Silk Fibroin Hydrogels Research Articles

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.

Optical Fiber Magnetic Field Sensor Based on Silk Fibroin Hydrogel

In this work, an optical fiber magnetic field sensor is proposed and demonstrated based on silk fibroin hydrogel (SFH). The primary function of SFH is the carrier of material (Streptavidin magnetic nanoparticles, SMN) that is susceptible to the magnetic field. The multimode interference spectrum is obtained by configuring a single-mode -multimode -single-mode (SMS) structure. The SMS structure is consisted by sequentially connecting a single-mode fiber (SMF), a no-core fiber (NCF), and a SMF. In addition, the SMN responds to a magnetic field, and SMN will modify the effective refractive index (RI) of SFH. Hence, magnetic field sensing can be performed by coating a mixture of SMN and SFH on the SMS structure. Our experimental results show that the maximum sensitivity of the proposed sensor reaches 673.53pm/Oe, and the average sensitivity of 263pm/Oe in the magnetic field strength range from 0 to 115Oe. The proposed optical fiber sensor of the magnetic field, featuring the benefits of environmental friendliness, low cost, easy manufacturing, and simplicity of structure, can find potential application prospects in various areas such as the industry, environment, and military.

Enhanced transdermal insulin basal release from silk fibroin (SF) hydrogels via iontophoresis

Insulin is the peptide hormone used to treat the diabetes patient. The hormone is normally taken by injection. The transdermal drug delivery system (TDDS) is an alternative route. The silk fibroin (SF) hydrogels were fabricated via solution casting as the insulin matrix. The release and release-permeation experiments of the insulin loaded SF hydrogels were carried out using a modified Franz-diffusion cell at 37 °C for 36 h, under the effects of SF concentrations, pH, and electric field. The release-permeation mechanism through the pig skin was from the Case-II transport with the constant release rate. The diffusion coefficient (D) increased with decreasing SF concentration due to a larger mesh size, and with increasing electric field due to the electroreplusive forces between the insulin and the SF hydrogels against the negatively-charged electrode, and the induced SF hydrogel expansion. The rate and amount of insulin release-permeation became relatively lower as it required a longer time to generate aqueous pathways through the pig skin. The present SF hydrogels are demonstrated here deliver insulin with the required constant release rate, and the suitable amount within a prescribed duration.

Silk Fibroin Hydrogel Reinforced With Magnetic Nanoparticles as an Intelligent Drug Delivery System for Sustained Drug Release.

Due to the well-known biocompatibility, tunable biodegradability, and mechanical properties, silk fibroin hydrogel is an exciting material for localized drug delivery systems to decrease the therapy cost, decrease the negative side effects, and increase the efficiency of chemotherapy. However, the lack of remote stimuli response and active drug release behavior has yet to be analyzed comparatively. In this study, we developed magnetic silk fibroin (SF) hydrogel samples through the facile blending method, loaded with doxorubicin hydrochloride (DOX) and incorporated with different concentrations of iron oxide nanoparticles (IONPs), to investigate the presumable ability of controlled and sustained drug release under the various external magnetic field (EMF). The morphology and rheological properties of SF hydrogel and magnetic SF hydrogel were compared through FESEM images and rheometer analysis. Here, we demonstrated that adding magnetic nanoparticles (MNPs) into SFH decreased the complex viscosity and provided a denser porosity with a bigger pore size matrix structure, which allowed the drug to be released faster in the absence of an EMF. Release kinetic studies show that magnetic SF hydrogel could achieve controlled release of DOX in the presence of an EMF. Furthermore, the drug release from magnetic SF hydrogel decreased in the presence of a static magnetic field (SMF) and an alternating magnetic field (AMF), and the release rate decreased even more with the higher MNPs concentration and magnetic field strength. Subsequently, Wilms' tumor and human fibroblast cells were cultured with almost the same concentration of DOX released in different periods, and cell viability was investigated using MTT assay. MTT results indicated that the Wilms' tumor cells were more resistant to DOX than the human fibroblasts, and the IC50 values were calculated at 1.82 0.001 and 2.73 0.004 (μg/ml) for human fibroblasts and Wilms' tumor cells, respectively. Wilms' tumor cells showed drug resistance in a higher DOX concentration, indicating the importance of controlled drug delivery. These findings suggest that the developed magnetic SFH loaded with DOX holds excellent potential for intelligent drug delivery systems with noninvasive injection and remotely controlled abilities.

Enzymatic Crosslinked Silk Fibroin Hydrogel for Biodegradable Electronic Skin and Pulse Waveform Measurements.

The development of a portable, controllable, and environmentally friendly electronic skin (e-skin) is highly desirable; however, it presents a major challenge. Herein, a biocompatible, biodegradable, and easily usable hydrogel was designed and fabricated as e-skin to enable the transmission of information regarding the spatial pressure distribution. Silk fibroin (SF) was used as the hydrogel skeleton, which endowed the hydrogel with intelligent mechanical sensitivity. During its conditioning in weakly acidic media, the density of the enzymatic crosslink increased and a dense network was formed due to the formation of covalent/hydrogen bonds. Additionally, a conductive SF/polyvinyl alcohol (PVA) hybrid film was molded as a flexible electrode after graphite deposition. The above SF sensing unit based on SF hydrogels and SF/PVA hybrid films showed high strain sensitivity (4.78), fast responsiveness (<0.1 s), good cycling stability (≥10,000), excellent biocompatibility, and biodegradability. Importantly, a coplanar 8 × 8 pixel SF-based e-skin array was successfully fabricated and applied for 3D signal transmission of the object. The SF-based e-skin was capable of precisely tracking the changes in the pulse pressure, the movement of the finger joint, and the vibrations of the vocal cord. Therefore, the current findings provide a solid foundation for future studies exploring the next generation of electronic devices.

Nanosilicate-Reinforced Silk Fibroin Hydrogel for Endogenous Regeneration of Both Cartilage and Subchondral Bone.

Osteochondral defects are characterized by injuries to both cartilage and subchondral bone, which is a result of trauma, inflammation, or inappropriate loading. Due to the unique biological properties of subchondral bone and cartilage, developing a tissue engineering scaffold that can promote dual-lineage regeneration of cartilage and bone simultaneously remains a great challenge. In this study, a microporous nanosilicate-reinforced enzymatically crosslinked silk fibroin (SF) hydrogel is fabricated by introducing montmorillonite (MMT) nanoparticles via intercalation chemistry. In vitro studies show that SF-MMT nanocomposite hydrogel has improved mechanical properties and hydrophilicity, as well as the bioactivities to promote the osteogenic differentiation of bone marrow mesenchymal stem cells and maintain chondrocyte phenotype compared with SF hydrogel. Global proteomic analysis verifies the dual-lineage bioactivities of SF-MMT nanocomposite hydrogel, which are probably regulated by multiple signaling pathways. Furthermore, it is observed that the biophysical interaction of cells and SF-MMT nanocomposite hydrogel is partially mediated by clathrin-mediated endocytosis and its downstream processes. In vivo, the SF-MMT nanocomposite hydrogel effectively promotes osteochondral regeneration as evidenced by macroscopic, micro-CT, and histological evaluation. In conclusion, a functionalized SF-MMT nanocomposite hydrogel is developed with dual-lineage bioactivity for osteochondral regeneration, indicating its potential in osteochondral tissue engineering.

Intraarticularly injectable silk hydrogel microspheres with enhanced mechanical and structural stability to attenuate osteoarthritis

A silk fibroin (silk) hydrogel was prepared by using diglycidyl ether (BDDE), a chemical crosslinker commonly used to generate Food and Drug Administration (FDA)-approved hyaluronic acid (HA) medical products. The silk/BDDE hydrogels exhibited high elasticity (compressive modulus of 166 ± 15.0 kPa), anti-fatigue properties, and stable structure and mechanical strength in aqueous solution. Chemical crosslinking was conducted in a high concentration (9.3 M) of lithium bromide (LiBr) solution, a salt that is commonly used to dissolve degummed silk fibers during silk solubilization. The unfolded and extended structure of silk molecules with these reaction conditions, as well as the unique ionic environment provided by LiBr facilitated a high degree of crosslinking in the hydrogel. Similar hydrogels were not obtained when the silk was dissolved in other silk fiber-dissolving reagents (e.g., Ajisawa's, formic acid (FA)/LiBr, FA/CaCl2 solutions), likely because partially folded silk structures and the ionic conditions with these reagents were less favorable for the crosslinking reaction. Based on these findings, silk/BDDE hydrogel spheres were prepared using an oil/water (o/w) emulsification method and biocompatibility and biodegradation were evaluated in vivo, along with other silk gel control systems (e.g., enzyme-catalyzed di-tyrosine and pulverized silk/BDDE gel particles with irregular shapes). Histological and immunohistochemical analyses demonstrated that the silk/BDDE hydrogel spheres were biocompatible and served as a bio-lubricant to treat osteoarthritis (OA). The intra-articular injection of the gel spheres reduced pain as measured with OA rats, reduced cartilage damage and resisted the digestive environment in the articular cavity for extended time frames (>4 weeks), suggesting utility for pain relief and sustained drug release for future OA treatments.