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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. Replicating natural silk spinning is of interest for its eco-friendly process. Here, the authors find that calcium ions trigger phase separation in silkworm fibroin under crowded conditions, forming fibrils with acid and shear. Based on these findings, they produce silk fiber by pultrusion-based biomimetic spinning. [ABSTRACT FROM AUTHOR]

An antimicrobial peptide motif (Cys-KR12) originating from human cathelicidin peptide (LL37) was immobilized onto electrospun SF nanofiber membranes using EDC/NHS and thiol-maleimide click chemistry to confer the various bioactivities of LL37 onto the membrane for wound care purposes. Surface characterizations revealed that the immobilization density of Cys-KR12 on SF nanofibers could be precisely controlled with a high reaction yield. The Cys-KR12-immobilized SF nanofiber membrane exhibited antimicrobial activity against four pathogenic bacterial strains ( Staphylococcus aureus , Staphylococcus epidermidis , Escherichia coli , and Pseudomonas aeruginosa ) without biofilm formation on the membrane surface. It also facilitated the proliferation of keratinocytes and fibroblasts and promoted the differentiation of keratinocytes with enhanced cell-cell attachment. In addition, immobilized Cys-KR12 significantly suppressed the LPS-induced TNF-α expression of monocytes (Raw264.7) cultured on the membrane. These results suggest that a Cys-KR12-immobilized SF nanofiber membrane, which has multiple biological activities, would be a promising candidate as a wound dressing material. Statement of Significance This research article reports various bioactivities of an antimicrobial peptide on electrospun silk fibroin nanofiber membrane. Recently, human cathelicidin peptide LL37 has been extensively explored as an alternative antibiotic material. It has not only a great antimicrobial activity but also a wide variety of bioactivities which can facilitate wound healing process. Especially, many studies on immobilization of LL37 or its analogues have shown the immobilization technique could improve performance of wound dressing materials or tissue culture matrices. Nevertheless, so far studies have only focused on the bactericidal effect of immobilized peptide on material surface. On the other hand, we tried to evaluate multi-biofunction of immobilized antimicrobial peptide Cys-KR12, which is the shortest peptide motif as an analogue of LL37. We fabricated silk fibroin nanofiber membrane as a model wound dressing by electrospinning and immobilized the antimicrobial peptide. As a result, we confirmed that the immobilized peptide can play multi-role in wound healing process, such as antimicrobial activity, facilitation of cell proliferation and keratinocyte differentiation, and inhibition of inflammatory cytokine expression. These findings have not been reported and can give an inspiration in wound-care application. [ABSTRACT FROM AUTHOR]

ABSTRACT Silk fibroin (SF) nanofiber scaffold containing microalgae Spirulina extract were prepared by electrospinning and the performance and functionality of the scaffold were evaluated. The viscosity and conductivity of the dope solution of Spirulina containing SF were examined for electrospinability and we found that the morphological structure of SF nanofiber is affected by the concentration of Spirulina extract added. The platelet adhesion and coagulation time test confirmed that the Spirulina containing SF nanofiber scaffold had excellent ability to prevent blood clotting or antithrombogenicity that is comparable to heparin. Low cytotoxicity and excellent cell adhesion and proliferation were also observed for Sprulina containing SF nanofiber scaffold by methylthiazolyldiphenyl-tetrazolium bromide assay and confocal fluorescence microscope using fibroblast and human umbilical vein endothelial cells. Based on these results, we believe SF nanofiber scaffold containing Spirulina extract has the potential to be used as tissue engineering scaffold that requires high hemocompatibility. © 2013 Wiley Periodicals, Inc. Biopolymers 101: 307-318, 2014. [ABSTRACT FROM AUTHOR]

Abstract: Silk fibroin (SF)/poly(vinyl alcohol) (PVA) blend filaments were prepared by a wet spinning process. Regenerated SF and PVA were dissolved in formic acid and the dope solution exhibited good fiber formation in a methanol coagulation bath. Due to the miscibility of SF/PVA in formic acid, the filament had a smooth surface and dense structure with a circular cross-section. The crystalline structure and thermal properties were varied with different SF/PVA ratios. The mechanical properties of the filament were also controlled by blending PVA with SF. Especially, the knot strength of the SF filament, which is a very important suture property, could be significantly improved by blending with PVA. [Copyright &y& Elsevier]

In the present study, the effects of different degumming methods on the structural characteristics and properties of regenerated silk fibroin (SF) were examined. The crystallinity index of the degummed silk increased with the degumming ratio. The crystallinity index at any given degumming ratio differed depending on the degumming method. The soda method and the soap/soda method using sodium carbonate resulted in a higher crystallinity index than the other methods The degumming method strongly affects the molecular weight (MW) and solution viscosity of the regenerated SF. The MW and viscosity of the regenerated SF, according to the degumming method, was in the order of urea method > HTHP method ≈ acid method > soap/soda method ≈ soda method. The turbidity of a silk formic acid solution decreased as a result of increasing the degumming ratio and was a minimum at a degumming ratio of around 26%. However, it was not affected by the degumming method. The mechanical properties of a regenerated SF film were strongly affected by the degumming method and the trend in the strength and elongation with the various degumming methods was the same as that of the MW and viscosity of the regenerated SF. [ABSTRACT FROM AUTHOR]

Silk fibroin (SF) is known to be a biocompatible material, and different forms of SF are used for various applications. However, the application of SF in particle form is rarely reported, compared to other forms. In this study, SF microparticles with a diameter of approximately 250 μm were prepared by the electrospray method, using 1 M LiCl/DMSO as a solvent. The dissolution time of SF in the CaCl 2 /CH 3 CH 2 OH/H 2 O solution and the concentration of the SF dope solution affected the final morphology of the microparticles. A long dissolution time and a low SF concentration led to the formation of irregular microparticles, but a short dissolution time and a high concentration produced sphere-like microparticles. The shear viscosity of the SF dope solution was the main parameter that affected the morphology of the SF microparticles. Regardless of the dissolution time in the CaCl 2 /CH 3 CH 2 OH/H 2 O solution and the concentration of the SF dope solution, the shear viscosity of the dope solution must be higher than 0.33 Pa s to produce sphere-like microparticles. Finally, cell adhesion experiments demonstrated that these SF microparticles show potential for use as cell carriers. [ABSTRACT FROM AUTHOR]

Silk has attracted the attention of biomedical researchers because of its good biocompatibility. Although various characteristics of silk are needed for its successful application in biomedical fields, the performance of silk material is limited. Although there are many varieties of Bombyx mori silkworm, the effect of different silkworm varieties on regenerated silk has not been considered in detail. That is, the use of a diverse variety of silkworms has not been considered in non-textile applications resulting in limited performance of silk materials. In this study, the effects of different silkworm varieties on the structural characteristics and properties of silk cocoon and regenerated silk fibroin (SF) were examined. Structural characteristics of silk cocoon including color, fiber diameter, and porosity, differed depending on the silkworm variety. Furthermore, molecular weight, solution viscosity, and mechanical properties of regenerated SF were influenced by the variety of silkworm, while the amino acid composition, β-sheet crystallization by formic acid, and cyto-compatibility of regenerated SF did not differ between the samples from different varieties of silkworm. These results imply that diverse performance of silk can be obtained by controlling the silkworm variety, and that the use of different varieties of silkworm might be a good way to strengthen the performance of silk in biomedical fields. [ABSTRACT FROM AUTHOR]

Silk has recently been exploitedin various fields due to its superiormechanical properties. However, this material’s lack of biologicalfunctions and relatively poor biodegradation have hindered its wideuse in applications related to cells and tissues. Here, we improvedthe overall characteristics of silkworm silk fibroin (SF) by introductionof RGD peptide-fused recombinant mussel adhesive protein (MAP-RGD).Simple blending of MAP-RGD provided not only bulk-scale adhesive abilitybut also microscale adhesiveness to cells and various biomolecules.MAP-RGD-blended SF fibers supported enhanced adhesion, proliferation,and spreading of mammalian cells as well as the efficient attachmentof biomolecules, including carbohydrate and protein. In addition,the hydrophilicity, swelling, and biodegradability of the MAP-RGD-blendedSF material were improved without notable hampering of the originalmechanical properties of SF. Therefore, the adhesive silk fibrousscaffold could be successfully used in diverse biomedical engineeringapplications. [ABSTRACT FROM AUTHOR]

Abstract: In this study, we elucidated the effect of residual silk sericin (SS) on structure and mechanical properties of regenerated silk filament as well as on fiber formation. The dope viscosity markedly increased with increasing residual SS content in dope solution which was prepared by dissolving the silk protein in formic acid. As a result of FTIR, 13C NMR, and XRD, a small amount of SS (9.6%) contained in the filament showed highest content of β-sheet conformation and maximum crystallinity. It seems that the SS affects the structural change of SF up to a certain level by inducing the β-transition easily. The tenacity of the filaments, containing 9.6–18.9% SS, was in the range of 2.1–2.4gf/d, which was about 50% higher than the filament without SS (pure SF). Consequently, with the enhancement of spinnability in wet spinning process, the SS can play an important role for developing the crystalline structure of SF as well as for improving mechanical properties of the regenerated silk fiber. [Copyright &y& Elsevier]