[Heterologous expression of bacterial tyrosinase and its applications in biological hair dyeing and DOPA modification of hydrolyzed silk fibroin].

Tyrosinase is a copper-containing polyphenol oxidase widely applied in the food, cosmetics, pharmaceutical, and other industries. Currently, the production of commercial tyrosinase primarily relies on extraction from fungi, which has high costs, low purity, low specific activity, and poor stability. The objective of this study is to obtain highly expressed bacterial tyrosinase with potential for industrial applications. The bacterial tyrosinases from five different sources were heterologously expressed in Escherichia coli BL21(DE3), and the tyrosinases TyrBm and TyrVs derived from Bacillus megaterium and Verrucomicrobium spinosum were obtained with the enzyme activities of (16.1±0.2) U/mL and (48.6±0.9) U/mL, respectively. After protein purification, we compared the enzymatic properties of TyrBm and TyrVs, which revealed that TyrVs exhibited better thermal stability and higher substrate specificity than TyrBm. On the basis of characterizing TyrVs with high catalytic performance, we established a biological hair dyeing system based on TyrVs catalysis to achieve in-situ catalytic hair dyeing. The color washing fastness test measured the ∆E value less than 7.38±0.64 after simulated 14-day cleaning. To facilitate the rapid separation of catalytic products and enzymes, we successfully constructed an immobilized enzyme TyrVs-CipA dependent on self-assembly label CipA and applied this enzyme in the DOPA modification of hydrolyzed silk fibroin (HSF). The immobilized enzyme continuously catalyzed HSF for more than seven cycles, resulting in a single DOPA modification degree exceeding 70.00%. Further investigations demonstrated that DOPA modification enhances the scavenging activity of HSF towards DPPH and O2- radicals by 507.80% and 78.23%, respectively. This study provides a technical foundation for the development of environmentally friendly biological hair dye based on tyrosinase and biomaterials for tissue engineering.

Study on Printability Evaluation of Alginate/Silk Fibroin/Collagen Double-Cross-Linked Inks and the Properties of 3D Printed Constructs.

In recent years, biological 3D printing has garnered increasing attention for tissue and organ repair. The challenge with 3D-printing inks is to combine mechanical properties as well as biocompatibility. Proteins serve as vital structural components in living systems, and utilizing protein-based inks can ensure that the materials maintain the necessary biological activity. In this study, we incorporated two natural biomaterials, silk fibroin (SF) and collagen (COL), into a low-concentration sodium alginate (SA) solution to create novel composite inks. SF and COL were modified with glycidyl methacrylate (GMA) to impart photo-cross-linking properties. The UV light test and 1H NMR results demonstrated successful curing of silk fibroin (SF) and collagen (COL) after modification and grafting. Subsequently, the printability of modified silk fibroin (RSFMA)/SA with varying concentration gradients was assessed using a set of three consecutive printing models, and the material's properties were tested. The research results prove that the addition of RSFMA and ColMA enhances the printability of low-concentration SA solutions, with the Pr values increasing from 0.85 ± 0.02 to 0.90 ± 0.03 and 0.92 ± 0.02, respectively, and the mechanical strength increasing from 0.19 ± 0.01 to 0.28 ± 0.01 and 0.38 ± 0.01 MPa; cytocompatibility has also been improved. Furthermore, rheological tests indicated that all of the inks exhibited shear thinning properties. CCK-8 experiments demonstrated that the addition of ColMA increased the cytocompatibility of the ink system. Overall, the utilization of SF and COL-modified SA materials as inks represents a promising advancement in 3D-printed ink technology.

Efficient Inhibition of Ice Recrystallization During Frozen Storage: Based on the Diffusional Suppression Effect of Silk Fibroin.

Effectively controlling ice recrystallization (IR) during the frozen storage of food remains highly challenging. Inspired by the structural characteristics of antifreeze proteins in nature, silk fibroin (SF) derived from silk fibers has been developed. Through dual validation using the "splat" assay and "sucrose sandwich" assay, the IR inhibition activity of SF at various concentrations was confirmed, revealing that its regular alternating hydrophilic/hydrophobic domains endow SF with the potential to inhibit the axial growth of single ice crystal and significantly reduce the average maximum crystal size by approximately 67%. Additionally, the quality stability of frozen muscle foods treated with SF was comprehensively evaluated. In stark contrast to traditional commercial antifreeze agents (4% sucrose and 4% sorbitol), prepared steaks with the addition of 2% SF maintained rich juiciness and excellent color acceptability over a three-month frozen storage period. Thus, SF holds promise as a potential protective agent for frozen muscle foods, enhancing their quality during storage.

Physical-chemical double crosslinked silk fibroin hydrogel for high-performance bone screws.

Hydrogels are widely used due to their exceptional biocompatibility and adaptability, but their weak mechanical properties limit their application in biomedical engineering. Herein, we rapidly attained a comprehensive enhancement of silk fibroin hydrogels in mechanical properties by employing a physical-chemical double crosslinking strategy. The SF was ultrasonicated and simultaneously photo-crosslinked to form a di-tyrosine network interspersed with ß-sheet blocks, resulting in a SF hydrogel network structure with both rigid and flexible domains. The SF hydrogels exhibited a maximum breaking strength of 59.7 kPa and a Young's modulus of 82.2 MPa, demonstrating significant rigidity and flexibility. Subsequently, the silk screws prepared by this double crosslinking strategy showed extraordinary compressive strength and Young's modulus of 41.8 MPa and 10.9 MPa, respectively. The silk screws cocultured with osteoblasts showed optimal biocompatibility, and the rate of biodegradation could be matched to the rate of osteogenesis. The screw also exhibited high adaptability in the requirements of bone screws. In this study, the SF hydrogels prepared by physical-chemical double crosslinking have extraordinary mechanical properties and biocompatibility, which provides a new avenue for the preparation of high-performance hydrogels and has great potential in bone tissue engineering.

A Nontoxic and Biocompatible Method for Augmenting Mechanical Strength of Acellular Matrix by Silk Fibroin Impregnation.

Biological scaffolds are plagued by poor biomechanical properties and untimely degradation. These limitations have yet to be addressed without compromising their biocompatibility. It is desirable to avoid inflammation and have degradation with concomitant host collagen deposition or even site-appropriate in situ regeneration for the successful outcome of an implanted biological scaffold. This work aims to achieve this by utilizing a biocompatible method to modify acellular scaffolds by impregnating alkaline-catalyzed citric acid (CA) cross-linking between the extracellular matrix proteins and silk fibroin (SF)/SF-gelatin (SFG) blends. Combinatorial detergent decellularization was employed to prepare a decellularized porcine liver scaffold (DPL). After proving the decellularization efficiency, the scaffold underwent modification by vacuum impregnation with CA containing SF (SF100DPL) and SFG blends (SFG5050DPL and SFG3070DPL) following pre-cross-linking, drying, and post-cross-linking. The subsequent strength augmentation was demonstrated by significant improvement in tensile strength from 2.4 ± 0.4 MPa (DPL) to, 3.8 ± 0.7 MPa (SF100DPL), 3.4 ± 0.7 MPa (SFG5050DPL), and 3.5 ± 0.2 MPa (SFG3070DPL); Young's modulus from 8.7 ± 1.8 MPa (DPL) to 20 ± 1.9 MPa (SF100DPL), 13.3 ± 2.6 MPa (SFG5050DPL), and 16 ± 1.2 MPa (SFG3070DPL); and suture retention strength from 0.9 ± 0.08 MPa (DPL) to 2.3 ± 0.2 MPa (SF100DPL), 2.8 ± 1.2 MPa (SFG5050DPL), and 2.6 ± 0.9 MPa (SFG3070DPL). The degradation resistance of the modified scaffolds was also markedly improved. Being cytocompatible, its ability to incite tolerable inflammatory and immune responses was confirmed by rat subcutaneous implantation for 14, 30, and 90 days, in terms of inflammatory cell infiltration, neoangiogenesis, and in vitro cytokine release to assess B-cell and T-cell activation. Such ECM composite scaffolds with appropriate strength and biocompatibility offer great promise in soft tissue repair applications such as skin grafting.

Electrospun Silk-ICG Composite Fibers and the Application toward Hemorrhage Control.

In trauma and surgery, efficient hemorrhage control is crucial to avert fatal blood loss and increase the likelihood of survival. There is a significant demand for novel biomaterials capable of promptly and effectively managing bleeding. This study aimed to develop flexible biocomposite fibrous scaffolds with an electrospinning technique using silk fibroin (SF) and indocyanine green (ICG). The FDA-approved ICG dye has unique photothermal properties. The water permeability, degradability, and biocompatibility of Bombyx mori cocoon-derived SF make it promising for biomedical applications. While as-spun SF-ICG fibers were dissolvable in water, ethanol vapor treatment (EVT) effectively induced secondary structural changes to promote ß-sheet formation. This resulted in significantly improved aqueous stability and mechanical strength of the fibers, thereby increasing their fluid uptake capability. The enhanced SF-ICG interaction effectively prevented ICG leaching from the composite fibers, enabling them to generate heat under NIR irradiation due to ICG's photothermal properties. Our results showed that an SF-ICG 0.4% fibrous matrix can uptake 473% water. When water was replaced by bovine blood, a 25 s NIR irradiation induced complete blood coagulation. However, pure silk did not have the same effect. Additionally, NIR irradiation of the SF-ICG fibers successfully stopped the flow of blood in an in vitro model that mimicked a damaged blood vessel. This novel breakthrough offers a biotextile platform poised to enhance patient outcomes across various medical scenarios, representing a significant milestone in functional biomaterials.

Silk Fibroin Film Decorated with Ultralow FeCo Content by Sputtering Deposition Results in a Flexible and Robust Biomaterial for Magnetic Actuation.

Magnetically responsive soft biomaterials are at the forefront of bioengineering and biorobotics. We have created a magnetic hybrid material by coupling silk fibroin─i.e., a natural biopolymer with an optimal combination of biocompatibility and mechanical robustness─with the FeCo alloy, the ferromagnetic material with the highest saturation magnetization. The material is in the form of a 6 µm-thick silk fibroin film, coated with a FeCo layer (nominal thickness: 10 nm) grown by magnetron sputtering deposition. The sputtering deposition technique is versatile and eco-friendly and proves effective for growing the magnetic layer on the biopolymer substrate, also allowing one to select the area to be decorated. The hybrid material is biocompatible, lightweight, flexible, robust, and water-resistant. Electrical, structural, mechanical, and magnetic characterization of the material, both as-prepared and after being soaked in water, have provided information on the adhesion between the silk fibroin substrate and the FeCo layer and on the state of internal mechanical stresses. The hybrid film exhibits a high magnetic bending response under a magnetic field gradient, thanks to an ultralow fraction of the FeCo component (less than 0.1 vol %, i.e., well below 1 wt %). This reduces the risk of adverse health effects and makes the material suitable for bioactuation applications.

Multi-parametric Photoacoustic Imaging Combined with Acoustic Radiation Force Impulse Imaging for Applications in Tissue Engineering.

Tissue engineering is a dynamic field focusing on the creation of advanced scaffolds for tissue and organ regeneration. These scaffolds are customized to their specific applications and are often designed to be complex, large structures to mimic tissues and organs. This study addresses the critical challenge of effectively characterizing these thick, optically opaque scaffolds that traditional imaging methods fail to fully image due to their optical limitations. We introduce a novel multi-modal imaging approach combining ultrasound, photoacoustic, and acoustic radiation force impulse imaging. This combination leverages its acoustic-based detection to overcome the limitations posed by optical imaging techniques. Ultrasound imaging is employed to monitor the scaffold structure, photoacoustic imaging is employed to monitor cell proliferation, and acoustic radiation force impulse imaging is employed to evaluate the homogeneity of scaffold stiffness. We applied this integrated imaging system to analyze melanoma cell growth within silk fibroin protein scaffolds with varying pore sizes and therefore stiffness over different cell incubation periods. Among various materials, silk fibroin was chosen for its unique combination of features including biocompatibility, tunable mechanical properties, and structural porosity which supports extensive cell proliferation. The results provide a detailed mesoscale view of the scaffolds' internal structure, including cell penetration depth and biomechanical properties. Our findings demonstrate that the developed multimodal imaging technique offers comprehensive insights into the physical and biological dynamics of tissue-engineered scaffolds. As the field of tissue engineering continues to advance, the importance of non-ionizing and non-invasive imaging systems becomes increasingly evident, and by facilitating a deeper understanding and better characterization of scaffold architectures, such imaging systems are pivotal in driving the success of future tissue-engineering solutions.

Flexible Neural Interface From Non-Transient Silk Fibroin With Outstanding Conformality, Biocompatibility, and Bioelectric Conductivity.

Silk fibroin (SF) with good biocompatibility can enable an efficient and safe implementation of neural interfaces. However, it has been difficult to achieve a robust integration of patterned conducting materials (multichannel electrodes) on flexible SF film substrates due to the absence of some enduring interactions. In this study, a thermo-assisted pattern-transfer technique is demonstrated that can facilely transfer a layer of pre-set poly(3,4-ethylenedioxythiophene) (PEDOT) onto the flexible SF substrate through an interpenetrating network of 2 polymer chains, achieving a desired substrate/conductor intertwined interface with good flexibility (≈33 MPa), conductivity (386 S cm-1) and stability in liquid state over 4 months simultaneously. Importantly, this technique can be combined with ink-jet printing to prepare a multichannel SF-based neural interface for the electrocorticogram (ECoG) recording and inflammation remission in rat models. The SF-based neural interface with satisfied tissue conformability, biocompatibility, and bioelectric conductivity is a promising ECoG acquisition tool, where the demonstrated approach can also be useful to develop other SF-based flexible bioelectronics.

In-Situ and Green Synthesis of Silk Fibroin-Silver Nanoparticles Composite Microfibers for Enhanced Antibacterial Applications.

The antibacterial properties of modified silk fibroin microfibers (SF MFs) have been widely studied. Among various modifications, integration of silver nanoparticles (Ag NPs) and SF MFs has garnered significant attention due to the broad-spectrum antibacterial activities and long-term antibacterial effect of Ag nanomaterials. However, the traditional introduction of reducing agents or other additives during the synthesis of Ag-SF composite MFs potentially affects their structure and antibacterial properties. Facile, green and effective methods for the preparation of Ag-SF MFs with enhanced antibacterial properties are therefore highly desired. In this study, Ag NPs were uniformly in-situ deposited onto the optimized SF MFs by adjusting the pH and duration conditions under the guidance of green chemistry. The loaded Ag NPs have a good dispersibility and an average size of ~10 nm. The stability of SF MFs after the deposition of Ag NPs and the crystalline features of the loaded Ag NPs have been carefully investigated. Moreover, antibacterial experiments confirmed that Ag-SF MFs exhibited superior antibacterial activities. After co-incubating Ag-SF MFs with L929 cells, the cell viability reached 90%, demonstrating the great biocompatibility of the modified fibers. This green in-situ synthetic method will promote the further medical use of Ag-SF MFs in antibacterial fields.

Preparation and osteogenesis of a multiple crosslinking silk fibroin/carboxymethyl chitosan/sodium alginate composite scaffold loading with mesoporous silica/poly (lactic acid-glycolic acid) microspheres.

Large bone defect repair is a striking challenge in orthopedics. Currently, inorganic-organic composite scaffolds are considered as a promising approach to these bone regeneration. Silicon ions (Si4+) are bioactive and beneficial to bone regeneration and Si4+-containing inorganic mesoporous silica (MS) can effectively load drugs for bone repair. To better control the release of drug, we prepared biodegradable MS/PLGA (MP) microspheres. MP loaded organic silk fibroin/carboxymethyl chitosan/sodium alginate (MP/SF/CMCS/SA) composite scaffolds were further constructed by genipin and Ca2+ crosslinking. All MP/SF/CMCS/SA scaffolds had good swelling ability, degradation rate and high porosity. The incorporation of 1% MP significantly enhanced the compressive strength of composite scaffolds. Besides, MP loaded scaffold showed a sustained release of Si4+ and Ca2+. Moreover, the release rate of rhodamine (a model drug) of MP/SF/CMCS/SA scaffolds was obviously lower than that of MP. When culturing with rat bone marrow mesenchymal stem cells, scaffolds with 1% MP displayed good proliferation, adhesion and enhanced osteogenic differentiation ability. Based on the results above, the addition of 1% MP in SF/CMCS/SA scaffolds is a prospective way for drug release in bone regeneration and is promising for further in vivo bone repair applications.

Silk fibroin/chitosan/montmorillonite sponge dressing: Enhancing hemostasis, antimicrobial activity, and angiogenesis for advanced wound healing applications.

Open wounds present a significant challenge in healthcare, requiring careful management to prevent infection and promote wound healing. Advanced wound dressings are critical need to enhance their hemostatic capabilities, antimicrobial properties, and ability to support angiogenesis and sustained moisture for optimal healing. This study introduces a flexible hemostatic dressing designed for open wounds, integrating chitosan (CS) for hemostasis and biocompatibility, silk fibroin (SF) for mechanical strength, and montmorillonite (MMT) for enhanced drug transport. The CSSF@MMT dressings showed promising mechanical strength and swift hemostasis. The CIP-loaded CSSF@MMT demonstrated sustained release for up to one week, exhibiting antibacterial properties against both Gram-positive and Gram-negative bacteria. In vitro cell migration assay demonstrated that erythropoietin-loaded CSSF@MMT dressings promoted the proliferation and migration of endothelial cells. Similarly, the chick embryo chorioallantoic membrane study indicated the same dressings exhibited a significant increase in vascular regeneration. This research suggests that the CSSF@MMT sponge dressing, incorporated with CIP and erythropoietin, holds promise in effectively halting bleeding, creating a protective environment, diminishing inflammation, and fostering wound tissue regeneration. This potential makes it a significant advancement in open wound care, potentially lowering the need for limb amputation and decreasing wound care burden worldwide.

Meticulously engineered three-dimensional-printed scaffold with microarchitecture and controlled peptide release for enhanced bone regeneration.

The repair of large load-bearing bone defects requires superior mechanical strength, a feat that a single hydrogel scaffold cannot achieve. The objective is to seamlessly integrate optimal microarchitecture, mechanical robustness, vascularisation, and osteoinductive biological responses to effectively address these critical load-bearing bone defects. To confront this challenge, three-dimensional (3D) printing technology was employed to prepare a polycaprolactone (PCL)-based integrated scaffold. Within the voids of 3D printed PCL scaffold, a methacrylate gelatin (GelMA)/methacrylated silk fibroin (SFMA) composite hydrogel incorporated with parathyroid hormone (PTH) peptide-loaded mesoporous silica nanoparticles (PTH@MSNs) was embedded, evolving into a porous PTH@MSNs/GelMA/SFMA/PCL (PM@GS/PCL) scaffold. The feasibility of fabricating this functional scaffold with a customised hierarchical structure was confirmed through meticulous chemical and physical characterisation. Compression testing unveiled an impressive strength of 17.81 ± 0.83 MPa for the composite scaffold. Additionally, in vitro angiogenesis potential of PM@GS/PCL scaffold was evaluated through Transwell and tube formation assays using human umbilical vein endothelium, revealing the superior cell migration and tube network formation. The alizarin red and alkaline phosphatase staining assays using bone marrow-derived mesenchymal stem cells clearly illustrated robust osteogenic differentiation properties within this scaffold. Furthermore, the bone repair potential of the scaffold was investigated on a rat femoral defect model using micro-computed tomography and histological examination, demonstrating enhanced osteogenic and angiogenic performance. This study presents a promising strategy for fabricating a microenvironment-matched composite scaffold for bone tissue engineering, providing a potential solution for effective bone defect repair.

Silk films with distinct surface topography modulate plasma membrane curvature to polarize macrophages.

The physical properties of a biomaterial play a vital role in modulating macrophage polarization. However, discerning the specific effects of individual parameters can be intricate due to their interdependencies, limiting the mechanism underlying a specific parameter on the polarization of macrophages. Here, we engineered silk fibroin (SF) films with tunable surface roughness while maintaining similar physical properties by combining casting and salting out techniques. We demonstrate that increased surface roughness in SF films promotes M2-like macrophage polarization, characterized by enhanced secretion of anti-inflammatory cytokines. Transcriptomic analysis unveils the modulation of genes associated with extracellular matrix-cell interactions, highlighting the role of surface topography in regulating cellular processes. Mechanistically, we show that surface roughness induces macrophage membrane curvature, facilitating integrin αv endocytosis and thereby inhibiting the integrin-NF-kB signaling pathway. In vivo implantation assays corroborate that rough SF films substantially mitigate early inflammatory responses. This work establishes a direct link between surface roughness and intracellular signaling in macrophages, adding to our understanding of the biomaterial surface effect at the material-cell interface and bringing insights into material design.

A new method to immobilize urease in silk fibroin membrane by unidirectional nanopore dehydration.

The immobilization of free enzymes is crucial for enhancing their stability in different environments, enabling reusability, and expanding their applications. However, the development of a straightforward immobilization method that offers stability, high efficiency, biocompatibility, and modifiability remains a significant challenge. Silk fibroin (SF) is a good carrier for immobilized enzymes and drugs. Here, we employed urease as a model enzyme and utilized our developed technology called unidirectional nanopore dehydration (UND) to efficiently dehydrate a regenerated SF solution containing urease in a single step, resulting in the preparation of a highly functionalized SF membrane immobilizing urease (UI-SFM). The preparation process of UI-SFM is based on an all-water system, which is mild, green and able to efficiently and stably immobilize urease in the membranes, maintaining 92.7% and 82.8% relative enzyme activity after 30 days of storage in dry and hydrated states, respectively. Additionally, we performed additional post-treatments, including stretching and cross-linking with polyethylene glycol diglycidyl ether (PEGDE), to obtain two more robust immobilized urease membranes (UI-SFMs and UI-SFMc). The thermal and storage stability of these two membranes were significantly improved, and the recovery ratio of enzyme activity reached more than 90%. After 10 repetitions of the enzymatic reaction, the activity recovery of UI-SFMs and UI-SFMc remained at 92% and 88%, respectively. The results suggest that both UND-based and post-treatment-developed membranes exhibit excellent urease immobilization capabilities. Furthermore, the enzyme immobilization method offers a straightforward and versatile approach for efficient and stable enzyme immobilization, while its flexible modifiability caters to diverse application requirements.

Ultra-Strong Protein-Based Hydrogels via Promoting Intermolecular Entanglement of the Amorphous Region.

Proteins are classified as biopolymers which share similar structural features with semi-crystalline polymers. Although their unique biocompatibility facilitates the universal applications of protein-based hydrogels in the biomedical field, the mechanical performances of protein-based hydrogels fall short of practical requirements. Conventional strategies for enhancing mechanical properties focus on forming regularly folded secondary structures as analogs of crystalline regions. This concept is based on proteins as the analogy of semi-crystalline polymers, in which crystalline regions profoundly contribute to the mechanical performances. Even though the contribution of the amorphous region is equally weighted for semi-crystalline polymers, their capacity to improve the mechanical performances of protein-based structures is still undervalued. Herein, the potential of promoting the mechanical performances is explored by controlling the state of amorphous regions in protein-based hydrogels. A fibril protein is chosen, regenerated silk fibroin (RSF), as a model molecule for its similar viscoelasticity with a semi-crystalline polymer. The amorphous regions in the RSF hydrogels are transformed from extended to entangled states through a double-crosslinking method. The formation of entanglement integrates new physically crosslinked points for remarkable improvement in mechanical performances. A robust hydrogel is not only developed but also intended to provide new insights into the structural-property relationship of protein-based hydrogels.

Silk protein: A novel antifungal and edible coating for strawberry preservation.

In this study, an antimicrobial component, silk protease inhibitors (SPIs), was extracted from discarded silkworm cocoons, and a suitable degumming method for obtaining regenerated silk fibroin (SF) was screened. An edible antimicrobial coating was prepared by mixing SPIs with SF for evaluation of potential in strawberries preservation. Results demonstrated that SPI could effectively inhibit mycelial growth and spore germination. The alkaline protease method exhibited the highest degumming rate of 24.4 %. The SPI-SF coating exhibited excellent mechanical properties, high water vapor permeability, and easy washability. Within 10 days, seedlings treatment with SPI-SF coating solution showed a germination rate of 94.3 %, and exhibited good biocompatibility with HepG2 cells. Coating with SPI-SF led to increase in the storage period of strawberries to 10-14 days, concurrent with considerable reduction in decay rate at room temperature. Conclusively, this study demonstrates the potential of SPI-SF edible coating in strawberries preservation.

Development of stopped-flow hyper-CEST NMR method on recirculating hyperpolarization system as applied to void space analysis in polymers.

129Xe NMR spectroscopy of polymers can provide important information on void spaces, sometimes called free volume, in polymers. Unfortunately, the spectroscopy's low sensitivity has limited its widespread use in both academic and industrial research. In order to overcome such a difficult situation, hyper-CEST method which employs hyperpolarization and CEST techniques, is examined after the introduction of recirculation and subtraction modes. Alongside the incorporated stopped-flow technique, these modes were very efficient in detecting very weak hidden signals from cellulose nanofiber (CNF) and silk fibroin (SF) films and in discussing the void space in these polymers. From the analysis of detailed saturation frequency dependence in the increment of 100 Hz, the chemical shifts of hidden peaks were successfully determined to give reasonable values for the size of void space in CNF and SF. Application on thermoplastic polyurethane film also supported our method of analysis. The subtraction mode was very efficient in judging the presence or absence of any peak at a fixed saturation frequency. These facts support that the mode will surely be useful in the future exploratory study of very weak hidden signals.

A review on the use of composites of a natural protein, silk fibroin with Mxene/carbonaceous materials in biomedical science.

Silk fibroin (SF), a natural biodegradable and biocompatible protein, has garnered significant attention in biomedical applications due to its impressive properties, including excellent biocompatibility, biodegradability, and mechanical resilience. Nevertheless, its broader usage faces obstacles by its insufficient mechanical strength and electrical conductivity. In order to address these constraints, recent studies have concentrated on combining SF with cutting-edge nanomaterials like MXene and carbon-based materials. This review comprehensively examines the applications and potential of silk fibroin-MXene/carbon-based nanocomposites in biomedical fields. The unique properties of SF, MXene, and carbon-based materials are explored, emphasizing how their combination enhances mechanical strength, conductivity, and biocompatibility. These composites show substantial enhancements in performance for several biomedical applications by utilising the excellent conductivity and mechanical capabilities of MXene and carbonaceous elements. The innovative potential of these nanocomposites is highlighted by critically discussing key applications such as tissue engineering, drug delivery, and biosensing. In addition, the work discusses the latest research progress, difficulties, and future prospects in the sector, providing valuable insights into possible breakthroughs and uses. This review seeks to comprehensively analyse the existing information on silk fibroin-MXene/carbon based nanocomposites in healthcare.

A PLGA/Silk Fibroin Nanofibre Membrane Loaded with Natural Flavonoid Compounds Extracted from Green Cocoons for Wound Healing.

The use of biodegradable materials combined with natural metabolites in wound dressings has received much attention. Flavonoids (FLs) from green cocoons, as metabolites, have antibacterial, antioxidant, anti-inflammatory, and other pharmacological effects. In this study, composite membranes of FL-loaded polylactic glycolic acid (PLGA)/silk fibroin (SF) were prepared by an electrospinning method. The prepared membranes, including SF, exhibited a good slow-release effect and cytocompatibility. An in vitro evaluation of the FL-loaded PLGA/SF membranes demonstrated good antioxidant, antibacterial, and anti-inflammatory properties. Animal experiments showed that the wound healing rate of PLGA/SF-2.5FL membranes within 15 days was 97.3%, and that of the control group was 72.5%. The PLGA/SF-2.5FL membranes shortened the inflammatory period of a full-layer wound model and promoted skin regeneration and wound healing by downregulating expression of the pro-inflammatory cytokines IL-1ß and TNF-α and promoting expression of the growth factors VEGF, TGF-ß, and EGF. In summary, the PLGA/SF-2.5FL composite nanofibre membrane with anti-inflammatory properties is an ideal wound dressing to promote acute wound healing.