Silk Fibroin Hydrogels Research Articles

Potential benefits of using chitosan and silk fibroin topical hydrogel for managing wound healing and coagulation

Background & ObjectivesThe intricate process of wound healing involves replacing the cellular or tissue structure that has been destroyed. In recent years various wound dressings were launched but reported several limitations. The topical gel preparations are intended for certain skin wound conditions for local action. Chitosan-based hemostatic materials are the most effective in halting acute hemorrhage, and naturally occurring silk fibroin is widely utilized for tissue regeneration. So, this study was conducted to evaluate the potential of chitosan hydrogel(CHI-HYD) and chitosan silk fibroin hydrogel (CHI-SF-HYD) on blood clotting and wound healing. MethodsHydrogel was prepared using various concentrations of silk fibroin with guar gum as a gelling agent. The optimized formulations were evaluated for visual appearance, Fourier transforms infrared spectroscopy (FT-IR), pH, spreadability, viscosity, antimicrobial activity, HR-TEM analysis, ex vivo skin permeation, skin irritation, stability studies, and in vivo studies by using adult male Wistar albino rats. ResultsBased on the outcome of FT-IR, no chemical interaction between the components was noticed. The developed hydrogels exhibited a viscosity of 79.2 ± 4.2 Pa.s (CHI-HYD), 79.8 ± 3.8 Pa.s (CHI-SF-HYD), and pH of 5.87 ± 0.2 (CHI-HYD), 5.96 ± 0.1 (CHI-SF-HYD). The prepared hydrogels were sterile and non-irritant to the skin. The in vivo study outcomes show that the CHI-SF-HYD treated group has significantly shortened the span of tissue reformation than other groups. This demonstrated that the CHI-SF-HYD could consequently accelerate the regeneration of the damaged area. Interpretation & ConclusionOverall, the positive outcomes revealed improved blood coagulation and re-epithelialization. This indicates that the CHI-SF-HYD could be used to develop novel wound-healing devices.

Biocompatibility of silk methacrylate/gelatin-methacryloyl composite hydrogel and its feasibility as a vascular tissue engineering scaffold

Silk methacrylate (SilMA) has been studied extensively due to its ability to modify Silk fibroin (SF) by increasing the water solubility and enhancing the mechanical properties of SF hydrogels. However, SilMA hydrogels are generally soft with weak mechanical properties. In order to enhance the mechanical properties of hydrogel scaffolds, we used liquid nitrogen to modify SilMA to obtain a novel N2-SilMA/gelatin-methacryloyl (GelMA) composite hydrogel. N2-SilMA was successfully detected by Fourier transform infrared (FTIR) spectroscopy and 1H nuclear magnetic resonance. Scanning electron microscope showed that the composite hydrogel still had certain arrangement characteristics of SF and dense pores which met the necessary conditions for the cell scaffold. The mechanical tests showed that the mechanical properties of SilMA were greatly enhanced after modification at ultra-low temperature. We evaluated its cytocompatibility and biocompatibility, and the results showed that the composite scaffold promoted the growth of cells. Different types of composite hydrogels were injected into ICR mice and the results showed a stable scaffold structure in vivo, suggesting their ability to promote angiogenesis. In conclusion, the N2-SilMA/GelMA composite hydrogel had better mechanical properties, excellent cytocompatibility, and biological properties compared to the other groups.

Synergistic Effect of Co-Culturing Breast Cancer Cells and Fibroblasts in the Formation of Tumoroid Clusters and Design of In Vitro 3D Models for the Testing of Anticancer Agents.

Breast cancer is still the leading cause of women's death due to relapse and metastasis. In vitro tumor models are considered reliable tools for drug screening and understanding cancer-driving mechanisms due to the possibility of mimicking tumor heterogeneity. Herein, a 3D breast cancer model (3D-BCM) is developed based on enzymatically-crosslinked silk fibroin (eSF) hydrogels. Human MCF7 breast cancer cells are encapsulated into eSF hydrogels, with and without human mammary fibroblasts. The spontaneously occurring conformational change from random coil to β-sheet is correlated with increased eSF hydrogels' stiffness over time. Moreover, mechanical properties analysis confirms that the cells can modify the stiffness of the hydrogels, mimicking the microenvironment stiffening occurring in vivo. Fibroblasts support cancer cells growth and assembly in the eSF hydrogels up to 14 days of culture. Co-cultured 3D-BCM exhibits an upregulated expression of genes related to extracellular matrix remodeling and fibroblast activation. The 3D-BCM is subjected to doxorubicin and paclitaxel treatments, showing differential drug response. Overall, these results suggest that the co-culture of breast cancer cells and fibroblasts in eSF hydrogels allow the development of a mimetic in vitro platform to study cancer progression. This opens up new research avenues to investigate novel molecular targets for anti-cancer therapy.

Cell Development Enhanced Bionic Silk Hydrogel on Remodeling Immune Pathogenesis of Spinal Cord Injury via M2 Polarization of Microglial

AbstractDue to the complex spatial‐temporal pathophysiology of spinal cord injury (SCI), effective modulation of SCI‐specific inflammatory pathogenesis to achieve desirable therapeutic effects on functional recovery still remains challenging. Herein, cell‐enhanced photocrosslinked silk fibroin hydrogels with extracellular matrix‐mimicking cues of mechanical properties and RGD (Arg‐Gly‐Asp) signals are gelled in situ to fill the lesion site to modulate injury‐induced neuroinflammation and promote neurite regrowth after SCI. The bionic hydrogel system provides biomimetic mechanical cues to promote neuronal differentiation of neural stem/progenitor cells (NPCs) and neurite growth by activating YAP nuclear expression. Importantly, favored by the strong capacity of silk fibroin hydrogels on macrophage/microglia recruitment, NPCs encapsulated hydrogel (NPCs@SFRGD0.1) effectively promotes recruited macrophages/microglia to M2 polarization in the lesion site by releasing S100A4 and thereby remodels the inflammatory microenvironment after SCI. Moreover, NPCs@SFRGD0.1 successfully reduces glial scar formation and accelerates corticospinal tract axon regrowth to improve locomotor recovery. Overall, this work contributes to illustrating the therapeutic mechanism of NPCs development based biomaterial therapies on modulating inflammatory microenvironment and this NPCs enhanced silk fibroin hydrogel provides a promising therapeutic strategy for SCI.

Fabrication of mulberry leaf extract (MLE)- and tasar pupal oil (TPO)-loaded silk fibroin (SF) hydrogels and their antimicrobial properties.

Biocomposites have gained tremendous advantages over synthetic composites due to their biocompatibility, sustainable degradation, and ability to easily combine with other substances. In the present study, we have prepared silk fibroin (SF) hydrogel, mulberry leaf extract (MLE), tasar pupal oil (TPO), and their composites, such as TPO-loaded SF hydrogel and MLE-loaded SF hydrogel, and characterized them by using a phase contrast microscope (PCM), scanning electron microscope (SEM) SEM- EDX, and Fourier transform infrared spectroscopy (FTIR). In addition, 1H-NMR was used for profiling of mulberry leaf extract and GC-MS was used to find tasar pupal oil composition. Further, the disc diffusion method evaluated their antimicrobial activities against S. aureus, E. coli, A. flavus, and A. brassicae. PCM, SEM, and FTIR results validated the conjugation of MLE and SF hydrogel composite; 1H-NMR confirmed the 41 metabolites in MLE, and GC-MS established the composition of tasar pupal oil. Since both composites, such as TPO-loaded SF hydrogel and MLE-loaded SF hydrogel, reduced the S. aureus and E. coli activities at all tested concentrations, the antibacterial results were unambiguous in their conclusion. S. aureus could only be inhibited by SF hydrogel at a high concentration (300g/ml), despite suppressing E. coli growth at all tested concentrations. At 300g/ml, MLE demonstrated antibacterial action against S. aureus. Furthermore, at a dosage of 300g/ml, TPO inhibited both S. aureus and E. coli. Both mulberry leaf extract (at 200 and 300g/ml) and the MLE-loaded SF hydrogel composite displayed antifungal activity against A. flavus at all tested concentrations (100, 200, and 300g/ml).

Pro-angiogenic photo-crosslinked silk fibroin hydrogel: a potential candidate for repairing alveolar bone defects.

This study aimed to develop a pro-angiogenic hydrogel with in situ gelation ability for alveolar bone defects repair. Silk fibroin was chemically modified by Glycidyl Methacrylate (GMA), which was evaluated by proton nuclear magnetic resonance (1H-NMR). Then, the photo-crosslinking ability of the modified silk fibroin was assessed. Scratch and transwell-based migration assays were conducted to investigate the effect of the photo-crosslinked silk fibroin hydrogel on the migration of human umbilical vein endothelial cells (HUVECs). In vitro angiogenesis was conducted to examine whether the photo-crosslinked silk fibroin hydrogel would affect the tube formation ability of HUVECs. Finally, subcutaneous implantation experiments were conducted to further examine the pro-angiogenic ability of the photo-crosslinked silk fibroin hydrogel, in which the CD31 and α-smooth muscle actin (α-SMA) were stained to assess neovascularization. The tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were also stained to evaluate inflammatory responses after implantation. GMA successfully modified the silk fibroin, which we verified by our 1H-NMR and in vitro photo-crosslinking experiment. Scratch and transwell-based migration assays proved that the photo-crosslinked silk fibroin hydrogel promoted HUVEC migration. The hydrogel also enhanced the tube formation of HUVECs in similar rates to Matrigel®. After subcutaneous implantation in rats for one week, the hydrogel enhanced neovascularization without triggering inflammatory responses. This study found that photo-crosslinked silk fibroin hydrogel showed pro-angiogenic and inflammation inhibitory abilities. Its photo-crosslinking ability makes it suitable for matching irregular alveolar bone defects. Thus, the photo-crosslinkable silk fibroin-derived hydrogel is a potential candidate for constructing scaffolds for alveolar bone regeneration.

Silk fibroin microgels as a platform for cell microencapsulation

Cell microencapsulation has been utilized for years as a means of cell shielding from the external environment while facilitating the transport of gases, general metabolites, and secretory bioactive molecules at once. In this light, hydrogels may support the structural integrity and functionality of encapsulated biologics whereas ensuring cell viability and function and releasing potential therapeutic factors once in situ. In this work, we describe a straightforward strategy to fabricate silk fibroin (SF) microgels (µgels) and encapsulate cells into them. SF µgels (size ≈ 200 µm) were obtained through ultrasonication-induced gelation of SF in a water-oil emulsion phase. A thorough physicochemical (SEM analysis, and FT-IR) and mechanical (microindentation tests) characterization of SF µgels were carried out to assess their nanostructure, porosity, and stiffness. SF µgels were used to encapsulate and culture L929 and primary myoblasts. Interestingly, SF µgels showed a selective release of relatively small proteins (e.g., VEGF, molecular weight, MW = 40 kDa) by the encapsulated primary myoblasts, while bigger (macro)molecules (MW = 160 kDa) were hampered to diffusing through the µgels. This article provided the groundwork to expand the use of SF hydrogels into a versatile platform for encapsulating relevant cells able to release paracrine factors potentially regulating tissue and/or organ functions, thus promoting their regeneration.Graphical

Performance of Colombian Silk Fibroin Hydrogels for Hyaline Cartilage Tissue Engineering.

The development and evaluation of scaffolds play a crucial role in the engineering of hyaline cartilage tissue. This work aims to evaluate the performance of silk fibroin hydrogels fabricated from the cocoons of the Colombian hybrid in the in vitro regeneration of hyaline cartilage. The scaffolds were physicochemically characterized, and their performance was evaluated in a cellular model. The results showed that the scaffolds were rich in random coils and β-sheets in their structure and susceptible to various serine proteases with different degradation profiles. Furthermore, they showed a significant increase in ACAN, COL10A1, and COL2A1 expression compared to pellet culture alone and allowed GAG deposition. The soluble portion of the scaffold did not affect chondrogenesis. Furthermore, they promoted the increase in COL1A2, showing a slight tendency to differentiate towards fibrous cartilage. The results also showed that Colombian silk could be used as a source of biomedical devices, paving the way for sericulture to become a more diverse economic activity in emerging countries.

Silk fibroin hydrogels for biomedical applications.

Silk fibroin hydrogels occupy an essential position in the biomedical field due to their remarkable biological properties, excellent mechanical properties, flexible processing properties, as well as abundant sources and low cost. Herein, we introduce the unique structures and physicochemical characteristics of silk fibroin, including mechanical properties, biocompatibility, and biodegradability. Then, various preparation strategies of silk fibroin hydrogels are summarized, which can be divided into physical cross-linking and chemical cross-linking. Emphatically, the applications of silk fibroin hydrogel biomaterials in various biomedical fields, including tissue engineering, drug delivery, and wearable sensors, are systematically summarized. At last, the challenges and future prospects of silk fibroin hydrogels in biomedical applications are discussed.

An all-silk-derived bilayer hydrogel for osteochondral tissue engineering.

Osteochondral repair remains a challenge in clinical practice nowadays despite extensive advances in tissue engineering. The insufficient recruitment of endogenous cells in the early stage and incomplete cell differentiation in the later stage constitute the major difficulty of osteochondral repair. Here, a novel all-silk-derived multifunctional biomaterial platform for osteochondral engineering is reported. The bilayer methacrylated silk fibroin (SilMA) hydrogel was fabricated through stratified photocuring as the basic provisional matrix for tissue regeneration. Platelet-rich plasma (PRP) incorporation promoted the migration and pre-differentiation of the bone marrow mesenchymal stem cells (BMSCs) in the early stage of implantation. The long-term regulation of BMSCs chondrogenesis and osteogenesis was realized by the stratified anchoring of the silk fibroin (SF) microspheres respectively loaded with Kartogenin (KGN) and berberine (BBR) in the hydrogel. The composite hydrogels were further demonstrated to promote BMSCs chondrogenic and osteogenic differentiation under an inflammatory microenvironment and to achieve satisfying cartilage and subchondral bone regeneration with great biocompatibility after 8 weeks of implantation. Since all the components used are readily available and biocompatible and can be efficiently integrated via a simple process, this composite hydrogel scaffold has tremendous potential for clinical use in osteochondral regeneration.

Surface-fill H2S-releasing silk fibroin hydrogel for brain repair through the repression of neuronal pyroptosis

Traumatic brain injury (TBI) remains the major cause of disability and mortality worldwide due to the persistent neuroinflammation and neuronal death induced by TBI. Among them, pyroptosis, a specific type of programmed cell death (PCD) triggered by inflammatory signals, plays a significant part in the pathological process after TBI. Inhibition of neuroinflammation and pyroptosis is considered a possible strategy for the treatment of TBI. In our previous study, exogenous hydrogen sulfide(H2S) exerted a neuroprotective effect after TBI. Here, we developed a surface-fill H2S-releasing silk fibroin (SF) hydrogel (H2S@SF hydrogel) to achieve small-dose local administration and avoid volatile and toxic side effects. We used a controlled cortical impact (CCI) to establish a mild TBI model in mice to examine the effect of H2S@SF hydrogel on TBI-induced pyroptosis. We found that H2S@SF hydrogel inhibited the expression of H2S synthase in neurons after TBI and significantly inhibited TBI-induced neuronal pyroptosis. In addition, immunofluorescence staining results showed that the necroptosis protein receptor-interacting serine/threonine-protein kinase 1 (RIPK1) partially colocalized with the pyroptosis protein Gasdermin D (GSDMD) in the same cells. H2S@SF hydrogel can also inhibit the expression of the necroptosis protein. Moreover, H2S@SF hydrogel also alleviates brain edema and the degree of neurodegeneration in the acute phase of TBI. The neuroprotective effect of H2S@SF hydrogel was further confirmed by wire-grip test, open field test, Morris water maze, beam balance test, radial arm maze, tail suspension, and forced swimming test. Lastly, we also measured spared tissue volume, reactive astrocytes and activated microglia to demonstrate H2S@SF hydrogel impacts on long-term prognosis in TBI. Our study provides a new theoretical basis for the treatment of H2S after TBI and the clinical application of H2S@SF hydrogel. Statement of significanceSilk fibroin (SF) hydrogel controls the release of hydrogen sulfide (H2S) to inhibit neuronal pyroptosis and neuroinflammation in injured brain tissue. In this study, we synthesized a surface-fill H2S-releasing silk fibroin hydrogel, which could slowly release H2S to reshape the homeostasis of endogenous H2S in injured neurons and inhibit neuronal pyroptosis in a mouse model of traumatic brain injury. Meanwhile, H2S@SF hydrogel could alleviate brain edema and the degree of neurodegeneration, improve motor dysfunction, anxious behavior and memory impairment caused by TBI, reduce tissue loss and ameliorate neuroinflammation. Our study provides a new theoretical basis for the treatment of H2S after TBI and the clinical application of H2S@SF hydrogel.

Structural-Functional Pluralistic Modification of Silk Fibroin via MOF Bridging for Advanced Wound Care.

Silk fibroin (SF) is widely used to fabricate biomaterials for skin related wound caring or monitoring, and its hydrogel state are preferred for their adaptability and easy to use. However, in-depth development of SF hydrogel is restricted by their limited mechanical strength, increased risk of infection, and inability to accelerate tissue healing. Therefore, a structure-function pluralistic modification strategy using composite system of metal organic framework (MOF) as bridge expanding SF's biomedical application is proposed. After developing the photocuring and bonding SF hydrogel, a MOF drug-loading system is utilized to enhance hydrogel's structural strength while endowing its antibacterial and angiogenic properties, yielding a multifunctional SF hydrogel. The synergy between the MOF and SF proteins at the secondary structure level gives this hydrogel reliable mechanical strength, making it suitable for conventional wound treatment, whether for closing incisions quickly or acting as adhesive dressings (five times the bonding strength of ordinary fibrin glue). Additionally, with the antibacterial and angiogenic functions getting from MOF system, this modified SF hydrogel can even treat ischemic trauma with cartilage exposure. This multiple modification should contribute to the improvement of advanced wound care, by promoting SF application in the production of tissue engineering materials.

Gallium ions incorporated silk fibroin hydrogel with antibacterial efficacy for promoting healing of Pseudomonas aeruginosa-infected wound

The continual resistance to antibiotics and the generation of a series of bacterial infections has emerged as a global concern, which requires appropriate measures and therapeutics to address such a menace. Herein, we report on Silk fibroin (SF) hydrogel with good biocompatibility and biodegradability fabricated through the crosslinking of the SF of different concentrations with Gallium nitrate (Ga (NO3)3) against Pseudomonas aeruginosa. However, the SF: Ga = 500: 1 (w/w) (SF/Ga) demonstrated a good bactericidal and wound healing effect as a result of the moderate and prolonged release of the Ga3+ following the gradual degradation of the hydrogel. The Ga3+, known for its innovative nature acted as a crosslinked agent and a therapeutic agent employing the "Trojan horse" strategy to effectively deal with the bacteria. Also, the Ga3+, which is positively charged neutralizes the negative potential value of the SF particles to reduce the charge and further induce the β-sheet formation in the protein structure, a characteristic of gelation in SF. The morphology showed a fabricated homogenous structure with greater storage modulus- G' with low loss modulus- G'' modulus demonstrating the mechanical performance and the ability of the SF/Ga hydrogel to hold their shape, at the same time allowing for the gradual release of Ga3+. A demonstration of biocompatibility, biodegradability, bactericidal effect and wound healing in in vitro and in vivo present the SF/Ga hydrogel as an appropriate platform for therapeutic and for antibacterial wound dressing.

A hydrogel-fiber-hydrogel composite scaffold based on silk fibroin with the dual-delivery of oxygen and quercetin.

Supplying sufficient oxygen within the scaffolds is one of the essential hindrances in tissue engineering that can be resolved by oxygen-generating biomaterials (OGBs). Two main issues related to OGBs are controlling oxygenation and reactive oxygen species (ROS). To address these concerns, we developed a composite scaffold entailing three layers (hydrogel-electrospun fibers-hydrogel) with antioxidant and antibacterial properties. The fibers, the middle layer, reinforced the composite structure, enhancing the mechanical strength from 4.27 ± 0.15 to 8.27 ± 0.25 kPa; also, this layer is made of calcium peroxide and silk fibroin (SF) through electrospinning, which enables oxygen delivery. The first and third layers are physical SF hydrogels to control oxygen release, containing quercetin (Q), a nonenzymatic antioxidant. This composite scaffold resulted in almost more than 40 mmHg of oxygen release for at least 13 days, and compared with similar studies is in a high range. Here, Q was used for the first time for an OGB to scavenge the possible ROS. Q delivery not only led to antioxidant activity but also stabilized oxygen release and enhanced cell viability. Based on the given results, this composite scaffold can be introduced as a safe and controllable oxygen supplier, which is promising for tissue engineering applications, particularly for bone.

Osteogenic differentiation of the MSCs on silk fibroin hydrogel loaded Fe3O4@PAA NPs in static magnetic field environment

The combination of static magnetic field and magnetic materials for bone tissue repair provides a remote, minimally invasive, and uncomplicated treatment method. Although materials based on Fe3O4 nanoparticles (Fe3O4 NPs) have been widely applied in bone tissue regeneration, the high mobility of Fe3O4 NPs and the oxidative stress involving hydrogen peroxide (H2O2) limit its applications. In this work, silk fibroin (SF) hydrogel was selected to astrict the flowability of Fe3O4 NPs. SF has strong biocompatibility and inducement to the osteogenic differentiation of mesenchymal stem cells (MSCs). In order to reduce the influence of oxidative stress, polyacrylic acid (PAA) was employed to modify the Fe3O4 NPs. The results indicated that Fe3O4@PAA nanoparticles (Fe3O4@PAA NPs) eliminated about 40 % H2O2 in 3 hrs and reduced hydroxyl radicals produced by Fenton reaction. Intracellular studies have shown that SF hydrogel contained Fe3O4@PAA NPs reduced intracellular ROS-induced damage and thus improved cell activity. Compared with other groups, the ALP activity, mineralization ability and collagen secretion level of MSCs on SF hydrogel with Fe3O4@PAA NPs were higher when magnetic field exists.

High Concentration Crystalline Silk Fibroin Solution for Silk-Based Materials.

As a functional biomaterial, silk fibroin has been widely used in drug release, cell encapsulation and tissue regeneration. To meet the requirements of these applications, the properties of silk fibroin-based materials should be finely tunable. Many useful properties of biomaterials emerge from the collective interactions among ordered and disordered domains. Thus, increasing subtle control of silk hierarchical structures is required. As a characteristic of ordered silk fibroin, crystalline silk fibroin (CSF) is an important part of silk fibroin-based biomaterials, but the preparation of CSF solution, especially high concentration CSF solution, remains a challenge. Here, a solution composed of β-sheet-rich silk fibroin is reported. These CSF were obtained by the sonication of silk fibroin hydrogel, destroying the hydrogel network, and turning silk fibroin hydrogels into CSF solution. These β-sheet-rich CSF solutions were stable enough for several days or even weeks. In addition, they were typically ordered crystalline domains, which could be mixed with disordered domains and fabricated into porous scaffolds, films, hydrogels and other silk fibroin-based scaffolds with different properties.

Green tea catechin-grafted silk fibroin hydrogels with reactive oxygen species scavenging activity for wound healing applications

BackgroundOverproduction of reactive oxygen species (ROS) is known to delay wound healing by causing oxidative tissue damage and inflammation. The green tea catechin, (–)-Epigallocatechin-3-O-gallate (EGCG), has drawn a great deal of interest due to its strong ROS scavenging and anti-inflammatory activities. In this study, we developed EGCG-grafted silk fibroin hydrogels as a potential wound dressing material.MethodsThe introduction of EGCG to water-soluble silk fibroin (SF-WS) was accomplished by the nucleophilic addition reaction between lysine residues in silk proteins and EGCG quinone at mild basic pH. The resulting SF-EGCG conjugate was co-crosslinked with tyramine-substituted SF (SF-T) via horseradish peroxidase (HRP)/H2O2 mediated enzymatic reaction to form SF-T/SF-EGCG hydrogels with series of composition ratios.ResultsInterestingly, SF-T70/SF-EGCG30 hydrogels exhibited rapid in situ gelation (< 30 s), similar storage modulus to human skin (≈ 1000 Pa) and superior wound healing performance over SF-T hydrogels and a commercial DuoDERM® gel dressings in a rat model of full thickness skin defect.ConclusionThis study will provide useful insights into a rational design of ROS scavenging biomaterials for wound healing applications.

Silk Bionanocomposites for Organic Dye Absorption and Degradation

Organic dyes are extensively used in the textile, paper and paint industries, among others. However, the lack of efficient treatment of disposals leads to the release of these toxic molecules into the environment, which has an enormous impact on living organisms. Dye absorption is the most common approach used to tackle this problem. However, the ideal solution should include dye degradation and absorbent regeneration, reducing the environmental impact of the procedure. Dye degradation can be achieved by catalysis. Recently, silk fibroin (SF) has been shown to have incredible absorbent properties. Herein, we characterized the capacity of SF hydrogels to absorb methylene blue (MB), an extensively used cationic organic dye. Moreover, the effect of a pretreatment of the SF hydrogel at different pH and ionic environments is also studied. Interestingly, opposite behaviors are observed when absorbing MB or brilliant blue (an anionic dye), suggesting an electrostatic-based interaction. Furthermore, the regeneration of a MB-saturated SF hydrogel by immersion in acidic pH and its further reuse were evaluated. Finally, the SF hydrogel was coupled with a gold nanoparticle catalyst, which resulted in a material able to absorb and catalyze the MB reduction by sodium borohydride in situ, leading to dye degradation. Overall, this work presents a biodegradable reusable material able to absorb and reduce MB in aqueous media.

Biomineralized hydrogel DC vaccine for cancer immunotherapy: A boosting strategy via improving immunogenicity and reversing immune-inhibitory microenvironment

The postoperative recurrence and metastasis of triple negative breast cancer (TNBC) remain one fatal reason for the failure of clinical treatments. Although the rise of immunotherapy has brought hopes for reducing postoperative recurrence and potential metastasis, the low immune response and immunosuppression of tumor microenvironment (TME) still restrain its extensive application. Herein, we reported a boosting strategy by improving immunogenicity and reversing suppressive TME to realize efficient immunotherapy of TNBC. In this work, a CaCO3 biomineralized hydrogel DC vaccine was synthesized by fixing the membrane proteins of 4T1 cells-DCs fusion cells (FP) into biomineralized silk fibroin hydrogel. On one side, the FP-containing biomineralized hydrogel vaccine (SH@FP@CaCO3) has increased immunogenicity by providing a wide variety of tumor-associated antigens (TAAs) and realizing long-term protein release for DCs maturation and T cell activation. On the other side, the introduction of CaCO3 would increase the pH of TME and promote the polarization of M2-type macrophages to M1-type macrophages, thus reversing the immune-inhibitory microenvironment and relieving the immunosuppressive effect on T cells. The results indicate that the biomineralized hydrogel vaccine shows excellent immune activation effects by simultaneously enhancing the immunogenicity and reversing the immunosuppression TME, which provides a promising strategy for cancer immunotherapy.

Conformational Transition-Driven Self-Folding Hydrogel Based on Silk Fibroin and Gelatin for Tissue Engineering Applications.

Self-folding is a rapidly evolving method for converting flat objects into three-dimensional(3D) structures. However, because there are few materials with suitable properties, the application of self-foldingin tissue engineering has been hindered greatly. Herein, a novel self-folding hydrogel using conformational transition mechanism is developed by employing photocrosslinkable silk fibroin and gelatin composite hydrogel. It is hypothesized that differences in the amount of beta-sheet(β-sheet) formation between the upper and lower layers will supply additional folding stress and drive the self-folding behavior of a bilayer patch, which canimprove the mechanical properties and long-term stability of the self-folded structure. In this study, the impact of various proportions of β-sheets in composite hydrogels on their swelling, mechanics, and internal microstructures are investigated. Subsequently, the folding process parameters are optimized, and diffusion through the folded tubular structure is studied with a perfusion test. Finally, it is proven that the self-folding hydrogel system is cytocompatible and can be utilized to build a 3D coculture system of "endothelial cells-smooth muscle cells". These findings suggest that the self-folding hydrogel can be a promising candidate for applications in blood vessel tissue engineering and regenerative medicine.