Injectable tissue prosthesis for instantaneous closed-loop rehabilitation.

To construct tissue-like prosthetic materials, soft electroactive hydrogels are the best candidate owing to their physiological mechanical modulus, low electrical resistance and bidirectional stimulating and recording capability of electrophysiological signals from biological tissues1,2. Nevertheless, until now, bioelectronic devices for such prostheses have been patch type, which cannot be applied onto rough, narrow or deep tissue surfaces3-5. Here we present an injectable tissue prosthesis with instantaneous bidirectional electrical conduction in the neuromuscular system. The soft and injectable prosthesis is composed of a biocompatible hydrogel with unique phenylborate-mediated multiple crosslinking, such as irreversible yet freely rearrangeable biphenyl bonds and reversible coordinate bonds with conductive gold nanoparticles formed in situ by cross-coupling. Closed-loop robot-assisted rehabilitation by injecting this prosthetic material is successfully demonstrated in the early stage of severe muscle injury in rats, and accelerated tissue repair is achieved in the later stage.

Injection-on-Skin Granular Adhesive for Interactive Human-Machine Interface.

Realization of interactive human-machine interfaces (iHMI) is improved with development of soft tissue-like strain sensors beyond hard robotic exosuits, potentially allowing cognitive behavior therapy and physical rehabilitation for patients with brain disorders. Here, this study reports on a strain-sensitive granular adhesive inspired by the core-shell architectures of natural basil seeds for iHMI as well as human-metaverse interfacing. The granular adhesive sensor consists of easily fragmented hydropellets as a core and tissue-adhesive catecholamine layers as a shell, satisfying great on-skin injectability, ionic-electrical conductivity, and sensitive resistance changes through reversible yet robust cohesion among the hydropellets. Particularly, it is found that the ionic-electrical self-doping of the catecholamine shell on hydrosurfaces leads to a compact ion density of the materials. Based on these physical and electrical properties of the sensor, it is demonstrated that successful iHMI integration with a robot arm in both real and virtual environments enables robotic control by finger gesture and haptic feedback. This study expresses benefits of using granular hydrogel-based strain sensors for implementing on-skin writable bioelectronics and their bridging into the metaverse world.

Functional significance of serine 13 in the active site of glutathione S-transferase F3 from Oryza sativa.

Plant glutathione S-transferase (GST, EC 2.5.1.18) is an enzyme that detoxifies various electrophilic compounds including herbicides and organic pollutants by catalyzing the formation of conjugates with reduced glutathione (GSH). Although the structure and function of the GST subunits in rice, an important food in Asia, are not well understood, they are crucial for herbicide development. To investigate the role of active site residues in rice Phi-class GSTF3 (OsGSTF3), evolutionarily conserved serine residues were replaced with alanine using site-directed mutagenesis to obtain the mutants S13A, S38A, S69A, and S169A. These four mutants were expressed in Escherichia coli and purified to electrophoretic homogeneity using immobilized GSH affinity chromatography. Mutation of Ser13 to Ala resulted in substantial reductions in specific activities and kcat/Km values for the GSH-[1-chloro-2,4-dinitrobenzene (CDNB)] conjugation reaction. In contrast, mutations of Ser38, Ser69, and Ser169 to Ala had little effect on the activities and kinetic parameters. Additionally, the mutation of Ser13 to Ala significantly affected the KmGSH and I50 values of S-hexylglutathione and S-(2,4-dinitrophenyl)glutathione, which compete with GSH and the product of GSH-CDNB conjugation, respectively. A pH-log (kcat/KmCDNB) plot was used to estimate the pKa value of GSH in the enzyme-GSH complex of the wild-type enzyme, which was approximately 6.9. However, the pKa value of GSH in the enzyme-GSH complex of the S13A mutant was approximately 8.7, which was about 1.8 pK units higher than that of the wild-type enzyme. OsGSTF3 was also crystallized for crystallographic study, and the structure analyses revealed that Ser13 is located in the active site and that its side chain is in close proximity to the thiol group of glutathione bound in the enzyme. Based on these substitution effects on kinetic parameters, the dependence of kinetic parameters on the pH and 3-dimensional structure, it was suggested that Ser13 in rice OsGSTF3 is the residue responsible for catalytic activity by lowering the pKa of GSH in the enzyme-GSH complex and enhancing the nucleophilicity of the GSH thiol in the active site.

Balanced Coexistence of Reversible and Irreversible Covalent Bonds in a Conductive Triple Polymeric Network Enables Stretchable Hydrogels with High Toughness and Adhesiveness.

The application of soft hydrogels to stretchable devices has attracted increasing attention in deformable bioelectronics owing to their unique characteristic, "modulus matching between materials and organs". Despite considerable progress, their low toughness, low conductivity, and absence of tissue adhesiveness remain substantial challenges associated with unstable skin-interfacing, where body movements undesirably disturb electrical signal acquisitions. Herein, we report a material design of a highly tough strain-dissipative and skin-adhesive conducting hydrogel fabricated through a facile one-step sol-gel transition and its application to an interactive human-machine interface. The hydrogel comprises a triple polymeric network where irreversible amide linkage of polyacrylamide with alginate and dynamic covalent bonds entailing conjugated polymer chains of poly(3,4-ethylenedioxythiophene)-co-(3-thienylboronic acid) are simultaneously capable of high stretchability (1300% strain), efficient strain dissipation (36,209 J/m2), low electrical resistance (590 Ω), and even robust skin adhesiveness (35.0 ± 5.6 kPa). Based on such decent characteristics, the hydrogel was utilized as a multifunctional layer for successfully performing either electrophysiological cardiac/muscular on-skin sensors or an interactive stretchable human-machine interface.

Tyramine-Functionalized Alginate-Collagen Hybrid Hydrogel Inks for 3D-Bioprinting.

Extrusion-based 3D-bioprinting using hydrogels has exhibited potential in precision medicine; however, researchers are beset with several challenges. A major challenge of this technique is the production of constructs with sufficient height and fidelity to support cellular behavior in vivo. In this study, we present the 3D-bioprinting of cylindrical constructs with tunable gelation kinetics by controlling the covalent crosslinking density and gelation time of a tyramine-functionalized alginate hydrogel (ALG-TYR) via enzymatic reaction by horseradish peroxidase (HRP) and hydrogen peroxide (H2O2). The extruded filament was crosslinked for a second time on a support bath containing H2O2 to increase fidelity after printing. The resulting tubular construct, with a height of 6 mm and a wall thickness of 2 mm, retained its mechanical properties and had a maximum 2-fold swelling after 2 d. Furthermore, collagen (COL) was introduced into the ALG-TYR hydrogel network to increase the mechanical modulus and cell cytocompatibility, as the encapsulated fibroblast cells exhibited a higher cell viability in the ALG-TYR/COL construct (92.13 ± 0.70%) than in ALG-TYR alone (68.18 ± 3.73%). In summary, a vascular ECM-mimicking scaffold was 3D-bioprinted with the ALG-TYR/COL hybrid hydrogel, and this scaffold can support tissue growth for clinical translation in regenerative and personalized medicine.

Punicalagin-Loaded Alginate/Chitosan-Gallol Hydrogels for Efficient Wound Repair and Hemostasis.

For recently devised wound-healing materials, a variety of acute application systems with sustainable therapeutic effects on wound sites have been suggested. For example, hydrogel-type healing agents with porous structures and high drug encapsulation efficiencies have been developed for wound repair. However, challenges remain about the poor mechanical and adhesive properties of hydrogels. Herein, we propose a punicalagin (PC)-containing wound-healing hydrogel in adhesive form that is mechanically stable and has sustainable wound-healing therapeutic efficiency. The APC hydrogel, composed of alginate (ALG), PC, and chitosan-gallol (CHI-G), exhibits significant mechanical and self-healing properties, thus indicating that PC increases cross-linking in ALG/CHI-G as macromolecule. The PC-containing mechanically enhanced hydrogel demonstrates high tissue adhesiveness. Sustainable PC release for 192 h, which indicates high therapeutic effect of the released PC, and great blood compatibility are evaluated based on rapid blood coagulation and minimal hemolysis. The cytocompatibility and wound-healing abilities of the PC-containing APC hydrogel are greater than those of the non-PC hydrogel, as verified by cell compatibility and wound scratch assays. These results indicate that a suitable concentration of PC-containing hydrogel with sustainable moisture condition and PC release may inspire further polyphenol-agent-containing hydrogels as wound-healing agents with structural stability and therapeutic efficiency.

Tissue Adhesive, Conductive, and Injectable Cellulose Hydrogel Ink for On-Skin Direct Writing of Electronics.

Flexible and soft bioelectronics used on skin tissue have attracted attention for the monitoring of human health. In addition to typical metal-based rigid electronics, soft polymeric materials, particularly conductive hydrogels, have been actively developed to fabricate biocompatible electrical circuits with a mechanical modulus similar to biological tissues. Although such conductive hydrogels can be wearable or implantable in vivo without any tissue damage, there are still challenges to directly writing complex circuits on the skin due to its low tissue adhesion and heterogeneous mechanical properties. Herein, we report cellulose-based conductive hydrogel inks exhibiting strong tissue adhesion and injectability for further on-skin direct printing. The hydrogels consisting of carboxymethyl cellulose, tannic acid, and metal ions (e.g., HAuCl4) were crosslinked via multiple hydrogen bonds between the cellulose backbone and tannic acid and metal-phenol coordinate network. Owing to this reversible non-covalent crosslinking, the hydrogels showed self-healing properties and reversible conductivity under cyclic strain from 0 to 400%, as well as printability on the skin tissue. In particular, the on-skin electronic circuit printed using the hydrogel ink maintained a continuous electrical flow under skin deformation, such as bending and twisting, and at high relative humidity of 90%. These printable and conductive hydrogels are promising for implementing structurally complicated bioelectronics and wearable textiles.

Addressing the Shortcomings of Polyphenol-Derived Adhesives: Achievement of Long Shelf Life for Effective Hemostasis.

For rapid and effective hemostasis of uncontrollable bleeding, versatile hemostatic agents have been emerging. Among them, polyphenol-derived adhesives have attracted those hemostatic materials due to instantaneous formation of sticky barriers by robust interactions between the material and the serum proteins from wound. However, a critical challenge in such phenolic materials lies in long-term storage due to spontaneous oxidation under humid environments, leading to changes in hemostatic capability and adhesive strength. Here, we report a transparent hemostatic film consisting of gallol-conjugated chitosan (CHI-G) for minimizing the phenolic oxidation even for 3 months and maintaining strong tissue adhesiveness and its hemostatic ability. The film undergoes a phase transition from solid to injectable hydrogels at physiological pH for efficiently stopping internal and external hemorrhage. Interestingly, the hemostatic capability of the CHI-G hydrogels after 3 month storage depends on (i) the folded microstructure of the polymer with optimal gallol modification and (ii) an initial phase of either a solution state or a solid film. When the hydrogels are originated from the dehydrated film, their successful hemostasis is observed in a liver bleeding model. Our finding would provide an insight for design rationale of hemostatic formulations with long shelf-life.

Lead-Sealed Stretchable Underwater Perovskite-Based Optoelectronics via Self-Recovering Polymeric Nanomaterials.

To harness the full potential of halide perovskite based optoelectronics, biological safety, compatibility with flexible/stretchable platforms, and operational stability must be guaranteed. Despite substantial efforts, none has come close to providing a solution that encompasses all of these requirements. To address these issues, we devise a multifunctional encapsulation scheme utilizing hydrogen bond-based self-recovering polymeric nanomaterials as an alternative for conventional glass-based encapsulation. We show that Pb in physically damaged halide perovskite solar cells can be completely contained within the self-recovering encapsulation upon submersion in a simulated rain bath, as indicated by in vitro cytotoxicity tests. In addition, self-recovering encapsulation accommodates stable device operation upon casual bending and even stretching, which is in stark contrast to conventional glass-based encapsulation schemes. We also demonstrate the concept of assembling user-defined scalable modular optoelectronics based on halide perovskite solar cells and light emitting diodes through the use of self-recovering conductive nanocomposites. Finally, long-term operational stability of over 1000 h was achieved under harsh accelerated conditions (50 °C/50% RH and 85 °C/0% RH) with the incorporation of an ultrathin atomic layer deposited TiO2 barrier underneath the multifunctional encapsulation. In light of these merits, the encapsulation scheme based on self-recovering polymeric nanomaterials is proposed as a simple, but practical solution to a multifaceted challenge in the field of halide perovskites.

Identification of Downstream Genes of the mTOR Pathway that Predict Recurrence and Progression in Non-Muscle Invasive High-Grade Urothelial Carcinoma of the Bladder.

Microarray analysis was used to investigate the lack of identified mammalian target of rapamycin (mTOR) pathway downstream genes to overcome cross-talk at non-muscle invasive high-grade (HG)-urothelial carcinoma (UC) of the bladder, gene expression patterns, gene ontology, and gene clustering by triple (p70S6K, S6K, and eIF4E) small interfering RNAs (siRNAs) or rapamycin in 5637 and T24 cell lines. We selected mTOR pathway downstream genes that were suppressed by siRNAs more than 2-fold, or were up-regulated or down-regulated by rapamycin more than 2-fold. We validated mTOR downstream genes with immunohistochemistry using a tissue microarray (TMA) of 125 non-muscle invasive HG-UC patients and knockout study to evaluate the synergistic effect with rapamycin. The microarray analysis selected mTOR pathway downstream genes consisting of 4 rapamycin up-regulated genes (FABP4, H19, ANXA10, and UPK3A) and 4 rapamycin down-regulated genes (FOXD3, ATP7A, plexin D1, and ADAMTS5). In the TMA, FABP4, and ATP7A were more expressed at T1 and FOXD3 was at Ta. ANXA10 and ADAMTS5 were more expressed in tumors ≤ 3 cm in diameter. In a multivariate Cox regression model, ANXA10 was a significant predictor of recurrence and ATP7A was a significant predictor of progression in non-muscle invasive HG-UC of the bladder. In an ATP7A knock-out model, rapamycin treatment synergistically inhibited cell viability, wound healing, and invasion ability compared to rapamycin only. Activity of the ANXA10 and ATP7A mTOR pathway downstream genes might predict recurrence and progression in non-muscle invasive HG-UC of the bladder. ATP7A knockout overcomes rapamycin cross-talk.