/creatinine) concentration at the "blood" inlet and outlet. For dialyzer A, local clearance of urea and creatinine were significantly higher in the outer ring than in the inner ring. With increasing Qd, local solute clearance increased significantly for all rings. For dialyzer B, at any given Qd, solutes local clearance also increased from the inner to outer rings. In comparison, the effect of increasing Qd on solute clearance was greater for dialyzer B than for dialyzer A. In conclusion, using the new experimental method, the authors quantitatively evaluated the solute clearance contributed by the hollow fibers at different locations (concentric rings) in dialyzers. Hollow fibers at different locations did contribute differently to the solute clearance, which may be caused by the channeling of dialysate flow. A careful design of the dialyzer to minimize the channeling is needed.     

Copolymers of 2-methacryloyloxyethyl phosphorylcholine (MPC) as biomaterials

Copolymers of 2-methacryloyloxyethyl phosphorylcholine (MPC) showed good hemocompatibility as hypothesized. The hypothesis was surfaces having phosphorylcholine groups by polymerization of MPC could accumulate phospholipids from blood stream and show good blood compatibility. We designed and prepared a methacylate having a phosphorylcholine group. While it was possible to introduce them by polymer reactions, polymer reaction is not always good method to prepare the desired pure surface. This must be very important point to consider biomaterials, as we have to apply them in our body without any adverse effects.The hypothesis was confirmed by changing copolymer composition. The adsorption amount of phospholipids on the surfaces increased with increasing the MPC units in the copolymers. On the other hand, increasing MPC units in MPC copolymers decreased adsorption amount of peptides. There is limitation in blood compatibility tests in vitro due to unstable characteristics of blood itself. We evaluated them with series of blood compatibility tests, in vitro, ex vivo and in vivo, on coated PMMA beads, modified hollow fibers for hemodialysis and 2 mm small diameter blood vessels, respectively. These data suggested MPC is a promising methacrylate to develop good blood contacting devises, which may not require systemic anticoagulation. Conventional blood compatible biomaterials were not suitable to make permeable membranes. But MPC is soluble in water and we could prepare permeable membranes to various solutes by the copolymerization. Introduction of MPC copolymers on cellulose and polysulfone hollow fiber membranes gave them nonthrombogenicity but it did not give adverse effect on their permeability. These data suggested that it is possible to apply them to hemodialyzers, oxygenators and percutaneous glucose sensors to keep diabetic patients easier. MPC surfaces are good hydrogel to minimize damage on tissues by lubricating between organs and the coated devices. They do not induce denaturation of peptides, which is beneficial to keep activities of enzymes longer. And poly-MPC dissolved is applicable to stabilize several bioactive peptides in aqueous phase. So MPC polymers are useful to minimize fouling by inhibiting the adsorption of bioactive proteins. MPC has high potential to develop many varieties of new biomaterials useful in so-called biotechnology. MPC and their copolymers are commercially available from NOF (Tokyo, Japan) and Biocompatibles (UK, as PC technology).     

Modification of polysulfone with phospholipid polymer for improvement of the blood compatibility. Part 2. Protein adsorption and platelet adhesion.

Protein adsorption and platelet adhesion from human plasma on polysulfone (PSf) membranes modified with 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer were studied. The modification was carried out by blending of the MPC polymer in the PSf. The amount of protein adsorbed on the PSf/MPC polymer blend membrane was significantly decreased with an increase in the composition of the blended MPC polymer. The distribution of the specific proteins adsorbed on the membrane surface was also determined by a gold-colloid immunoassay. Albumin, gamma-globulin and fibrinogen were observed on every membrane surface after contact with plasma. However, in the case of the blended membrane, the density of the adsorbed proteins decreased compared with that of original PSf membrane. That is, the MPC polymer blended in the membrane could function as a protein-adsorption-resistant additive. The number of platelets adhered on the PSf membrane was reduced, and change in the morphology of adherent platelets was also suppressed by the modification with the MPC polymer. Therefore, the PSf/MPC polymer blend membrane had improved blood compatibility compared with the PSf membrane.     

Toward the treatment for Alzheimer’s disease: adsorption is primary mechanism of removing amyloid β protein with hollow-fiber dialyzers of the suitable materials,polysulfone and polymethyl methacrylate

The accumulation of amyloid β protein (Aβ) in the brain reflects cognitive impairment in Alzheimer’s disease. We hypothesized that the rapid removal of Aβ from the blood by an extracorporeal system may act as a peripheral Aβ sink from the brain. The present study aimed to determine the optimal materials and modality for Aβ removal by hemodialyzers. In a batch analysis, hollow-fiber fragments of polysulfone (PSf) and polymethyl methacrylate (PMMA) showed greater removal efficiency of Aβ than did other materials, such as cellulose-triacetates and ethylene–vinyl alcohol copolymer (PSf:PMMA at 30 min, 98.6 ± 2.4 %:97.8 ± 0.4 % for Aβ_1–40 and 96.6 ± 0.3 %:99.0 ± 1.0 % for Aβ_1–42). In a modality study, the Aβ solution was applied to PSf dialyzers and circulated in the dialysis and Air-filled adsorption-mode (i.e., the outer space of the hollow fibers was filled with air) or phosphate-buffered saline (PBS)-filled adsorption modes. The Aβ_1–40 removal efficiency of the pre/post dialyzer in the Air-filled adsorption-mode was the highest (62.4 ± 12.6 %, p = 0.007). In a flow rate study in the Air-filled adsorption-mode, 200 mL/min showed the highest Aβ_1–40 reduction rate of pool solution (97.3 ± 0.8 % at 15 min) compared with 20 mL/min (p = 0.00001) and 50 mL/min (p = 0.00382). PMMA dialyzers showed similar high reduction rates. Thus, the optimal modality for Aβ removal was the adsorption-mode with PSf or PMMA hollow fibers at around 50 mL/min flow rate, which seems to be suitable for clinical use.     

A modified equivalent annulus model for the hollow fiber hemodialyzer

Experimental approaches to optimize hollow fiber hemodialyzer design are expensive and time-consuming. Computer modeling is an effective way to study mass transfer in the hemodialyzer because a substantial reduction in experimental time and cost can be achieved. This paper presents a two-dimensional modified "equivalent annulus" model, which employs Navier-Stokes (N-S) equations to describe blood and dialysate flow, and Kedem-Katchalsky (K-K) equations to calculate transmembrane flow. N-S equations and K-K equations must be coupled together in the process of computing. The corresponding experiments were designed to validate this model, and experimental results agreed well with numerical results. The distribution of velocity, pressure and solute concentration were investigated in detail, presenting a clear insight into dialyzer mass transfer. This model can be applied to help optimize hemodialyzer design.     

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.     

细胞膜渗透系数的研究 

提出了一个全血传质的两相模型和新的透析器清除率表达式，通过测定不同时刻血浆和细胞内溶质浓度，获得了尿素、肌酐和尿酸的细胞膜渗透系数，研究结果对医生准确地评价透析医疗效果和小型、高效透析器的开发具有重要意义。     

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.     

Hemocompatibility of human whole blood on polymers with a phospholipid polar group and its mechanism.

The hemocompatibility of a polymer containing a phospholipid polar group, poly(2-methacryloyloxyethyl phosphorylcholine (MPC)-co-n-butyl methacrylate(BMA)), with human whole blood was evaluated. When human whole blood without an anticoagulant was contacted with polymers, the blood cell adhesion and aggregation on the polymer without the MPC moiety was extensive, and considerable fibrin deposition was observed. This phenomenon was suppressed with an increase in the polymer MPC composition. Thus, the MPC moiety in the copolymer plays an important role in the nonthrombogenic behavior of the copolymer. These results were also confirmed by the whole blood coagulation time on the polymer surface which was determined by Lee-White method. The adsorption of phospholipids and proteins from human plasma on poly(MPC-co-BMA) was investigated to clarify the mechanism of the nonthrombogenicity observed with the polymer. The amount of phospholipids was increased; whereas, adsorbed proteins were decreased with an increase in the MPC composition. From these results, we concluded that the phospholipids adsorbed on poly(MPC-co-BMA) play the most important role in the nonthrombogenicity of the MPC copolymer.     

Computational modeling of effects of mechanical shaking on hemodynamics in hollow fibers

Introduction: Blood-membrane interaction during hemodialysis develops a secondary protein layer on the dialysis membrane surface, resulting in reduction of hemodialyzer performance. Wall shear stress at the surface of the hollow-fiber membrane is one of the determinant factors able to influence dialysis efficiency. Shaking of hemodialyzer during treatment could increase the wall shear stress of the membrane, which could enhance hemodialyzer performance. Methods: In this study, hemodynamic changes in hollow fibers were analyzed using computational fluid dynamics software for various shaking conditions of hemodialyzer (longitudinal, transverse, rotational motions). Results: Longitudinal motion induced reverse flow, while transverse motion induced symmetric swirling inside the hollow fiber. During rotational motions, nonuniform vortices were developed according to the rotational radius of the hollow fiber. These changes in flow pathlines induced by different shaking profiles increased the relative motion of blood, transmembrane pressure, and wall shear stress on dialysis membrane surfaces. Both longitudinal and transverse shaking profiles showed a linear relationship between shaking velocity (the product of amplitude and frequency) and wall shear stress. Conclusion: Performance of hemodialyzer can be enhanced with simple mechanical shaking motions, and optimal shaking profiles for clinical application can be investigated and predicted with the computational fluid dynamics model proposed in this study.     

Preparation of non-thrombogenic materials using 2-methacryloyloxyethyl phosphorylcholine

This review addresses the non-thrombogenic characteristics of copolymers based on 2-methacryloyloxyethyl phosphorylcholine (MPC), originally developed by Nakabayashi and colleagues. The hypothesis underlying these developments was that such materials would adsorb phospholipids from blood, yielding surfaces with good natural blood compatibility. Methacrylates were found to have excellent properties for this copolymerisation. The characteristics of the MPC copolymers relevant to the improved blood compatibility were minimisation of protein adsorption through an increase in the amount of free water in the MPC hydrogels, which prevents protein conformational change and increased protein stability in solution. Non-thrombogenicity has been evaluated by in vitro, ex vivo and in vivo procedures. Non-thrombogenic dialysis membranes and a durable glucose biosensor have been developed using this MPC copolymer.     

An interpretation of cell separation mechanism on polyamine graft copolymers

Interaction between polyamide microcapsules having different balances of negative and positive charges on their surface, that is, different isoelectric points and copolymers with different numbers of polyamine macromer of a definite chain length grafted on poly(2-hydroxyethyl methacrylate) backbone was studied by measuring the amount adhered of the microcapsules onto the surface of copolymer-coated glass beads at different pH values. Maximum microcapsule adhesion was observed for a proper combination of microcapsule and copolymer to suggest that separation of a specified cell population from others on the surface of copolymer-coated glass beads can be explained in terms of preferential adhesion through electrostatic interaction of the specified cells with the copolymer.     

Solute Washout Experiments for Characterizing Mass Transport in Hollow Fiber Immunoisolation Membranes

The transport characteristics of immunoisolation membranes can have a critical effect on the design of hybrid artificial organs and cell therapies. However, it has been difficult to quantitatively evaluate the desired transport properties of different hollow fiber membranes due to bulk mass transfer limitations in the fiber lumen and annular space. An attractive alternative to existing methodologies is to use the rate of solute removal or washout from the annular space during constant flow perfusion through the fiber lumen. Experimental washout curves were obtained for glucose and a 10 kD dextran in two different hollow fiber devices. Data were analyzed using a theoretical model which accounts for convective and diffusive transport in the lumen, membrane, and annular space. The model was in good agreement with the experimental results and provided an accurate measure of the effective membrane diffusion coefficient for both small and large solutes. This approach should prove useful in theoretical analyses of solute transport and performance of hollow fiber artificial organs. © 1998 Biomedical Engineering Society. PAC98: 8722Fy, 8790+y     

An interpretation of cell separation mechanism on polyamine graft copolymers.

Interaction between polyamide microcapsules having different balances of negative and positive charges on their surface, that is, different isoelectric points and copolymers with different numbers of polyamine macromer of a definite chain length grafted on poly(2-hydroxyethyl methacrylate) backbone was studied by measuring the amount adhered of the microcapsules onto the surface of copolymer-coated glass beads at different pH values. Maximum microcapsule adhesion was observed for a proper combination of microcapsule and copolymer to suggest that separation of a specified cell population from others on the surface of copolymer-coated glass beads can be explained in terms of preferential adhesion through electrostatic interaction of the specified cells with the copolymer.     

Transmission electron microscopic study of hepatocytes in bioartificial liver

A bioartificial liver (BAL) was prepared by simple inoculation of hepatocytes into the inner space of hollow fibers of a hemodialyzer and it was maintained in a closed circuit for in vitro culture. Morphology of hepatocytes in the hollow fibers was studied in detail using transmission electron microscopy (TEM). The hepatocytes formed three-dimensional, rod-shaped aggregates of 200 microm in diameter throughout the whole dimension of the hollow fibers after 1 day of culture. Approximately five hepatocyte layers existed from the surface to the center of the aggregate. The hepatocytes in the aggregate displayed mostly polygonal shapes and were surrounded by five to six cells. Abundant bile canaliculi were formed between the hepatocytes and were sealed by tight junctions. The distance between the adjacent hepatocytes except the bile canaliculus domain was approximately 20 nm, and interdigitation was observed between some hepatocytes. These observations indicate that the hepatocytes formed functionally associated aggregates, that is, organoids. Although the cells facing the inner surface of the hollow fiber lost their polygonal shape and became flattened during the following several-day culture, no drastic change was observed in the morphology of the hepatocytes located inside the aggregate. After 14 days of culture, the number of living cells decreased and most of these had a deformed nucleus, few numbers of organelles, and intermittent lipid droplets.     

Effect of hollow fiber length on solute removal and quantification of internal filtration rate by Doppler ultrasound

Renal replacement therapy with dialyzers capable of enhanced internal filtration (IF) can be an alternative to standard hemodiafiltration, as it provides convective solute removal comparable to that of hemodiafiltration by a simple procedure. In this study, we clinically evaluated the effect of the hollow fiber length in the dialyzer, a crucial factor influencing the rate of IF, by comparing two commercial dialyzers (BS-1.6U, BS-1.6UL, Toray, Japan) which differed in the fiber length, but had the same surface area and inner diameter of their hollow fibers. We showed that in the dialyzer with the longer fibers, the pressure profile along the dialyzer was significantly altered, and the solute clearance tended to be increased. In addition, we successfully quantified the IF rate with a Doppler ultrasound in the experimental circuit, by measuring the blood flow velocities along the bundle of fibers. We showed that the changes in the blood flow velocity were more marked in the dialyzer with the longer fibers; the calculated IF rates in the dialyzers with the shorter and longer fibers were 11.1 mL/min and 37.7 mL/min, respectively, which seemed to be compatible with the solute clearances. This simple and readily applicable method is expected to be useful in the development of modified dialyzers to fully exploit the benefits of IF in renal replacement therapy.     

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.     

Hollow fiber shape alters solute clearances in high flux hemodialyzers

The mass transfer properties of hemodialyzers containing hollow fiber membranes are known to be influenced by membrane chemical composition, surface area, and pore size; however, the effects of hollow fiber shape (or configuration) and packing density within the dialyzer housing have not been well characterized. We determined, both in vitro and ex vivo (clinical), solute clearances and mass transfer-area coefficients (KoA) for high flux dialyzers containing polysulfone hollow fibers of identical chemical composition but different shapes. Hemoflow F80A (1.8 m2 of membrane surface area) dialyzers contained hollow fibers with a conventional shape, but Optiflux F180A (1.8 m2), F200A (2.0 m2), and F200NR (2.0 m2) dialyzers contained hollow fibers with a wavy shape. Clearances and KoA values determined in vitro for urea and creatinine increased with increasing dialysate flow rate and were higher for Optiflux F180A and F200A dialyzers than for Hemoflow F80A dialyzers. In vitro clearances for lysozyme and myoglobin were also higher for Optiflux F180A and F200A dialyzers than for Hemoflow F80A dialyzers, suggesting that a wavy hollow fiber shape increases mass transfer by increasing effective membrane surface area, conceivably by altering dialysate flow patterns. Urea clearances and KoA values determined ex vivo were higher for Optiflux F200NR dialyzers than for Hemoflow F80A dialyzers, confirming that the in vitro results are applicable to clinical hemodialysis. These increases in mass transfer efficiency for dialyzers containing hollow fibers with a wavy shape are consistent with improved mass transfer within the dialysate compartment as evidenced by the manufacturer-reported dialysate pressure-flow relationships. We conclude that the mass transfer characteristics of high flux dialyzers can be altered by the shape of the hollow fibers.