Skeletal muscle regeneration with 3D bioprinted hyaluronate/gelatin hydrogels incorporating MXene nanoparticles.

There has been significant progress in the field of three-dimensional (3D) bioprinting technology, leading to active research on creating bioinks capable of producing structurally and functionally tissue-mimetic constructs. Ti3C2Tx MXene nanoparticles (NPs), promising two-dimensional nanomaterials, are being investigated for their potential in muscle regeneration due to their unique physicochemical properties. In this study, we integrated MXene NPs into composite hydrogels made of gelatin methacryloyl (GelMA) and hyaluronic acid methacryloyl (HAMA) to develop bioinks (namely, GHM bioink) that promote myogenesis. The prepared GHM bioinks were found to offer excellent printability with structural integrity, cytocompatibility, and microporosity. Additionally, MXene NPs within the 3D bioprinted constructs encouraged the differentiation of C2C12 cells into skeletal muscle cells without additional support of myogenic agents. Genetic analysis indicated that representative myogenic markers both for early and late myogenesis were significantly up-regulated. Moreover, animal studies demonstrated that GHM bioinks contributed to enhanced regeneration of skeletal muscle while reducing immune responses in mice models with volumetric muscle loss (VML). Our results suggest that the GHM hydrogel can be exploited to craft a range of strategies for the development of a novel bioink to facilitate skeletal muscle regeneration because these MXene-incorporated composite materials have the potential to promote myogenesis.

Highly Aligned Ternary Nanofiber Matrices Loaded with MXene Expedite Regeneration of Volumetric Muscle Loss.

Current therapeutic approaches for volumetric muscle loss (VML) face challenges due to limited graft availability and insufficient bioactivities. To overcome these limitations, tissue-engineered scaffolds have emerged as a promising alternative. In this study, we developed aligned ternary nanofibrous matrices comprised of poly(lactide-co-ε-caprolactone) integrated with collagen and Ti3C2Tx MXene nanoparticles (NPs) (PCM matrices), and explored their myogenic potential for skeletal muscle tissue regeneration. The PCM matrices demonstrated favorable physicochemical properties, including structural uniformity, alignment, microporosity, and hydrophilicity. In vitro assays revealed that the PCM matrices promoted cellular behaviors and myogenic differentiation of C2C12 myoblasts. Moreover, in vivo experiments demonstrated enhanced muscle remodeling and recovery in mice treated with PCM matrices following VML injury. Mechanistic insights from next-generation sequencing revealed that MXene NPs facilitated protein and ion availability within PCM matrices, leading to elevated intracellular Ca2+ levels in myoblasts through the activation of inducible nitric oxide synthase (iNOS) and serum/glucocorticoid regulated kinase 1 (SGK1), ultimately promoting myogenic differentiation via the mTOR-AKT pathway. Additionally, upregulated iNOS and increased NO- contributed to myoblast proliferation and fiber fusion, thereby facilitating overall myoblast maturation. These findings underscore the potential of MXene NPs loaded within highly aligned matrices as therapeutic agents to promote skeletal muscle tissue recovery.

Laser-Assisted Structuring of Graphene Films with Biocompatible Liquid Crystal Polymer for Skin/Brain-Interfaced Electrodes.

The work presented here introduces a facile strategy for the development of flexible and stretchable electrodes that harness the robust characteristics of carbon nanomaterials through laser processing techniques on a liquid crystal polymer (LCP) film. By utilizing LCP film as a biocompatible electronic substrate, control is demonstrated over the laser irradiation parameters to achieve efficient pattern generation and transfer printing processes, thereby yielding highly conductive laser-induced graphene (LIG) bioelectrodes. To enhance the resolution of the patterned LIG film, shadow masks are employed during laser scanning on the LCP film surface. This approach is compatible with surface-mounted device integration, enabling the circuit writing of LIG/LCP materials in a flexible format. Moreover, kirigami-inspired on-skin bioelectrodes are introduced that exhibit reasonable stretchability, enabling independent connections to healthcare hardware platforms for electrocardiogram (ECG) and electromyography (EMG) measurements. Additionally, a brain-interfaced LIG microelectrode array is proposed that combines mechanically compliant architectures with LCP encapsulation for stimulation and recording purposes, leveraging their advantageous structural features and superior electrochemical properties. This developed approach offers a cost-effective and scalable route for producing patterned arrays of laser-converted graphene as bioelectrodes. These bioelectrodes serve as ideal circuit-enabled flexible substrates with long-term reliability in the ionic environment of the human body.

Regulated Behavior in Living Cells with Highly Aligned Configurations on Nanowrinkled Graphene Oxide Substrates: Deep Learning Based on Interplay of Cellular Contact Guidance.

Micro-/nanotopographical cues have emerged as a practical and promising strategy for controlling cell fate and reprogramming, which play a key role as biophysical regulators in diverse cellular processes and behaviors. Extracellular biophysical factors can trigger intracellular physiological signaling via mechanotransduction and promote cellular responses such as cell adhesion, migration, proliferation, gene/protein expression, and differentiation. Here, we engineered a highly ordered nanowrinkled graphene oxide (GO) surface via the mechanical deformation of an ultrathin GO film on an elastomeric substrate to observe specific cellular responses based on surface-mediated topographical cues. The ultrathin GO film on the uniaxially prestrained elastomeric substrate through self-assembly and subsequent compressive force produced GO nanowrinkles with periodic amplitude. To examine the acute cellular behaviors on the GO-based cell interface with nanostructured arrays of wrinkles, we cultured L929 fibroblasts and HT22 hippocampal neuronal cells. As a result, our developed cell-culture substrate obviously provided a directional guidance effect. In addition, based on the observed results, we adapted a deep learning (DL)-based data processing technique to precisely interpret the cell behaviors on the nanowrinkled GO surfaces. According to the learning/transfer learning protocol of the DL network, we detected cell boundaries, elongation, and orientation and quantitatively evaluated cell velocity, traveling distance, displacement, and orientation. The presented experimental results have intriguing implications such that the nanotopographical microenvironment could engineer the living cells' morphological polarization to assemble them into useful tissue chips consisting of multiple cell types.

Mobile Point-of-Care Device Using Molecularly Imprinted Polymer-Based Chemosensors Targeting Interleukin-1ß Biomarker.

Molecularly imprinted polymers (MIPs) have garnered significant attention as a promising material for engineering specific biological receptors with superior chemical complementarity to target molecules. In this study, we present an electrochemical biosensing platform incorporating MIP films for the selective detection of the interleukin-1ß (IL-1ß) biomarker, particularly suitable for mobile point-of-care testing (POCT) applications. The IL-1ß-imprinted biosensors were composed of poly(eriochrome black T (EBT)), including an interlayer of poly(3,4-ethylene dioxythiophene) and a 4-aminothiophenol monolayer, which were electrochemically polymerized simultaneously with template proteins (i.e., IL-1ß) on custom flexible screen-printed carbon electrodes (SPCEs). The architecture of the MIP films was designed to enhance the sensor sensitivity and signal stability. This approach involved a straightforward sequential-electropolymerization process and extraction for leaving behind cavities (i.e., rebinding sites), resulting in the efficient production of MIP-based biosensors capable of molecular recognition for selective IL-1ß detection. The electrochemical behaviors were comprehensively investigated using cyclic voltammograms and electrochemical impedance spectroscopy responses to assess the imprinting effect on the MIP films formed on the SPCEs. In line with the current trend in in vitro diagnostic medical devices, our simple and effective MIP-based analytical system integrated with mobile POCT devices offers a promising route to the rapid detection of biomarkers, with particular potential for periodontitis screening.

Spontaneous Osteogenic Differentiation of Human Mesenchymal Stem Cells by Tuna-Bone-Derived Hydroxyapatite Composites with Green Tea Polyphenol-Reduced Graphene Oxide.

In recent years, bone tissue engineering (BTE) has made significant progress in promoting the direct and functional connection between bone and graft, including osseointegration and osteoconduction, to facilitate the healing of damaged bone tissues. Herein, we introduce a new, environmentally friendly, and cost-effective method for synthesizing reduced graphene oxide (rGO) and hydroxyapatite (HAp). The method uses epigallocatechin-3-O-gallate (EGCG) as a reducing agent to synthesize rGO (E-rGO), and HAp powder is obtained from Atlantic bluefin tuna (Thunnus thynnus). The physicochemical analysis indicated that the E-rGO/HAp composites had exceptional properties for use as BTE scaffolds, as well as high purity. Moreover, we discovered that E-rGO/HAp composites facilitated not only the proliferation, but also early and late osteogenic differentiation of human mesenchymal stem cells (hMSCs). Our work suggests that E-rGO/HAp composites may play a significant role in promoting the spontaneous osteogenic differentiation of hMSCs, and we envision that E-rGO/HAp composites could serve as promising candidates for BTE scaffolds, stem-cell differentiation stimulators, and implantable device components because of their biocompatible and bioactive properties. Overall, we suggest a new approach for developing cost-effective and environmentally friendly E-rGO/HAp composite materials for BTE application.

Strain-tunable optical microlens arrays with deformable wrinkles for spatially coordinated image projection on a security substrate.

As a new concept in materials design, a variety of strategies have been developed to fabricate optical microlens arrays (MLAs) that enable the miniaturization of optical systems on the micro/nanoscale to improve their characteristic performance with unique optical functionality. In this paper, we introduce a cost-effective and facile fabrication process on a large scale up to ~15 inches via sequential lithographic methods to produce thin and deformable hexagonally arranged MLAs consisting of polydimethylsiloxane (PDMS). Simple employment of oxygen plasma treatment on the prestrained MLAs effectively harnessed the spontaneous formation of highly uniform nanowrinkled structures all over the surface of the elastomeric microlenses. With strain-controlled tunability, unexpected optical diffraction patterns were characterized by the interference combination effect of the microlens and deformable nanowrinkles. Consequently, the hierarchically structured MLAs presented here have the potential to produce desirable spatial arrangements, which may provide easily accessible opportunities to realize microlens-based technology by tunable focal lengths for more advanced micro-optical devices and imaging projection elements on unconventional security substrates.

Surface-mediated high antioxidant and anti-inflammatory effects of astaxanthin-loaded ultrathin graphene oxide film that inhibits the overproduction of intracellular reactive oxygen species.

BACKGROUND: Astaxanthin (AST) is known as a powerful antioxidant that affects the removal of active oxygen and inhibits the production of lipid peroxide caused by ultraviolet light. However, it is easily decomposed by heat or light during production and storage because of the unsaturated compound nature with a structural double bond. The activity of AST can be reduced and lose its antioxidant capability. Graphene oxide (GO) is an ultrathin nanomaterial produced by oxidizing layered graphite. The chemical combination of AST with GO can improve the dispersion properties to maintain structural stability and antioxidant activity because of the tightly bonded functionalized GO surface. METHODS: Layered GO films were used as nanocarriers for the AST molecule, which was produced via flow-enabled self-assembly and subsequent controlled solution deposition of RGD peptide and AST molecules. Synthesis of the GO-AST complex was also carried out for the optimized concentration. The characterization of prepared materials was analyzed through transmission electron microscopy (TEM), scanning electron microscope (SEM), Fourier-transform infrared spectroscopy (FT-IR), atomic force microscope (AFM), and Raman spectroscopy. Antioxidant activity was tested by 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2.2-diphenyl-1-picrylhydrazyl (DPPH) assays. The antibacterial effect and antioxidant effects were monitored for the ultrathin GO/RGD/AST Film. Further, reactive oxygen species (ROS) assay was used to evaluate the anti-inflammatory effects on L-929 fibroblasts. RESULTS: Cotreatment of GO-AST solution demonstrated a high antioxidant combined effect with a high ABTS and DPPH radicals scavenging activity. The GO/RGD/AST film was produced by the self-assembly process exhibited excellent antibacterial effects based on physicochemical damage against E. coli and S. aureus. In addition, the GO/RGD/AST film inhibited H2O2-induced intracellular ROS, suppressed the toxicity of lipopolysaccharide (LPS)-induced cells, and restored it, thereby exhibiting strong antioxidant and anti-inflammatory effects. CONCLUSION: As GO nanocarrier-assisted AST exerted promising antioxidant and antibacterial reactions, presented a new concept to expand basic research into the field of tissue engineering.

Recent Advances of Point-of-Care Devices Integrated with Molecularly Imprinted Polymers-Based Biosensors: From Biomolecule Sensing Design to Intraoral Fluid Testing.

Recent developments of point-of-care testing (POCT) and in vitro diagnostic medical devices have provided analytical capabilities and reliable diagnostic results for rapid access at or near the patient's location. Nevertheless, the challenges of reliable diagnosis still remain an important factor in actual clinical trials before on-site medical treatment and making clinical decisions. New classes of POCT devices depict precise diagnostic technologies that can detect biomarkers in biofluids such as sweat, tears, saliva or urine. The introduction of a novel molecularly imprinted polymer (MIP) system as an artificial bioreceptor for the POCT devices could be one of the emerging candidates to improve the analytical performance along with physicochemical stability when used in harsh environments. Here, we review the potential availability of MIP-based biorecognition systems as custom artificial receptors with high selectivity and chemical affinity for specific molecules. Further developments to the progress of advanced MIP technology for biomolecule recognition are introduced. Finally, to improve the POCT-based diagnostic system, we summarized the perspectives for high expandability to MIP-based periodontal diagnosis and the future directions of MIP-based biosensors as a wearable format.

Phenotypic change of mesenchymal stem cells into smooth muscle cells regulated by dynamic cell-surface interactions on patterned arrays of ultrathin graphene oxide substrates.

The topographical interface of the extracellular environment has been appreciated as a principal biophysical regulator for modulating cell functions, such as adhesion, migration, proliferation, and differentiation. Despite the existed approaches that use two-dimensional nanomaterials to provide beneficial effects, opportunities evaluating their impact on stem cells remain open to elicit unprecedented cellular responses. Herein, we report an ultrathin cell-culture platform with potential-responsive nanoscale biointerfaces for monitoring mesenchymal stem cells (MSCs). We designed an intriguing nanostructured array through self-assembly of graphene oxide sheets and subsequent lithographical patterning method to produce chemophysically defined regions. MSCs cultured on anisotropic micro/nanoscale patterned substrate were spontaneously organized in a highly ordered configuration mainly due to the cell-repellent interactions. Moreover, the spatially aligned MSCs were spontaneously differentiated into smooth muscle cells upon the specific crosstalk between cells. This work provides a robust strategy for directing stem cells and differentiation, which can be utilized as a potential cell culture platform to understand cell-substrate or cell-cell interactions, further developing tissue repair and stem cell-based therapies.

Rotating Cylinder-Assisted Nanoimprint Lithography for Enhanced Chemisorbable Filtration Complemented by Molecularly Imprinted Polymers.

Rotating cylindrical stamp-based nanoimprint technique has many advantages, including the continuous fabrication of intriguing micro/nanostructures and rapid pattern transfer on a large scale. Despite these advantages, the previous nanoimprint lithography has rarely been used for producing sophisticated nanoscale patterns on a non-planar substrate that has many extended applications. Here, the simple integration of nanoimprinting process with a help of a transparent stamp wrapped on the cylindrical roll and UV optical source in the core to enable high-throughput pattern transfer, particularly on a fabric substrate is demonstrated. Moreover, as a functional resin material, this innovative strategy involves a synergistic approach on the synthesis of molecularly imprinted polymer, which are spatially organized free-standing perforated nanostructures such as nano/microscale lines, posts, and holes patterns on various woven or nonwoven blank substrates. The proposed materials can serve as a self-encoded filtration medium for selective separation of formaldehyde molecules. It is envisioned that the combinatorial fabrication process and attractive material paves the way for designing next-generation separation systems in use to capture industrial or household toxic substances.

Graphene Templated DNA Arrays and Biotin-Streptavidin Sensitive Bio-Transistors Patterned by Dynamic Self-Assembly of Polymeric Films Confined within a Roll-on-Plate Geometry.

Patterning of surfaces with a simple strategy provides insights into the functional interfaces by suitable modification of the surface by novel techniques. Especially, highly ordered structural topographies and chemical features from the wide range of interfaces have been considered as important characteristics to understand the complex relationship between the surface chemistries and biological systems. Here, we report a simple fabrication method to create patterned surfaces over large areas using evaporative self-assembly that is designed to produce a sacrificial template and lithographic etch masks of polymeric stripe patterns, ranging from micrometer to nanoscale. By facilitating a roll-on-plate geometry, the periodically patterned surface structures formed by repetitive slip-stick motions were thoroughly examined to be used for the deposition of the Au nanoparticles decorated graphene oxide (i.e., AuNPs, ~21 nm) and the formation of conductive graphene channels. The fluorescently labeled thiol-modified DNA was applied on the patterned arrays of graphene oxide (GO)/AuNPs, and biotin-streptavidin sensitive devices built with graphene-based transistors (GFETs, effective mobility of ~320 cm2 V-1 s-1) were demonstrated as examples of the platform for the next-generation biosensors with the high sensing response up to ~1 nM of target analyte (i.e., streptavidin). Our strategy suggests that the stripe patterned arrays of polymer films as sacrificial templates can be a simple route to creating highly sensitive biointerfaces and highlighting the development of new chemically patterned surfaces composed of graphene-based nanomaterials.

One-Step Laser Patterned Highly Uniform Reduced Graphene Oxide Thin Films for Circuit-Enabled Tattoo and Flexible Humidity Sensor Application.

The conversion of graphene oxide (GO) into reduced graphene oxide (rGO) is imperative for the electronic device applications of graphene-based materials. Efficient and cost-effective fabrication of highly uniform GO films and the successive reduction into rGO on a large area is still a cumbersome task through conventional protocols. Improved film casting of GO sheets on a polymeric substrate with quick and green reduction processes has a potential that may establish a path to the practical flexible electronics. Herein, we report a facile deposition process of GO on flexible polymer substrates to create highly uniform thin films over a large area by a flow-enabled self-assembly approach. The self-assembly of GO sheets was successfully performed by dragging the trapped solution of GO in confined geometry, which consisted of an upper stationary blade and a lower moving substrate on a motorized translational stage. The prepared GO thin films could be selectively reduced and facilitated from the simple laser direct writing process for programmable circuit printing with the desired configuration and less sample damage due to the non-contact mode operation without the use of photolithography, toxic chemistry, or high-temperature reduction methods. Furthermore, two different modes of the laser operating system for the reduction of GO films turned out to be valuable for the construction of novel graphene-based high-throughput electrical circuit boards compatible with integrating electronic module chips and flexible humidity sensors.