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The balance of crystalline and amorphous regions in the fibroin structure underpins the tensile strength of bagworm silk.

ABSTRACT: Protein-based materials are considered versatile biomaterials, and their biodegradability is an advantage for sustainable development. Bagworm produces strong silk for use in unique situations throughout its life stages. Rigorous molecular analyses of Eumeta variegata suggested that the particular mechanical properties of its silk are due to the coexistence of poly-A and GA motifs. However, little molecular information on closely related species is available, and it is not understood how these properties were acquired evolutionarily or whether the motif combination is a conserved trait in other bagworms. Here, we performed a transcriptome analysis of two other bagworm species (Canephora pungelerii and Bambalina sp.) belonging to the family Psychidae to elucidate the relationship between the fibroin gene and silk properties. The obtained transcriptome assemblies and tensile tests indicated that the motif combination and silk properties were conserved among the bagworms. Furthermore, our analysis showed that C. pungelerii produces extraordinarily strong silk (breaking strength of 1.4 GPa) and indicated that the cause may be the C. pungelerii -specific balance of crystalline/amorphous regions in the H-fibroin repetitive domain. This particular H-fibroin architecture may have been evolutionarily acquired to produce strong thread to maintain bag stability during the relatively long development period of Canephora species relative to other bagworms.

SUBMITTER: Kono N

PROVIDER: S-EPMC8314566 | biostudies-literature |

REPOSITORIES: biostudies-literature

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The bagworm genome reveals a unique fibroin gene that provides high tensile strength.

Project description:Arthropod silk is known as a versatile tool, and its variability makes it an attractive biomaterial. Eumeta variegata is a bagworm moth (Lepidoptera, Psychidae) that uses silk throughout all life stages. Notably, the bagworm-specific uses of silk include larval development in a bag coated with silk and plant materials and the use of silk attachments to hang pupae. An understanding at the molecular level of bagworm silk, which enables such unique purposes, is an opportunity to expand the possibilities for artificial biomaterial design. However, very little is known about the bagworm fibroin gene and the mechanical properties of bagworm silk. Here, we report the bagworm genome, including a silk fibroin gene. The genome is approximately 700 Mbp in size, and the newly found fibroin gene has a unique repetitive motif. Furthermore, a mechanical property test demonstrates a phylogenetic relationship between the unique motif and tensile strength of bagworm silk.

| S-EPMC6488591 | biostudies-literature

Recombinant Silk Fibroin Crystalline Regions as Biomaterial Alternatives to the Full-Length Protein.

Project description:Silk fibroin is a natural polymer with a unique repetitive structure that translates to extraordinary properties in terms of processability and mechanical properties. The Bombyx mori silk has a molecular weight of ∼415 kDa and consists of a light chain and a heavy chain. Its heavy chain is organized into 12 crystalline domains. Each of these crystalline domains contains subdomains of ∼70 amino acid containing blocks. It is well understood that the heavy chain of the protein is responsible for its processing versatility and excellent mechanical properties; however, the need for the high number of monomeric repeating units is unclear, and the individual properties of crystalline regions compared to those of the full-length protein are not understood. The work described herein assessed the possibility of using recombinant crystalline regions as alternative biomaterials for applications such as tissue adhesives. Our results indicate that while the two tested substructures do not fully recapitulate the native silk fibroin's properties, they appear to be a suitable alternative for the production of silk-based medical adhesives.

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High Concentration Crystalline Silk Fibroin Solution for Silk-Based Materials.

Project description:As a functional biomaterial, silk fibroin has been widely used in drug release, cell encapsulation and tissue regeneration. To meet the requirements of these applications, the properties of silk fibroin-based materials should be finely tunable. Many useful properties of biomaterials emerge from the collective interactions among ordered and disordered domains. Thus, increasing subtle control of silk hierarchical structures is required. As a characteristic of ordered silk fibroin, crystalline silk fibroin (CSF) is an important part of silk fibroin-based biomaterials, but the preparation of CSF solution, especially high concentration CSF solution, remains a challenge. Here, a solution composed of β-sheet-rich silk fibroin is reported. These CSF were obtained by the sonication of silk fibroin hydrogel, destroying the hydrogel network, and turning silk fibroin hydrogels into CSF solution. These β-sheet-rich CSF solutions were stable enough for several days or even weeks. In addition, they were typically ordered crystalline domains, which could be mixed with disordered domains and fabricated into porous scaffolds, films, hydrogels and other silk fibroin-based scaffolds with different properties.

| S-EPMC9571966 | biostudies-literature

Viscous friction between crystalline and amorphous phase of dragline silk.

Project description:The hierarchical structure of spider dragline silk is composed of two major constituents, the amorphous phase and crystalline units, and its mechanical response has been attributed to these prime constituents. Silk mechanics, however, might also be influenced by the resistance against sliding of these two phases relative to each other under load. We here used atomistic molecular dynamics (MD) simulations to obtain friction forces for the relative sliding of the amorphous phase and crystalline units of Araneus diadematus spider silk. We computed the coefficient of viscosity of this interface to be in the order of 10(2) Ns/m(2) by extrapolating our simulation data to the viscous limit. Interestingly, this value is two orders of magnitude smaller than the coefficient of viscosity within the amorphous phase. This suggests that sliding along a planar and homogeneous surface of straight polyalanine chains is much less hindered than within entangled disordered chains. Finally, in a simple finite element model, which is based on parameters determined from MD simulations including the newly deduced coefficient of viscosity, we assessed the frictional behavior between these two components for the experimental range of relative pulling velocities. We found that a perfectly relative horizontal motion has no significant resistance against sliding, however, slightly inclined loading causes measurable resistance. Our analysis paves the way towards a finite element model of silk fibers in which crystalline units can slide, move and rearrange themselves in the fiber during loading.

| S-EPMC4132047 | biostudies-literature

High-Strength, Durable All-Silk Fibroin Hydrogels with Versatile Processability toward Multifunctional Applications.

Project description:Hydrogels have been 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, we generate pristine silk fibroin (SF) hydrogels with excellent mechanical properties 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. ?-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-silk fibroin 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 filled 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.

| S-EPMC7241598 | biostudies-literature

Enhancing strength and ductility via crystalline-amorphous nanoarchitectures in TiZr-based alloys.

Project description:Crystalline-amorphous composite have the potential to achieve high strength and high ductility through manipulation of their microstructures. Here, we fabricate a TiZr-based alloy with micrometer-size equiaxed grains that are made up of three-dimensional bicontinuous crystalline-amorphous nanoarchitectures (3D-BCANs). In situ tension and compression tests reveal that the BCANs exhibit enhanced ductility and strain hardening capability compared to both amorphous and crystalline phases, which impart ultra-high yield strength (~1.80 GPa), ultimate tensile strength (~2.3 GPa), and large uniform ductility (~7.0%) into the TiZr-based alloy. Experiments combined with finite element simulations reveal the synergetic deformation mechanisms; i.e., the amorphous phase imposes extra strain hardening to crystalline domains while crystalline domains prevent the premature shear localization in the amorphous phases. These mechanisms endow our material with an effective strength-ductility-strain hardening combination.

| S-EPMC8906574 | biostudies-literature

Customized Flagelliform Spidroins Form Spider Silk-like Fibers at pH 8.0 with Outstanding Tensile Strength.

Project description:Spider flagelliform silk shows the best extensibility among various types of silk, but its biomimetic preparation has not been much studied. Herein, five customized flagelliform spidroins (FlSps: S and NTDFl-Sn-CTDFl, n = 1-4), in which the repetitive region (S) and N-/C- terminal domains (NTDFl and CTDFl) are from the same spidroin and spider species, were produced recombinantly. The recombinant spidroins with terminal domains were able to form silk-like fibers with diameters of ∼5 μm by manual pulling at pH 8.0, where the secondary structure transformation occurred. The silk-like fibers from NTDFl-S4-CTDFl showed the highest tensile strength (∼250 MPa), while those ones with 1-3 S broke at a similar stress (∼180 MPa), suggesting that increasing the amounts of the repetitive region can improve the tensile strength, but a certain threshold might need to be reached. This study shows successful preparation of flagelliform silk-like fibers with good mechanical properties, providing general insights into efficient biomimetic preparations of spider silks.

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Molecularly Imprinted Silk Fibroin Nanoparticles.

Project description:Nanosized biomimetics prepared by the strategy of molecular imprinting, that is, the stamping of recognition sites by means of a template-assisted synthesis, are demonstrating potential as plastic antibodies in medicine, proving effective for cell imaging and targeted therapies. Most molecularly imprinted nanoparticles (MIP-NPs) are currently made of soft matter, such as polyacrylamide and derivatives. Yet, MIP-NPs biocompatibility is crucial for their effective translation into clinical uses. Here, we propose the original idea to synthesize fully biocompatible molecularly imprinted nanoparticles starting from the natural polymer silk fibroin (MIP SF-NPs), which is nontoxic and highly biocompatible. The conditions to produce MIP SF-NPs of different sizes (dmean ∼ 50 nm; dmean ∼ 100 nm) were set using the response surface method. The stamping of a single, high affinity (KD = 57 × 10-9 M), and selective recognition site per silk fibroin nanoparticle was demonstrated, together with the confirmation of nontoxicity. Additionally, MIP SF-NPs were used to decorate silk microfibers and silk nanofibers, providing a general means to add entailed biofunctionalities to materials.

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Andrographolide-loaded silk fibroin nanoparticles.

Project description:Andrographolide (AP) is a diterpenoid separated from Andrographis paniculata with a wide spectrum of biological activities including anti-inflammatory, anticancer, hepatoprotective, and antihyperlipidemic. However, its poor water solubility and instability result in lower bioavailability, which seriously limit its pharmacological function. In this study, the attempt to use regenerated silk fibroin (RSF) as a drug-carrier to encapsulate AP was reported. The AP-loaded RSF nanoparticles were prepared by a facile and clean method without any toxic agents. Moreover, special attention was paid to the optimization of formulation. Finally, the sizes of the AP-loaded RSF nanoparticles ranged from 200 to 1000 nm, and the nanoparticles were spherically shaped, as seen by transmission electron microscopy. The drug loading and encapsulation efficiency were about 25.9% and 87.3%, respectively. Furthermore, the release time of AP-loaded RSF nanoparticles was about 3 days. The particle size and drug release behaviour could be adjusted by treating with glycol amine. The in vitro cytotoxicity studies demonstrated that the RSF nanoparticles showed negligible cytotoxicity to cells, and the anti-proliferative activity of AP-loaded RSF nanoparticles showed that the AP-loaded RSF nanoparticles can adhere to Hela cells and MDA-MB-231 cells easily. All these results imply that this biomacromolecule drug nanocarrier has great potential for chemotherapy in clinical applications.

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Impacts of Blended Bombyx mori Silk Fibroin and Recombinant Spider Silk Fibroin Hydrogels on Cell Growth.

Project description:Binary-blended hydrogels fabricated from Bombyx mori silk fibroin (SF) and recombinant spider silk protein eADF4(C16) were developed and investigated concerning gelation and cellular interactions in vitro. With an increasing concentration of eADF4(C16), the gelation time of SF was shortened from typically one week to less than 48 h depending on the blending ratio. The biological tests with primary cells and two cell lines revealed that the cells cannot adhere and preferably formed cell aggregates on eADF4(C16) hydrogels, due to the polyanionic properties of eADF4(C16). Mixing SF in the blends ameliorated the cellular activities, as the proliferation of L929 fibroblasts and SaOS-2 osteoblast-like cells increased with an increase of SF content. The blended SF:eADF4(C16) hydrogels attained the advantages as well as overcame the limitations of each individual material, underlining the utilization of the hydrogels in several biomedical applications.

| S-EPMC8659740 | biostudies-literature

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