Silk-reinforced composites (SRCs) manifest the unique properties of silkworm silk fibers, offering enhanced mechanical strength, biocompatibility, and biodegradability. These composites present an eco-friendly alternative to conventional synthetic materials, with applications expanding beyond biomedical engineering, flexible electronics, and environmental filtration. This review explores the diverse forms of silkworm silk fibers including fabrics, long fibers, and nanofibrils, for functional composites. It highlights advancements in composite design and processing techniques that allow precise engineering of mechanical and functional performance. Despite substantial progress, challenges remain in making optimally functionalized SRCs with multi-faceted performance and understanding the mechanics for reverse-design of SRCs. Future research should focus on the unique sustainable, biodegradable and biocompatible advantages and embrace advanced processing technology, as well as artificial intelligence-assisted material design to exploit the full potential of SRCs. This review on SRCs will offer a foundation for future advancements in multifunctional and high-performance silk-based composites.

Spider silk spidroins, nature's advanced polymers, have long hampered efficient in vitro production due to their considerable size, repetitive sequences, and aggregation-prone nature. This study harnesses the power of a cell-free protein synthesis (CFPS) system, presenting the first successful in vitro production and detailed characterization of recombinant spider silk major ampullate spidroins (MaSps) utilizing a reformulated and optimizedEscherichia coli based CFPS system. Through systematic optimization, including cell strain engineering via knockout generation, energy sources, crowding agents, and amino acid supplementation, we effectively addressed the specific challenges associated with recombinant spidroin biosynthesis, resulting in high yields of 0.61 mg/mL for MaSp1 (69 kDa) and 0.52 mg/mL for MaSp2 (73 kDa). The synthesized spidroins self-assembled into micelles, facilitating efficient purification compared to in vivo methods, and were further processed into prototype silk fiber products. The functional characterization demonstrated that the purified spidroins maintain essential natural properties, such as phase separation and fiber formation triggered by pH and ions. This tailored CFPS platform also facilitates versatile cosynthesis and serves as an accessible platform for studying the supramolecular coassembly and dynamic interactions among spidroins. This CFPS platform offers a viable alternative to conventional in vivo methods, facilitating innovative approaches for silk protein engineering and biomaterial development in a high-throughput, efficient manner.

This review is the first to offer a comprehensive exploration of the complex interplay between silk polymorphism, its unique self-assembly properties, and its versatility in forming various silk-based emulsions, with a particular emphasis on the interfacial dynamics at oil/water interfaces. Silk demonstrates remarkable potential as a stabilizing agent through diverse stabilization mechanisms, resulting in the formation of distinct emulsion types. Three primary classes of silk-based emulsions are highlighted: (1) Film-stabilized emulsions, where silk in solution forms an elastic, gel-like 2D film at the oil/water interface; (2) Pickering emulsions, achieved by the adsorption of silk nano-assemblies onto oil droplets; and (3) Emulsion-filled gels, where oil droplets are encapsulated within a gel network using silk's inherent gelling properties. Additionally, the interfacial behaviour of silk-encompassing interfacial rheology and conformational changes-at the oil/water interface is discussed. A detailed analysis of the mechanisms driving silk self-assembly, along with the factors influencing this process, is also provided. The complexity of silk's self-assembly adds significant challenges to the classification of silk-based emulsions and a thorough understanding of their stabilizing mechanisms. Finally, this review addresses existing knowledge gaps and offers potential future directions for advancing research in this area.

Nature's fibrous proteins, such as α-keratin, achieve remarkable mechanical properties by undergoing strain-induced α-to-β conformational transitions. Inspired by these materials, we report a strategy for designing synthetic polypeptides that undergo similar transformations at elevated temperatures far exceeding keratin's operational range. By employing helix-confined ring-opening polymerization (ROP) of N-carboxyanhydrides (NCAs) initiated by a short poly(γ-benzyl-l-glutamate) (PBLG) precursor, we synthesized poly(O-benzyl-l-serine) (PBLS) chains that adopt an α-helical structure yet transition into β-sheets upon heating. Compression molding at carefully chosen temperatures drives PBLS segments into an α-β intermediate state, characterized by relaxed intrachain hydrogen bonds and a hexagonal packing arrangement. Under mechanical strain, these intermediate states convert in situ into β-sheets, producing significant strain-hardening well below the spontaneous α-to-β temperature threshold. This approach extends to polypeptides bearing different side chains, such as poly(S-benzyl-l-cysteine), demonstrating robust mechanical reinforcement across a wide temperature window up to ∼ 200 °C. In situ synchrotron X-ray analysis confirms that chain alignment, β-sheet formation, and domain growth occur stepwise during deformation. By harnessing the intermediate states and the supramolecular cooperativity conferred by compression-molded films, our method provides a versatile platform for developing next-generation polypeptide materials with tunable mechanical resilience and responsiveness─surpassing the temperature limitations of natural fibrous proteins and enabling potential applications demanding broad-temperature mechanical adaptability.

Chitin and chitosan-based nanofibers (ChNFs), derived from renewable sources, have emerged as promising biomaterials due to their unique properties such as high surface area, porosity, biocompatibility, and biodegradability. This review provides a comprehensive overview of ChNF extraction and synthesis, focusing on both top-down and bottom-up approaches. A comparative analysis of these methods is presented, highlighting the challenges, opportunities, environmental impact, cost-effectiveness, and quality consistency associated with each. The advantages of ChNFs over similar nanomaterials are elucidated, emphasizing their diverse applications in biomedical and environmental fields. Biomedical applications include drug delivery, tissue engineering, cancer treatment, wound healing, and biosensing. Environmental applications encompass water treatment, air filtration, agriculture, and biodegradable packaging. Despite their potential, challenges remain, including low solubility, unstable mechanical properties, and inconsistent quality, which limit their widespread use. This review also examines recent advancements in ChNF research, aiming to guide the development of efficient and environmentally friendly synthesis methods. By encouraging innovation in ChNF-based nanotechnologies, this research contributes to a more sustainable future.

We report the successful moss-produced recombinant spider silk key protein component containing both the N- and the C-terminal domain. Spider dragline silk stands out as a remarkable biomaterial, representing one of nature's toughest fibres. Its strength rivals that of many synthetic fibres used commercially, rendering it applicable across various industrial and medical domains. However, its widespread utilisation requires cost-effective mass production. Biotechnology presents a promising avenue for achieving this goal, particularly through the production of recombinant dragline silk proteins in transgenic plant systems. This study aimed to assess the feasibility of producing one key protein component of dragline silk, MaSp1, from the western black widow spider, Latrodectus hesperus, the protein LhMaSp1, in the moss Physcomitrella (Physcomitrium patens). Here, we present the successful recombinant production of spider silk protein containing both the N- and C-terminal domains of LhMaSp1 in moss cells. The production of recombinant LhMaSp1 protein in Physcomitrella was performed in shake flasks and in five-litre photobioreactors and the correct synthesis of LhMaSp1 was proven via mass spectrometry. We estimate that the yield of recombinant spider silk protein in Physcomitrella bioreactors is above 0.82 mg/g fresh weight.

Dry eye disease (DED) is a multifactorial illness affecting tears and the ocular surface. The neurokinin 1 receptor (NK1R) is a target for controlling T helper 17 (Th17) and regulatory T cell (Treg) imbalances. This work creates a silk fibroin (SF) nanoparticle hydrogel that targets NK1R with CP-99,994 (CP). Combining CP and SF to generate stable nanoparticles while integrating a flexible hydrogel material results in a sustained-release ophthalmic drop formulation (SF@CP@Gel), which provides a long-lasting ocular formulation with anti-inflammatory and reparative properties. SF@CP@Gel could maintain a stable CP concentration for 25 h with detectable biological activity. The cell counting kit-8 and 2,7-DHL-DA results reveal that SF@CP@Gel has no cytotoxic effect on human corneal epithelial cells (HCECs) and decreases the reactive oxygen species level in oxidatively damaged HCECs. Cell scratch assays demonstrate that SF@CP@Gel can greatly increase HCEC migration and proliferation within 24 h. Furthermore, in vivo therapy with topical SF@CP@Gel twice daily markedly reduce clinical symptoms by reducing the amount of pathogenic Th17 cells while efficiently restoring Treg activity. In summary, this work reveals that SF@CP@Gel might attenuate DED by inhibiting NK1R-mediated SP signaling and thereby modulating the Th17/Treg ratio, a potential anti-inflammatory and repair treatment method for DED.

The remarkable toughness (>70 MJ m-3) of silkworm silk is largely attributed to its hierarchically arranged nanofibrillar nanostructure. Recreating such tough fibers through artificial spinning is often challenging, in part because degummed, dissolved silk is drastically different to the unspun native feedstock found in the spinning gland. The present work demonstrates a method to dissolve silk without degumming to produce a solution containing undegraded fibroin and sericin. This solution exhibits liquid-liquid phase separation above 10% (wt/wt), a behavior observed in the silk gland but not in degummed silk solutions to date. This partitioning enhances the stability of the undegummed solution, delaying gelation two-fold compared with degummed silk at the same concentration. When spun under identical conditions, undegummed solutions produces fibers 8× stronger and 218× tougher than degummed silk feedstocks. Through ultrasonication, undegummed wet spun fibers are seen to possess hierarchical structure of densely packed ≈20 nm nanofibrils, similar to native silks, although completely absent from fibers wet-spun from degummed silk solutions. This work demonstrates that the preservation of molecular weight, presence of sericin and stimulation of liquid-liquid phase separation underpin a new pathway to recreate a hierarchical fiber with structures akin to native silk.

Sericin has been characterized as demonstrating a variety of bioactivities, establishing it as a valuable resource for biomedical and pharmaceutical applications. The diverse biological activities of sericin are likely linked to its unique biochemical composition and properties. This study aimed to assess the effect of origin, seasonality, and amino acid composition on the bioactivity of sericin samples from two Portuguese regions compared to commercial sericin. The amino acid profile was analyzed using HPLC-FLD. Moreover, several bioactivities were assessed through in vitro assays, including antiproliferative effects, cell migration, antimicrobial activity, anticoagulant properties, antioxidant capacity, and anti-inflammatory effects. The results obtained in this work revealed that the origin and season affect the sericin amino acid profile. In its pure state, sericin exhibited a low content of free amino acids, with tyrosine being the most abundant (53.42-84.99%). In contrast, hydrolyzed sericin displayed a higher amino acid content dominated by serine (54.05-59.48%). Regarding bioactivities, the sericin tested did not demonstrate antioxidant or anti-inflammatory potential in the conducted tests. Notwithstanding, it showed antiproliferative activity in contact with human tumor cell lines at a minimum concentration of 0.52 mg/mL. Regarding antimicrobial activity, sericin had the capacity to inhibit the growth of the bacteria and fungi tested at concentrations between 5 and 10 mg/mL. Additionally, sericin demonstrated its capacity to prolong the coagulation time in pooled human plasma, indicating a potential anticoagulant activity. In addition, the origin and season also revealed their impact on biological activities, and sericin collected in Bragança in 2021 (S3) and 2022 (S4) demonstrated higher antiproliferative, antibacterial, and anticoagulant potentials. Future studies should focus on optimizing sericin's bioactivities and elucidating its molecular mechanisms for clinical and therapeutic applications.

We explore the development of biodegradable poly(vinyl alcohol) (PVA) films loaded with silk fibroin (SF) functionalized with copaiba oleoresin (SFCO) for potential use in active food packaging. The films were characterized, showing significant improvements in both their physicochemical and nanomorphological properties. Films containing 10% SFCO exhibited superior mechanical strength, with a Young modulus of 145 MPa and an elongation at break of 385%, compared to the control film with 42 MPa and 314%, respectively. The films also demonstrated barrier properties, with water vapor transmission rates (WVTRs) as low as 25.95 g/h·m2. Antimicrobial activity against Staphylococcus aureus and Escherichia coli was significantly improved, showing inhibition zones of up to 10 ± 1 mm and a minimum inhibitory concentration (MIC) of 100 µg∙mL-1. Three-dimensional nanomorphological analysis via atomic force microscopy (AFM) showed increased roughness in films with higher SFCO content, with root mean square (RMS) roughness values ranging from 2.70 nm to 11.5 nm. These results highlight the potential of SFCO-loaded PVA films as robust, eco-friendly alternatives to conventional packaging materials. They provide improved mechanical and antimicrobial properties, essential for extending the shelf life of perishable foods and advancing sustainability in the packaging industry.

Silk-fibroin hydrogels have gained considerable attention in recent years for their versatile biomedical applications. The physical properties of a complex hydrogel, comprising silk fibroin and riboflavin, surpass those of the silk fibroin-hydrogel without additives. This study investigates silk fibroin-riboflavin (silk-RIB) hydrogel at the atomistic level to uncover molecular structures and chemical characteristics specific to silk fibroin and riboflavin molecules in an aqueous medium. The interplay between hydrophilic riboflavin and hydrophobic silk fibroin polymers facilitates the formation of solubilized silk fiber, which subsequently evolves into a nano-scale hydrogel over time. Eventually, the interlinked RIB stacks form a scaffold that not only accommodates silk fibroin aggregates but also encloses water pockets, preserving the moisture level and enhancing the thermal conductivity of the hydrogel. To explore water retention properties and the role of ions, two sets of simulations of semi-hydrated hydrogel in the presence and absence of ions are conducted. The presence of ions significantly influences the dynamics of RIB and silk fibroin. Favorable interactions with the ions impede the unrestricted diffusion of these larger molecules, potentially leading to a stable structure capable of retaining water for a prolonged duration. The complete removal of water results in further shrinkage of the anhydrous silk-RIB hydrogel or xerogel (XG), yet its porosity and structural integrity remain intact. These findings offer valuable insights into the behavior of silk fibroin hydrogel and XG, paving the way for materials engineering in aqueous environments to develop biomedical devices with customized functional properties.

The increasing prevalence of electronic devices has led to a significant rise in electronic waste (e-waste), necessitating the development of sustainable materials for flexible electronics. In this study, silk fibroin ionic touch screen (SFITS) is introduced, a new platform integrating natural silk fibroin (SF) with ionic conductors to create highly elastic, environmentally stable, and multifunctional touch interfaces. Through a humidity-induced crystallization strategy, the molecular structure of SF is precisely controlled to achieve a balanced combination of mechanical strength, electrical conductivity, and biodegradability. The assembly and operational reliability of SFITS are demonstrated under various environmental conditions, along with their reusability through green recycling methods. Additionally, the intelligent design and application of SFITS are explored by incorporating Internet of Things (IoT) and artificial intelligence (AI) technologies. This integration enables real-time touch sensing, handwriting recognition, and advanced human-computer interactions. The versatility of SFITS is further exemplified through applications in remote control systems, molecular model generation, and virtual reality interfaces. The findings highlight the potential of sustainable ionic conductors in next-generation flexible electronics, offering a path toward greener and more intelligent device designs.

High-performance silk fibers, with their eco-friendly degradability and renewability, have long captivated researchers as an alternative to synthetic fibers. Spider dragline silk, renowned for its exceptional strength (>1 GPa), has an extremely low yield, hindering its widespread use. While domesticated silkworms (Bombyx mori) can produce silk fibers industrially, their moderate strength (≈0.5 GPa) pales in comparison to the formidable spider dragline silk. In this study, naturally produced strong silkworm silk fibers are reported with a tensile strength of ≈1.2 GPa achieved through combining feeding carbon nanotubes (CNTs) to silkworms and in situ forced reeling for alignment. Molecular dynamics simulations confirm the interaction between the CNTs and silk fibroin, while the forced reeling process aligns these reinforcing fillers and the silk fibroin β-sheet nanocrystals along the fiber axis. Structural analysis reveals a significant enhancement in the content and alignment of β-sheet nanocrystals within the silk fibers, accounting for their superior mechanical properties, including tensile strength of ≈1.2 GPa and Young's modulus of 24.4 GPa, surpassing various types of silkworm silk and spider silk. This advancement addresses the historical trade-off between the strength and scalability of silk, potentially paving the way for eco-friendly, biodegradable, and renewable alternatives to synthetic fibers in a variety of applications.

Many studies try to comprehend and replicate the natural silk spinning process due to its energy-efficient and eco-friendly process. In contrast to spider silk, the mechanisms of how silkworm silk fibroin (SF) undergoes liquid-liquid phase separation (LLPS) concerning the various environmental factors in the silk glands or how the SF coacervates transform into fibers remain unexplored. Here, we show that calcium ions, among the most abundant metal ions inside the silk glands, induce LLPS of SF under macromolecular crowded conditions by increasing both hydrophobic and electrostatic interactions between SF. Furthermore, SF coacervates assemble and further develop into fibrils under acidification and shear force. Finally, we prepare SF fiber using a pultrusion-based dry spinning, mirroring the natural silk spinning system. Unlike previous artificial spinning methods requiring concentrated solutions or harsh solvents, our process uses a less concentrated aqueous SF solution and minimal shear force, offering a biomimetic approach to fiber production.

Spider silk, known for its exceptional tensile strength, extensibility, and toughness, continues to inspire advancements in polymer and materials science. Despite extensive research, synthesizing materials that encompass all these properties remains a significant challenge. This study addresses this challenge by developing high molecular-weight multiblock synthetic copolypeptides that mimic the hierarchical structure and mechanical properties of spider silk. Using autoaccelerated ring-opening polymerization of N-carboxyanhydrides, we synthesized copolypeptides featuring transformable β-sheet blocks. These blocks retain a helical structure during synthesis but transition into β-sheet nanocrystals in situ during solvent-free thermal mechanical processing. Compression molding was employed to induce hierarchical ordering within the copolypeptide films, resulting in a solid "liquid crystalline" structure that undergoes a temperature-induced α-to-β structural transformation. This transformation integrates β-sheet nanocrystals throughout the helical block matrix, significantly enhancing the material's mechanical performance. Our innovative synthesis and processing strategy, which involves alternating sequences of α-helical and β-sheet blocks with various β-sheet-forming NCAs, enables the customization of diverse mechanical characteristics. These advancements not only deepen our understanding of the fundamental design principles of spider silk but also pave the way for a new generation of high-performance, silk-inspired synthetic copolypeptides with broad application potential.

Water-stable proteins may offer a new field of applications in smart materials for buildings and infrastructures where hydraulic reactions are involved. In this study, cement mortars modified through water-soluble silk fibroin (SF) are proposed. Water-soluble SF obtained by redissolving SF films in phosphate buffer solution (PBS) showed the formation of a gel with the β sheet features of silk II. Electrical measurements of SF indicate that calcium ions are primarily involved in the conductivity mechanism. By exploiting the water solubility properties of silk II and Ca2+ ion transport phenomena as well as their trapping effect on water molecules, SF provides piezoresistive and piezocapacitive properties to cement mortars, thus enabling self-sensing of mechanical strain, which is quite attractive in structural health monitoring applications. The SF/cement-based composite introduces a capacitive gauge factor which surpasses the traditional resistive gauge factor reported in the literature by threefold. Cyclic voltammetry measurements demonstrated that the SF/cement mortars possessed memcapacitive behavior for positive potentials near +5 V, which was attributed to an interfacial charge build-up modulated by the SF concentration and the working electrode. Electrical square-biphasic excitation combined with cyclic compressive loads revealed memristive behavior during the unloading stages. These findings, along with the availability and sustainability of SF, pave the way for the design of novel multifunctional materials, particularly for applications in masonry and concrete structures.

Liquid-liquid phase separation (LLPS) of proteins can be considered an intermediate solubility regime between disperse solutions and solid fibers. While LLPS has been described for several pathogenic amyloids, recent evidence suggests that it is similarly relevant for functional amyloids. Here, we review the evidence that links spider silk proteins (spidroins) and LLPS and its role in the spinning process. Major ampullate spidroins undergo LLPS mediated by stickers and spacers in their repeat regions. During spinning, the spidroins droplets shift from liquid to crystalline states. Shear force, altered ion composition, and pH changes cause micelle-like spidroin assemblies to form an increasingly ordered liquid-crystalline phase. Interactions between polyalanine regions in the repeat regions ultimately yield the characteristic β-crystalline structure of mature dragline silk fibers. Based on these findings, we hypothesize that liquid-liquid crystalline phase separation (LLCPS) can describe the molecular and macroscopic features of the phase transitions of major ampullate spidroins during spinning and speculate whether other silk types may use a similar mechanism to convert from liquid dope to solid fiber.

As a result of their hierarchical structure and biological processing, silk fibers rank among nature's most remarkable materials. The biocompatibility of silk-based materials and the exceptional mechanical properties of certain fibers has inspired the use of silk in numerous technical and medical applications. In recent years, computational modeling has clarified the relationship between the molecular architecture and emergent properties of silk fibers and has demonstrated predictive power in studies on novel biomaterials. Here, we review advances in modeling the structure and properties of natural and synthetic silk-based materials, from early structural studies of silkworm cocoon fibers to cutting-edge atomistic simulations of spider silk nanofibrils and the recent use of machine learning models. We explore applications of modeling across length scales: from quantum mechanical studies on model peptides, to atomistic and coarse-grained molecular dynamics simulations of silk proteins, to finite element analysis of spider webs. As computational power and algorithmic efficiency continue to advance, we expect multiscale modeling to become an indispensable tool for understanding nature's most impressive fibers and developing bioinspired functional materials.

Recombinantly produced collagens present a sustainable, ethical, and safe substitute for collagens derived from natural sources. However, controlling the folding of the recombinant collagens, crucial for replicating the mechanical properties of natural materials, remains a formidable task. Collagen-like proteins from willow sawfly are relatively small and contain no hydroxyprolines, presenting an attractive alternative to the large and post-translationally modified mammalian collagens. Utilizing CD spectroscopy and analytical ultracentrifugation, we demonstrate that recombinant willow sawfly collagen assembles into collagen triple helices in a concentration-dependent manner. Interestingly, we observed that the lower concentration threshold for the folding can be overcome by freezing or adding crowding agents. Microscopy data show that both freezing and the addition of crowding agents induce phase separation. We propose that the increase in local protein concentration during phase separation drives the nucleation-step of collagen folding. Finally, we show that freezing also induces the folding of recombinant human collagen fragments and accelerates the folding of natural bovine collagen, indicating the potential to apply phase separation as a universal mechanism to control the folding of recombinant collagens. We anticipate that the results provide a method to induce the nucleation of collagen folding without any requirements for genetic engineering or crosslinking.

Nanopatterning on biomaterials has attracted significant attention as it can lead to the development of biomedical devices capable of performing diagnostic and therapeutic functions while being biocompatible. Among various nanopatterning techniques, electron-beam lithography (EBL) enables precise and versatile nanopatterning in desired shapes. Various biomaterials are successfully nanopatterned as bioresists by using EBL. However, the use of high-energy electron beams (e-beams) for high-resolutive patterning has incorporated functional materials and has caused adverse effects on biomaterials. Moreover, the scattering of electrons not absorbed by the bioresist leads to proximity effects, thus deteriorating pattern quality. Herein, EBL-based nanopatterning is reported by inducing molecular degradation of amorphous silk fibroin, followed by selectively inducing secondary structures. High-resolution EBL nanopatterning is achievable, even at low-energy e-beam (5 keV) and low doses, as it minimizes the proximity effect and enables precise 2.5D nanopatterning via grayscale lithography. Additionally, integrating nanophotonic structures into fluorescent material-containing silk allows for fluorescence amplification. Furthermore, this post-exposure cross-linking way indicates that the silk bioresist can maintain nanopatterned information stored in silk molecules in the amorphous state, utilizing for the secure storage of nanopatterned information as a security patch. Based on the fabrication technique, versatile biomaterial-based nanodevices for biomedical applications can be envisioned.

Silk-producing animals use spigots to generate natural silk fibers for various purposes. These natural looms must be able to withstand prolonged silk extrusion. To gain insight into the functional basis of spigots, we report on the structural design of the spigot of the silkworm Bombyx mori. The B. mori spigot exhibits a unique triple-ridged strip surface structure, consisting of cuticle proteins, resilin, chitin, and metal ions (such as K and Ca). This multi-microstructure endows the spigot with superior hierarchical mechanical properties, enabling it to function as a spinning tool for silk formation, thereby influencing the structure and performance of the silk. These findings demonstrate new pathways for achieving specialized functions in confined spaces, providing theoretical support for understanding the natural spinning mechanism and inspiring new directions for developing innovative biomimetic materials.

Silk fibroin is a fiber-forming protein derived from the thread of Bombyx mori silkworm cocoons. This biocompatible protein, under the kosmotropic influence of potassium phosphate, can undergo supramolecular self-assembly driven by a random coil to β-sheet secondary structure transition. By leveraging concurrent nonspecific adsorption and self-assembly of silk fibroin, we demonstrate an interfacial phenomenon that yields adherent, defect-free nanothin protein coatings that grow continuously in time, without observable saturation in mass deposition. This noncovalent growth of silk fibroin coatings is a departure from traditionally studied protein adsorption phenomena, which generally yield adsorbed layers that saturate in mass with time and often do not completely cover the surface. Here, we explore the fundamental mechanisms of coating growth by examining the effects of coating solution parameters that promote or inhibit silk fibroin self-assembly. Results show a strong dependence of coating kinetics and structure on solution pH, salt species, and salt concentration. Moreover, coating growth was observed to occur in two stages: an early stage driven by protein-surface interactions and a late stage driven by protein-protein interactions. To describe this phenomenon, we developed a kinetic adsorption model with Langmuir-like behavior at early times and a constant steady-state growth rate at later times. Structural analysis by FTIR and photoinduced force microscopy show that small β-sheet-rich structures serve as anchoring sites for absorbing protein nanoaggregates, which is critical for coating formation. Additionally, β-sheets are preferentially located at the interface between protein nanoaggregates in the coating, suggesting their role in forming stable, robust coatings.

This is a Perspective on nature as a story-teller, where inputs of evolution drove the remarkable protein designs found in silks. This selection process has resulted in silk materials with novel chemistry and properties to support organism survival in nature, yet with newfound utility in everything from comic books and automobiles to medicine. With growing global concerns related to environmental health, silks also serve as an invaluable instructional guide to the future of sustainable material designs.

The endowment of guided bone regeneration (GBR) membranes with the ability to activate the endogenous regenerative capability of bone to regenerate bone defects is of clinical significance. Herein we explored the preparation of the calcium compound (CC) (calcium sulfate (CaSL), calcium hydrophosphate (CaHP), or tricalcium phosphate (TCaP)) loaded ultrathin silk fibroin (SF)/gelatin (G) fibre membranes via electrospinning as the GBR membranes to regenerate the calvarial bone defects. The in vitro experiments demonstrated that the CaSL-loaded ultrathin fibrous membranes could simulate optimally the bone remodelling microenvironment in comparison with the CaHP- and TCaP-loaded fibrous membranes, displaying the highest activity to regulate the migration, proliferation, and differentiation of mesenchymal stem cells (MSCs). Also, the in vivo experiments demonstrated that the CaSL-loaded fibrous membranes presented the highest intrinsic osteoinduction to guide in situ regeneration of bone. Furthermore, the in vivo experiments demonstrated that the as-prepared composite fibrous membranes possessed good degradability. In summary, our results suggested that the CaSL-loaded fibrous membranes with high intrinsic osteoinduction and good degradability have potential to translate into clinical practice.

Organohydrogels (OHGs) are intriguing materials due to their unique composition of both hydrophilic and hydrophobic domains. This antagonistic nature endows the OHGs with several remarkable properties, making them highly versatile for various applications. We present here a simple and inexpensive approach to fabricate silk fibroin (SF)-based OHGs with multistage switching mechanics and viscoelasticity. The continuous hydrophilic phase of the OHG precursor consists of an aqueous SF solution, while the hydrophobic droplet phase consists of a crystallizable n-octadecyl acrylate (C18A) monomer and several long-chain saturated hydrocarbons (HCs) with various chain lengths between 14 and 32 carbon atoms, namely, n-tetradecane, n-octadecane, n-docosane, n-dotriacontane, and 1-docosanol. After the addition of a C18A/HC mixture containing Irgacure photoinitiator into the continuous aqueous SF phase under stirring, a stable oil-in-water emulsion was obtained, which was then photopolymerized at 23 ± 2 °C to obtain nonswelling OHGs with multiple-shape-memory behavior. By changing the chain length and mass proportion of HCs, a series of OHGs with tunable transition temperatures could be obtained, meeting various applications. OHGs containing dimer, trimer, and quadruple combinations of in situ-formed poly(C18A) and HC microinclusions exhibit effective triple- or quintuple-shape memory whose shape-recovery temperatures could be adjusted over a wide range, e.g., between 7 and 70 °C.

Water-based processing plays a crucial role in high technology, especially in electronics, material sciences and life sciences, with important implications in the development of high-quality reliable devices, fabrication efficiency, safety and sustainability. At the micro- and nanoscale, water is uniquely enabling as a bridge between biological and technological systems. However, new approaches are needed to overcome fundamental challenges that arise from the high surface tension of water, which hinders wetting and, thus, fabrication at the bio-nano interface. Here we report the use of silk fibroin as a surfactant to enable water-based processing of nanoscale devices. Even in minute quantities (for example, 0.01 w/v%), silk fibroin considerably enhances surface coverage and outperforms commercial surfactants in precisely controlling interfacial energy between water-based solutions and hydrophobic surfaces. This effect is ascribed to the amphiphilic nature of the silk molecule and its adaptive adsorption onto substrates with diverse surface energy, facilitating intermolecular interactions between unlikely pairs of materials. The approach's versatility is highlighted by manufacturing water-processed nanodevices, ranging from transistors to photovoltaic cells. Its performance is found to be equivalent to analogous vacuum-processed devices, underscoring the utility and versatility of this approach for water-based nanofabrication.

Ultrasound-assisted regulation of biomaterial properties has attracted increasing attention due to the unique reaction conditions induced by ultrasound cavitation. In this study, we explored the fabrication of wild tussah silk nanofiber membranes via ultrasound spray spinning from an ionic liquid system, characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), X-ray powder diffraction (XRD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), atomic force microscopy (AFM), water contact angle, cytocompatibility tests, and enzymatic degradation studies. We investigated the effects of ultrasound propagation in an ionic liquid on the morphology, structure, thermal and mechanical properties, surface hydrophilicity, biocompatibility, and biodegradability of the fabricated fibers. The results showed that as ultrasound treatment time increased from 0 to 60 min, the regenerated silk fiber diameter decreased by 0.97 μm and surface area increased by 30.44 μm2, enhancing the fiber surface smoothness and uniformity. Ultrasound also promoted the rearrangement of protein molecular chains and transformation of disordered protein structures into β-sheets, increasing the β-sheet content to 54.32 %, which significantly improved the materials' thermal stability (with decomposition temperatures rising to 256.38 °C) and mechanical properties (elastic modulus reaching 0.75 GPa). In addition, hydrophilicity, cytocompatibility, and biodegradability of the fiber membranes all improved with longer ultrasound exposure, highlighting the potential of ultrasound technology in advancing the properties of natural biopolymers for applications in sustainable materials science and tissue regeneration.

Silk fibers' unique mechanical properties have made them desirable materials, yet their formation mechanism remains poorly understood. While ions are known to support silk fiber production, their exact role has thus far eluded discovery. Here, we use cryo-electron microscopy coupled with elemental analysis to elucidate the changes in the composition and spatial localization of metal ions during silk evolution inside the silk gland. During the initial protein secretion and storage stages, ions are homogeneously dispersed in the silk gland. Once the fibers are spun, the ions delocalize from the fibroin core to the sericin-coating layer, a process accompanied by protein chain alignment and increased feedstock viscosity. This change makes the protein more shear-sensitive and initiates the liquid-to-solid transition. Selective metal ion doping modifies silk fibers' mechanical performance. These findings enhance our understanding of the silk fiber formation mechanism, laying the foundations for developing new concepts in biomaterial design.

Three-dimensional (3D) bioprinting technology enables the controlled deposition of cells and biomaterials (i.e., bioink) to easily create complex 3D biological microenvironments. Silk fibroin (SF) has recently emerged as a compelling bioink component due to its advantageous mechanical and biological properties. This study reports on the development and optimization of a novel bioink for extrusion-based 3D bioprinting and compares different bioink formulations based on mixtures of alginate methacrylate (ALMA), gelatin and SF. The rheological parameters of the bioink were investigated to predict printability and stability, and the optimal concentration of SF was selected. The bioink containing a low amount of SF (0.002% w/V) was found to be the best formulation. Light-assisted gelation of ALMA was exploited to obtain the final hydrogel matrix. Rheological analyses showed that SF-enriched hydrogels exhibited greater elasticity than SF-free hydrogels and were more tolerant to temperature fluctuations. Finally, MG-63 cells were successfully bioprinted and their viability and proliferation over time were analyzed. The SF-enriched bioink represents an excellent biomaterial in terms of printability and allows high cell proliferation over a period of up to 3 weeks. These data confirm the possibility of using the selected formulation for the successful bioprinting of cells into extracellular matrix-like microenvironments.

3D printing of water stable proteins with elastic properties offers a broad range of applications including self-powered biomedical devices driven by piezoelectric biomaterials. Here, we present a study on water-soluble silk fibroin (SF) films. These films were prepared by mixing degummed silk fibers and calcium chloride (CaCl2) in formic acid, resulting in a silk I-like conformation, which was then converted into silk II by redissolving in phosphate buffer (PBS). Circular dichroism, Raman and infrared (IR) spectroscopies were used to investigate the transitions of secondary structure in silk I and silk II as the pH of the solvent and the sonication time were changed. We showed that a solvent with low pH (e.g. 4) maintains the silk I β-turn structure; in contrast solvent with higher pH (e.g. 7.4) promotes β-sheet features of silk II. Ultrasonic treatment facilitates the transition to water stable silk II only for the SF redissolved in PBS. SF from pH 7.4 solution has been printed using extrusion-based 3D printing. A self-powered memristor was realized, comprising an SF-based electric generator and an SF 3D-printed memristive unit connected in series. By exploiting the piezoelectric properties of silk II with higher β-sheet content and Ca2+ ion transport phenomena, the application of an input voltage driven by a SF generator to SF 3D printed holey structures induces a variation from an initial low resistance state (LRS) to a high resistance state (HRS) that recovers in a few minutes, mimicking the transient memory, also known as short-term memory. Thanks to this holistic approach, these findings can contribute to the development of self-powered neuromorphic networks based on biomaterials with memory capabilities.

Efforts to engineer high-performance protein-based materials inspired by nature have mostly focused on altering naturally occurring sequences to confer the desired functionalities, whereas de novo design lags significantly behind and calls for unconventional innovative approaches. Here, using partially disordered elastin-like polypeptides (ELPs) as initial building blocks this work shows that de novo engineering of protein materials can be accelerated through hybrid biomimetic design, which this work achieves by integrating computational modeling, deep neural network, and recombinant DNA technology. This generalizable approach involves incorporating a series of de novo-designed sequences with α-helical conformation and genetically encoding them into biologically inspired intrinsically disordered repeating motifs. The new ELP variants maintain structural conformation and showed tunable supramolecular self-assembly out of thermal equilibrium with phase behavior in vitro. This work illustrates the effective translation of the predicted molecular designs in structural and functional materials. The proposed methodology can be applied to a broad range of partially disordered biomacromolecules and potentially pave the way toward the discovery of novel structural proteins.

The sophisticated, elegant protein-polymers designed by nature can serve as inspiration to redesign and biomanufacture protein-based materials using synthetic biology. Historically, petro-based polymeric materials have dominated industrial activities, consequently transforming our way of living. While this benefits humans, the fabrication and disposal of these materials causes environmental sustainability challenges. Fortunately, protein-based biopolymers can compete with and potentially surpass the performance of petro-based polymers because they can be biologically produced and degraded in an environmentally friendly fashion. This paper reviews four groups of protein-based polymers, including fibrous proteins (collagen, silk fibroin, fibrillin, and keratin), elastomeric proteins (elastin, resilin, and wheat glutenin), adhesive/matrix proteins (spongin and conchiolin), and cyanophycin. We discuss the connection between protein sequence, structure, function, and biomimetic applications. Protein engineering techniques, such as directed evolution and rational design, can be used to improve the functionality of natural protein-based materials. For example, the inclusion of specific protein domains, particularly those observed in structural proteins, such as silk and collagen, enables the creation of novel biomimetic materials with exceptional mechanical properties and adaptability. This review also discusses recent advancements in the production and application of new protein-based materials through the approach of synthetic biology combined biomimetics, providing insight for future research and development of cutting-edge bio-inspired products. Protein-based polymers that utilize nature's designs as a base, then modified by advancements at the intersection of biology and engineering, may provide mankind with more sustainable products.

Hair, or hair-like fibrillar structures, are ubiquitous in biology, from fur on the bodies of mammals, over trichomes of plants, to the mastigonemes on the flagella of single-celled organisms. While these long and slender protuberances are passive, they are multifunctional and help to mediate interactions with the environment. They provide thermal insulation, sensory information, reversible adhesion, and surface modulation (e.g., superhydrophobicity). This review will present various functions that biological hairs have been discovered to carry out, with the hairs spanning across six orders of magnitude in size, from the millimeter-thick fur of mammals down to the nanometer-thick fibrillar ultrastructures on bateriophages. The hairs are categorized according to their functions, including protection (e.g., thermal regulation and defense), locomotion, feeding, and sensing. By understanding the versatile functions of biological hairs, bio-inspired solutions may be developed across length scales.

Fibroin is a structural protein derived from silk cocoons, which may be used in a variety of biomedical applications due to its high biocompatibility and controllable material properties. Conversely, fibroin solution is inherently unstable in solution, which limits its potential utility. Fibroin hydrolysates possess enhanced aqueous solubility and stability, with known anti-inflammatory bioactivity. Here, silk-derived protein (SDP) was produced through controlled time, temperature, and pressure conditions to generate a novel and reproducible hydrolysate population. Both regenerated fibroin and SDP solution stability were characterized for MWD, amino acid content, solubility, viscosity, surface interaction, secondary structure formation, and in vitro assessment of NF-kB pathway activity. Mechanistic studies indicate that hydrolysis processing is required to enhance material stability by abolishing fibroin's ability to self-associate. In vitro assays using HCLE cells indicate SDP has dose dependent potency for inhibiting NF-kB driven gene expression of TNF-α and MMP-9. Collectively, the results support SDP's use as an anti-inflammatory wetting agent compatible with a wide range of both biomedical and industrial applications. Furthermore, the conditions used to generate SDP hydrolysates are readily accessible, produce a highly consistent material from batch-to-batch, and permit widespread investigation of this novel population for these purposes.

Spider silk is the self-assembling product of silk proteins each containing multiple repeating units. Each repeating unit is entirely intrinsically disordered or contains a small disordered domain. The role of the disordered domain/unit in conferring silk protein storage and self-assembly is not fully understood yet. Here, we used biophysical and biochemical techniques to investigate the self-assembly of a miniature version of a minor ampullate spidroin (denoted as miniMiSp). miniMiSp consists of two identical intrinsically disordered domains, one folded repetitive domain, and two folded terminal domains. Our data indicated that miniMiSp self-assembles into oligomers and further into liquid droplets. The oligomerization is attributed to the aggregation-prone property of both the disordered domains and the folded repetitive domain. Our results support the model of micellar structure for silk proteins at high protein concentrations. The disordered domain is indispensable for liquid droplet formation via liquid-liquid phase separation, and tyrosine residues located in the disordered domain make dominant contributions to stability of the liquid droplets. As the same tyrosine residues are also critical to fibrillation, the liquid droplets are likely an intermediate state between the solution state and the fiber state. Additionally, the terminal domains contribute to the pH- and salt-dependent self-assembly properties.

Silk nanofibrils (SNFs), the fundamental building blocks of silk fibers, endow them with exceptional properties. However, the intricate mechanism governing SNF assembly, a process involving both protein conformational transitions and protein molecule conjunctions, remains elusive. This lack of understanding has hindered the development of artificial silk spinning techniques. In this study, we address this challenge by employing a graphene plasmonic infrared sensor in conjunction with multi-scale molecular dynamics (MD). This unique approach allows us to probe the secondary structure of nanoscale assembly intermediates (0.8-6.2 nm) and their morphological evolution. It also provides insights into the dynamics of silk fibroin (SF) over extended molecular timeframes. Our novel findings reveal that amorphous SFs undergo a conformational transition towards β-sheet-rich oligomers on graphene. These oligomers then connect to evolve into SNFs. These insights provide a comprehensive picture of SNF assembly, paving the way for advancements in biomimetic silk spinning.

Salinity gradient energy, often referred to as the Gibbs free energy difference between saltwater and freshwater, is recognized as "blue energy" due to its inherent cleanliness, renewability, and continuous availability. Reverse electrodialysis (RED), relying on ion-selective membranes, stands as one of the most prevalent and promising methods for harnessing salinity gradient energy to generate electricity. Nevertheless, conventional RED membranes face challenges such as insufficient ion selectivity and transport rates and the difficulty of achieving the minimum commercial energy density threshold of 5 W/m2. In contrast, two-dimensional nanostructured materials, featuring nanoscale channels and abundant functional groups, offer a breakthrough by facilitating rapid ion transport and heightened selectivity. This comprehensive review delves into the mechanisms of osmotic power generation within a single nanopore and nanochannel, exploring optimal nanopore dimensions and nanochannel lengths. We subsequently examine the current landscape of power generation using two-dimensional nanostructured materials in laboratory-scale settings across various test areas. Furthermore, we address the notable decline in power density observed as test areas expand and propose essential criteria for the industrialization of two-dimensional ion-selective membranes. The review concludes with a forward-looking perspective, outlining future research directions, including scalable membrane fabrication, enhanced environmental adaptability, and integration into multiple industries. This review aims to bridge the gap between previous laboratory-scale investigations of two-dimensional ion-selective membranes in salinity gradient energy conversion and their potential large-scale industrial applications.

Spider dragline (major ampullate) silk is one of the toughest known fibers in nature and exhibits an excellent combination of high tensile strength and elasticity. Increasing evidence has indicated that preassembly plays a crucial role in facilitating the proper assembly of silk fibers by bridging the mesoscale gap between spidroin molecules and the final strong fibers. However, it remains challenging to control the preassembly of spidroins and investigate its influence on fiber structural and mechanical properties. In this study, we explored to bridge this gap by modulating the polyalanine (polyA) motifs in repetitive region of spidroins to tune their preassemblies in aqueous dope solutions. Three biomimetic silk proteins with varying numbers of alanine residues in polyA motif and comparable molecular weights were designed and biosynthesized, termed as N16C-5A, N15C-8A, and N13C-12A, respectively. It was found that all three proteins could form nanofibril assemblies in the concentrated aqueous dopes, but the size and structural stability of the fibrils were distinct from each other. The silk protein N15C-8A with 8 alanine residues in polyA motif allowed for the formation of stable nanofibril assemblies with a length of approximately 200 nm, which were not prone to disassemble or aggregate as that of N16C-5A and N13C-12A. More interestingly, the stable fibril assembly of N15C-8A enabled spinning of simultaneously strong (623.3 MPa) and tough (107.1 MJ m-3) synthetic fibers with fine molecular orientation and close interface packing of fibril bundles. This work highlights that modulation of polyA motifs is a feasible way to tune the morphology and stability of the spidroin preassemblies in dope solutions, thus controlling the structural and mechanical properties of the resulting fibers.

Flexible electronics, applicable to enlarged health, AI big data medications, etc., have been one of the most important technologies of this century. Due to its particular mechanical properties, biocompatibility, and biodegradability, cocoon silk (or SF, silk fibroin) plays a key role in flexible electronics/photonics. The review begins with an examination of the hierarchical meso network structures of SF materials and introduces the concepts of meso reconstruction, meso doping, and meso hybridization based on the correlation between the structure and performance of silk materials. The SF meso functionalization was developed according to intermolecular nuclear templating. By implementation of the techniques of meso reconstruction and functionalization in the refolding of SF materials, extraordinary performance can be achieved. Relying on this strategy, particularly designed flexible electronic and photonic components can be developed. This review covers the latest ideas and technologies of meso flexible electronics and photonics based on SF materials/meso functionalization. As silk materials are biocompatible and human skin-friendly, SF meso flexible electronic/photonic components can be applied to wearable or implanted devices. These devices are applicable in human physiological signals and activities sensing/monitoring. In the case of human-machine interaction, the devices can be applicable in in-body information transmission, computation, and storage, with the potential for the combination of artificial intelligence and human intelligence.

The toughness of silk naturally obtained from spiders and silkworms exceeds that of all other natural and man-made fibers. These insects transform aqueous protein feedstocks into mechanically specialized materials, which represents an engineering phenomenon that has developed over millions of years of natural evolution. Silkworms have become a new research hotspot due to the difficulties in collecting spider silk and other challenges. According to continuous research on the natural spinning process of the silkworm, it is possible to divide the main aspects of bionic spinning into two main segments: the solvent and behavior. This work focuses on the various methods currently used for the spinning of artificial silk fibers to replicate natural silk fibers, providing new insights based on changes in the fiber properties and production processes over time.

Silk is one of the toughest fibrous materials known despite spun at ambient temperature and pressure with water as a solvent. It is a great challenge to reproduce high-performance artificial fibers comparable to natural silk by bionic for the incomplete understanding of silkworm spinning in vivo. Here, we found that amphipol and digitonin stabilized the structure of natural silk fibroin (NSF) by a large-scale screening in vitro, and then studied the close-to-native ultrastructure and hierarchical assembly of NSF in the silk gland lumen. Our study showed that NSF formed reversible flexible nanofibrils mainly composed of random coils with a sedimentation coefficient of 5.8 S and a diameter of about 4 nm, rather than a micellar or rod-like structure assembled by the aggregation of globular NSF molecules. Metal ions were required for NSF nanofibril formation. The successive pH decrease from posterior silk gland (PSG) to anterior silk gland (ASG) resulted in a gradual increase in NSF hydrophobicity, thus inducing the sol-gelation transition of NSF nanofibrils. NSF nanofibrils were randomly dispersed from PSG to ASG-1, and self-assembled into anisotropic herringbone patterns at ASG-2 near the spinneret ready for silkworm spinning. Our findings reveal the controlled self-assembly mechanism of the multi-scale hierarchical architecture of NSF from nanofibrils to herringbone patterns programmed by metal ions and pH gradient, which provides novel insights into the spinning mechanism of silk-secreting animals and bioinspired design of high-performance fibers.

This review will present the latest research related to the production and application of spider silk and silk-based materials in reconstructive and regenerative medicine and tissue engineering, with a focus on musculoskeletal tissues, and including skin regeneration and tissue repair of bone and cartilage, ligaments, muscle tissue, peripheral nerves, and artificial blood vessels. Natural spider silk synthesis is reviewed, and the further recombinant production of spider silk proteins. Research insights into possible spider silk structures, like fibers (1D), coatings (2D), and 3D constructs, including porous structures, hydrogels, and organ-on-chip designs, have been reviewed considering a design of bioactive materials for smart medical implants and drug delivery systems. Silk is one of the toughest natural materials, with high strain at failure and mechanical strength. Novel biomaterials with silk fibroin can mimic the tissue structure and promote regeneration and new tissue growth. Silk proteins are important in designing tissue-on-chip or organ-on-chip technologies and micro devices for the precise engineering of artificial tissues and organs, disease modeling, and the further selection of adequate medical treatments. Recent research indicates that silk (films, hydrogels, capsules, or liposomes coated with silk proteins) has the potential to provide controlled drug release at the target destination. However, even with clear advantages, there are still challenges that need further research, including clinical trials.

Achieving ultra-high tensile strength and exceptional toughness is a longstanding goal for structural materials. However, previous attempts using covalent and non-covalent bonds have failed, leading to the belief that these two properties are mutually exclusive. Consequently, commercial fibers have been forced to compromise between tensile strength and toughness, as seen in the differences between nylon and Kevlar. To address this challenge, we drew inspiration from the disparate tensile strength and toughness of nylon and Kevlar, both of which are polyamide fibers, and developed an innovative approach that combines specific intermolecular disulfide bonds and reversible hydrogen bonds to create ultra-strong and ultra-tough polyamide spider silk fibers. Our resulting Supramolecular polyamide spider silk, which has a maximum molecular weight of 1084 kDa, exhibits high tensile strength (1180 MPa) and extraordinary toughness (433 MJ/m3), surpassing Kevlar's toughness 8-fold. This breakthrough presents a new opportunity for the sustainable development of spider silk as an environmentally friendly alternative to synthetic commercial fibers, as spider silk is composed of amino acids. Future research could explore the use of these techniques and fundamental knowledge to develop other super materials in various mechanical fields, with the potential to improve people's lives in many ways. STATEMENT OF SIGNIFICANCE: • By emulating synthetic commercial fibers such as nylon and polyethylene, we have successfully produced supramolecular-weight polyamide spider silk fibers with a molecular weight of 1084 kDa through a unique covalent bond-mediated linear polymerization reaction of spider silk protein molecules. This greatly surpasses the previous record of a maximum molecular weight of 556 kDa. • We obtained supramolecular polyamide spider silk fibers with both high-tensile strength and toughness. The stress at break is 1180 MPa, and the toughness is 8 times that of kevlar, reaching 433 MJ/m3. • Our results challenge the notion that it is impossible to manufacture fibers with both ultra-high tensile strength and ultra-toughness, and provide theoretical guidance for developing environmentally friendly and sustainable structural materials that meet industrial needs.

To realize the full potential of emerging nucleic acid therapies, there is a need for effective delivery agents to transport cargo to cells of interest. Protein materials exhibit several unique properties, including biodegradability, biocompatibility, ease of functionalization via recombinant and chemical modifications, among other features, which establish a promising basis for therapeutic nucleic acid delivery systems. In this review, we highlight progress made in the use of non-viral protein-based nanoparticles for nucleic acid delivery in vitro and in vivo, while elaborating on key physicochemical properties that have enabled the use of these materials for nanoparticle formulation and drug delivery. To conclude, we comment on the prospects and unresolved challenges associated with the translation of protein-based nucleic acid delivery systems for therapeutic applications.

Spider silk protein, renowned for its excellent mechanical properties, biodegradability, chemical stability, and low immune and inflammatory response activation, consists of a core domain with a repeat sequence and non-repeating sequences at the N-terminal and C-terminal. In this review, we focus on the relationship between the silk structure and its mechanical properties, exploring the potential applications of spider silk materials in the detection of energetic materials.