Abstract: Silk fibroin/chitosan blend films were examined through IR spectroscopy to determine the conformational changes of silk fibroin. The effects of the fibroin/chitosan blend ratios (chitosan content) on the physical and mechanical properties were investigated to discover the feasibility of using these films as biomedical materials such as artificial skin and wound dressing. The mechanical properties of the blend films containing 10–40% chitosan were found to be excellent. The tensile strength, breaking elongation, and Young's modulus were affected by the chitosan contents of the blend films, which were also related to the density and degree of swelling. The coefficient of water vapor permeability of the blend films increased linearly with the chitosan content, and the values of 1000–2000 g m−2 day−1 were comparable to those of commercial wound dressings. Silk fibroin/chitosan blend films had good oxygen and water vapor permeabilities, making them useful as biomaterials. In particular, the blend film containing 40–50% chitosan showed very high oxygen permeability. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 80: 928–934, 2001

Abstract: The near-surface structure and the wettability of silk fibroin films cast from aqueous solutions on hydrophobic polystyrene substrates at various temperatures is investigated by Fourier transform infrared attenuated total reflection spectroscopy (FTIR-ATR) and measurement of contact angle. The FTIR data reveal that the near-surface region of the films is enriched in random coil conformations of the protein at the expense of a reduced fraction of α-helix and β-sheet conformations. The relative random coil/β-sheet content shows a marked dependence on the casting temperature, displaying a minimum at 50 °C. The minimum occurs concurrently with a maximum in the wettability of film surfaces by polar liquids. In the lower wettability region, the film surfaces of this hydrophilic protein are hydrophobic, whereas in the enhanced wettability range they are slightly hydrophilic. The experimental data indicate that during formation of fibron films, α-helix and β-sheet structures are rejected by the interface because ...

Abstract: Silk fibroin solution was prepared by dissolving the silk fibroin in triad solvent CaCl2 · CH3CH2OH · H2O. In this article we tested and analyzed the state of frozen silk fibroin solution and fine structure of freeze dried porous silk fibroin materials. The results indicated that the glass transition temperature of frozen silk fibroin solution ranges from −34 to −20°C, and the initial melting temperature of ice in frozen solution is about −8.5°C. When porous silk fibroin materials are prepared by means of freeze drying, if freezing temperature is below −20°C, the structure of silk fibroin is mainly amorphous with a little silk II crystal structure, and if freezing temperature is above −20°C, quite a lot of silk I crystal structure forms. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 79: 2185–2191, 2001

Abstract: Porous silk fibroin materials, with average pore size 10 ∼ 300 μm, pore density 1 ∼ 2000/mm2, and porosity 35 ∼ 70%, were prepared by freeze drying aqueous solution of silk fibroin obtained by dissolving silk fibroin in ternary solvent CaCl2 · CH3CH2OH · H2O. Pore size distribution of such materials mostly accorded with logarithmic normal distribution. It is possible to control the aforementioned structural parameters and the physical properties of moisture permeability, compressibility, strength, elongation, etc., by adjusting freezing temperature and concentration of silk fibroin solution. Above glass transition zone (−34 ∼ −20°C) of silk fibroin, the freezing temperature has more significant effect on the structure and properties of porous silk fibroin materials. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 79: 2192–2199, 2001

Abstract: Dissolution of Antheraea pernyi silk fiber was carried out in a calcium nitrate solution with various dissolving conditions. The solubility was significantly dependent on the concentration of calcium nitrate, dissolving temperature, and time. The proper conditions of dissolution were found as 7M calcium nitrate, 100°C temperature, and 3 h dissolving time. The aqueous solution of A. pernyi silk fibroin was composed of a mixture of polypeptides with several molecular weights above 14 kDa. FTIR and XRD showed that regenerated A. pernyi silk fibroin was composed of an α-helix as well as a random-coil conformation while silk fiber had a traditional (3-sheet structure. The endo-exo transition in the temperature ranges of 228-232°C also supports these conformations of regenerated silk fibroin film. TGA and DTG curves showed that the thermal decomposition of regenerated A. pernyi silk fibroin proceeded by three steps, not dependent on the conformation. The mechanical damping peaks (tan δ) appeared about 227°C with a minor shoulder maximum about 240°C, which were somewhat lower than those of tussah silk fiber.

Abstract: Silk fibroin (SF) is a highly promising protein for its surface and structural properties, associated with a good bio- and hemo-compatibility. However, its mechanical properties and architecture cannot be easily tailored to meet the requirements of specific applications. In this work, SF was used to modify the surface properties of polyurethanes (PUs), thus obtaining 2D and 3D scaffolds for tissue regeneration. PUs were chosen for their well known advantageous properties and versatility; they can be obtained either as 2D (films) or 3D (foams) substrates. Films of a medical-grade poly-carbonate-urethane were prepared by solvent casting; PU foams were purposely designed and prepared with a morphology (porosity and cell size) adequate for cell growth. PU substrates were coated with fibroin by a dipping technique. To stabilize the coating layer, a conformational change of the protein from the α-form (water soluble) to the β-form (not water soluble) was induced. Novel methodology in UV spectroscopy were developed for quantitatively analyzing the SF-concentration in dilute solutions. Pure fibroin was used as standard, as an alternative to the commonly used albumin, allowing real concentration values to be obtained. SF-coatings showed good stability in physiological-like conditions. A treatment with methanol further stabilized the coating. Preliminary results with human fibroblasts indicated that SF coating promote cell adhesion and growth, suggesting that SF-modified PUs appear to be suitable scaffolds for tissue engineering applications. © 2001 Kluwer Academic Publishers

Abstract: Fibroin gene expression during the larval developmental stages of the Saturniid silkworm, Antheraea mylitta, was analyzed. Northern blot analysis of larval silk gland total RNA using the fibroin gene as a probe showed that fibroin is expressed in the intermoult stages and repressed during the moulting stages. Abundance of fibroin transcripts gradually increased from the third to fifth intermoult stage, reaching a peak in the fifth intermoult. Transcripts declined during the early spinning stage. Western blot analysis of fibroin protein production with anti-fibroin antibody confirmed the differential fibroin expression, in accordance with fibroin mRNA synthesis. Dot blot hybridization of genomic DNA isolated from each larval developmental stage with the labelled fibroin gene showed that at the genomic level, the relative concentration of the fibroin gene was constant throughout the developmental stages. Our data confirm that fibroin gene expression in A. mylitta, like in B. mori, is transcriptionally controlled and shows differential temporal variations.

Abstract: A series of novel human-made functional fibers (biofibers) based on chitin and chitosan are prepared by the wet-spinning and the post chemical modification of chitosan fiber. The wet-spinning gives rise to a series of biofibers: chitin, chitosan, chitin-cellulose, chitosan-cellulose, chitin-silk fibroin, chitin-glycosaminoglycans, chitin-cellulose-silk fibroin, chitosan-tropocollagen, and chitin-cellulose-silk fibroin. The post chemical modification of chitosan fiber gives rise to a series of chemically modified fibers: N-acylchitosans, N-arylidene- and N-alkylidene-chitosans, N-acetylchitosan (chitin)-tropocollagen, and chitosan-transition metal complexes. Some of the current and potential applications of these biofibers are described.

Abstract: A process for producing non-woven silk fiber fabrics comprises the following steps: a) obtaining silk fibroin, for example either from silk cocoons, or silk textiles or waste silk; b) removing the sericin layer covering the silk fibroin fibers, when present; c) breaking the disulfide bonds between heavy (350 kDa) and light (27kDa) chains of silk fibroin in order to obtain the production of chain fragments which serve as a specific cellular recognition sites promoting the attachment and growth of cells. d) homogenising of the material resulting from step c).

Abstract: Semi-interpenetrating polymer networks (SIPNs) composed of silk fibroin (SF) and a poly(ethylene glycol) (PEG) diacrylate macromer were synthesized and characterized. The conformation of SF in SIPNs was changed from a random coil to a β-sheet structure by formation of SIPNs with PEG, while the crystalline structures of SF and PEG in SIPNs was not changed. The decomposition temperature of SF in the SIPNs was inner-shifted, an indication of SIPN formation between the SF and the PEG macromer. The tensile strength and elongation of the SIPNs were much higher than were those of SF itself or the SF/PEG blend and increased with an increase of the PEG content. The equilibrium water content of SF was remarkably increased by formation of SIPNs with PEG due to the hydrophilic property of PEG. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 80: 1848–1853, 2001

Abstract: A method for producing a silk or silk-like fiber, characterized in that it is spun from a solution of silk fibroin and/or a silk-like material in hexafluoroacetone hydrate and optionally is then stretched, or in hat it is casted from a solution of silk fibroin and/or a silk-like material in hexafluoroacetone hydrate followed by drying, and optionally is then stretched.

Abstract: In the presence of fine silk cocoon (silk fibroin, SF) powder, a low viscosity sol of nano-hydroxyapatite (HAp, Ca10(PO4)6(OH)2)—SF was synthesized by a wet mechanochemical reaction. Nano crystals of HAp are oriented along their c-axis. The secondary structure of SF was changed by milling. A uniform thin gel film was obtained by a simple dip coating on the glass substrate precoated by chitosan.

Abstract: The nucleotide sequences containing an entire genomic region and 5′ upstream region of Antheraea yamamai fibroin gene have been determined. The gene consists of an initial exon encoding 14 amino acids, an intron (150 bp), and a long second exon coding for 2641 amino acids. The fibroin coding sequence shows a specialized organization with a highly repetitive region flanked by non repetitive 5′ and 3′ ends. Northern blot analyses confirmed that fibroin gene is actively expressed in the posterior silk gland of the final instar larvae of Antheraea yamamai.

Abstract: Regenerated Antheraea pernyi silk fibroin films prepared from calcium nitrate solution were treated with heat for crystallization. The structural and conformational changes caused by heat treatment were investigated with X-ray diffraction, infrared spectroscopy, and differential scanning calorimetry. The temperature and treatment time of heat greatly influenced the conformation of the regenerated films. The conformational change was caused by heat treatment over 230°C, and the transition was enhanced by increasing time and temperature. The contents of α-helix, β-sheet, and random-coil conformations were calculated and examined to investigate the effect of heat treatment on the conformational changes. The β-sheet structure could be transformed from a random-coil conformation on heat treatment, whereas the content of α-helix structure was nearly unchanged, regardless of heat-treatment conditions. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 81: 2271–2276, 2001

Abstract: Poly(e-caprolactone-co-D,L-lactide) copolymers with 10, 30, and 50% by weight of silk particles (size range: 5-250 μm) derived from Bombyx mori were blended in acetone solution. After evaporation of the solvent, the morphology, thermal behavior, and mechanical properties of the composites were examined. The composites were transparent and the silk fibroin particles were homogeneously distributed within the composite structure. The particles appeared as bright reflected images under the optical microscope, suggesting that they were in a crystalline state. DSC thermograms of the composites revealed that the glass transition of the matrix was at ca. -18°C. Degradation of the silk fibroin occurred beyond 270°C. The decomposition temperatures and degradation rate decreased with increasing silk fibroin content as revealed by TGA analysis. FTIR spectra of the composites showed absorption bands at 1730 and 1088 cm-1 for the copolymer and at 3273 and 1617 cm-1 for the silk fibroin. Although the characteristic line...

Abstract: Silk fibroin is a simple protein expected to have functional applications in medicine and bioelectronics. The primary structure of this protein is quite simple, and the main secondary structures are β-sheet crystals and amorphous random coils. In the present study, we investigated pulsed laser deposition (PLD) of fibroin with the β-sheet structures as targets. The primary and secondary structures in films deposited were analyzed using infrared spectroscopy. Normal laser deposition at 351 nm using neat fibroin targets produced thin films of fibroin with a random coiled structure. Ablation was triggered by two-photonic excitation of the peptide chains, which resulted in the destruction of β-sheet structure in PLD. In order to avoid the two-photonic excitation, we adopted a PLD method utilizing anthracene (5–0.1 wt %) in a photosensitized reaction involving doped fibroin targets. Laser light (351 or 355 nm) was absorbed only by anthracene, which plays an important role converting photon energy to thermal ene...

Abstract: Surface characterization of materials has been considered critical in the development of biomaterials, as many unfavorable responses from the body occur at the interface between a material and the body component. The contact angle measurement is one means to characterize the surface properties and to correlate them to the biocompatibility of materials. In this paper, surface characteristics of silk fibroin/S-carboxymethyl kerateine, representative fibrous proteins, were investigated by contact angle measurements and ESCA. The biocompatibility of the blends was evaluated based on minimal interfacial free energy concept, and compared with other potential biomaterials. It was also hypothesized that the enhanced surface polarity of the blends was generated from the conformational transition of proteins. This approach to evaluate the biocompatibility of materials based on surface characteristics may find wide utility in many biomedical applications.

Abstract: In this study, the synergistic aminolysis (sodium hydroxide and ethylene diamine in one bath) mechanism of polyethylene terephthalate (PET) microfiber crepe fabric was preliminarily investigated. The results showed that not only active groups (i.e., NHR, COOH, and OH) but also considerable cracks and craters were introduced onto PET fibers, which provided locations for a subsequent crosslinking reaction and mechanical attachment and so facilitated the silk-fibroin finishing. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 81: 1467–1473, 2001

Abstract: Silk protein has been investigated by many researchers to apply to biomedical field. We reviewed biomedical applications of silk protein such as matrix of wound dressing and drug delivery system. Since silk fibroin/ poly (ethylene glycol) (PEG) semi-interpenetrating polymer networks showed good mechanical properties and wound healing phenomena, it can be used as wound dressing materials. Sericin nanoparticles pre- pared by conjugation with PEG and silk protein/ poloxamer mixture gel are expected to become a deliv- ery as matrix for hydrophobic drug.

Abstract: An emulsifier for cosmetics which comprises an aqueous solution of silk protein containing non-crystalline silk fibroin as the main component, wherein the aqueous solution is a solution of silk protein in water obtained by solubilizing unrefined, partly refined or refined fiber spun by silkworm with the use of a neutral salt and then dialyzing. The emulsifier for cosmetics comprising an aqueous solution or an aqueous gel of silk protein containing non-crystalline silk fibroin as the main component is effective not only in improving emulsifying ability and texture in using but also promoting the growth of skin cells. Owing to these effects, emulsions prepared by emulsifying oily components with the use of this emulsifier are usable in oil-water type emulsified cosmetics and highly efficacious in skin care.

Abstract: The crystal structure, thermal properties and growth rates of spherulites of the Tussah silk fibroin, produced upon drying of the silk taken directly from the lumen which is essentially a poly(L-alanine)polypeptide, are investigated. Depending on casting conditions, spherulites with either αhelical chain conformation or β parallel sheet structure are produced. The growth rates display a strong positive temperature coefficient, with an apparent transition, which however cannot be related with the formation of two different crystal structures at this stage.

Abstract: Several ice nucleating substances have been identified, which exist in vivo or can be extracted from biological materials. Spider silk, which has a strong ability for water condensation, has also been found to possess an ice nucleation activity. The freezing temperature of water droplets was higher in the presence than in the absence of spider silk. Moreover, by means of environmental scanning electron microscopy, it was observed that the activity is not due to foreign matter attached to the silk but to the silk fibroin itself.