The removal of protein-bound uremic toxins (PBUTs) from the blood of kidney failure patients with conventional dialysis is limited. However, as their harmful effects and association with morbidity and mortality in dialysis patients are increasingly recognized, PBUTs have become important therapeutic targets. In this review, PBUT removal with current state-of-the-art dialysis technologies and future perspectives are discussed. Strategies to enhance PBUT clearance include methods that interfere with PBUT–albumin binding, such as chemical displacers, high ionic strength, pH changes, or electromagnetic fields, thereby increasing the free fraction available for dialysis. While these methods have shown promise in vitro, and some also in vivo, long-term safety data are lacking. PBUT removal can also be increased by adsorption, either directly via hemoperfusion, or indirectly, e.g., via sorbents incorporated in a mixed-matrix membrane or dissolved in the dialysate. In the kidney, PBUTs are secreted in the proximal tubules; hence, a cell-based bioartificial kidney (BAK) that secretes PBUTs is proposed as an add-on to current dialysis. Yet both PBUT adsorption strategies and, in particular, BAKs face considerable challenges in upscaling and mass production at acceptable costs. In conclusion, many novel technologies are under development, all requiring further (pre)clinical testing and upscaling before these strategies can be applied in the clinic.

Hemodialysis (HD) is a critical treatment for patients with end-stage kidney disease (ESKD). The effectiveness of conventional dialyzers used there could be compromised during extended use due to limited blood compatibility of synthetic polymeric membranes and sub-optimal dialyzer design. In fact, blood flow in the hollow fiber (HF) membrane could trigger inflammatory responses and thrombus formation, leading to reduced filtration efficiency and limiting therapy duration, a consequence of flowing the patients’ blood through the lumen of each fiber while the dialysate passes along the inter-fiber space (IOF, inside-out filtration). This study investigates the development of HF membranes for “outside-in filtration” (OIF) in HD. In OIF, blood flows through the inter-fiber space while dialysate flows within the fiber lumens, reducing the risk of fiber clogging and potentially extending treatment duration. For the OIF mode, the membrane should have a blood-compatible outer selective layer in contact with the patient’s blood. We develop HFs for OIF via liquid-induced phase separation using PES/PVP (polyethersulphone/polyvinylpyrrolidone) blends. The fibers’ surface morphology (SEM, scanning electron microscopy), chemistry (ATR-FTIR—attenuated total reflection-Fourier transform infrared spectroscopy, XPS—X-ray photoelectron spectroscopy), transport properties, and uremic toxin removal from human plasma are evaluated and compared to commercial HFs. These membranes feature a smooth, hydrophilic outer layer, porous lumen, ultrafiltration coefficient of 13–34 mL m2 h−1 mmHg−1, adequate mechanical properties, low albumin leakage, and toxin removal performance on par with commercial membranes in IOF and OIF. They offer potential for more efficient long-term HD by reducing clogging and systemic anticoagulation needs and enhancing treatment time and toxin clearance.

Most patients in need of renal replacement therapy use peritoneal or hemodialysis (HD) therapy. Major drawbacks of HD are the incomplete removal of uremic solutes (especially middle-sized uremic solutes and proteinbound uremic solutes, PBUTs) as well as the non-continuous therapy (3 times per week for 4 h), causing large fluctuations in water balance and uremic waste, potassium, and phosphate. For achieving better patient outcomes, more continuous therapies are required including the application of home HD or portable HD. The latter require membranes with excellent long-term biocompatibility. Heparin-modified dialysis membranes can offer improved biocompatibility, but systemic anticoagulation is still needed, whereas potential heparin-induced thrombocytopenia (HIT) may occur. Glycosaminoglycans (GAGs) contribute to the kidney glomerular filtration barrier and provide natural antifouling properties. Here, we hypothesized that the incorporation of GAGs within polymeric membranes, either via membrane coating, post membrane fabrication, or via blending GAGs into the membrane forming polymer matrix, would provide membranes with improved hemocompatibility. We implemented these strategies to mixed matrix membranes (MMMs) which combine diffusion and adsorption for removing a broad range of uremic solutes. Firstly, we fabricated flat sheet MMMs with various GAG sources, including heparan sulphate from bovine kidney (HSBK), heparinase III-digested HSBK, HS isolated from cultured glomerular endothelial glycocalyx, GAGs from porcine intestinal mucosa (danaparoid, DA), or heparin. The flat sheet MMMs with DA blended within the membrane selective layer have excellent blood compatibility, based on a panel of anti-coagulation and immune activation assays, combined to high water transport and very low albumin leakage. Besides, these MMMs had low clot formation, did not activate immune cells or the complement system and had low platelet adhesion. Based on these findings, we also developed first hollow fiber MMMs with DA blended in the selective layer, which also had high water transport, no leakage of protein, and achieved very good removal of various uremic solutes, comparable to commercial HD membranes.

A comprehensive review of known uremic retention molecules goes back to more than 10 years ago and did not consider metabolomic analyses. The present analysis searches for as of yet unclassified solutes retained in chronic kidney disease (CKD) by analyzing metabolites associated with relevant outcomes of CKD. This untargeted metabolomics-based approach is compared with a conventional targeted literature search. For the selected molecules, the literature was screened for arguments regarding toxic (harmful), beneficial, or neutral effects in experimental or clinical studies. Findings were independently crosschecked. In total, 103 molecules were selected. No literature on any effect was found for 55 substances, 3 molecules had no significant effect, and 13 others showed beneficial effects. For the remaining 32 compounds, we found at least one report of a toxic effect. Whereas 62.5% of the compounds with at least one study on a toxic effect was retrieved via the bottom-up approach, 69.2% of the substances originating from metabolomics-based approaches showed a beneficial effect. Our results suggest that untargeted metabolomics offer a more balanced view of uremic retention than the targeted approaches, with higher chances of revealing the beneficial potential of some of the metabolites.

Patients with end-stage kidney disease (ESKD) suffer from high levels of protein-bound uremic toxins (PBUTs) that contribute to various comorbidities. Conventional dialysis methods are ineffective in removing these PBUTs. A potential solution could be offered by a bioartificial kidney (BAK) composed of porous membranes covered by proximal tubule epithelial cells (PTECs) that actively secrete PBUTs. However, BAK development is currently being hampered by a lack of knowledge regarding the cytocompatibility of the dialysis fluid (DF) that comes in contact with the PTECs. Here, we conducted a comprehensive functional assessment of the DF on human conditionally immortalized PTECs (ciPTECs) cultured as monolayers in well plates, on Transwell® inserts, or on hollow fiber membranes (HFMs) that form functional units of a BAK. We evaluated cell viability markers, monolayer integrity, and PBUT clearance. Our results show that exposure to DF did not affect ciPTECs' viability, membrane integrity, or function. Seven anionic PBUTs were efficiently cleared from the perfusion fluid containing a PBUTs cocktail or uremic plasma, an effect which was enhanced in the presence of albumin. Overall, our findings support that the DF is cytocompatible and does not compromise ciPTECs function, paving the way for further advancements in BAK development and its potential clinical application.

Haemodialysis is life sustaining but expensive, provides limited removal of uraemic solutes, is associated with poor patient quality of life and has a large carbon footprint. Innovative dialysis technologies such as portable, wearable and implantable artificial kidney systems are being developed with the aim of addressing these issues and improving patient care. An important challenge for these technologies is the need for continuous regeneration of a small volume of dialysate. Dialysate recycling systems based on sorbents have great potential for such regeneration. Novel dialysis membranes composed of polymeric or inorganic materials are being developed to improve the removal of a broad range of uraemic toxins, with low levels of membrane fouling compared with currently available synthetic membranes. To achieve more complete therapy and provide important biological functions, these novel membranes could be combined with bioartificial kidneys, which consist of artificial membranes combined with kidney cells. Implementation of these systems will require robust cell sourcing; cell culture facilities annexed to dialysis centres; large-scale, low-cost production; and quality control measures. These challenges are not trivial, and global initiatives involving all relevant stakeholders, including academics, industrialists, medical professionals and patients with kidney disease, are required to achieve important technological breakthroughs. Portable, wearable and implantable artificial kidney systems require compact and efficient dialysate regeneration systems and novel membranes for improved toxin removal and long-term patency. Here, the authors discuss efforts to overcome these challenges and future perspectives for achieving miniaturized dialysis.

The world faces a dramatic man-made ecologic disaster and healthcare is a crucial part of this problem. Compared with other therapeutic areas, nephrology care, and especially dialysis, creates an excessive burden via water consumption, greenhouse gas emission and waste production. In this advocacy article from the European Kidney Health Alliance we describe the mutual impact of climate change on kidney health and kidney care on ecology. We propose an array of measures as potential solutions related to the prevention of kidney disease, kidney transplantation and green dialysis. For dialysis, several proactive suggestions are made, especially by lowering water consumption, implementing energy-neutral policies, waste triage and recycling of materials. These include original proposals such as dialysate regeneration, dialysate flow reduction, water distillation systems for dialysate production, heat pumps for unit climatization, heat exchangers for dialysate warming, biodegradable and bio-based polymers, alternative power sources, repurposing of plastic waste (e.g. incorporation in concrete), registration systems of ecologic burden and platforms to exchange ecologic best practices. We also discuss how the European Green Deal offers real potential for supporting and galvanizing these urgent environmental changes. Finally, we formulate recommendations to professionals, manufacturers, providers and policymakers on how this correction can be achieved.

Within the fields of regenerative medicine and tissue engineering, the development of tough hydrogel biomaterials is a challenging topic that has received much attention over the past few years. Recently, a method was developed to synthesize tough combinatorial poly(urethane-isocyanurate) (PUI)-type hydrogels by the trimerization of mixtures of NCO-functionalized prepolymers. As this synthesis approach allows a large degree of freedom in terms of polymer network design with a high level of control over the polymer network structure, the resulting systems are ideally suited for studying structure-property relations in tough hydrogel systems. In this work, we aim to systematically investigate the influence of introducing a hydrophobic component into a PUI polymer network on the mechanical properties of the resulting PUI hydrogels. Additionally, the effect of (the degree of) crystallinity of the hydrophobic network component is investigated. For this, two series of combinatorial PUI hydrogels are synthesized, based on a hydrophilic poly(ethylene glycol) prepolymer and increasing amounts of either a crystallizable hydrophobic prepolymer (poly(epsilon-caprolactone)) or an amorphous hydrophobic prepolymer (poly(propylene glycol)). It is shown that the toughness of amorphous PUI hydrogels is hardly influenced by the hydrophobic content, whereas the toughness of semi-crystalline PUI hydrogels strongly increases with increasing hydrophobic content. Also, the toughness of the latter hydrogels increases further with increasing degree of crystallinity of the hydrogel. Finally, it is shown that the semi-crystalline PUI hydrogels are promising materials for biomedical adhesive and coating applications, as well as for load-bearing biomedical applications within the fields of tissue engineering and regenerative medicine. (c) 2022 Covestro Deutschland AG. Polymer International published by John Wiley & Sons Ltd on behalf of Society of Industrial Chemistry.

A comparatively straightforward approach to accomplish more physiological realism in organ-on-a-chip (OoC) models is through substrate geometry. There is increasing evidence that the strongly, microscale curved surfaces that epithelial or endothelial cells experience when lining small body lumens, such as the alveoli or blood vessels, impact their behavior. However, the most commonly used cell culture substrates for modeling of these human tissue barriers in OoCs, ion track-etched porous membranes, provide only flat surfaces. Here, we propose a more realistic culture environment for alveolar cells based on biomimetically microcurved track-etched membranes. They recreate the mainly spherical geometry of the cells' native microenvironment. In this feasibility study, the membranes were given the shape of hexagonally arrayed hemispherical microwells by an innovative combination of three-dimensional (3D) microfilm (thermo)forming and ion track technology. Integrated in microfluidic chips, they separated a top from a bottom cell culture chamber. The microcurved membranes were seeded by infusion with primary human alveolar epithelial cells. Despite the pronounced topology, the cells fully lined the alveoli-like microwell structures on the membranes' top side. The confluent curved epithelial cell monolayers could be cultured successfully at the air-liquid interface for 14 days. Similarly, the top and bottom sides of the microcurved membranes were seeded with cells from the Calu-3 lung epithelial cell line and human lung microvascular endothelial cells, respectively. Thereby, the latter lined the interalveolar septum-like interspace between the microwells in a network-type fashion, as in the natural counterpart. The coculture was maintained for 11 days. The presented 3D lung-on-a-chip model might set the stage for other (micro)anatomically inspired membrane-based OoCs in the future.

EDITORIAL article Front. Bioeng. Biotechnol., 22 November 2022Sec. Regenerative Medicine Volume 10 - 2022 | https://doi.org/10.3389/fbioe.2022.1090317

Common methods for fabricating membrane-based scaffolds for tissue engineering with (hydrophobic) polymers include thermal or liquid-phase inversion, sintering, particle leaching, electrospinning and stereolithography. However, these methods have limitations, such as low resolution and pore interconnectivity and may often require the application of high temperatures and/or toxic porogens, additives or solvents. In this work, we aim to overcome some of these limitations and propose a one-step method to produce large porous membrane-based scaffolds formed by air-water interfacial phase separation using water as a pore-forming agent and casting substrate. Here, we provide proof of concept using poly (trimethylene carbonate), a flexible and biocompatible hydrophobic polymer. Membrane-based scaffolds were prepared by dropwise addition of the polymer solution to water. Upon contact, rapid solvent–non-solvent phase separation took place on the air-water interface, after which the scaffold was cured by UV irradiation. We can tune and control the morphology of these scaffolds, including pore size and porosity, by changing various parameters, including polymer concentration, solvent type and temperature. Importantly, human hepatic stellate cells cultured on these membrane-based scaffolds remained viable and showed no signs of pro-inflammatory stress. These results indicate that the proposed air-water interfacial phase separation represents a versatile method for creating porous membrane-based scaffolds for tissue engineering applications.

The General Data Protection Regulation (GDPR) became binding law in the European Union Member States in 2018, as a step toward harmonizing personal data protection legislation in the European Union. The Regulation governs almost all types of personal data processing, hence, also, those pertaining to biomedical research. The purpose of this article is to highlight the main practical issues related to data and biological sample sharing that biomedical researchers face regularly, and to specify how these are addressed in the context of GDPR, after consulting with ethics/legal experts. We identify areas in which clarifications of the GDPR are needed, particularly those related to consent requirements by study participants. Amendments should target the following: (1) restricting exceptions based on national laws and increasing harmonization, (2) confirming the concept of broad consent, and (3) defining a roadmap for secondary use of data. These changes will be achieved by acknowledged learned societies in the field taking the lead in preparing a document giving guidance for the optimal interpretation of the GDPR, which will be finalized following a period of commenting by a broad multistakeholder audience. In parallel, promoting engagement and education of the public in the relevant issues (such as different consent types or residual risk for re-identification), on both local/national and international levels, is considered critical for advancement. We hope that this article will open this broad discussion involving all major stakeholders, toward optimizing the GDPR and allowing a harmonized transnational research approach.

The General Data Protection Regulation (GDPR) became binding law in the European Union Member States in 2018, as a step toward harmonizing personal data protection legislation in the European Union. The Regulation governs almost all types of personal data processing, hence, also, those pertaining to biomedical research. The purpose of this article is to highlight the main practical issues related to data and biological sample sharing that biomedical researchers face regularly, and to specify how these are addressed in the context of GDPR, after consulting with ethics/legal experts. We identify areas in which clarifications of the GDPR are needed, particularly those related to consent requirements by study participants. Amendments should target the following: (1) restricting exceptions based on national laws and increasing harmonization, (2) confirming the concept of broad consent, and (3) defining a roadmap for secondary use of data. These changes will be achieved by acknowledged learned societies in the field taking the lead in preparing a document giving guidance for the optimal interpretation of the GDPR, which will be finalized following a period of commenting by a broad multistakeholder audience. In parallel, promoting engagement and education of the public in the relevant issues (such as different consent types or residual risk for re-identification), on both local/national and international levels, is considered critical for advancement. We hope that this article will open this broad discussion involving all major stakeholders, toward optimizing the GDPR and allowing a harmonized transnational research approach.

Urea removal is one of the biggest challenges in dialysate regeneration in Wearable Artificial Kidney (WAK) devices. In this work, a new Mixed Matrix Membrane (MMM) is developed for urea removal in WAK applications. The MMM consists of polystyrene-based ninhydrin particles within a polyethersulfone/polyvinylpyrmlidone polymer blend matrix. The MMM is prepared via dry-wet spinning technique and characterized in terms of its morphology via electron microscopy and clean water permeance. Urea removal is studied both in static and in dynamic conditions. Thanks to the good dispersion of small size ninhydrin particles (size < 63 mu m), the MMM removed under static conditions, at 70 degrees C, 2.1 +/- 0.1 mmol of urea per grams of particles at 24 h, while urea removal by the particles in suspension reached 1.7 +/- 0.1 mmol/g under the same conditions. Importantly, in continuous recirculation experiments, performed at 70 degrees C using a laboratory scale module, the MMM removed 3.4 +/- 0.3 mmol of urea per grams of particles, in 4 h, due to the high particle accessibility by urea within the membrane. Based on these results it is estimated that only 215 g of MMM are needed for removing the daily produced urea from spent dialysate (400 mmol) making MMM suitable for application to WAK, where miniaturization and lightweight are required.

Due to the continuing high impact of lung diseases on society and the emergence of new respiratory viruses, such as SARS-CoV-2, there is a great need for in vitro lung models that more accurately recapitulate the in vivo situation than current models based on lung epithelial cell cultures on stiff membranes. Therefore, we developed an in vitro airway epithelial–endothelial cell culture model based on Calu-3 human lung epithelial cells and human lung microvascular endothelial cells (LMVECs), cultured on opposite sides of flexible porous poly(trimethylene carbonate) (PTMC) membranes. Calu-3 cells, cultured for two weeks at an air–liquid interface (ALI), showed good expression of the tight junction (TJ) protein Zonula Occludens 1 (ZO-1). LMVECs cultured submerged for three weeks were CD31-positive, but the expression was diffuse and not localized at the cell membrane. Barrier functions of the Calu-3 cell cultures and the co-cultures with LMVECs were good, as determined by electrical resistance measurements and fluorescein isothiocyanate-dextran (FITC-dextran) permeability assays. Importantly, the Calu-3/LMVEC co-cultures showed better cell viability and barrier function than mono-cultures. Moreover, there was no evidence for epithelial- and endothelial-to-mesenchymal transition (EMT and EndoMT, respectively) based on staining for the mesenchymal markers vimentin and α-SMA, respectively. These results indicate the potential of this new airway epithelial–endothelial model for lung research. In addition, since the PTMC membrane is flexible, the model can be expanded by introducing cyclic stretch for enabling mechanical stimulation of the cells. Furthermore, the model can form the basis for biomimetic airway epithelial–endothelial and alveolar–endothelial models with primary lung epithelial cells.

Islet encapsulation in membrane-based devices could allow for transplantation of donor islet tissue in the absence of immunosuppression. To achieve long-term survival of islets, the device should allow rapid exchange of essential nutrients and be vascularized to guarantee continued support of islet function. Recently, we have proposed a membrane-based macroencapsulation device consisting of a microwell membrane for islet separation covered by a micropatterned membrane lid. The device can prevent islet aggregation and support functional islet survivalin vitro. Here, based on previous modeling studies, we develop an improved device with smaller microwell dimensions, decreased spacing between the microwells and reduced membrane thickness and investigate its performancein vitroandin vivo. This improved device allows for encapsulating higher islet numbers without islet aggregation and by applying anin vivoimaging system we demonstrate very good perfusion of the device when implanted intraperitoneally in mice. Besides, when it is implanted subcutaneously in mice, islet viability is maintained and a vascular network in close proximity to the device is developed. All these important findings demonstrate the potential of this device for islet transplantation.

Hemodialysis mainly removes small water-soluble uremic toxins but cannot effectively remove middle molecules and protein-bound uremic toxins. Besides, the therapy is intermittent leading to fluctuating blood values and fluid status which adversely impacts patients’ health. Prolonged hemodialysis could improve the removal of toxins and the development of portable and wearable artificial kidneys could offer more flexibility in the dialysis scheme. This would enhance patients’ overall health, autonomy, mobility and flexibility, allowing patients to participate in social and economic life. However, during prolonged hemodialysis, blood clots could obstruct the fiber lumen, resulting in a decrease of the effective membrane surface area available for toxin removal. The outside-in filtration (OIF) mode, wherein blood flows through the inter-fiber space instead of through the fiber lumina, has been applied widely in blood oxygenators to prevent fiber clotting, but not in hemodialysis. In this study, we present for the first time the development of a mixed matrix membrane (MMM) for OIF of human blood plasma. This MMM combines diffusion and adsorption and consists of a polymeric membrane matrix with activated carbon (AC) particles on the inside layer, and a polymeric particle-free layer on the outer fiber layer. Our results show that in vitro MMM fibers for OIF demonstrate superior removal of the protein-bound uremic toxins, indoxyl sulfate and hippuric acid, compared to both earlier MMM fibers designed for inside-out filtration mode and commercial high-flux fibers.

Haemodialysis is a life-saving therapy. However, in comparison with the healthy kidney, it removes only a small fraction of the uraemic toxins produced, does not function continuously and cannot replicate biological kidney functions. Innovations in membrane design hold promise to overcome these limitations with potential to improve patient outcomes.

As increasing demand for hemodialysis (HD) treatment incurs significant financial burden to healthcare systems and ecological burden as well, novel therapeutic approaches as well as innovations and technological advances are being sought that could lead to the development of purification devices such as dialyzers with improved characteristics and wearable technology. Novel knowledge such as the development of more accurate kinetic models, the development of novel HD membranes with the use of nanotechnology, novel manufacturing processes, and the latest technology in the science of materials have enabled novel solutions already marketed or on the verge of becoming commercially available. This collaborative article reviews the latest advances in HD as they were presented by the authors in a recent symposium titled "Frontiers in Haemodialysis," held on 12th December 2019 at the Royal Society of Medicine in London.