Albumin Infusion in Critically Ill COVID-19 Patients: Hemodilution and Anticoagulation

Hypercoagulation is one of the major risk factors for ICU treatment, mechanical ventilation, and death in critically ill patients infected with SARS-CoV-2. At the same time, hypoalbuminemia is one risk factor in such patients, independent of age and comorbidities. Especially in patients with severe SARS-CoV-2-infection, albumin infusion may be essential to improve hemodynamics and to reduce the plasma level of the main marker of thromboembolism, namely, the D-dimer plasma level, as suggested by a recent report. Albumin is responsible for 80% of the oncotic pressure in the vessels. This is necessary to keep enough water within the systemic circulatory system and for the maintenance of sufficient blood pressure, as well as for sufficient blood supply for vital organs like the brain, lungs, heart, and kidney. The liver reacts to a decrease in oncotic pressure with an increase in albumin synthesis. This is normally possible through the use of amino acids from the proteins introduced with the nutrients reaching the portal blood. If these are not sufficiently provided with the diet, amino acids are delivered to the liver from muscular proteins by systemic circulation. The liver is also the source of coagulation proteins, such as fibrinogen, fibronectin, and most of the v WF VIII, which are physiological components of the extracellular matrix of the vessel wall. While albumin is the main negative acute-phase protein, fibrinogen, fibronectin, and v WF VIII are positive acute-phase proteins. Acute illnesses cause the activation of defense mechanisms (acute-phase reaction) that may lead to an increase of fibrinolysis and an increase of plasma level of fibrinogen breakdown products, mainly fibrin and D-dimer. The measurement of the plasma level of the D-dimer has been used as a marker for venous thromboembolism, where a fourfold increase of the D-dimer plasma level was used as a negative prognostic marker in critically ill SARS-CoV-2 hospitalized patients. Increased fibrinolysis can take place in ischemic peripheral sites, where the mentioned coagulation proteins can become part of the provisional clot (e.g., in the lungs). Although critically ill SARS-CoV-2-infected patients are considered septic shock patients, albumin infusions have not been considered for hemodynamic resuscitation and as anticoagulants. The role of coagulation factors as provisional components of the extracellular matrix in case of generalized peripheral ischemia due to hypoalbuminemia and hypovolemia is discussed in this review.     

Air‐dried 3D‐collagen–chitosan biocomposite scaffold for tissue engineering application

Collagen–chitosan scaffolds of different compositions were developed using emulsion air‐drying method. The scaffolds prepared adding 10–30 wt% of chitosan to collagen improved the mechanical properties of the composite scaffold, and 7:3 ratio (collagen :chitosan) was found to be a better composite having a tensile strength of 13.57 MPa with 9% elongation at break. The water‐uptake characteristics were performed at different pH and found to be ameliorated for the composite scaffolds compared to pure collagen and chitosan scaffold, respectively. The pores ranging from 100 to 300 μm were well interconnected, and their distribution was fairly homogeneous in the scaffold as observed through scanning electron microscopy. Furthermore, the scaffold decreased the bacterial counts and supported fibroblasts attachment and proliferation, thus demonstrating this composite to be a good substrate for biomedical application.POLYM. COMPOS., 33:2029–2035, 2012. © 2012 Society of Plastics Engineers     

Microporosities in 3D-Printed Tricalcium-Phosphate-Based Bone Substitutes Enhance Osteoconduction and Affect Osteoclastic Resorption

Additive manufacturing is a key technology required to realize the production of a personalized bone substitute that exactly meets a patient’s need and fills a patient-specific bone defect. Additive manufacturing can optimize the inner architecture of the scaffold for osteoconduction, allowing fast and reliable defect bridging by promoting rapid growth of new bone tissue into the scaffold. The role of scaffold microporosity/nanoarchitecture in osteoconduction remains elusive. To elucidate this relationship, we produced lithography-based osteoconductive scaffolds from tricalcium phosphate (TCP) with identical macro- and microarchitecture, but varied their nanoarchitecture/microporosity by ranging maximum sintering temperatures from 1000 °C to 1200 °C. After characterization of the different scaffolds’ microporosity, compression strength, and nanoarchitecture, we performed in vivo studies that showed that ingrowth of bone as an indicator of osteoconduction significantly decreased with decreasing microporosity. Moreover, at the 1200 °C peak sinter temperature and lowest microporosity, osteoclastic degradation of the material was inhibited. Thus, even for wide-open porous TCP-based scaffolds, a high degree of microporosity appears to be essential for optimal osteoconduction and creeping substitution, which can prevent non-unions, the major complication during bone regeneration procedures.     

Design of Artificial Enzymes: Insights into Protein Scaffolds

The design of artificial enzymes has emerged as a promising tool for the generation of potent biocatalysts able to promote new-to-nature reactions with improved catalytic performances, providing a powerful platform for wide-ranging applications and a better understanding of protein functions and structures. The selection of an appropriate protein scaffold plays a key role in the design process. This review aims to give a general overview of the most common protein scaffolds that can be exploited for the generation of artificial enzymes. Several examples are discussed and categorized according to the strategy used for the design of the artificial biocatalyst, namely the functionalization of natural enzymes, the creation of a new catalytic site in a protein scaffold bearing a wide hydrophobic pocket and de novo protein design. The review is concluded by a comparison of these different methods and by our perspective on the topic.     

The incorporation and controlled release of platelet‐rich plasma‐derived biomolecules from polymeric tissue engineering scaffolds

Platelet‐rich plasma (PRP) has been gaining popularity in recent years as a cost‐effective material capable of stimulating healing in a number of different clinical applications. As the clinical role of PRP has been growing so too has its prevalence in the fields of tissue engineering and regenerative medicine, particularly in the field of extracellular matrix (ECM) analogue scaffold fabrication. As polymeric scaffold fabrication techniques strive to create structures that ever more closely replicate the native ECM's form and function, the need for increased scaffold bioactivity becomes more pronounced. PRP, which has been shown to contain over 300 bioactive molecules, has the potential to deliver a combination of growth factors and cytokines capable of stimulating cellular activity through enhanced chemotaxis, proliferation and ECM production. The ability to incorporate such a potent bioactive milieu into a polymeric tissue engineering scaffold, which lacks intrinsic cell signaling molecules, may help to promote scaffold integration with native tissues and increase the overall patency of polymeric ECM analogue structures. This mini‐review briefly discusses the physiological basis of PRP and its current clinical use, as well as the potential role that PRP may play in the future of polymeric tissue engineering scaffold design. Copyright © 2012 Society of Chemical Industry     

Superior Alignment of Human iPSC-Osteoblasts Associated with Focal Adhesion Formation Stimulated by Oriented Collagen Scaffold

Human-induced pluripotent stem cells (hiPSCs) can be applied in patient-specific cell therapy to regenerate lost tissue or organ function. Anisotropic control of the structural organization in the newly generated bone matrix is pivotal for functional reconstruction during bone tissue regeneration. Recently, we revealed that hiPSC-derived osteoblasts (hiPSC-Obs) exhibit preferential alignment and organize in highly ordered bone matrices along a bone-mimetic collagen scaffold, indicating their critical role in regulating the unidirectional cellular arrangement, as well as the structural organization of regenerated bone tissue. However, it remains unclear how hiPSCs exhibit the cell properties required for oriented tissue construction. The present study aimed to characterize the properties of hiPSCs-Obs and those of their focal adhesions (FAs), which mediate the structural relationship between cells and the matrix. Our in vitro anisotropic cell culture system revealed the superior adhesion behavior of hiPSC-Obs, which exhibited accelerated cell proliferation and better cell alignment along the collagen axis compared to normal human osteoblasts. Notably, the oriented collagen scaffold stimulated FA formation along the scaffold collagen orientation. This is the first report of the superior cell adhesion behavior of hiPSC-Obs associated with the promotion of FA assembly along an anisotropic scaffold. These findings suggest a promising role for hiPSCs in enabling anisotropic bone microstructural regeneration.     

FEA evaluation of material stiffness changes for a polymer assisted 3D polycaprolactone/β-tricalcium phosphate scaffold in a mandibular defect reconstruction model

In bone regenerative engineering, the biomechanical performance of the scaffold at the bone-tissue interface is a key consideration. The evaluation of this parameter is a crucial step in designing, optimizing and manufacturing of bone substitute materials for clinical application, but is as yet comparatively unexplored. To this end, we utilized a novel polymer-assisted method to fabricate a three-dimensional (3D) Polycaprolactone/β-Tricalcium phosphate scaffold. The compressive modulus was measured and the data used to inform finite element analysis. The scaffolds achieved a maximum compressive moduli of 151 MPa, close to that of cortical bone. Further computational simulations were performed to determine the stresses and local scaffold adaptation profile, using data from computer tomography scans of the mandible. Local stresses were simulated based on the density changes in new bone forming in the scaffold at different stages of healing. The stress distribution in the mandible, scaffold center and scaffold interface were explored for a static load of 200 N, which corresponds to the load of adult mastication near the incisors. The analysis revealed that the maximum cross-sectional stress at the scaffold center and at the scaffold interface was 2.7 and 4.12 MPa respectively. The majority of the stress was localized in the bone of the mandible, with the scaffold bearing minimal loading at the start, but more over time as infiltration of more new bone progressed.     

Rational design of highly active and selective ligands for the alpha5beta1 integrin receptor

The inhibition of integrin function is a major challenge in medicinal chemistry. Potent ligands are currently in different stages of clinical trials for the antiangiogenic therapy of cancer and age-related macula degeneration (AMD). The subtype alpha5beta1 has recently been drawn into the focus of research because of its genuine role in angiogenesis. In our previous work we could demonstrate that the lack of structural information about the receptor could be overcome by a homology model based on the X-ray structure of the alphavbeta3 integrin subtype and the sequence similarities between both receptors. In this work, we describe the rational design and synthesis of high affinity alpha5beta1 binders, and the optimisation of selectivity against alphavbeta3 by means of extensive SAR studies and docking experiments. A first series of compounds based on the tyrosine scaffold resulted in affinities in the low and even subnanomolar range and selectivities of 400-fold against alphavbeta3. The insights about the structure-activity relationship gained from tyrosine-based ligands could be successfully transferred to ligands that bear an aza-glycine scaffold to yield alpha5beta1 ligands with affinities of approximately 1 nm and selectivities that exceed 10(4)-fold. The ligands have already been successfully employed as selective alpha5beta1 ligands in biological research and might serve as lead structures for antiangiogenic cancer therapy.     

Solvent effect in phase separation for fabrication of micropatterned porous scaffold sheets

Cellular orientation control is important for tissue regeneration. Design of oriented structures for cells with suitable features can be used in tissue engineering. One of the methods of cellular orientation with the aim of regenerating which damaged tissues is utilizing oriented biocompatible substrates. This paper reports a one-step method with different solvents to fabricate porous micropatterned polyhydroxybutyrate scaffold sheets. The results indicated that the porosity and pore morphology of the scaffolds are viable with respect to proliferation rate, and a micropattern for cell alignment. Stem cells culturing proved that the scaffold sheets are suitable for cell culturing. Preliminary experiments indicate that the 2-D scaffold sheets are very promising as basis for building 3-D scaffolds.     

Synthesizing and Characterizing of Gelatin-Chitosan-Bioactive Glass (58s) Scaffolds for Bone Tissue Engineering

Nowadays repairing and regenerating of lost or damaged tissue still remain an important challenge in clinical techniques. Due to the variety of available bone grafts, different types of biodegradable materials are being utilized as a scaffold implant. The basic structure of the bone is an excellent natural composite which contains varieties of polymers and ceramics; therefore, it is important to manufacture a bone scaffold featuring sufficient mechanical strength, a good degree of biocompatibility, biodegradation and an increased rate of formation of new tissue. Bioactive glass has an appealing characteristic which can be utilized for repairing purposes as well as to cause a rapid response from the bone graft. In this study, a composite scaffold based on polymer matrix (gelatin-chitosan) and bioactive glass 58s was synthesized in the laboratory. Five samples of polymer scaffold with different proportions of bioactive glass were designed and investigated. The scaffolds were dried with freeze dryer, and a spongy structure was generated. The composite survey was carried out through FTIR technique to examine the crystallization of the structure, XRD to examine the morphology of the porosities, and SEM to examine the size of porosities and formation of apatite. This study reveals that the size of porosities is about 170–320 μm, which is suitable for angiogenesis and cell growth in the bone. The combination of enhanced properties and the formation of apatite on the surface of the scaffolds make them an ideal option as a bone substitute.     

Alginate/TiO2@LDH microspheres: A promising bioactive scaffold with cytocompatibility and antibacterial activity

In this study, a hybrid of TiO₂@LDH was applied to fabricate a bone scaffold to take the advantages of both nanostructures. In addition to the role of LDH in the biomineralization process, it was used to provide a suitable bed for the better dispersion of TiO₂ nanoparticles and prevent them from aggregations. The main limitation in the usage of these nanoceramics in the preparation of bone scaffolds is their powder state, which prevents their fixation in the desired place. So, alginate (Alg) was selected as the matrix to embed this nanohybrid and support its stability and fixation. Bioactivity of the prepared Alg/TiO₂@LDH scaffolds was assessed by incubation in the simulated body fluid at 37 °C for 28 days. Field emission scanning electron microscopy as well as energy dispersive X-ray were used to prove the biomineralization on the scaffolds. MTT test showed that the scaffold containing 6 wt% of the TiO₂@LDH do not have any toxicity effects on the MG-63 cell line. Also, this scaffold presented outstanding antibacterial ability against Staphylococcus aureus, which is known as an important infecting agent in the bone scaffolds. The proposed Alg/TiO₂@LDH scaffold in this study could be a proper candidate to be used in bone tissue engineering.     

A Conformationally Stable Acyclic β-Hairpin Scaffold Tolerating the Incorporation of Poorly β-Sheet-Prone Amino Acids

The β-hairpin is a structural element of native proteins, but it is also a useful artificial scaffold for finding lead compounds to convert into peptidomimetics or non-peptide structures for drug discovery. Since linear peptides are synthetically more easily accessible than cyclic ones, but are structurally less well-defined, we propose XWXWXpPXK(/R)X(R) as an acyclic but still rigid β-hairpin scaffold that is robust enough to accommodate different types of side chains, regardless of the secondary-structure propensity of the X residues. The high conformational stability of the scaffold results from tight contacts between cross-strand cationic and aromatic side chains, combined with the strong tendency of the d -Pro-l -Pro dipeptide to induce a type II′ β-turn. To demonstrate the robustness of the scaffold, we elucidated the NMR structures and performed molecular dynamics (MD) simulations of a series of peptides displaying mainly non-β-branched, poorly β-sheet-prone residues at the X positions. Both the NMR and MD data confirm that our acyclic β-hairpin scaffold is highly versatile as regards the amino-acid composition of the β-sheet face opposite to the cationic−aromatic one.     

角蛋白基药物载体的发展及应用探讨

从毛发中提取角蛋白，开发具有独特功能的角蛋白基药物载体材料，提高药物的活性和生物利用率，既能够增加毛发的利用价值，又能够实现节能减排，符合可持续发展战略要求。对毛发中提取角蛋白进行二硫键重建、自由基聚合、多糖改性和烷基化等改性处理，赋予角蛋白基药物载体优异地功能特性，进一步拓宽其作为角蛋白基药物载体的应用范围。角蛋白基药物载体可被加工成纳米粒子、纤维支架、水凝胶等各种形式用于药物的递送。作为一种高硫含量且具有良好细胞相容性的结构蛋白，角蛋白基药物载体在组织工程、再生医学等领域扮演着重要的角色。     

Negatively charged purification tags for selective anion-exchange recovery

A novel strategy for the highly selective purification of recombinant fusion proteins using negatively charged protein domains, which were constructed by protein design, is described. A triple alpha-helical domain of 58 amino acids was used as scaffold. Far-ultraviolet circular dichroism measurements showed that the designed domains had very low alpha-helicity in a low-conductivity environment in contrast to the scaffold. The secondary structure could be induced by adding salt, giving a structure comparable to the parental molecule. Further studies showed that the new domains were able to bind to an anion exchanger even at pH values down to 5 and 6. Gene fusions between one of the designed domains and different target proteins, such as green fluorescent protein (GFP), maltose binding protein (MBP) and firefly luciferase, were also constructed. These gene products could be efficiently purified from whole cell lysates at pH 6 using anion-exchange chromatography.     

纳米纤维应用于组织工程的研究进展

组织工程支架的关键作用是起到引导细胞繁殖、生长,促进组织修复的一个过程。纳米纤维支架由于具有特殊的纳米效应,而更有利于细胞的黏附、增殖、功能化,因而被广泛应用于组织工程。本文分别介绍了纳米纤维支架在各类组织工程中应用,包括皮肤和创伤敷料组织工程、血管组织工程、神经组织工程、骨组织工程、软骨组织工程中应用的研究进展,同时介绍了纳米纤维支架药物控制释放中的应用。指出目前纳米技术还不成熟,需要从制备工艺的优化、基因工程的引入及纳米材料安全性能的科学评价等几方面解决纳米纤维支架在组织修复工程中面临的问题。     

Preparation and characterization of polyhydroxybutyrate‐co‐hydroxyvalerate/silk fibroin nanofibrous scaffolds for skin tissue engineering

Nanofibrous scaffolds were obtained by co‐electrospinning poly (3‐hydroxybuty‐rate‐co‐3‐hydroxyvalerate) (PHBV) and fibroin regenerated from silk in different proportions using 1,1,1,3,3,3‐hexafluoro‐2‐isopropanol (HFIP) as solvent. Field emission scanning electron microscope (FESEM) investigation showed that the fiber diameters of the nanofibrous scaffolds ranged from 190 to 460 nm. X‐ray diffraction (XRD) and Fourier transform infrared spectroscopy analysis (FT‐IR) showed that the main structure of silk fibroin (SF) in the nanofibrous scaffold was β‐sheet. Compared to the PHBV nanofibrous scaffold, the surface hydrophilicity and water‐uptake capability of the PHBV/SF nanofibrous scaffold with 50/50 were improved. The results of cell adhesion experiment showed that the fibroblasts adhered more to the PHBV/SF nanofibrous scaffold with 50/50 than the pure PHBV nanofibrous scaffold. The proliferation of fibroblast on the PHBV/SF nanofibrous scaffold with 50/50 was higher than that on the pure PHBV nanofibrous scaffold. Our results indicated that the PHBV/SF nanofibrous scaffold with 50/50 may be a better candidate for biomedical applications such as skin tissue engineering and wound dressing. POLYM. ENG. SCI., 55:907–916, 2015. © 2014 Society of Plastics Engineers     

Assembly of a DNA Origami Chinese Knot by Only 15% of the Staple Strands

As a giant leap in DNA self-assembly, DNA origami has exhibited an unprecedented ability to construct nanostructures with arbitrary shapes and sizes. In typical DNA origami, hundreds of short DNA staple strands fold a long, single-stranded (ss) DNA scaffold cooperatively into designed nanostructures. However, large numbers of DNA strands are expensive and would hinder applications such as pharmaceutical investigations because of the complicated components. Therefore, one challenge is how to reduce the number of staple strands needed to construct DNA origami. For a DNA origami structure, the scale-free folding pattern of the scaffold strand is determined by staple strands at the branching vertexes. Simple duplex regions help to define the size-related features of the origami geometry. In this study, we hypothesized that a scaffold strand can be correctly folded into a designed topology by using only staple strands involved in branching vertexes. After assembly, any remaining, flexible, single-stranded regions of the scaffold could be converted into rigid duplexes by DNA polymerase to achieve the designed geometric structures. To demonstrate the concept, we used only 18 staple strands (covering 15 % of the scaffold strand) to assemble a porous DNA nanostructure, which was visualized by atomic force microscopy (AFM). This study helps understanding of the role of cooperativity in origami folding, and provides a cost-effective approach for small-scale prototyping DNA origami.     

Design and Synthesis of Tricyclic JAK3 Inhibitors with Picomolar Affinities as Novel Molecular Probes

The Janus kinase (JAK) signaling pathway is of particular importance in the pathology of inflammatory diseases and oncological disorders, and the inhibition of Janus kinase 3 (JAK3) with small molecules has proven to provide therapeutic immunosuppression. A novel class of tricyclic JAK inhibitors derived from the 3‐methyl‐1,6‐dihydrodipyrrolo[2,3‐b:2′,3′‐d]pyridine scaffold was designed based on the tofacitinib–JAK3 crystal structure by applying a rigidization approach. A convenient synthetic strategy to access the scaffold via an intramolecular Heck reaction was developed, and a small library of inhibitors was prepared and characterized using in vitro biochemical as well as cellular assays. IC₅₀ values as low as 220 pM could be achieved with selectivity for JAK3 over other JAK family members. Both activity and selectivity were confirmed in a cellular STAT phosphorylation assay, providing also first‐time data for tofacitinib. Our novel inhibitors may serve as tool compounds and useful probes to explore the role of JAK3 inhibition in pharmacodynamics studies.     

Physico‐optical properties of a crosslinked hyaluronic acid scaffold for biomedical applications

Optical techniques are increasingly employed for monitoring cell–matrix interactions in suitably prepared 3D scaffolds. The ability of designing and realizing synthetic extracellular matrix with well‐controlled optical properties is a crucial need in this field. For this purpose, a crosslinked hyaluronic acid (HA) scaffold is prepared. Fourier transform infrared and ultraviolet–visible spectroscopies enable to monitor the scaffold preparation process and to evidence scaffold high transparency and low fluorescence in the visible range. 3D optical characteristics of the HA scaffold are tested by two‐photon microscopy (TPM) imaging of embedded fluorescent microbeads and alive keratinocytes labeled with vital PKH67 dye at different depths from the scaffold surface. Some useful indications about the potentiality of TPM measurements for the determination of attenuation coefficient of turbid media are also reported. Moreover, the use of the presented HA scaffold for preparing tissue phantoms for fluorescence imaging or diffuse imaging is proposed. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017 , 134, 45243.     

Single and Binary Adsorption of Food Dyes on Chitosan/Activated Carbon Hydrogels

The removal of dye contaminants from water is an important issue and different adsorbents have been developed for this purpose. Here, chitosan hydrogel, scaffold‐chitosan hydrogel, and scaffold‐chitosan hydrogel with activated carbon were developed and used in the adsorption of food blue 2 and food red 17, from single and binary aqueous systems. The adsorbents presented good mechanical properties, high efficiency, high adsorption capacity, and fast kinetics for single and binary systems. Chitosan hydrogel presented excellent reusability capacity and can be reused for five cycles. The use of these hydrogels enables the application of adsorbent materials with three‐dimensional frameworks for dye adsorption in single or binary systems.