Search

Continuous monitoring of biochemical analytes like oxygen is of interest in biomedicine to provide insight into physiology and health. Silk‐protein biomaterials are particularly useful as the scaffold material in oxygen sensors due to silk's unique amphiphilic chemistry, which promotes noncovalent stabilization of the protein and additives in aqueous environments. Silk films containing a water‐insoluble oxygen‐sensing chromophore, Pd (II) tetramethacrylated benzoporphyrin (PdBMAP), are evaluated as optical oxygen sensors in vitro and in vivo. These silk‐chromophore composites are stabilized by the self‐assembled, physically crosslinked protein network. The deaerated phosphorescence lifetime (τm,0 ≈300 µs) of the chromophore in vitro is quenched to 50% of its initial value at ≈31 µm dissolved oxygen, indicating sensing functionality within physiological ranges of oxygen. In vitro enzymatic degradation of the silk films with and without the chromophore is demonstrated. The silk‐chromophore composite films are cytocompatible in vitro, biocompatible in vivo upon implantation in rats, and displayed mechanical properties suitable for subcutaneous implantation. Further, the films maintain oxygen‐sensing function in vivo and demonstrate real‐time sensing capabilities throughout various physiological states (i.e., hyperoxia, normoxia, and hypoxia). [ABSTRACT FROM AUTHOR]

Abstract: Hydrogels are the focus of extensive research due to their potential use in fields including biomedical, pharmaceutical, biosensors, and cosmetics. However, the general weak mechanical properties of hydrogels limit their utility. Here, pristine silk fibroin (SF) hydrogels with excellent mechanical properties are generated via a binary‐solvent‐induced conformation transition (BSICT) strategy. In this method, the conformational transition of SF is regulated by moderate binary solvent diffusion and SF/solvent interactions. β‐sheet formation serves as the physical crosslinks that connect disparate protein chains to form continuous 3D hydrogel networks, avoiding complex chemical and/or physical treatments. The Young's modulus of these new BSICT–SF hydrogels can reach up to 6.5 ± 0.2 MPa, tens to hundreds of times higher than that of conventional hydrogels (0.01–0.1 MPa). These new materials fill the “empty soft materials' space” in the elastic modulus/strain Ashby plot. More remarkably, the BSICT–SF hydrogels can be processed into different constructions through different polymer and/or metal‐based processing techniques, such as molding, laser cutting, and machining. Thus, these new hydrogel systems exhibit potential utility in many biomedical and engineering fields. [ABSTRACT FROM AUTHOR]

The production of structural and functional materials with enhanced mechanical properties through the integration of soft and hard components is a common approach to Nature's material design. However, directly mimicking these optimized design routes in the lab for practical applications remains challenging. For example, graphene and silk are two materials with complementary mechanical properties that feature ultrahigh stiffness and toughness, respectively. Yet, no simple and controllable approach is developed to homogeneously integrate these two components into functional composites, mainly due to the hydrophobicity and chemical inertness of graphene. In this study, well-dispersed and highly stable graphene/silk fibroin (SF) suspension systems are developed, which are suitable for processing to fabricate polymorphic materials, such as films, fibers, and coatings. The obtained graphene/SF nanocomposites maintain the electronic advantages of graphene, and they also allow tailorable mechanical performance to form including ultrahigh stretchable (with a strain to failure to 611 ± 85%), or high strength (339 MPa) and high stiffness (7.4 GPa) material systems. More remarkably, the electrical resistances of these graphene/SF materials are sensitive to material deformation, body movement, as well as humidity and chemical environmental changes. These unique features promise their utility as wearable sensors, smart textiles, intelligent skins, and human-machine interfaces. [ABSTRACT FROM AUTHOR]

Elastomeric, robust, and biocompatible hydrogels are rare, while the need for these types of biomaterials in biomedical-related uses remains high. Here, a new family of genetically engineered silk-elastin-like proteins (SELPs) with encoded enzymatic crosslinking sites is developed for a new generation of stimuli-responsive yet robust hydrogels. Input into the designs is guided by simulation and realized via genetic engineering strategies. The avoidance of gamma irradiation or chemical crosslinking during gel fabrication, in lieu of an enzymatic process, expands the versatility of these new gels for the incorporation of labile proteins and cells. In the present study, the new SELP hydrogels offer sequence-dependent, reversible stimuli-responsive features. Their stiffness covers almost the full range of the elasticity of soft tissues. Further, physical modification of the silk domains provides a secondary control point to fine-tune mechanical stiffness while preserving stimuli-responsive features, with implications for a variety of biomedical material and device needs. [ABSTRACT FROM AUTHOR]

Functional vascularization is critical for the clinical regeneration of complex tissues such as kidney, liver, or bone. The immobilization or delivery of growth factors has been explored to improve vascularization capacity of tissue-engineered constructs; however, the use of growth factors has inherent problems such as the loss of signaling capability and the risk of complications including immunological responses and cancer. Here, a new method of preparing water-insoluble silk protein scaffolds with vascularization capacity using an all-aqueous process is reported. Acid is added temporally to tune the self-assembly of silk in the lyophilization process, resulting in water-insoluble scaffold formation directly. These biomaterials are mainly noncrystalline, offering improved cell proliferation than previously reported silk materials. These systems also have an appropriate softer mechanical property that could provide physical cues to promote cell differentiation into endothelial cells, and enhance neovascularization and tissue ingrowth in vivo without the addition of growth factors. Therefore, silk-based degradable scaffolds represent an exciting biomaterial option, with vascularization capacity for soft tissue engineering and regenerative medicine. [ABSTRACT FROM AUTHOR]

Elastomeric, fully degradable, and biocompatible biomaterials are rare, with current options presenting significant limitations in terms of ease of functionalization and tunable mechanical and degradation properties. A new method for covalently crosslinking tyrosine residues in silk proteins, via horseradish peroxidase and hydrogen peroxide, to generate highly elastic hydrogels with tunable properties, is reported. These materials offer tunable mechanical properties, gelation kinetics, and swelling properties. In addition, these new polymers withstand shear strains on the order of 100%, compressive strains greater than 70% and display stiffness between 200-10 000 Pa, covering a significant portion of the properties of native soft tissues. Molecular weight and solvent composition allow control of material mechanical properties over several orders of magnitude while maintaining high resilience and resistance to fatigue. Encapsulation of human bone marrow derived mesenchymal stem cells (hMSC) shows long term survival and exhibits cell-matrix interactions reflective of both silk concentration and gelation conditions. Further biocompatibility of these materials is demonstrated with in vivo evaluation. These new protein-based elastomeric and degradable hydrogels represent an exciting new biomaterials option, with a unique combination of properties, for tissue engineering and regenerative medicine. [ABSTRACT FROM AUTHOR]

The need for dynamic, elastomeric polymeric biomaterials remains high, with few options with tunable control of mechanical properties, and environmental responses. Yet the diversity of these types of protein polymers pursued for biomaterials-related needs remains limited. Robust high-throughput synthesis and characterization methods will address the need to expand options for protein-polymers for a range of applications. Here, a combinatorial library approach with high throughput screening is used to select specific examples of dynamic protein silk-elastin-like polypeptides (SELPs) with unique stimuli responsive features, including tensile strength and adhesion. Using this approach, 64 different SELPs with different sequences and molecular weights are selected out of over 2000 recombinant E. coli colonies. New understanding of sequence-function relationships with this family of proteins is gained through this combinatorial-screening approach and can provide a guide to future library designs. Further, this approach yields new families of SELPs to match specific material functions. [ABSTRACT FROM AUTHOR]

There is great need for soft biomaterials that match the stiffness of human tissues for tissue engineering and regeneration. Hydrogels are frequently employed for extracellular matrix functionalization and to provide appropriate mechanical cues. It is challenging, however, to achieve structural integrity and retain bioactive molecules in hydrogels for complex tissue formation that may take months to develop. This work aims to investigate mechanical and biochemical characteristics of silk hydrogels for soft tissue engineering, specifically for the nervous system. The stiffness of 1 to 8% silk hydrogels, measured by atomic force microscopy, is 4 to 33 kPa. The structural integrity of silk gels is maintained throughout embryonic chick dorsal root ganglion (cDRG) explant culture over 4 days whereas fibrin and collagen gels decrease in mass over time. Neurite extension of cDRGs cultured on 2 and 4% silk hydrogels exhibit greater growth than softer or stiffer gels. Silk hydrogels release <5% of neurotrophin-3 (NT-3) over 2 weeks and 11-day old gels show maintenance of growth factor bioactivity. Finally, fibronectin- and NT-3-functionalized silk gels elicit increased axonal bundling suggesting their use in bridging nerve injuries. These results support silk hydrogels as soft and sustainable biomaterials for neural tissue engineering. [ABSTRACT FROM AUTHOR]

In the field of regenerative medicine there is a need for scaffolds that support large, critically-sized tissue formation. Major limitations in reaching this goal are the delivery of oxygen and nutrients throughout the bulk of the engineered tissue as well as host tissue integration and vascularization upon implantation. To address these limitations, the development of a porous scaffold platform made from biodegradable silk protein that contains an array of vascular-like structures that extend through the bulk of the scaffold was previously reported. Here, the hollow channels play a pivotal role in enhancing cell infiltration, delivering oxygen and nutrients to the scaffold bulk, and promoting in vivo host tissue integration and vascularization. The unique features of this protein biomaterial system, including the vascular structures and tunable material properties, render this scaffold a robust and versatile tool for implementation in a variety of tissue engineering, regenerative medicine, and disease modeling applications. [ABSTRACT FROM AUTHOR]

Neural engineering provides promise for cell therapy by integrating the host brain with brain-machine-interface technologies in order to externally modulate functions. Long-term interfaces with the host brain remain a critical challenge due to insufficient graft cell survivability and loss of brain electrode sensitivity over time. Here, integrated neuron-electrode interfaces are developed on thin flexible and transparent silk films as brain implants. Mechanical properties and surface topography of silk films are optimized to promote cell survival and alignment of primary rat cortical cells. Compartmentalized neural cultures and co-patterned electrode arrays are incorporated on the silk films with built-in wire connections. Electrical stimulation via electrodes embedded in the films activated surrounding neurons to produce evoked calcium responses. In mice brains, silk film implants show conformal contact capable of modulating host brain cells with minimal inflammatory response and stable indwelling for weeks. The approach of combining cell therapy and brain electrodes could provide sustained functional interfaces with ex vivo control with spatial precision. [ABSTRACT FROM AUTHOR]

There is critical clinical demand for tissue-engineered (TE), 3D constructs for tissue repair and organ replacements. Current efforts toward this goal are prone to necrosis at the core of larger constructs because of limited oxygen and nutrient diffusion. Therefore, critically sized 3D TE constructs demand an immediate vascular system for sustained tissue function upon implantation. To address this challenge the goal of this project was to develop a strategy to incorporate microchannels into a porous silk TE scaffold that could be fabricated reproducibly using microfabrication and soft lithography. Silk is a suitable biopolymer material for this application because it is mechanically robust, biocompatible, slowly degrades in vivo, and has been used in a variety of TE constructs. Here, the fabrication of a silk-based TE scaffold that contains an embedded network of porous microchannels is reported. Enclosed porous microchannels support endothelial lumen formation, a critical step toward development of the vascular niche, while the porous scaffold surrounding the microchannels supports tissue formation, demonstrated using human mesenchymal stem cells. This approach for fabricating vascularized TE constructs is advantageous compared to previous systems, which lack porosity and biodegradability or degrade too rapidly to sustain tissue structure and function. The broader impact of this research will enable the systemic study and development of complex, critically-sized engineered tissues, from regenerative medicine to in vitro tissue models of disease states. [ABSTRACT FROM AUTHOR]

The reliability of chronic, brain-penetrating electrodes must be improved for these -neural recording technologies to be viable in widespread clinical applications. One approach to improving electrode reliability is to reduce the foreign body response at the probe-tissue interface. In this work, silk fibroin is investigated as a candidate material for fabricating mechanically dynamic neural probes with enhanced biocompatibility compared to traditional electrode materials. Silk coatings are applied to flexible cortical electrodes to produce devices that transition from stiff to flexible upon hydration. Theoretical modeling and in vitro testing show that the silk coatings impart mechanical properties sufficient for the electrodes to penetrate brain tissue. Further, it is demonstrated that silk coatings may reduce some markers of gliosis in an in vitro model and that silk can encapsulate and release the gliosis-modifying enzyme chondroitinase ABC. This work establishes a basis for future in vivo studies of silk-based brain-penetrating electrodes, as well as the use of silk materials for other applications in the central nervous system where gliosis must be controlled. [ABSTRACT FROM AUTHOR]

Animal silks, consisting of pure protein components, offer an extraordinary combination of strength, elongation, and toughness, exceeding most engineered materials. The secret to this success is their unique nanoarchitectures formed through the hierarchical self‐assembly of silk proteins. This natural process contrasts the production of artificial silk materials, which usually are directly constructed as bulk structures from silk fibroin (SF) molecules. A variety of fabrication strategies to control nanostructures of silks or to create functional materials from silk nanoscale building blocks have been developed in the recent years. These emerging fabrication strategies offer an opportunity to tailor the structure of SF at the nanoscale and provide a promising route to produce structurally and functionally optimized silk nanomaterials. Herein, the critical roles of silk nanoarchitectures in property and function of natural silk fibers are reviewed and the strategies of utilization of these silk nanobuilding blocks are outlined. Further, the state‐of‐the‐art techniques to create silk nanoarchitectures and to generate silk‐based nanocomponents are summarized. An effective approach to constructing sophisticated silk functional nanocomposites with promising applications in regenerative medicine, drug delivery, and optical and electronic device designs is provided. Further, such insights suggest templates to consider for other material systems. A critical summary of the state of the art in the field of design, fabrication, and function of silk‐based nanomaterials is provided in this review. The structure–property–function relationships of nanobuilding blocks in silk fibers and the emerging strategies for fabrication and use of natural and regenerated silk fibroin–based nanomaterials are highlighted. [ABSTRACT FROM AUTHOR]

A ubiquitous feature of natural silk fibers is the presence of well‐organized mesostructures, including microfibrils, nanofibrils, and nanoparticles. These mesoscale building blocks are typically well organized into sophisticated arrangements and contribute robust mechanical performance and functions as part of natural silk fibers. However, it remains a major challenge to directly isolate these mesostructures for engineering applications. Here, an environmentally friendly and scalable "partial dissolution and physical dispersion" strategy is developed to exfoliate silk fibers into different mesostructures, including microfibrils, nanofibrils, nanorods, and nanoparticles. On the basis of the advantages of these mesosilks in tunable sizes, sharp size distributions, high modulus, excellent redispersibility, as well as versatile processability, the applications of these mesosilks in electronic and environmental fields are further explored, including water treatment, recycling organic solvent, paper sensors, and nanofertilizers. These explorations open a new avenue for silk fiber applications while also providing a pathway to help address critical issues in electronic and environmental fields. An environmentally friendly and scalable "partial dissolution and physical dispersion" strategy is designed to exfoliate silk fibers into different mesostructures, including microfibrils, nanofibrils, nanorods, and nanoparticles. These silk mesostructures can be further assembled into polymorphic materials, showing promising applications in electronic and environmental fields. [ABSTRACT FROM AUTHOR]