Emerging 4D fabrication of next-generation nerve guiding conduits: a critical perspective.

The latest advancements in the field of manufacturing for biomedicine, digital health, targeted therapy, and personalized medicine have fuelled the fabrication of smart medical devices. Four-dimensional (4D) fabrication strategies, which combine the manufacturing of three-dimensional (3D) parts with smart materials and/or design, have proved beneficial in creating customized and self-fitting structures that change their properties on demand with time. These frontier techniques that yield dynamic implants can indeed alleviate various drawbacks of current clinical practices, such as the use of sutures and complex microsurgeries and associated inflammation, among others. Among various clinical applications, 4D fabrication has lately made remarkable progress in the development of next-generation nerve-guiding conduits for treating peripheral nerve injuries (PNIs) by improving the end-to-end co-aptation of transected nerve endings. The current perspective highlights the relevance of 4D fabrication in developing state-of-the-art technologies for the treatment of PNIs. Various 4D fabrication/bio-fabrication techniques for PNI treatment are summarized while identifying the challenges and opportunities for the future. Such advancements hold immense promise for improving the quality of life of patients suffering from nerve damage and the potential for extending the treatment of many other disorders. Although the techniques are being described for PNIs, they will lend themselves suitably to certain cases of cranial nerve injuries as well.

"Affimer" synthetic protein scaffolds block oxidized LDL binding to the LOX-1 scavenger receptor and inhibit ERK1/2 activation.

In multicellular organisms, a variety of lipid-protein particles control the systemic flow of triacylglycerides, cholesterol, and fatty acids between cells in different tissues. The chemical modification by oxidation of these particles can trigger pathological responses, mediated by a group of membrane proteins termed scavenger receptors. The lectin-like oxidized low-density lipoprotein (LOX-1) scavenger receptor binds to oxidized low-density lipoprotein (oxLDL) and mediates both signaling and trafficking outcomes. Here, we identified five synthetic proteins termed Affimers from a phage display library, each capable of binding recombinant LOX-1 extracellular (oxLDL-binding) domain with high specificity. These Affimers, based on a phytocystatin scaffold with loop regions of variable sequence, were able to bind to the plasma membrane of HEK293T cells exclusively when human LOX-1 was expressed. Binding and uptake of fluorescently labeled oxLDL by the LOX-1-expressing cell model was inhibited with subnanomolar potency by all 5 Affimers. ERK1/2 activation, stimulated by oxLDL binding to LOX-1, was also significantly inhibited (p < 0.01) by preincubation with LOX-1-specific Affimers, but these Affimers had no direct agonistic effect. Molecular modeling indicated that the LOX-1-specific Affimers bound predominantly via their variable loop regions to the surface of the LOX-1 lectin-like domain that contains a distinctive arrangement of arginine residues previously implicated in oxLDL binding, involving interactions with both subunits of the native, stable scavenger receptor homodimer. These data provide a new class of synthetic tools to probe and potentially modulate the oxLDL/LOX-1 interaction that plays an important role in vascular disease.

Trends in Photopolymerizable Bioinks for 3D Bioprinting of Tumor Models.

Three-dimensional (3D) bioprinting technologies involving photopolymerizable bioinks (PBs) have attracted enormous attention in recent times owing to their ability to recreate complex structures with high resolution, mechanical stability, and favorable printing conditions that are suited for encapsulating cells. 3D bioprinted tissue constructs involving PBs can offer better insights into the tumor microenvironment and offer platforms for drug screening to advance cancer research. These bioinks enable the incorporation of physiologically relevant cell densities, tissue-mimetic stiffness, and vascularized channels and biochemical gradients in the 3D tumor models, unlike conventional two-dimensional (2D) cultures or other 3D scaffold fabrication technologies. In this perspective, we present the emerging techniques of 3D bioprinting using PBs in the context of cancer research, with a specific focus on the efforts to recapitulate the complexity of the tumor microenvironment. We describe printing approaches and various PB formulations compatible with these techniques along with recent attempts to bioprint 3D tumor models for studying migration and metastasis, cell-cell interactions, cell-extracellular matrix interactions, and drug screening relevant to cancer. We discuss the limitations and identify unexplored opportunities in this field for clinical and commercial translation of these emerging technologies.

A Global Challenge: Sustainability of Submicrometer PEO and PVP Fiber Production.

The field of submicrometer polymeric production currently has a predominant research focus on morphology and application. In comparison, the sustainability of the manufacture of submicrometer polymeric fibers, specifically the energy efficiency, is less explored. The principles of Green Chemistry and Green Engineering outline frameworks for the manufacture of "greener" products, where the most significant principles in the two frameworks are shown to be centered on energy efficiency, material wastage, and the use of non-hazardous materials. This study examines the power consumption during the production of Polyethylene oxide (PEO) and Polyvinylpyrrolidone (PVP) submicrometer fibers under magnitudes of the key forming parameters to generate fibers via pressure spinning. The energy consumption, along with the fiber diameter, and production rate during the manufacture of fibers is predominantly attributed to the characteristics of polymeric solutions utilized.

Pressure-Spun Fibrous Surgical Sutures for Localized Antibacterial Delivery: Development, Characterization, and In Vitro Evaluation.

Surgical sutures designed to prevent infection are critical in addressing antibiotic-resistant pathogens that cause surgical site infections. Instead of antibiotics, alternative materials such as biocides have been assessed for coating commercially used sutures due to emerging antibiotic resistance concerns worldwide. This study has a new approach to the development of fibrous surgical sutures with the ability to deliver localized antibacterial agents. A new manufacturing process based on pressure spinning was used for the first time in the production of fibrous surgical sutures by physically blending antibacterial triclosan (Tri) agent with poly(lactic-co-glycolic acid) (PLGA) and poly(ethylene oxide) (PEO) polymers. Fibrous surgical sutures with virgin PLGA, virgin PEO, different ratios of PLGA-PEO, and different ratios of Tri-loaded PLGA-PEO fibrous sutures were produced to mimic the FDA- and NICE-approved PLGA-based sutures available in the market and compared for their characteristics. They were also tested simultaneously with commercially available sutures to compare their in vitro biodegradation, antibacterial, drug release, and cytotoxicity properties. After in vitro antibacterial testing for 24 h, sutures having 285 ± 12 µg/mg Tri loading were selected as a model for further testing as they exhibited antibacterial activity against all tested bacteria strains. The selected model of antibacterial fibrous sutures exhibited an initial burst of Tri release within 24 h, followed by a sustained release for the remaining time until the sutures completely degraded within 21 days. The cell viability assay showed that these surgical sutures had no cytotoxic effect on mammalian cells.

Casein fibres for wound healing.

The name casein is given to a family of phosphoproteins which is commonly found in milk. Until recently, this was a constituent of milk that was commonly discarded; however today, it is widely used in health supplements all over the world. In this work, a high loading (50 wt%) of casein is mixed with a solution of polycaprolactone (PCL) to produce bandage-like fibres with an average fibre diameter of 1.4 ± 0.5 µm, which would be used to cover wounds in a series of tests with diabetic rats. Mouse fibroblast cell viability tests show that the casein-loaded fibres had little cytotoxicity with over 90% observed viability. A 14-day in vivo trial involving three groups of rats, used as control (no treatment), pure PCL fibres and casein-loaded fibres, showed that the casein within the fibres contributed to a significantly more extensive healing process. Histological analysis showed increased development of granulation tissue and follicle regrowth for the casein-loaded fibres. Further analysis showed that casein-loaded fibres have significantly lower levels of TNF-α, TGF-ß IL-1ß, NF-κB and IL-6, contributing to superior healing. The results presented here show an economical and simple approach to advanced wound healing.

Beneficial Effects of Insulin on Ischemia Reperfusion Injury in Human Skeletal Muscle.

Purpose: Exaggerated leucocyte activity is a crucial step in the pathophysiology of skeletal muscle ischemia-reperfusion injury (IRI). We tested the hypothesis that insulin, via its' anti-leukocyte activity, attenuates skeletal muscle IRI in humans. Materials and Methods: This randomized, blinded, placebo-controlled trial was conducted in patients with skeletal muscle ischemia who required revascularization. Treatment protocols were similar among them except for the insulin group, which received an infusion of insulin at 2.5 U/h. The degree of endothelial adhesiveness; leukocyte activity and pro-inflammatory status via P-selectin, tumor necrosis factor (TNF)-alpha, and myeloperoxidase (MPO) levels in the venous effluent; and clinical outcomes were measured. Results: Twenty-four consenting patients were randomized to the insulin or control group. There were no significant differences between the two groups except for the median serum insulin level, which was higher in the insulin group (P<0.01). No serious intervention-related adverse events were observed. P-selectin (55.04-99.86 pg/mL; P<0.001), MPO (110.8-160.6 pg/mL; P<0.001), and TNF-alpha (12.16-36.01 pg/mL; P<0.001) levels demonstrated a significant increase post-reperfusion in the 'control' group, reaching a peak value at 2 hours post-reperfusion. The increase in all three markers from baseline was significantly diminished in the insulin group at the two-hour (P-selectin, P=0.001; MPO, P=0.001; TNF-alpha, P=0.005) and four-hour (P-selectin, P=0.003; MPO, P=0.002; TNF-alpha, P=0.01) intervals. The differences in clinical outcomes between the insulin and control groups were not statistically significant. Conclusion: In clinical practice, insulin has the potential to attenuate the severity of skeletal muscle IRI inhibiting P-selectin, MPO, and TNF-alpha levels.

Emerging trends in biliary stents: a materials and manufacturing perspective.

Biliary stent technology has come a long way since its inception. There have been significant advancements in the materials used, and design and deployment strategies. Options have expanded from plastic and metallic stents to a wider variety of materials and manufacturing technologies to offer several options to clinicians, including self-expandable metallic stents and bioresorbable stents. Bioresorbable biliary stents are still in the early stages of clinical adoption. This review encompasses the materials currently used for biliary stents and the significant developments in the past few years in the resorbable materials for use as biliary stents. We critically discuss the emerging trends in the development of new resorbable materials for fabricating biliary stents. We then assess the developments in drug-eluting stents and advanced manufacturing technologies that could be leveraged for biliary stents. Challenges in the paths for translation for the future, such as pre-clinical and clinical trials, are highlighted. Finally, we present future directions that could drive the biliary stent market to meet the increasingly complex and diverse clinical needs of patients.

Purification and Analysis of Circulating Lipid Particles.

Lipid particles found in circulating extracellular fluids such as blood or lymph are essential for cellular homeostasis, metabolism and survival. Such particles provide essential lipids and fats which enable cells to synthesize new membranes and regulate different biochemical pathways. Imbalance in lipid particle metabolism can cause pathological states such as atherosclerosis. Here, elevated low-density lipoprotein (LDL) accumulation leads to fat-filled lesions or plaques in arterial walls. In this chapter, we provide a detailed set of protocols for the rapid and safe purification of lipid particles from human blood using high-speed ultracentrifugation. We provide a detailed set of assays for further analysis of the biochemical and cellular properties of these lipid particles. By combining these assays, we can better understand the complex roles of different lipid particles in normal physiology and disease pathology.

Facile One-Pot Method for All Aqueous Green Formation of Biocompatible Silk Fibroin-Poly(Ethylene Oxide) Fibers for Use in Tissue Engineering.

Silk fibroin (SF) fibers are highly regarded in tissue engineering because of their outstanding biocompatibility and tunable properties. A challenge remains in overcoming the trade-off between functioning and biocompatible fibers and the use of cytotoxic, environmentally harmful organic solvents in their processing and formation. The aim of this research was to produce biocompatible SF fibers without the use of cytotoxic solvents, via pressurized gyration (PG). Aqueous SF was blended with poly(ethylene oxide) (PEO) in ratios of 80:20 (labeled SF-PEO 80:20) and 90:10 (labeled SF-PEO 90:10) and spun into fibers using PG, assisted by a range of applied pressures and heat. Pure PEO (labeled PEO-Aq) and SF solubilized in hexafluoro-isopropanol (HFIP) (labeled SF-HFIP) and aqueous SF (labeled SF-Aq) were also prepared for comparison. The resulting fibers were characterized using SEM, TGA, and FTIR. Their in vitro cell behavior was analyzed using a Live/Dead assay and cell proliferation studies with the SaOS-2 human bone osteosarcoma cell line (ATCC, HTB-85) and human fetal osteoblast cells (hFob) (ATCC, CRL-11372) in 2D culture conditions. Fibers in the micrometer range were successfully produced using SF-PEO blends, SF-HFIP, and PEO-Aq. The fiber thickness ranged from 0.71 ± 0.17 µm for fibers produced using SF-PEO 90:10 with no applied pressure to 2.10 ± 0.78 µm for fibers produced using SF-PEO 80:10 with 0.3 MPa applied pressure. FTIR confirmed the presence of SF via amide I and amide II bands in the blend fibers because of a change in structural conformation. No difference was observed in thermogravimetric properties among varying pressures and no significant difference in fiber diameters for pressures. SaOS-2 cells and hFOb cell studies demonstrated higher cell densities and greater live cells on SF-PEO blends when compared to SF-HFIP. This research demonstrates a scalable and green method of producing SF-based constructs for use in bone-tissue engineering applications.

Experimental evaluation of the patient-specific haemodynamics of an aortic dissection model using particle image velocimetry.

Aortic Dissection (AD) is a complex pathology that affects the aorta. Diagnosis, management and treatment remain a challenge as it is a highly patient-specific pathology and there is still a limited understanding of the fluid-mechanics phenomena underlying clinical outcomes. Although in vitro models can allow the accurate study of AD flow fields in physical phantoms, they are currently scarce and almost exclusively rely on over simplifying assumptions. In this work, we present the first experimental study of a patient-specific case of AD. An anatomically correct phantom was produced and combined with a state-of-the-art in vitro platform, informed by clinical data, employed to accurately reproduce personalised conditions. The complex AD haemodynamics reproduced by the platform was characterised by flow rate and pressure acquisitions as well as Particle Image Velocimetry (PIV) derived velocity fields. Clinically relevant haemodynamic indices, that can be correlated with AD prognosis - such as velocity, shear rate, turbulent kinetic energy distributions - were extracted in two regions of interest in the aortic domain. The acquired data highlighted the complex nature of the flow (e.g. recirculation regions, low shear rate in the false lumen) and was in very good agreement with the available clinical data and the CFD results of a study conducted alongside, demonstrating the accuracy of the findings. These results demonstrate that the described platform constitutes a powerful, unique tool to reproduce in vitro personalised haemodynamic conditions, which can be used to support the evaluation of surgical procedures, medical devices testing and to validate state-of-the-art numerical models.

Severe Acute Respiratory Syndrome Type 2-Causing Coronavirus: Variants and Preventive Strategies.

COVID-19 vaccines have constituted a substantial scientific leap in countering severe acute respiratory syndrome type 2-causing coronavirus (SARS-CoV-2), and worldwide implementation of vaccination programs has significantly contributed to the global pandemic effort by saving many lives. However, the continuous evolution of the SARS-CoV-2 viral genome has resulted in different variants with a diverse range of mutations, some with enhanced virulence compared with previous lineages. Such variants are still a great concern as they have the potential to reduce vaccine efficacy and increase the viral transmission rate. This review summarizes the significant variants of SARS-CoV-2 encountered to date (December 2021) and discusses a spectrum of possible preventive strategies, with an emphasis on physical and materials science.

3D direct-write printing of water soluble micromoulds for high-resolution rapid prototyping.

Direct-write printing has contributed tremendously to additive manufacturing; in particular extrusion based printing where it has extended the range of materials for 3D printing and thus enabled use across many more sectors. The printing inks for direct-write printing however, need careful synthesis and invariably undergo extensive preparation before being able to print. Hence, new ink synthesis efforts are required every time a new material is to be printed; this is particularly challenging for low storage modulus (G') materials like silicones, especially at higher resolutions (under 10 µm). Here we report the development of a precise (< 10 µm) 3D printable polymer, with which we 3D print micromoulds which are filled with standard silicones like polydimethylsiloxane (PDMS) and left to cure at room temperature. The proof of concept is demonstrated using a simple water soluble polymer as the mould material. The approach enables micrometre scale silicone structures to be prototyped with ease, away from the cleanroom.

Co-Axial Gyro-Spinning of PCL/PVA/HA Core-Sheath Fibrous Scaffolds for Bone Tissue Engineering.

The present study aspires towards fabricating core-sheath fibrous scaffolds by state-of-the-art pressurized gyration for bone tissue engineering applications. The core-sheath fibers comprising dual-phase poly-ε-caprolactone (PCL) core and polyvinyl alcohol (PVA) sheath are fabricated using a novel "co-axial" pressurized gyration method. Hydroxyapatite (HA) nanocrystals are embedded in the sheath of the fabricated scaffolds to improve the performance for application as a bone tissue regeneration material. The diameter of the fabricated fiber is 3.97 ± 1.31 µm for PCL-PVA/3%HA while pure PCL-PVA with no HA loading gives 3.03 ± 0.45 µm. Bead-free fiber morphology is ascertained for all sample groups. The chemistry, water contact angle and swelling behavior measurements of the fabricated core-sheath fibrous scaffolds indicate the suitability of the structures in cellular activities. Saos-2 bone osteosarcoma cells are employed to determine the biocompatibility of the scaffolds, wherein none of the scaffolds possess any cytotoxicity effect, while cell proliferation of 94% is obtained for PCL-PVA/5%HA fibers. The alkaline phosphatase activity results suggest the osteogenic activities on the scaffolds begin earlier than day 7. Overall, adaptations of co-axial pressurized gyration provides the flexibility to embed or encapsulate bioactive substances in core-sheath fiber assemblies and is a promising strategy for bone healing.

Surface interactions and viability of coronaviruses.

The recently emerged coronavirus pandemic (COVID-19) has become a worldwide threat affecting millions of people, causing respiratory system related problems that can end up with extremely serious consequences. As the infection rate rises significantly and this is followed by a dramatic increase in mortality, the whole world is struggling to accommodate change and is trying to adapt to new conditions. While a significant amount of effort is focused on developing a vaccine in order to make a game-changing anti-COVID-19 breakthrough, novel coronavirus (SARS-CoV-2) is also developing mutations rapidly as it transmits just like any other virus and there is always a substantial chance of the invented antibodies becoming ineffective as a function of time, thus failing to inhibit virus-to-cell binding efficiency as the spiked protein keeps evolving. Hence, controlling the transmission of the virus is crucial. Therefore, this review summarizes the viability of coronaviruses on inanimate surfaces under different conditions while addressing the current state of known chemical disinfectants for deactivation of the coronaviruses. The review attempts to bring together a wide spectrum of surface-virus-cleaning agent interactions to help identify material selection for inanimate surfaces that have frequent human contact and cleaning procedures for effective prevention of COVID-19 transmission.

Structural Basis for Vascular Endothelial Growth Factor Receptor Activation and Implications for Disease Therapy.

Vascular endothelial growth factors (VEGFs) bind to membrane receptors on a wide variety of cells to regulate diverse biological responses. The VEGF-A family member promotes vasculogenesis and angiogenesis, processes which are essential for vascular development and physiology. As angiogenesis can be subverted in many disease states, including tumour development and progression, there is much interest in understanding the mechanistic basis for how VEGF-A regulates cell and tissue function. VEGF-A binds with high affinity to two VEGF receptor tyrosine kinases (VEGFR1, VEGFR2) and with lower affinity to co-receptors called neuropilin-1 and neuropilin-2 (NRP1, NRP2). Here, we use a structural viewpoint to summarise our current knowledge of VEGF-VEGFR activation and signal transduction. As targeting VEGF-VEGFR activation holds much therapeutic promise, we examine the structural basis for anti-angiogenic therapy using small-molecule compounds such as tyrosine kinase inhibitors that block VEGFR activation and downstream signalling. This review provides a rational basis towards reconciling VEGF and VEGFR structure and function in developing new therapeutics for a diverse range of ailments.

Scavenger Receptors as Biomarkers and Therapeutic Targets in Cardiovascular Disease.

The process of atherosclerosis leads to the formation of plaques in the arterial wall, resulting in a decreased blood supply to tissues and organs and its sequelae: morbidity and mortality. A class of membrane-bound proteins termed scavenger receptors (SRs) are closely linked to the initiation and progression of atherosclerosis. Increasing interest in understanding SR structure and function has led to the idea that these proteins could provide new routes for cardiovascular disease diagnosis, management, and treatment. In this review, we consider the main classes of SRs that are implicated in arterial disease. We consider how our understanding of SR-mediated recognition of diverse ligands, including modified lipid particles, lipids, and carbohydrates, has enabled us to better target SR-linked functionality in disease. We also link clinical studies on vascular disease to our current understanding of SR biology and highlight potential areas that are relevant to cardiovascular disease management and therapy.

A Combined In Vivo, In Vitro, In Silico Approach for Patient-Specific Haemodynamic Studies of Aortic Dissection.

The optimal treatment of Type-B aortic dissection (AD) is still a subject of debate, with up to 50% of the cases developing late-term complications requiring invasive intervention. A better understanding of the patient-specific haemodynamic features of AD can provide useful insights on disease progression and support clinical management. In this work, a novel in vitro and in silico framework to perform personalised studies of AD, informed by non-invasive clinical data, is presented. A Type-B AD was investigated in silico using computational fluid dynamics (CFD) and in vitro by means of a state-of-the-art mock circulatory loop and particle image velocimetry (PIV). Both models not only reproduced the anatomical features of the patient, but also imposed physiologically-accurate and personalised boundary conditions. Experimental flow rate and pressure waveforms, as well as detailed velocity fields acquired via PIV, are extensively compared against numerical predictions at different locations in the aorta, showing excellent agreement. This work demonstrates how experimental and numerical tools can be developed in synergy to accurately reproduce patient-specific AD blood flow. The combined platform presented herein constitutes a powerful tool for advanced haemodynamic studies for a range of vascular conditions, allowing not only the validation of CFD models, but also clinical decision support, surgical planning as well as medical device innovation.

Generation of Core-Sheath Polymer Nanofibers by Pressurised Gyration.

The ability to generate core-sheath bicomponent polymer nanofibers in a single-step with scale-up possibilities is demonstrated using pressurised gyration manufacturing. This is the first time that nanofiber containing more than one polymer having a core-sheath configuration has been generated in this way. Water-soluble polymers polyethylene oxide (PEO) and polyvinyl pyrrolidone (PVP) are used as the core and sheath layers, respectively. Core-sheath nanofibers with a diameter in the range of 331 to 998 nm were spun using 15 wt % PEO and 15 wt % PVP polymer solutions. The forming parameters, working pressure and rotating speed, had a significant influence on the size, size distribution and the surface morphology of the nanofibers generated. Overall, fibre size decreased with increasing working pressure and rotating speed. The fibre size was normally distributed in all cases, with 0.2 MPa working pressure in particular showing narrower distribution. The fibre size distributions for 0.1 and 0.3 MPa working pressure were broader and a mean fibre size of 331 nm was obtained in the latter case. The fibre size was evenly distributed and narrower for rotating speeds of 2000 and 4000 RPMs. The distribution was broader for rotating speed of 6000 RPM with a mean value obtained at 430 nm. Continuous, smooth and bead-free fibre morphologies were obtained in each case. The fibre cross-section analysis using a focused ion beam machine showed a solid core surrounded by a sheath layer. Our findings demonstrate that the pressurised gyration could be used to produce core-sheath polymer nanofibers reliably and cost-effectively with scale-up possibilities (~4 kg h-1).

Multiscale, patient-specific computational fluid dynamics models predict formation of neointimal hyperplasia in saphenous vein grafts.

Stenosis due to neointimal hyperplasia (NIH) is among the major causes of peripheral graft failure. Its link to abnormal hemodynamics in the graft is complex, and isolated use of hemodynamic markers is insufficient to fully capture its progression. Here, a computational model of NIH growth is presented, establishing a link between computational fluid dynamics simulations of flow in the lumen and a biochemical model representing NIH growth mechanisms inside the vessel wall. For all three patients analyzed, NIH at proximal and distal anastomoses was simulated by the model, with values of stenosis comparable to the computed tomography scans.