High-performance Silk Fibers Research Articles

Strong Silkworm Silk Fibers through CNT-Feeding and Forced Reeling.

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.

High mechanical property silk produced by transgenic silkworms expressing the Drosophila Dumpy.

Extensive research has been conducted on utilizing transgenic silkworms and their natural spinning apparatus to produce high-performance spider silk fibers. However, research on using non-spider biological proteins to optimize the molecular structure of silk protein and improve the mechanical performance of silk fibers is still relatively scarce. Dumpy, a massive extracellular matrix polypeptide, is essential for preserving the shape and structural integrity of the insect cuticle due to its remarkable tension and elasticity. Here, we constructed two transgenic donor plasmids containing the fusion genes of FibH-Dumpy and FibL-Dumpy. The results indicated the successful integration of two exogenous gene expression cassettes, driven by endogenous promoters, into the silkworm genome using piggyBac-mediated transgenic technology. Secondary structure analysis revealed a 16.7% and 13.6% increase in the β-sheet content of transgenic silks compared to wild-type (WT) silk fibers. Mechanical testing demonstrated that, compared to the WT, HDUY and LDUY transgenic silk fibers exhibited respective increases of 39.54% and 21.45% in maximum stress, 44.43% and 45.02% in toughness, and 24.91% and 28.51% in elastic recovery rate. These findings suggest that Drosophila Dumpy significantly enhanced the mechanical properties of silk, positioning it as an excellent candidate for the development of extraordinary-performance fibers. This study provides rich inspiration for using other biological proteins to construct high-performance silk fibers and expands the possibilities for designing and applying novel biomaterials.

Ultrafine and High-Strength Silk Fibers Secreted by Bimolter Silkworms.

Ultrafine fibers are widely employed because of their lightness, softness, and warmth retention. Although silkworm silk is one of the most applied natural silks, it is coarse and difficult to transform into ultrafine fibers. Thus, to obtain ultrafine high-performance silk fibers, we employed anti-juvenile hormones in this study to induce bimolter silkworms. We found that the bimolter cocoons were composed of densely packed thin fibers and small apertures, wherein the silk diameter was 54.9% less than that of trimolter silk. Further analysis revealed that the bimolter silk was cleaner and lighter than the control silk. In addition, it was stronger (739 MPa versus 497 MPa) and more stiffness (i.e., a higher Young’s modulus) than the trimolter silk. FTIR and X-ray diffraction results revealed that the excellent mechanical properties of bimolter silk can be attributed to the higher β-sheet content and crystallinity. Chitin staining of the anterior silk gland suggested that the lumen is narrower in bimolters, which may lead to the formation of greater numbers of β-sheet structures in the silk. Therefore, this study reveals the relationship between the structures and mechanical properties of bimolter silk and provides a valuable reference for producing high-strength and ultrafine silk fibers.

Characterization of the second type of aciniform spidroin (AcSp2) provides new insight into design for spidroin-based biomaterials

Spiders spin a range of silks from different glands for distinct functions, and each silk type exhibits distinct material properties. Silk extruded by the aciniform gland is used for prey wrapping and egg case construction and displays high toughness and extensibility. So far, only the aciniform spidroin 1 (AcSp1) gene which was firstly identified as a silk gene in aciniform gland has been obtained. Here we present the gene sequence for the second type of full-length aciniform silk protein, AcSp2. Analysis of the AcSp2 primary sequence reveals relatively conserved terminal regions and a distinct repetitive sequence relative to AcSp1. A fraction of the gene can be expressed in recombinant systems. Secondary structure analysis of the recombinant AcSp2 protein in solution reveals that the protein adopts mainly an α-helical conformation. Artificial spinning of recombinant AcSp2 demonstrates that the spidroins can be spun into fine fibers which display up to 142% extensibility. The silk fibers are dominated by β-sheet and β-turn secondary structures. Moreover, the mechanical data collected from these synthetic fibers revealed that the mechanical properties are partly correlated with the molecular weights. Overall, our studies enrich our knowledge of spidroin gene family members and provide a new insight into creation of high-performance silk fibers for next generation biomaterials. Statement of SignificanceIn this study, we presented the second type of aciniform silk protein (AcSp2) gene sequence of orb-weaving spider Araneus ventricosus, expanding the spider silk gene family members. The primary structure revealed the central repetitive sequence of the new spidroin gene is distinctly different from other AcSp1 genes. Characterization of the recombinant minispidroin fibers of AcSp2 revealed the mechanical properties are partly correlate with the molecular weights, and the spidroins can be spun into fine fibers which display up to 142% extensibility. Overall, our studies enrich our knowledge of spidroin gene family members and provide a new insight into creation of high-performance silk fibers for next generation biomaterials.

Combination of Amorphous Silk Fiber Spinning and Postspinning Crystallization for Tough Regenerated Silk Fibers.

An artificial spinning system using regenerated silk fibroin solutions is adopted to produce high-performance silk fibers. In previous studies, alcohol-based agents, such as methanol or ethanol, were used to coagulate silk dope solutions, producing silk fiber with poor mechanical properties compared with those of native silk fibers. The alcohol-based coagulation agents induce rapid β-sheet crystallization of the silk molecules, which inhibits subsequent alignment of the β-sheet crystals. Here, we induce gradual β-sheet formation to afford adequate β-sheet alignment similar to that of native silk fiber. To this aim, we developed an amorphous silk fiber spinning process that prevents fast β-sheet formation in silk molecules by using tetrahydrofuran (THF) as a coagulation solvent. In addition, we apply postdrawing to the predominantly amorphous silk fibers to induce β-sheet formation and orientation. The resultant silk fibers showed a 2.5-fold higher extensibility, resulting in 1.5-fold tougher silk fibers compared with native Bombyx mori silk fiber. The amorphous silk fiber spinning process developed here will pave the way to the production of silk fibers with desired mechanical properties.

Production of High Performance Bioinspired Silk Fibers by Straining Flow Spinning.

In the last years, there has been an increasing interest in bioinspired approaches for different applications, including the spinning of high performance silk fibers. Bioinspired spinning is based on the natural spinning system of spiders and worms and requires combining changes in the chemical environment of the proteins with the application of mechanical stresses. Here we present the novel straining flow spinning (SFS) process and prove its ability to produce high performance fibers under mild, environmentally friendly conditions, from aqueous protein dopes. SFS is shown to be an extremely versatile technique which allows controlling a large number of processing parameters. This ample set of parameters allows fine-tuning the microstructure and mechanical behavior of the fibers, which opens the possibility of adapting the fibers to their intended uses.

Microdissection of Black Widow Spider Silk-producing Glands

Modern spiders spin high-performance silk fibers with a broad range of biological functions, including locomotion, prey capture and protection of developing offspring. Spiders accomplish these tasks by spinning several distinct fiber types that have diverse mechanical properties. Such specialization of fiber types has occurred through the evolution of different silk-producing glands, which function as small biofactories. These biofactories manufacture and store large quantities of silk proteins for fiber production. Through a complex series of biochemical events, these silk proteins are converted from a liquid into a solid material upon extrusion. Mechanical studies have demonstrated that spider silks are stronger than high-tensile steel. Analyses to understand the relationship between the structure and function of spider silk threads have revealed that spider silk consists largely of proteins, or fibroins, that have block repeats within their protein sequences. Common molecular signatures that contribute to the incredible tensile strength and extensibility of spider silks are being unraveled through the analyses of translated silk cDNAs. Given the extraordinary material properties of spider silks, research labs across the globe are racing to understand and mimic the spinning process to produce synthetic silk fibers for commercial, military and industrial applications. One of the main challenges to spinning artificial spider silk in the research lab involves a complete understanding of the biochemical processes that occur during extrusion of the fibers from the silk-producing glands. Here we present a method for the isolation of the seven different silk-producing glands from the cobweaving black widow spider, which includes the major and minor ampullate glands [manufactures dragline and scaffolding silk], tubuliform [synthesizes egg case silk], flagelliform [unknown function in cob-weavers], aggregate [makes glue silk], aciniform [synthesizes prey wrapping and egg case threads] and pyriform [produces attachment disc silk]. This approach is based upon anesthetizing the spider with carbon dioxide gas, subsequent separation of the cephalothorax from the abdomen, and microdissection of the abdomen to obtain the silk-producing glands. Following the separation of the different silk-producing glands, these tissues can be used to retrieve different macromolecules for distinct biochemical analyses, including quantitative real-time PCR, northern- and western blotting, mass spectrometry (MS or MS/MS) analyses to identify new silk protein sequences, search for proteins that participate in the silk assembly pathway, or use the intact tissue for cell culture or histological experiments.

Microdissection of Black Widow Spider Silk-producing Glands

Modern spiders spin high-performance silk fibers with a broad range of biological functions, including locomotion, prey capture and protection of developing offspring 1,2. Spiders accomplish these tasks by spinning several distinct fiber types that have diverse mechanical properties. Such specialization of fiber types has occurred through the evolution of different silk-producing glands, which function as small biofactories. These biofactories manufacture and store large quantities of silk proteins for fiber production. Through a complex series of biochemical events, these silk proteins are converted from a liquid into a solid material upon extrusion. Mechanical studies have demonstrated that spider silks are stronger than high-tensile steel 3. Analyses to understand the relationship between the structure and function of spider silk threads have revealed that spider silk consists largely of proteins, or fibroins, that have block repeats within their protein sequences 4. Common molecular signatures that contribute to the incredible tensile strength and extensibility of spider silks are being unraveled through the analyses of translated silk cDNAs. Given the extraordinary material properties of spider silks, research labs across the globe are racing to understand and mimic the spinning process to produce synthetic silk fibers for commercial, military and industrial applications. One of the main challenges to spinning artificial spider silk in the research lab involves a complete understanding of the biochemical processes that occur during extrusion of the fibers from the silk-producing glands. Here we present a method for the isolation of the seven different silk-producing glands from the cobweaving black widow spider, which includes the major and minor ampullate glands [manufactures dragline and scaffolding silk] 5,6, tubuliform [synthesizes egg case silk] 7,8, flagelliform [unknown function in cob-weavers], aggregate [makes glue silk], aciniform [synthesizes prey wrapping and egg case threads] 9 and pyriform [produces attachment disc silk] 10. This approach is based upon anesthetizing the spider with carbon dioxide gas, subsequent separation of the cephalothorax from the abdomen, and microdissection of the abdomen to obtain the silk-producing glands. Following the separation of the different silk-producing glands, these tissues can be used to retrieve different macromolecules for distinct biochemical analyses, including quantitative real-time PCR, northern- and western blotting, mass spectrometry (MS or MS/MS) analyses to identify new silk protein sequences, search for proteins that participate in the silk assembly pathway, or use the intact tissue for cell culture or histological experiments.