Effect of solvent on ionic liquid dissolved regenerated antheraea assamensis silk fibroin

We report on the dissolution of semi-domestic silk type Antheraea assamensis using ionic liquids. We investigated the impact of different coagulating solvents, including isopropanol and water on the structure and the morphology of the regenerated silk. We found that the water regenerated silk film showed a high β-sheet content and a native silk-like XRD pattern.

Relationship between processing, surface energy and bulk properties of ultrafine silk particles

Silk particles of different sizes and shapes were produced by milling and interactions with a series of polar and non-polar gaseous probes were investigated using an inverse gas chromatography technique. The surface energy of all silk materials is mostly determined by long range dispersive interactions such as van der Waals forces. The surface energy increases and surface energy heterogeneity widens after milling. All samples have amphoteric surfaces and the concentration of acidic groups increases after milling while the surfaces remain predominantly basic. We also examined powder compression and flow behaviours using a rheometer. Increase in surface energy, surface area, and static charges in sub-micron air jet milled particles contributed to their aggregation and therefore improved flowability. However they collapse under large pressures and form highly cohesive powder. Alkaline hydrolysis resulted in more crystalline fibres which on milling produced particles with higher density, lower surface energy and improved flowability. The compressibility, bulk density and cohesion of the powders depend on the surface energy as well as on particle size, surface area, aggregation state and the testing conditions, notably the consolidated and unconsolidated states. The study has helped in understanding how surface energy and flowability of particles can be changed via different fabrication approaches.

Microstructure and mechanical properties of silk from different components of the Antheraea pernyi cocoon

Silk fibres from different components of the Antheraea pernyi silkworm cocoon, namely peduncle, outer floss, and cocoon shells (outermost layer and pelade layer) were studied in detail to gain insights into the structure-property-function relationship. Among the fibres from different components, peduncle fibres are the softest with the largest viscoelastic lag, which may reduce the oscillation amplitude when a cocoon hangs on a twig. Fibres from the outermost layer are the toughest and have the largest breaking energy. Outer floss fibres have the highest content of sericin (about 11.98%) but their hardness and elasticity are intermediate. Pelade fibres are shape - preservable and stable with superior hardness and elasticity. The understanding of the properties of different silk fibres is essential for understanding their respective roles in the function of a silk cocoon and will also inspire new designs of protective materials under stringent environmental conditions.

Comparison of milling and solution approach for production of silk particles

Silk particles were produced by regenerating from silk solution, and using a milling method. In the regenerated silk particle production, two methods which are reported to render submicron silk particles were selected. Their particle sizes and structures were compared with particles of milling method already developed by us. The volume median average particle sizes (d(0.5)) of regenerated particles were much higher than what was reported previously. In contrast, milling method could produce particles with adjustable particle sizes ranging from micron to submicron level. All the milled particles had advantage of at least 15. °C higher thermal decomposition temperature than regenerated particles. They had silk II structure, and the crystallinity reduced as particle fineness increased, but remained higher than regenerated particles of similar sizes. © 2014 Elsevier B.V.

Green electrospun pantothenic acid/silk fibroin composite nanofibers: fabrication, characterization and biological activity

Silk fibroin (SF) from Bombyx mori has many established excellent properties and has found various applications in the biomedical field. However, some abilities or capacities of SF still need improving to meet the need for using practically. Indeed, diverse SF-based composite biomaterials have been developed. Here we report the feasibility of fabricating pantothenic acid (vitamin B5, VB5)-reinforcing SF nanofibrous matrices for biomedical applications through green electrospinning. Results demonstrated the successful loading of D-pantothenic acid hemicalcium salt (VB5-hs) into resulting composite nanofibers. The introduction of VB5-hs did not alter the smooth ribbon-like morphology and the silk I structure of SF, but significantly decreased the mean width of SF fibers. SF conformation transformed into β-sheet from random coil when composite nanofibrous matrices were exposed to 75% (v/v) ethanol vapor. Furthermore, nanofibers still remained good morphology after being soaked in water environment for five days. Interestingly, as-prepared composite nanofibrous matrices supported a higher level of cell viability, especially in a long culture period and significantly assisted skin cells to survive under oxidative stress compared with pure SF nanofibrous matrices. These findings provide a basis for further extending the application of SF in the biomedical field, especially in the personal skin-care field.

Structure and characteristics of milled silk particles

This study examined the structure, thermal property, and ion adsorption of silk particles. The particles were prepared by attritor-bead mill combination, using alkaline (pH10) charge repulsion and surfactant steric repulsion methods. Both methods produced particles with a dominant β-sheet structure, similar to the silk fibre. There was no significant difference in the decomposition temperatures for either the silk fibre or the micro/nano silk particles. An important finding from this study is clear evidence of reduction of amorphous content during the final stage of powdering using the bead mill. As a result, despite reduction in β-sheet crystallites with the progressive milling, the relative β-sheet content actually increased during this process. However, intermolecular forces between the β-sheets reduced significantly and hence the XRD results showed significant reduction in crystallinity in nano silk particles but crystal forming segments remained with β-sheet conformations after milling. The structural change influenced the ion-adsorption property where particle-size reduction resulted in a significant increase in both the rate and volume of HCrO4- adsorption. © 2014 Elsevier B.V.

Silk films as a repair material for perforations of the tympanic membrane

Large, chronic perforations of the tympanic membrane or eardrum can cause hearing loss as well as a range of secondary health problems. Current methods of repair usually involve grafting a material such as cartilage from another site on the body across the perforation. However, given problems such as possible infections at the graft donor site and the inability to see through the graft to assess infection within the middle ear, there is a need to develop an alternative material that is strong, readily available and transparent. Such a material would allow for less invasive surgery and potentially result in a superior hearing outcome for the patient. Our recent work has identified silk fibroin films as a promising material for this application. This paper reviews the repair of large perforations and compares the mechanical properties of silk with some existing graft materials. It also briefly discusses the difficulties in defining and comparing these properties with such different materials.

The impact of degumming conditions on the properties of silk films for biomedical applications

The degumming process to remove sericin decreases silk fiber strength; however, the impact of degumming on the mechanical properties of regenerated silk biomaterials has not been established. This study investigated the effect of degumming temperature, time, alkaline component and alkaline concentration on the mechanical properties of silk fibroin films. Sericin removal was estimated using weight loss; 10 samples with 12.2–29.4% weight loss were then further characterized in terms of fiber mechanical properties, fiber surface morphology, molecular weight distribution and film tensile strength. A negative correlation was found between weight loss and fiber tensile strength. This loss of fiber strength under harsher degumming conditions had a direct impact on the tensile strength of regenerated films. Mild degumming conditions (weight loss of 12.2%) led to higher film strength (8.9 MPa), whereas aggressive degumming conditions (with 29.4% weight loss) resulted in significantly weaker films (4.3 MPa). The presence of some residual sericin, after mild degumming, is likely to affect the mechanical properties of the regenerated silk films. These results will assist in the development of materials with mechanical and biocompatibility properties tuned to specific biomedical applications.

Silk cocoon: structure and properties

This study investigated the silk cocoon structure and its protective roles. It focussed mainly on three protection roles, viz mechanical protection (indentation), bacterial and UV protection. Silk cocoon absorbed UV-A and UV-B radiations and had the potential to be used as a photoprotective agent in sunscreens.

Milled non-mulberry silk fibroin microparticles as biomaterial for biomedical applications

Silk fibroin has been widely employed in various forms as biomaterials for biomedical applications due to its superb biocompatibility and tunable degradation and mechanical properties. Herein, silk fibroin microparticles of non-mulberry silkworm species (Antheraea assamensis, Antheraea mylitta and Philosamia ricini) were fabricated via a top-down approach using a combination of wet-milling and spray drying techniques. Microparticles of mulberry silkworm (Bombyx mori) were also utilized for comparative studies. The fabricated microparticles were physico-chemically characterized for size, stability, morphology, chemical composition and thermal properties. The silk fibroin microparticles of all species were porous (∼5μm in size) and showed nearly spherical morphology with rough surface as revealed from dynamic light scattering and microscopic studies. Non-mulberry silk microparticles maintained the typical silk-II structure with β-sheet secondary conformation with higher thermal stability. Additionally, non-mulberry silk fibroin microparticles supported enhanced cell adhesion, spreading and viability of mouse fibroblasts than mulberry silk fibroin microparticles (p<0.001) as evidenced from fluorescence microscopy and cytotoxicity studies. Furthermore, in vitro drug release from the microparticles showed a significantly sustained release over 3 weeks. Taken together, this study demonstrates promising attributes of non-mulberry silk fibroin microparticles as a potential drug delivery vehicle/micro carrier for diverse biomedical applications.

Drug loading and release studies for milled silk particles of different sizes

Milled silk particles with volume median particle size (d(0.5)) of 7 μm and 281 nm as well as silk snippets were used for loading of model drugs Orange G, Azophloxine, Rhodamine B, and Crystal Violet. Loading and release of these chemicals depended on the size of silk particles, pH, and the structure and properties of model drugs. Both types of silk particles reached equilibrium loading in less than 10 min due to high surface area whereas silk fibres needed more than 2-3 days to reach equilibrium, depending on the drug type. The uptake rate in fibres could be improved by increasing temperature. Both fibres and particles could slowly release the drugs over many days at 37 °C without a significant initial burst. As particle size decreased, the amount of model drug release also decreased. The release of drugs by the silk fibres was quicker than the silk particles.

Self-assembly of monolayered lipid membranes for surface-coating of a nanoconfined Bombyx mori silk fibroin film

Regenerated Bombyx mori (B. mori) silk fibroin is a type of widely used biomaterial. The β-sheet structure of it after methanol treatment provides water-insolubility and mechanical stability while on the other side leads to a hydrophobic surface which is less preferred by biological systems. In this work we prepare a novel type of nanoconfined silk fibroin film with a thickness below 100 nm. The film has a flat while hydrophobic surface because of its β-sheet structure due to the z-direction confinement during formation. Different types of lipid monolayers, DOPC, DPPC and MO, are assembled on the silk film surface. The lipid coating, especially the DPPC membrane, provides a much smoother and more hydrophilic surface due to the gel phase tails of the lipids, in comparison with the DOPC and MO ones which are in a liquid phase and have a much stronger interfacial association between silk film surface and lipid tails. Such a lipid coating preserves the biocompatibility and cellular affinity of the silk film which promises potential applications as surface coatings for materials for biological use.