Cross-linking Methodology Research Articles

Rapid and Facile Light-Based Approach to Fabricate Protease-Degradable Poly(ethylene glycol)-norbornene Microgels for Cell Encapsulation.

Thiol-norbornene photoclickable poly (ethylene glycol) (PEG)-based (PEG-NB) hydrogels are attractive biomaterials for cell encapsulation, drug delivery, and regenerative medicine applications. Although many crosslinking strategies and chemistries have been developed for PEG-NB bulk hydrogels, fabrication approaches of PEG-NB microgels have not been extensively explored. Here, a fabrication strategy for 4-arm amide-linked PEG-NB (PEG-4aNB) microgels using flow-focusing microfluidics for human mesenchymal stem/stromal cell (hMSCs) encapsulation is presented. PEG-4aNB photochemistry allows high-throughput, ultrafast generation, and cost-effective synthesis of monodispersed microgels (diameter 340 ± 18, 380 ± 24, and 420 ± 15µm, for 6, 8, and 10 wt% of PEG-4aNB, respectively) using an in situ crosslinking methodology in a microfluidic device. PEG-4aNB microgels show in vitro degradability due to the incorporation of a protease-degradable peptide during photocrosslinking and encapsulated cells show excellent viability and metabolic activity for at least 13 days of culture. Furthermore, the secretory profile (i.e., MMP-13, ICAM-1, PD-L1, CXCL9, CCL3/MIP-1, IL-6, IL-12, IL-17E, TNF-α, CCL2/MCP-1) of encapsulated hMSCs shows increased expression in response to IFN-γ stimulation. Collectively, this work shows a versatile and facile approach for the fabrication of protease-degradable PEG-4aNB microgels for cell encapsulation.

Mussel-inspired chemistry in producing mechanically robust and bioactive hydrogels as skin dressings

The development of hydrogels as skin dressings demonstrates a great potential in real life applications. To achieve this, the hydrogel has to conquer its natural poor mechanical strength, and to prolong its lifetime, antifatigue and self-healing properties originating from dynamic interactions are also required. As skin dressings, the hydrogel needs to maintain its ductility while pursuing the above mentioned properties. In this work, poly(ethylene glycol) diacrylate is used to produce skin dressings by reinforcing poly(ethylene glycol) diacrylate/alginate double network hydrogels with a crosslinker from mussel-inspired chemistry, which is 3,4-dihydroxy-l-phenylalanine. This crosslinking methodology significantly improved mechanical strength of the hydrogel, with 11,200% increase in compressive failure strength; it endowed the hydrogel with outstanding antifatigue and training strengthening properties that makes its mechanical strength increasing in a 50 cycles compressive test; the hydrogel showed excellent self-healing properties that in rheological characterization; it also displayed enhanced storage modulus after withstanding a shear strain up to 1100%; meanwhile, the hydrogel exhibited extreme ductility with an elastic modulus of only 10.90–16.53 kPa. 3,4-dihydroxy-l-phenylalanine also renders the hydrogel its inherent antioxidant activity, conductivity, and bioadhesiveness. Together with the highly transparent appearance, the hydrogels possess a great potential and practibility in the fields of skin dressings.

Crosslinking Methodology for Imidazole-Grafted Silicone Elastomers Allowing for Dielectric Elastomers Operated at Low Electrical Fields with High Strains.

For improved actuation at low voltages of dielectric elastomers, a high dielectric permittivity has been targeted for several years but most successful methods then either increase the stiffness of the elastomer and/or introduce notable losses of both mechanical and dielectric nature. For polydimethylsiloxane (PDMS)-based elastomers, most high-permittivity moieties inhibit the sensitive platinum catalyst used in the addition curing scheme. In contrast to the classical addition curing pathway to prepare PDMS elastomers, here, an alternative strategy is reported to prepare PDMS elastomers via the crosslinking reaction between multifunctional imidazole-grafted PDMS with difunctional bis(1-ethylene-imidazole-3-ium) bromide ionic liquid (bis-IL). The prepared IL-elastomer entails uniformly dispersed IL and presents stable mechanical and dielectric properties due to the covalent nature of the crosslinking as opposed to previously reported physical mixing in of ILs. The relative permittivity was improved up to 200% by including the bis-IL in the elastomer, and Young's modulus was around 0.04 MPa. As a result of the excellent combination of properties, the dielectric actuator developed exhibits an area strain of 20% at 15 V/μm. The novel strategy to prepare PDMS elastomers provides a new paradigm for achieving high-performance dielectric elastomer actuators by a simple methodology.

Dual-Crosslinked Alginate-Based Hydrogels with Tunable Mechanical Properties for Cultured Meat.

Cultured meat refers to the production of animal tissue by utilizing the same techniques as tissue engineering through cell culture. Various biomaterials have been designed to serve as in vitro supports for cell viability, growth, and migration. In this study, visible light and dual-crosslinked alginate hydrogels were designed to enable control of the physical and mechanical properties needed for the fabrication of cultured meat scaffolds. We hypothesized that a difference in hydrogel stiffness would influence cell behavior, indicating the efficacy of our processing methods to benefit the cultured meat field. Herein, we synthesized and created: (1) methacrylated alginate (AlgMA) to enable covalent crosslinking via visible light exposure, (2) Methacrylated alginate and arginyl-glycyl-aspartic acid RGD conjugates (AlgMA-RGD), using carbodiimide chemistries to provide cell-binding sites on the material, and (3) designer hydrogels incorporating different crosslinking techniques. The material and mechanical properties were evaluated to determine the structural integrity of the hydrogels, and in vitro cell assays were conducted to verify cytocompatibility and cell adhesion. Gelation, swell ratio, and weight loss calculations revealed longer gelation times for the AlgMA scaffolds and similar physical properties for all hydrogel groups. We showed that by adjusting the polymer concentration and the crosslinking methodology, the scaffold’s mechanical properties can be controlled and optimized within physiological ranges. Incorporating dual crosslinking significantly increased the compressive moduli of the AlgMA hydrogels, compared to visible-light crosslinking alone. Moreover, the muscle satellite cells responded favorably to the AlgMA scaffolds, with clear differences in cell density when cultured on materials with significantly different mechanical properties. Our results indicate the usefulness of the dual-crosslinking alginate hydrogel system to support in vitro meat growth.

Ultrafast aquaporin-like multi-functionalized holey graphene membrane with tripartite nanochannels for organic solvent nanofiltration

Innovating next-generation two-dimensional (2D) membranes necessitates overcoming their selectivity/permeability trade-off limitations. Unlike most 2D multi-stacked membranes, where their water permeability always occurred through the single d-spacing channel, we engineered a novel aquaporin-like multi-functionalized holey graphene (HG) membrane with the potential of tripartite nanochannels. Through this strategy, high selectivity, superior permeability, and chemical stability were achieved. The utilized facile in-situ crosslinking methodology enabled the creation of a unique membrane featuring an aquaporin-like wide/tight interlayer d-spacing decorated with multifunctional SiO2 nanoparticles, in addition to direct nanochannels of HG. This unique structure boosted the solvent permeability by more than ten times without sacrificing the selectivity. This distinctive membrane category displayed excellent selectivity with a performance higher than 96% for dye rejection. This novel membrane has great potential to be utilized in various organic solvent nanofiltration and wastewater purification applications with extraordinary ability to surpass the permeance/separation trade-off.

Synthesis, Electrochemistry, Spectroelectrochemistry, and Electrochromism of Metallophthalocyanines Substituted with Four (2,4,5‐Trimethylphenyl)ethynyl Groups

AbstractThe synthesis of the novel phthalonitrile containing (2,4,5‐trimethylphenyl)ethynyl moieties was performed using the palladium‐catalyzed Sonogashira crosslinking methodology. Cobalt phthalocyanine was obtained with the cyclotetramerization reaction of this dinitrile derivative, while the synthesis of zinc and manganese phthalocyanines was reported by the Sonogashira coupling reaction of the appropriate iodo phthalocyanines. Voltammetric and electrochromic analyses were applied to determine the possible applications of these complexes in different electrochemical/display technologies. The rich redox responses of the complexes had a good influence on the spectroelectrochemical characters of them. Electrochromic measurements of the complexes indicated fast, and reversible color changes with reasonable optical contrasts.

Alginate-based self-healing hydrogels assembled by dual cross-linking strategy: Fabrication and evaluation of mechanical properties

One way to enhance the poor mechanical properties of the self-healing hydrogels based on host-guest (HG) interaction is employing the dual cross-linking method. Here, the alginate-based hydrogels based on HG complexation were prepared through the modification of alginate (ALG) polysaccharide with beta-cyclodextrin (βCD) and adamantane (Ad) as host and guest groups with different grafting values, respectively. The porous structure was confirmed for all ALG-CD:ALG-Ad hydrogels. The average pore size of ALG-CD1:ALG-Ad1 hydrogel cross-linked by HG interactions was 288 μm. Mechanical properties of the alginate-based HG hydrogels were improved by incorporating Ca2+ ions in their structure through dual cross-linking methodology. The maximum modulus of the porous dual-crosslinked hydrogel was reached up to 6500 Pa. The healing time of less than 5 s was obtained for the alginate-based hydrogels. The fabricated hydrogels can be used in 3D printing, tissue engineering, and drug delivery systems due to their biocompatibility and shear-thinning behavior.

Chitosan-Derived Magnetic Nanomaterials: Synthesis, Characterization, and Nitrite Adsorption in Water

Nitrite is one of the main pollutants in the water worldwide. In this study, we have applied the reverse suspension crosslinking methodology based on chitosan (CS) and Fe3O4 (FeO) to synthesize the novel magnetic nanomaterial of chitosan (CS-FeO). The physical and chemical properties of CS-FeO were further characterized by scanning electron microscopy, particle size distribution, thermogravimetry, fluxgate magnetometer, Fourier transform infrared spectroscopy, X-ray diffraction spectroscopy, and energy dispersive spectroscopy. Results revealed that CS-FeO showed high thermal stability in the temperature ranging from 50 to 200°C. CS-FeO showed high crystallinity and magnetism and was easily and quickly separated from aqueous solution in the presence of an external magnetic field. The molecular structure of CS-FeO showed that the core-shell structure of CS-FeO was established with FeO as the core and CS as the shell. Furthermore, the adsorption rate of nitrite by CS-FeO reached 65.83 ± 0.76 % under optimal conditions. Moreover, CS-FeO showed high regeneration capability with Na2SO4 used as the eluent. Our study demonstrated evidently that CS-FeO can be potentially used to remove nitrite from drinking water sources and industrial wastewater, suggesting the promising future of the application of CS-derived magnetic nanomaterials in the areas of environmental protections.

Dynamically crosslinked polydimethylsiloxane-based polyurethanes with contact-killing antimicrobial properties as implantable alloplasts for urological reconstruction

A large population of patients is reported to suffer from urinary bladder-associated irreversible physiological disorders, rationalizing a continuous surge for structural and functional substitutes of urinary tissues, including ureters, bladder-wall, and urethra. The current gold standard for bladder reconstruction, an autologous gastrointestinal graft, is proven not to be an ideal substitute in the clinic. While addressing this unmet clinical need, a unique platform of antimicrobial polydimethyl siloxane-modified polyurethanes (TPU/PDMS) is designed and developed for its potential application as a urological implant. To the best of our knowledge, this study reports for the first time the successful integration of varying contents of PDMS within the molten polyurethane matrix using in situ crosslinking methodology. Thus, compatibilized binary blends possess clinically relevant viscoelastic properties to sustain high pressure, large distensions, and surgical manipulation. Furthermore, different chemical strategies are explored to covalently incorporate quaternized moieties, including 4-vinyl pyridine (4-VP), branched-polyethyleneimine (bPEI) as well as bPEI-grafted-(acrylic acid-co-vinylbenzyltriphenyl phosphonium chloride) (PAP), and counter urinary tract infections. The modified compositions, endowed with contact killing surfaces, reveal nearly three log reduction in bacterial growth in pathogenically infected artificial urine. Importantly, the antimicrobial TPU/PDMS blends support the uninhibited growth of mitochondrially viable murine fibroblasts, in a manner comparable to the medical-grade polyurethane. Collectively, the obtained results affirmed the newly developed polymers as promising biomaterials in reconstructive urology. Statement of significanceThe clinical procedure for end-stage bladder disease remains replacement or augmentation of the bladder wall with a section of the patient's gastrointestinal tract. However, the absorptive and mucus-producing epithelium of intestinal segment is liable to short- and long-term complications. The dynamically crosslinked polydimethyl siloxane-based polyurethanes proposed herein, and the associated synthesis strategies to induce polycation grafted non-exhaustive contact-killing surfaces against uropathogents, have a significant clinical prospect in reconstructive urology. As an ‘off-the-shelf’ available alloplastic substitute, these blends offer the potential to circumvent the challenges associated with non-urinary autografts or scaffold based regenerative engineering and, thereby, shorten as well as simplify the surgical treatment. The targeted application has been conceived for a bladder patch to assist in various urinary diseases including, bladder carcinoma, refractory overactive bladder, interstitial cystitis, etc. However, given the ease of fabrication, moldability and the wide spectrum of mechanical properties that could be encompassed, these blends also present the possibility to be manifested into artificial ureteral or urethral conduits.

Cross-Linking Methodology for Fully Atomistic Models of Hydroxyl-Terminated Polybutadiene and Determination of Mechanical Properties

A computational method for preserving the equilibrated structure of a fully atomistic model of a prepolymer melt while introducing cross-links is introduced. An atomistic model of hydroxyl-terminated polybutadiene (HTPB) is cured using isophorone diisocyanate (IPDI) to form an HTPB-IPDI elastomer. After cross-linking, the elastomer is quenched to several different temperatures and subsequently stressed via both compressive and tensile deformations to assess mechanical properties as a function of temperature. This study establishes the framework for investigating the degradation and embrittlement of HTPB-IPDI binders by molecular simulation.

Physically-crosslinked polyvinyl alcohol composite hydrogels containing clays, carbonaceous materials and magnetic nanoparticles as fillers

Polyvinyl alcohol (PVA)/ Bentonite, acid-modified Bentonite, magnetic nanoparticles and mesoporous carbonaceous composite hydrogels were synthesized via an eco-friendly freeze-thawing technique, which is a simple and non-toxic crosslinking methodology. After thorough characterisation using X-Ray Diffraction (XRD), Scanning Electron microscopy (SEM), Dynamic Scanning Calorimetry (DSC), Thermogravimetric Analysis (TGA) and swelling assessment.Furthermore, the prepared materials were tested in order to explore their cadmium adsorption potential, a heavy metal usually found as a pollutant in water bodies. The data obtained showed that PVA has no affinity for cadmium ions but the incorporation of the different fillers makes the materials develop important adsorbent characteristics. Under the selected working conditions, and for a contact time of 1440 min, the maximum amount of cadmium ions adsorbed was 50.9 mg/g, 33.9 mg/g, 42.6 mg/g and 30.5 mg/g for composites containing bentonite; acid-modified bentonite; magnetic nanoparticles and carbonaceous materials, respectively. For all cases the higher the filler amounts the higher the adsorption efficiency.

Furan Cross-Linking Technology for Investigating GPCR-Ligand Interactions.

Interactions between G protein-coupled receptors and their ligands hold extensive potential for drug discovery. Studying these interactions poses technical problems due to their transient nature and the inherent difficulties when working with G protein-coupled receptors (GPCR) that are only functional in a membrane setting. Here, we describe the use of a furan-based chemical cross-linking methodology to achieve selective covalent coupling between a furan-modified peptide ligand and its native GPCR present on the surface of living cells under normal cell culture conditions. This methodology relies on the oxidation of the furan moiety, which can be achieved by either addition of an external oxidation signal or by the reactive oxygen species produced by the cell. The cross-linked ligand-GPCR complex is subsequently detected by Western blotting based on the biotin label that is incorporated in the peptide ligand.

Self-reporting visible light-induced polymer chain collapse

We introduce a facile photoinduced self-reporting crosslinking methodology for the compaction of polymer chains in highly diluted solution.

Microstructured Elastomer-PEG Hydrogels via Kinetic Capture of Aqueous Liquid-Liquid Phase Separation.

Heterogeneous hydrogels with desired matrix complexity are studied for a variety of biomimetic materials. Despite the range of such microstructured materials described, few methods permit independent control over microstructure and microscale mechanics by precisely controlled, single‐step processing methods. Here, a phototriggered crosslinking methodology that traps microstructures in liquid–liquid phase‐separated solutions of a highly elastomeric resilin‐like polypeptide (RLP) and poly(ethylene glycol) (PEG) is reported. RLP‐rich domains of various diameters can be trapped in a PEG continuous phase, with the kinetics of domain maturation dependent on the degree of acrylation. The chemical composition of both hydrogel phases over time is assessed via in situ hyperspectral coherent Raman microscopy, with equilibrium concentrations consistent with the compositions derived from NMR‐measured coexistence curves. Atomic force microscopy reveals that the local mechanical properties of the two phases evolve over time, even as the bulk modulus of the material remains constant, showing that the strategy permits control of mechanical properties on micrometer length scales, of relevance in generating mechanically robust materials for a range of applications. As one example, the successful encapsulation, localization, and survival of primary cells are demonstrated and suggest the potential application of phase‐separated RLP‐PEG hydrogels in regenerative medicine applications.

Dynamic roles for the N-terminus of the yeast G protein-coupled receptor Ste2p

The Saccharomyces cerevisiae α-factor receptor Ste2p has been used extensively as a model to understand the molecular mechanism of signal transduction by G protein-coupled receptors (GPCRs). Single and double cysteine mutants of Ste2p were created and served as surrogates to detect intramolecular interactions and dimerization of Ste2p using disulfide cross-linking methodology. When a mutation was introduced into the phylogenetically conserved tyrosine residue at position 26 (Y26C) in the N-terminus of Ste2p, dimerization was increased greatly. The amount of dimer formed by this Y26C mutant was greatly reduced by ligand binding even though the ligand binding site is far removed from the N-terminus; the lowering of the dimer formation was consistent with a conformational change in the N-terminus of the receptor upon activation. Dimerization was decreased by double mutations Y26C/V109C or Y26C/T114C indicating that Y26 is in close proximity to V109 and T114 of extracellular loop 1 in native Ste2p. Combined with earlier studies, these results indicate previously unrecognized roles for the N-terminus of Ste2p, and perhaps of GPCRs in general, and reveal a specific N-terminus residue or region, that is involved in GPCR signaling, intrareceptor interactions, and receptor dimerization.

Hyperbranched polyethyleneimine induced polycationic membranes for improved fouling resistance and high RO performance

The study shows a new method for hyperbranched polyethyleneimine (HPEI) induced polycation membranes with an objective of achieving better fouling resistance and high reverse osmosis (RO) performance. Chemically crosslinked composite membrane (CCCM) was prepared with polyvinyl alcohol (PVA), maleic acid (MA) and Pluronic F127 via chemical crosslinking methodology. Polycation induced membranes were fabricated by surface coating of HPEI onto the CCCM (PVA/MA/PluronicF127) RO membranes. Functional group analysis, morphology and surface roughness of CCCM and HPEI-RO membranes were analyzed by Fourier transform infrared spectroscopy, scanning electron microscopy and atomic force microscopy respectively. The hydrophilicity and water permeability of CCCM and HPEI-RO were examined with water contact angle measurement and RO performance test. Optimum RO performance was obtained for HPEI-RO3 membrane with NaCl and MgSO4, as for RNaCl=96.1%, JNaCl=58.2Lm−2h−1 and RMgSO4=99.6%, JMgSO4=15.8Lm−2h−1 (testing with 3.28wt.% aqueous NaCl and MgSO4 solution at 25°C and 55bar). Moreover, cetyl trimethylammonium bromide (CTAB) as standard foulants presented a smaller fouling tendency for the modified HPEI-RO3 membrane. HPEI-RO3 compared to the unmodified membrane, was able to decrease the stickiness of charged bacteria, Gram negative Escherichia coli and Gram positive Staphylococcus epidermidis. Therefore, the permeation flux, salt rejection and fouling resistant property of membrane (HPEI-RO3) were significantly improved with polycation induction into the membranes.

(Invited) Crosslinked High Temperature Polymer Electrolyte Membranes

High temperature polymer electrolyte membranes containing basic units that are able to interact with strong protic acids providing ionically conductive composite membrane have shown respectable performance and long operational stability. Further increase of the operation temperature and even an increase on the obtained conductivity values would facilitate the construction of efficient systems providing both electricity and heat, increasing thus the total efficiency. Initial attempts to stabilize the HTPEM electrolyte membranes through covalent crosslinking have proven that the operation temperature can be shifted up to 220oC 1,2. Different crosslinking methodologies have been tested to stabilize either PBI or aromatic polyethers bearing pyridine units3,4. In the present work we focused on the optimization of the crosslinking methodology using side double bonds and we also develop new crosslinking methodologies leading to wholly aromatic crosslinked structures. Different type of crosslinkers have been used, controlling thus the acid doping ability of the final membranes while the use of dopable crosslinkers that enable the high acid uptake and retention. Moreover, depending on the chemical structure of the crosslinked membranes, a significant increase of the conductivity values of the final acid doped composite membranes was obtained. High temperature operation as the one achieved here enables the use of liquid feed like methanol in a compact methanol reformer-high temperature PEM fuel Cell setup that is under construction and testing5.

Antifogging Surface Facilitated by Nanoscale Coatings with Controllable Hydrophobicity and Cross‐Linking Density

Antifogging and frost-resistant coatings can be used in a wide range of applications and enable high light transmission through substrates even under changes in environmental conditions. In this study, surface confined and cross-linked antifogging thin films are fabricated on glass slides via catalyst induced cross-linking (CIC), which has been recently introduced as an easy and efficient cross-linking methodology. Four different poly(ethylene glycol) (PEG)-based polymers with different hydrophilicity are synthesized and used to prepare films via CIC. Films prepared from the most hydrophilic PEG-based polymers display the best antifogging performances when exposed to a temperature change from −20 to 22 °C. Furthermore, several parameters including cross-linking density, surface roughness, hydrophobicity, and exposure time are also evaluated in terms of film transparency. Through these measurements, it is determined that, more loosely cross-linked films retain antifogging ability for longer time periods due to higher film swellability as compared to, more highly cross-linked films. This study signifies the crucial role of the film cross-linking density and hydrophilicity on the antifogging function.

Fabrication of gallium hexacyanoferrate modified carbon ionic liquid paste electrode for sensitive determination of hydrogen peroxide and glucose

Gallium hexacyanoferrate (GaHCFe) and graphite powder were homogeneously dispersed into n-dodecylpyridinium hexafluorophosphate and paraffin to fabricate GaHCFe modified carbon ionic liquid paste electrode (CILPE). Mixture experimental design was employed to optimize the fabrication of GaHCFe modified CILPE (GaHCFe-CILPE). A pair of well-defined redox peaks due to the redox reaction of GaHCFe through one-electron process was observed for the fabricated electrode. The fabricated GaHCFe-CILPE exhibited good electrocatalytic activity towards reduction and oxidation of H2O2. The observed sensitivities for the electrocatalytic oxidation and reduction of H2O2 at the operating potentials of +0.8 and −0.2V were about 13.8 and 18.3mAM−1, respectively. The detection limit (S/N=3) for H2O2 was about 1μM. Additionally, glucose oxidase (GOx) was immobilized on GaHCFe-CILPE using two methodology, entrapment into Nafion matrix and cross-linking with glutaraldehyde and bovine serum albumin, in order to fabricate glucose biosensor. Linear dynamic rage, sensitivity and detection limit for glucose obtained by the biosensor fabricated using cross-linking methodology were 0.1–6mM, 0.87mAM−1 and 30μM, respectively and better than those obtained (0.2–6mM, 0.12mAM−1 and 50μM) for the biosensor fabricated using entrapment methodology.

A Synthetic Oligonucleotide Model for Evaluating the Oxidation and Crosslinking Propensities of Natural Furan‐Modified DNA

We have previously developed a crosslinking methodology for oligonucleotides based on the incorporation of furan moieties, which can be selectively oxidised to reactive intermediates that will quickly react with the opposite bases in DNA, forming toxic interstrand crosslinks (ICLs). Furan moieties also occur in natural DNA, as a result of oxidative stress. Moreover, the furan-containing degradation product of this modified DNA-kinetin-has been found to display beneficial anti-ageing effects. To investigate the apparent discrepancy between the effects of the synthetic and the natural furan modifications in DNA, a quick and easy postsynthetic method providing access to the natural modification in short synthetic oligonucleotides was developed. On checking for potential crosslinking propensity, we found that the furan moiety does indeed undergo oxidation, in this way functioning as an important scavenger for oxidative stress. The reactive intermediate, however, was shown to degrade without producing toxic crosslinked products.