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. We report 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. The tunable mechanical properties, gelation kinetics and swelling properties of these new protein polymers, in addition to their ability to 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 allowed 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) showed long term survival and exhibited cell-matrix interactions reflective of both silk concentration and gelation conditions. Further biocompatibility of these materials were 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.

Traditional nanofabrication techniques often require complex lithographic steps and the use of toxic chemicals. To move from the laboratory scale to large scales, nanofabrication should be carried out using alternative procedures that are simple, inexpensive and use non-toxic solvents. Recent efforts have focused on nanoimprinting and the use of organic resists (such as quantum dot-polymer hybrids, DNA and poly(ethylene glycol)), which still require, for the most part, noxious chemicals for processing. Significant advances have been achieved using 'green' resists that can be developed with water, but so far these approaches have suffered from low electron sensitivity, line edge roughness and scalability constraints. Here, we present the use of silk as a natural and biofunctional resist for electron-beam lithography. The process is entirely water-based, starting with the silk aqueous solution and ending with simple development of the exposed silk film in water. Because of its polymorphic crystalline structure, silk can be used either as a positive or negative resist through interactions with an electron beam. Moreover, silk can be easily modified, thereby enabling a variety of 'functional resists', including biologically active versions. As a proof of principle of the viability of all-water-based silk electron-beam lithography (EBL), we fabricate nanoscale photonic lattices using both neat silk and silk doped with quantum dots, green fluorescent proteins (GFPs) or horseradish peroxidase (HRP).

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In natural systems, directed self-assembly of structural proteins produces complex, hierarchical materials that exhibit a unique combination of mechanical, chemical and transport properties. This controlled process covers dimensions ranging from the nano- to the macroscale. Such materials are desirable to synthesize integrated and adaptive materials and systems. We describe a bio-inspired process to generate hierarchically defined structures with multiscale morphology by using regenerated silk fibroin. The combination of protein self-assembly and microscale mechanical constraints is used to form oriented, porous nanofibrillar networks within predesigned macroscopic structures. This approach allows us to predefine the mechanical and physical properties of these materials, achieved by the definition of gradients in nano- to macroscale order. We fabricate centimetre-scale material geometries including anchors, cables, lattices and webs, as well as functional materials with structure-dependent strength and anisotropic thermal transport. Finally, multiple three-dimensional geometries and doped nanofibrillar constructs are presented to illustrate the facile integration of synthetic and natural additives to form functional, interactive, hierarchical networks.

A custom-made co-flow capillary device is used to synthesize monodisperse silk fibroin micro- and submicron-spheres with diameters tunable over a wide range of sizes. A model drug release is examined and control of degradation kinetics is obtained by changing sphere diameter.

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Silk fibroin from the Bombyx mori caterpillar has been processed into many material forms, with potential applications in areas ranging from optoelectronics to tissue engineering. As a hydrogel, silk fibroin has been engineered as a substrate for the regeneration of soft tissues where hydration and mechanical compatibility are necessary. Current fabrication of silk fibroin hydrogels produces microstructured materials that lack transparency and limits the ability to fully exploit the hydrogel form. Transparency is the main characteristic of some human tissues (e.g., cornea) where silk fibroin in the film format has shown potential as scaffolding material, however, lacking the necessary hydration and successful attachment of cells without biochemical functionalization. Additionally, detection using light is an important method to translate information for instruction, sensing, and visualization of biological entities and light sensitive molecules. Here, we introduce a method for the fabrication of transparent silk hydrogels by driving the formation of nanostructures in the silk fibroin material. These nanostructures are formed by exposing silk solution (concentration below 15 mg/mL) to organic solvents that induce the amorphous to crystalline transition of the protein and indeed the sol–gel transition of the material. We have also explored a process to modulate the mechanical properties of silk fibroin hydrogel within the physiological range by controlling the amount of metal ions present in the protein structure. Nanostructured silk fibroin hydrogels are biocompatible and allow for attachment and proliferation of human dermal fibroblasts without any biochemical functionalization. In addition, seeding of human cornea epithelial cells (HCECs) on the hydrogel surface results in the formation of an epithelium, which does not alter the gels’ transparency and shows biological properties that challenge the performances of HCECs seeded in collagen hydrogels, the current standard material for the engineering of corneal tissue.

High resolution bioimaging is not only critical to the study of cellular structures and processes but it also has important applications in drug delivery and therapeutics. Fluorescent nanodiamonds (NDs) are excellent candidates for long-term bioimaging and tracking of biological structures at the nanoscale. Encapsulating NDs in natural biopolymers like silk fibroin (SF) widens their biomedical applications. Here we report the synthesis, structural and optical characterization of ND incorporated SF nanospheres. The photoluminescence from optical defects within the NDs is found to increase when encapsulated in the SF spheres. The encapsulated NDs are applied in vitro to investigate the intracellular mobility compared to bare NDs. The diffusion rate of encapsulated NDs is shown to improve due to SF coating. These ND-SF spheres are envisioned as highly suitable candidates for bioinjectable imaging and drug release carriers for targeted drug delivery applications.