Dually-crosslinked ionic conductive hydrogels reinforced through biopolymer gellan gum for flexible sensors to monitor human activities.

Human-machine interactions, monitoring of health equipment, and gentle robots all depend considerably on flexible strain sensors. However, making strain sensors have better mechanical behavior and an extensive sensing range remains an urgent difficulty. In this study, poly acrylamide-co-butyl acrylate with gellan gum (poly(AAm-co-BA)@GG) hydrophobic association networks and intermolecular hydrogen bonding interactions are used to fabricate dual cross-linked hydrogels for wearable resistive-type strain sensors. This could be an acceptable way to minimize the limitations in hydrogels previously identified. The robust fracture strength (870 kPa) and exceptional stretchability (1297 %) of the hydrogel arise from the collaborative action of intermolecular hydrogen bonding and hydrophobic associations. It also demonstrates exceptional resilience to repeated cycles of uninterrupted stretching and relaxation, retaining its structural integrity. The response and restoration times are 110 and 120 ms respectively. Furthermore, a wide sensing range (0-900 %), notable sensitivity across various strain levels, and an impressive gauge factor (GF) of 31.51 with high durability were observed by the dual cross-linked (DC) hydrogel-based strain sensors. The measured conductivity of the hydrogel was 0.32 S/m which is due to the incorporation of NaCl. Therefore, the hydrogels can be tailored to function as wearable strain sensors that can detect subtle human gestures like speech patterns, distinguish between distinct words, and recognize vibrations of the larynx during drinking, as well as large joint motions like wrist, finger, and elbow. Furthermore, these hydrogels are capable of reliably distinguishing and reproducing various printed text. These findings imply that any electronic device that demands strain-sensing functionality might make use of these developed materials.

Dual-Cross-Linked Chitosan-Based Antibacterial Hydrogels with Tough and Adhesive Properties for Wound Dressing.

Biocompatible chitosan-based hydrogels have attracted extensive attention in wound dressing due to their human skin-like tissue characteristics. However, it is a crucial challenge to fabricate chitosan-based hydrogels with versatile properties, including flexibility, stretchability, adhesivity, and antibacterial activity. In this work, a kind of chitosan-based hydrogels with integrated functionalities are facilely prepared by solution polymerization of acrylamide (AAm) and sodium p-styrene sulfonate (SS) in the presence of quaternized carboxymethyl chitosan (QCMCS). Due to the dual cross-linking between QCMCS and P(AAm-co-SS), the optimized QCMCS/P(AAm-co-SS) hydrogel exhibits tough mechanical properties (0.767 MPa tensile stress and 1100% fracture strain) and moderate tissue adhesion (11.4 kPa). Moreover, biological evaluation in vitro illustrated that as-prepared hydrogel possesses satisfactory biocompatibility, hemocompatibility, and excellent antibacterial ability (against S. aureus and E. coli are 98.8% and 97.3%, respectively). Then, the hydrogels are tested in a rat model for bacterial infection incision in vivo, and the results show that they can significantly accelerate epidermal regeneration and wound closure. This is due to their ability to reduce the inflammatory response, promote the formation of collagen deposition and granulation tissue. The proposed chitosan-based antibacterial hydrogels have the potential to be a highly effective wound dressing in clinical wound healing.

Physicochemical Dual Cross-Linking Conductive Polymeric Networks Combining High Strength and High Toughness Enable Stable Operation of Silicon Microparticle Anodes.

The poor interfacial stability and insufficient cycling performance caused by undesirable stress hinder the commercial application of silicon microparticles (µSi) as next-generation anode materials for high-energy-density lithium-ion batteries. Herein, a conceptionally novel physicochemical dual cross-linking conductive polymeric network is designed combining high strength and high toughness by coupling the stiffness of poly(acrylic acid) and the softness of carboxyl nitrile rubber, which includes multiple H-bonds, by introducing highly branched tannic acid as a physical cross-linker. Such a design enables effective stress dissipation by folded molecular chains slipping and sequential cleavage of H-bonds, thus stabilizing the electrode interface and enhancing cycle stability. As expected, the resultant electrode (µSi/PTBR) delivers an unprecedented high capacity retention of ≈97% from 2027.9 mAh g-1 at the 19th to 1968.0 mAh g-1 at the 200th cycle at 2 A g-1 . Meanwhile, this unique stress dissipation strategy is also suitable for stabilizing SiOx anodes with a much lower capacity loss of ≈0.012% per cycle over 1000 cycles at 1.5 A g-1 . Atomic force microscopy analysis and finite element simulations reveal the excellent stress-distribution ability of the physicochemical dual cross-linking conductive polymeric network. This work provides an efficient energy-dissipation strategy toward practical high-capacity anodes for energy-dense batteries.

Dual-Crosslinked Degradable Elastomeric Networks With Self-Healing Properties: Bringing Multi(catechol) Star-Block Copolymers into Play.

In the biomedical field, degradable chemically crosslinked elastomers are interesting materials for tissue engineering applications, since they present rubber-like mechanical properties matching those of soft tissues and are able to preserve their three-dimensional (3D) structure over degradation. Their use in biomedical applications requires surgical handling and implantation that can be a source of accidental damages responsible for the loss of properties. Therefore, their inability to be healed after damage or breaking can be a major drawback. In this work, biodegradable dual-crosslinked networks that exhibit fast and efficient self-healing properties at 37 °C are designed. Self-healable dual-crosslinked (chemically and physically) elastomeric networks are prepared by two methods. The first approach is based on the mix of hydrophobic poly(ethylene glycol)-poly(lactic acid) (PEG-PLA) star-shaped copolymers functionalized with either catechol or methacrylate moieties. In the second approach, hydrophobic bifunctional PEG-PLA star-shaped copolymers with both catechol and methacrylate on their structure are used. In the two systems, the supramolecular network is responsible for the self-healing properties, thanks to the dynamic dissociation/reassociation of the numerous hydrogen bonds between the catechol groups, whereas the covalent network ensures mechanical properties similar to pure methacrylate networks. The self-healable materials display mechanical properties that are compatible with soft tissues and exhibit linear degradation because of the chemical cross-links. The performances of networks from mixed copolymers versus bifunctional copolymers are compared and demonstrate the superiority of the latter. The biocompatibility of the materials is also demonstrated, confirming the potential of these degradable and self-healable elastomeric networks to be used for the design of temporary medical devices.

Preparation and Properties of Egg White Dual Cross-Linked Hydrogel with Potential Application for Bone Tissue Engineering.

In this study, an egg white dual cross-linked hydrogel was developed based on the principle that the external stimulus can denature proteins and cause them to aggregate, forming hydrogel. The sodium hydroxide was used to induce gelation of the egg white protein, subsequently introducing calcium ions to cross-link with protein chains, thereby producing a dual cross-linked hydrogel. The characteristics of the dual cross-linked hydrogels-including the secondary structure, stability, microstructure, swelling performance, texture properties, and biosafety-were investigated to determine the effects of calcium ion on the egg white hydrogel (EWG) and evaluate the potential application in the field of tissue engineering. Results showed that calcium ions could change the ß-sheet content of the protein in EWG after soaking it in different concentrations of CaCl2 solution, leading to changes in the hydrogen bonds and the secondary structure of polypeptide chains. It was confirmed that calcium ions promoted the secondary cross-linking of the protein chain, which facilitated polypeptide folding and aggregation, resulting in enhanced stability of the egg white dual cross-linked hydrogel. Furthermore, the swelling capacity of the EWG decreased with increasing concentration of calcium ions, and the texture properties including hardness, cohesiveness and springiness of the hydrogels were improved. In addition, the calcium cross-linked EWG hydrogels exhibited biocompatibility and cell-surface adhesion in vitro. Hence, this work develops a versatile strategy to fabricate dual cross-linked protein hydrogel with biosafety and cell-surface adhesion, and both the strategy and calcium-egg white cross-linked hydrogels have potential for use in bone tissue engineering.

Chromatin Immunoprecipitation and Circadian Rhythms.

Organisms exhibit daily changes of physiology and behavior to exert homeostatic adaptations to day-night cycles. The cyclic fluctuation also takes place at transcriptional levels, giving rise to rhythmic gene expression. Central to this oscillatory transcription is the core clock machinery which constitutes a circuit of transcriptional-translational feedback and achieves circadian functions accordingly. Chromatin immunoprecipitation provides understanding of such mechanisms that clock and non-clock transcription factors along with co-regulators and chromatin modifications dictate circadian epigenome through cyclic alterations of chromatin structures and molecular functions in a concerted fashion. Besides, innovation of high-throughput sequencing technology has broadened our horizon and renewed perspectives in circadian research. This article summarizes the methodology of a chromatin immunoprecipitation experiment in light of circadian rhythm research.

Tough, Highly Oriented, Super Thermal Insulating Regenerated All-Cellulose Sponge-Aerogel Fibers Integrating a Graded Aligned Nanostructure.

Thermal insulating fibers can effectively regulate the human body temperature and decrease indoor energy consumption. However, designing super thermal insulating fibers integrating a sponge and aerogel structure based on biomass resources is still a challenge. Herein, a flow-assisted dynamic dual-cross-linking strategy is developed to realize the steady fabrication of regenerated all-cellulose graded sponge-aerogel fibers (CGFs) in a microfluidic chip. The chemically cross-linked cellulose solution is used as the core flow, which is passed through two sheath flow channels, containing either a diffusion solvent or a physical cross-linking solvent, resulting in CGFs with a porous sponge outer layer and a dense aerogel inner layer. By regulating and simulating the flow process in the microfluidic chip, CGFs with adjustable sponge thicknesses, excellent toughness (26.20 MJ m-3), and ultralow thermal conductivity (0.023 W m-1 K-1) are fabricated. This work provides a new method for fabricating graded biomass fibers and inspires attractive applications for thermal insulation in textiles.

Lignocellulosic nanofibril aerogel via gas phase coagulation and diisocyanate modification for solvent absorption.

Cellulose-based aerogels are considered to be carriers that can absorb oils and organic solvents owing to the merits of low density and high surface area. However, the natural hydrophility and poor mechanical strength often obstruct their widespread applications. In this work, Miscanthus-based dual cross-linked lignocellulosic nanofibril (LCNF) aerogels were prepared by gas phase coagulation and methylene diphenyl dissocyanate (MDI) modification. Due to physical and chemical cross-linking strategies, the optimally 4 M-LCNF aerogels had high surface area of 157.9 m2/g, water contact angle of 138.1°, and enhanced compression properties. Moreover, the modified aerogels exhibited absorption performance for various organic solvents, and the maximal absorption capacity of chloroform was 42 g/g aerogel. Because LCNF was directly produced from Miscanthus without using bleaching reagents, this research provided a more sustainable methodology to utilize lignocelluloses to design robust aerogels to deal with the leakage of oil and organic solvents in industrial applications.

Three-dimensional bioprinting of polysaccharide-based self-healing hydrogels with dual cross-linking.

Three-dimensional (3D) bioprinting technique is useful to fabricate constructs with functional and biological structures for various biomedical applications. Oxidized hyaluronate (OHA) and glycol chitosan (GC) can form autonomous self-healing hydrogels when adipic acid dihydrazide (ADH) is used. We demonstrate that hyaluronate-alginate hybrid (HAH) polymers can be used for secondary physical cross-linking of OHA/GC/ADH hydrogel with calcium ions after 3D printing. The molecular weight of hyaluronate can be varied while keeping the molecular weight of alginate in HAH. The mechanical stiffness and stability of gels after 3D printing are strongly dependent on the molecular weight of HAH at the same cross-linking density. In vitro chondrogenic differentiation of ATDC5 cells encapsulated in 3D-printed constructs is dependent on the molecular weight of HAH in gels. This dual cross-linking system consisting of naturally occurring biocompatible polysaccharides may have potential in the 3D bioprinting of custom-made scaffolds for tissue engineering applications.

Water-Based Dual-Cross-Linked Polymer Binders for High-Energy-Density Lithium-Sulfur Batteries.

High-mass-loading electrodes with long-term stability have long been a great challenge for lithium-sulfur batteries (LSBs), since the conventional binders are unable to cope with the shuttling of lithium polysulfides and the structural damage in an electrode. Here, a novel water-based polymer, polyphosphate acid cross-linked chitosan ethylamide carbamide (PACEC), is developed as a binder to construct high-energy-density sulfur cathode and flexible LSBs. With a dual-cross-linked network, the PACEC shows excellent affinity with lithium polysulfides to relieve the shuttle effect and robust mechanical properties to stabilize the electrode. The sulfur cathode based on PACEC demonstrates a high sulfur loading of 14.8 mg cm-2, the areal initial capacity of 17.5 mAh cm-2, and Coulombic efficiency of 99.3%, while the amount of electrolyte is strictly limited to 6 µL mg-1. More importantly, a robust pouch cell with an area of 6 cm2 and only 177% oversized lithium can successfully integrate the energy density of 6.5 mAh cm-2 with the cycling retention per cycle of 99.74% during 270 cycles and flexibility at a curvature of 3 mm. This study provides inspirations for the design of eco-friendly polymer binders and paving new ways for the development of LSBs.

Chitosan composite hydrogels cross-linked by multifunctional diazo resin as antibacterial dressings for improved wound healing.

Skin lesions and injuries can increase the risk of pathogen infections. Developing efficacious wound dressings could effectively prevent bacterial infection and accelerate wound healing. Herein, we developed chitosan composite hydrogels cross-linked by multifunctional diazo resin (DR) as antibacterial dressings for improved wound healing. The composite hydrogels were in situ formed by electrostatic interactions, chelation interactions, and covalent bonds between carboxylated chitosan and DR under ultraviolet assisted without small photosensitizer. The resultant hydrogels (noted as DR-CCH) showed good stability at different DR concentrations in physiological buffers. The antibacterial assays showed the DR-CCH could inhibit and kill Escherichia coli and Staphylococcus aureus. What is more, our hydrogels could accelerate wound healing in vivo. The present study demonstrates this composite DR-CCH with trace zinc has potential for accelerated wound healing.

Dual Cross-Linking Chromatin Immunoprecipitation Protocol for Next-Generation Sequencing (ChIPseq) in Macrophages.

Macrophages arise from distinct progenitor cell populations throughout development and are one of the most diverse cell types, capable of performing discrete functions, undergoing distinct modes of activation, and infiltrating or residing in numerous niches in the body. In adapting to their environments, macrophages display high levels of plasticity which is associated with profound epigenomic and transcriptional changes. Understanding these changes has been greatly facilitated by the next-generation sequencing (NGS)-based approaches such as RNAseq and chromatin immunoprecipitation (ChIP)seq. Despite the recent advances, obtaining quality ChIPseq data in macrophages for endogenous factors and especially coregulators recruited to DNA indirectly has proved to be extremely challenging. Here, we describe a dual crosslinking protocol for ChIPseq in macrophages that we developed for difficult-to-ChIP transcription factors, coregulators, and their posttranslational modifications. Further, we provide guidance on crucial optimization steps throughout this protocol. Although our experience has been predominantly in murine and human macrophages, we believe our protocols can be modified and optimized to study signal-induced epigenomic changes in any cell type of choice.

Covalent cross-links in polyampholytic chitosan fibers enhances bone regeneration in a rabbit model.

Chitosan fibers were prepared in citric acid bath, pH 7.4 and NaOH solution at pH 13, to form ionotropically cross-linked and uncross-linked fibers, respectively. The fibers formed in citric acid bath were further cross-linked via carbodiimide chemistry; wherein the pendant carboxyl moieties of citric acid were used for new amide bond formation. Moreover, upon covalent cross-linking in the ionically gelled citrate-chitosan fibers, incomplete conversion of the ion pairs to amide linkages took place resulting in the formation of a dual network structure. The dual cross-linked fibers displayed improved mechanical property, higher stability against enzymatic degradation, hydrophobicity and superior bio-mineralization compared to the uncross-linked and native citrate cross-linked fibers. Additionally, upon cyclic loading, the ion pairs in the dual cross-linked fibers dissociated by dissipating energy and reformed during the relaxation period. The twin property of elasticity and energy dissipation mechanism makes the dual cross-linked fiber unique under dynamic mechanical conditions. The differences in the physico-chemical characteristics were reflected in protein adsorption, which in turn influenced the cellular activities on the fibers. Compared to the uncross-linked and ionotropically cross-linked fibers, the dual cross-linked fibers demonstrated higher proliferation and osteogenic differentiation of the MSCs in vitro as well as better osseous tissue regeneration in a rabbit model.

Dual cross-linking ribonucleoprotein immunoprecipitation assay.

Ribonucleoprotein immunoprecipitation (RIP) is an antibody-based method to detect RNA-protein interactions in situ. In the assay, UV cross-linking is commonly used to preserve RNA-protein interactions for subsequent target identification. UV light is a zero-length cross linker and thus identifies proteins directly bound to RNAs. Here, we describe a dual cross-linking RIP method that involves sequential protein-protein cross-linking step with a protein-protein cross-linker, followed by protein-RNA fixation by UV irradiation. In this way, proteins that indirectly bound to RNA can be analyzed.