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Sample records for silk gland fibroin

  1. Preparation of Porous Scaffolds from Silk Fibroin Extracted from the Silk Gland of Bombyx mori (B. mori

    Liangjun Zhu

    2012-06-01

    Full Text Available In order to use a simple and ecofriendly method to prepare porous silk scaffolds, aqueous silk fibroin solution (ASF was extracted from silk gland of 7-day-old fifth instar larvae of Bombyx mori (B. mori. SDS-page analysis indicated that the obtained fibroin had a molecular weight higher than 200 kDa. The fabrication of porous scaffolds from ASF was achieved by using the freeze-drying method. The pore of porous scaffolds is homogenous and tends to become smaller with an increase in the concentration of ASF. Conversely, the porosity is decreased. The porous scaffolds show impressive compressive strength which can be as high as 6.9 ± 0.4 MPa. Furthermore, ASF has high cell adhesion and growth activity. It also exhibits high ALP activity. This implies that porous scaffolds prepared from ASF have biocompatibility. Therefore, the porous scaffolds prepared in this study have potential application in tissue engineering due to the impressive compressive strength and biocompatibility.

  2. Self-assembly of silk fibroin under osmotic stress

    Sohn, Sungkyun

    The supramolecular self-assembly behavior of silk fibroin was investigated using osmotic stress technique. In Chapter 2, a ternary phase diagram of water-silk-LiBr was constructed based on X-ray results on the osmotically stressed regenerated silk fibroin of Bombyx mori silkworm. Microscopic data indicated that silk I is a hydrated structure and a rough estimate of the number of water molecules lost by the structure upon converting from silk I to silk II has been made, and found to be about 2.2 per [GAGAGS] hexapeptide. In Chapter 3, wet-spinning of osmotically stressed, regenerated silk fibroin was performed, based on the prediction that the enhanced control over structure and phase behavior using osmotic stress method helps improve the physical properties of wet-spun regenerated silk fibroin fibers. The osmotic stress was applied in order to pre-structure the regenerated silk fibroin molecule from its original random coil state to more oriented state, manipulating the phase of the silk solution in the phase diagram before the start of spinning. Monofilament fiber with a diameter of 20 microm was produced. In Chapter 4, we investigated if there is a noticeable synergistic osmotic pressure increase between co-existing polymeric osmolyte and salt when extremely highly concentrated salt molecules are present both at sample subphase and stressing subphase, as is the case of silk fibroin self-assembly. The equilibration method that measures osmotic pressure relative to a reference with known osmotic pressure was introduced. Osmotic pressure of aqueous LiBr solution up to 2.75M was measured and it was found that the synergistic effect was insignificant up to this salt concentration. Solution parameters of stressing solutions and Arrhenius kinetics based on time-temperature relationship for the equilibration process were derived as well. In Chapter 5, self-assembly behavior of natural silk fibroin within the gland of Bombyx mori silkworm was investigated using osmotic stress technique. Microscopic and thermodynamic details of this self-assembly process along the spinline have been assessed. Formation of a needle-shaped molecular lath under appropriate osmotic stress was found. Silk I degree of hydration of silk gland was quantitatively estimated by image analysis of optical micrographs and the numbers varied from 2.2 to 2.7 depending on the region in the gland. Osmotic pressure in the gland was also estimated by equilibration method.

  3. Effect of degumming ph value on electrospining of silk fibroin

    Lu Shen-Zhou

    2014-01-01

    Full Text Available Regenerated silk fibroin fibers show properties dependent on the molecular weight of fibroin. The cocoon-degumming approaches had great impact on the degradation of silk fibroin. The effect of degumming pH value to electrospining of fibroin was studied in this paper. The viscosity and molecular weight of regenerated silk fibroin were studied using rheometer and gel electrophoresis. The results showed that the weaker the alkalinity of degumming reagent, there was the milder the effect on silk fibroin molecular. The fibroin fibers can be prepared by electrospining with low concentration of regenerated silk fibroin solution.

  4. Thermal crystallization mechanism of silk fibroin protein

    Hu, Xiao

    In this thesis, the thermal crystallization mechanism of silk fibroin protein from Bombyx mori silkworm, was treated as a model for the general study of protein based materials, combining theories from both biophysics and polymer physics fields. A systematic and scientific path way to model the dynamic beta-sheet crystallization process of silk fibroin protein was presented in the following sequence: (1) The crystallinity, fractions of secondary structures, and phase compositions in silk fibroin proteins at any transition stage were determined. Two experimental methods, Fourier transform infrared spectroscopy (FTIR) with Fourier self-deconvolution, and specific reversing heat capacity, were used together for the first time for modeling the static structures and phases in the silk fibroin proteins. The protein secondary structure fractions during the crystallization were quantitatively determined. The possibility of existence of a "rigid amorphous phase" in silk protein was also discussed. (2) The function of bound water during the crystallization process of silk fibroin was studied using heat capacity, and used to build a silk-water dynamic crystallization model. The fundamental concepts and thermal properties of silk fibroin with/without bound water were discussed. Results show that intermolecular bound water molecules, acting as a plasticizer, will cause silk to display a water-induced glass transition around 80C. During heating, water is lost, and the change of the microenvironment in the silk fibroin chains induces a mesophase prior to thermal crystallization. Real time FTIR during heating and isothermal holding above Tg show the tyrosine side chain changes only during the former process, while beta sheet crystallization occurs only during the latter process. Analogy is made between the crystallization of synthetic polymers according to the four-state scheme of Strobl, and the crystallization process of silk fibroin, which includes an intermediate precursor stage before crystallization. (3) The beta-sheet crystallization kinetics in silk fibroin protein were measured using X-ray, FTIR and heat flow, and the structure reveals the formation mechanism of the silk crystal network. Avrami kinetics theories, which were established for studies of synthetic polymer crystal growth, were for the first time extended to investigate protein self-assembly in multiblock silk fibroin samples. The Avrami exponent, n, was close to two for all methods, indicating formation of beta sheet crystals in silk proteins is different from the 3-D spherulitic crystal growth found in most synthetic homopolymers. A microphase separation pattern after chymotrypsin enzyme biodegradation was shown in the protein structures using scanning electron microscopy. A model was then used to explain the crystallization of silk fibroin protein by analogy to block copolymers. (4) The effects of metal ions during the crystallization of silk fibroin was investigated using thermal analysis. Advanced thermal analysis methods were used to analyze the thermal protein-metallic ion interactions in silk fibroin proteins. Results show that K+ and Ca2+ metallic salts play different roles in silk fibroin proteins, which either reduce (K+) or increase (Ca2+ ) the glass transition (Tg) of pure silk protein and affect the thermal stability of this structure.

  5. Silver nanoparticle containing silk fibroin bionanotextiles

    Development of new generation bionanotextiles is an important growing field, and they have found applications as wound dressings, bandages, tissue scaffolds, etc. In this study, silver nanoparticle (AgNP) containing silk-based bionanotextiles were fabricated by electrospinning, and processing parameters were optimized and discussed in detail. AgNPs were in situ synthesized within fibroin nanofibers by UV reduction of silver ions to metallic silver. The influence of post-treatments via methanol treatment and glutaraldehyde (GA) vapor exhibited changes in the secondary structure of silk. Methanol treatment increased the tensile properties of fibers due to supported crystalline silk structure, while GA vapor promoted amorphous secondary structure. AgNP containing silk fibroin bionanotextiles had strong antibacterial activity against gram-positive Staphylococcus aureus and gram-negative Pseudomonas aeruginosa

  6. Structural analysis and application to biomaterials of the silk fibroins

    Silk fibroin from Bombyx mori silkworm has outstanding mechanical properties despite being spun from aqueous solution. I have clarified two distinct structures in the solid state; silk I and silk II, which mean the structures before and after spinning, by using solid state NMR. Moreover, I have been developing several kinds of biomaterials, such as bone regeneration materials and vascular grafts. In this paper, I present two topics: one is the structural analyses of the silk fibroin in detail, the other is applications of silk fibroins to tissue engineering. In the case of vascular regeneration, I have developed the small diameter vascular grafts made by silk fibroins. The new grafts from silk fibroins have good patency, and these grafts were commonly covered with cells and platelets at 4 weeks after implantation. For bone tissue engineering, I performed structural analyses of a new silk-like peptide, En(AGSGAG)4, in order to consider the molecular design of biomaterials for bone regeneration. (author)

  7. Cytocompatibility of a silk fibroin tubular scaffold

    Regenerated silk fibroin (SF) materials are increasingly used for tissue engineering applications. In order to explore the feasibility of a novel biomimetic silk fibroin tubular scaffold (SFTS) crosslinked by poly(ethylene glycol) diglycidyl ether (PEG-DE), biocompatibility with cells was evaluated. The novel biomimetic design of the SFTS consisted of three distinct layers: a regenerated SF intima, a silk braided media and a regenerated SF adventitia. The SFTS exhibited even silk fibroin penetration throughout the braid, forming a porous layered tube with superior mechanical, permeable and cell adhesion properties that are beneficial to vascular regeneration. Cytotoxicity and cell compatibility were tested on L929 cells and human umbilical vein endothelial cells (EA.hy926). DNA content analysis, scanning electron and confocal microscopies and MTT assay showed no inhibitory effects on DNA replication. Cell morphology, viability and proliferation were good for L929 cells, and satisfactory for EA.hy926 cells. Furthermore, the suture retention strength of the SFTS was about 23 N and the Young's modulus was 0.2–0.3 MPa. Collectively, these data demonstrate that PEG-DE crosslinked SFTS possesses the appropriate cytocompatibility and mechanical properties for use as vascular scaffolds as an alternative to vascular autografts. - Highlights: • A PEG-DE cross-linked small caliber porous silk fibroin tubular scaffold (SFTS) • PEG-DE cross-linked SF film had no inhibitory effect on DNA replication of cells. • Cells cultured on the SFTS showed good morphology, cell viability and proliferative activity. • SFTS would be beneficial to endothelialization. • SFTS had good suture retention strength and flexibility

  8. Cytocompatibility of a silk fibroin tubular scaffold

    Wang, Jiannan, E-mail: wangjn@suda.edu.cn; Wei, Yali; Yi, Honggen; Liu, Zhiwu; Sun, Dan; Zhao, Huanrong

    2014-01-01

    Regenerated silk fibroin (SF) materials are increasingly used for tissue engineering applications. In order to explore the feasibility of a novel biomimetic silk fibroin tubular scaffold (SFTS) crosslinked by poly(ethylene glycol) diglycidyl ether (PEG-DE), biocompatibility with cells was evaluated. The novel biomimetic design of the SFTS consisted of three distinct layers: a regenerated SF intima, a silk braided media and a regenerated SF adventitia. The SFTS exhibited even silk fibroin penetration throughout the braid, forming a porous layered tube with superior mechanical, permeable and cell adhesion properties that are beneficial to vascular regeneration. Cytotoxicity and cell compatibility were tested on L929 cells and human umbilical vein endothelial cells (EA.hy926). DNA content analysis, scanning electron and confocal microscopies and MTT assay showed no inhibitory effects on DNA replication. Cell morphology, viability and proliferation were good for L929 cells, and satisfactory for EA.hy926 cells. Furthermore, the suture retention strength of the SFTS was about 23 N and the Young's modulus was 0.2–0.3 MPa. Collectively, these data demonstrate that PEG-DE crosslinked SFTS possesses the appropriate cytocompatibility and mechanical properties for use as vascular scaffolds as an alternative to vascular autografts. - Highlights: • A PEG-DE cross-linked small caliber porous silk fibroin tubular scaffold (SFTS) • PEG-DE cross-linked SF film had no inhibitory effect on DNA replication of cells. • Cells cultured on the SFTS showed good morphology, cell viability and proliferative activity. • SFTS would be beneficial to endothelialization. • SFTS had good suture retention strength and flexibility.

  9. Biological Properties of Regenerated Silk Fibroin Films

    L.А. Safonova

    2015-09-01

    Full Text Available The aim of the investigation was to study the biological and mechanical properties of silk fibroin films and composite silk fibroin films containing 30% collagen by weight. Materials and Methods. All films were prepared by casting method using water and formic acid as solvents. Scanning electron microscopy and atomic force microscopy were applied. Human hepatoblastoma cell line Hep-G2 was used to test film compatibility. Results. We studied surface roughness degree of the obtained films. Water-based films were found to have permeability for low molecular weight substances. Tensile strength and elasticity indices were measured for all types of films. Collagen added to film composition was revealed to have no significant effect on tensile strength, though it increased film elasticity. We studied the degradation of films. Collagen was shown to have no significant impact on film degradation rate both in phosphate-buffer saline and oxidizing media. We demonstrated with an example of cell line Hep-G2 that water-based films exhibit higher proliferative activity. Conclusion. Silk fibroin water solution has the best properties to develop films for tissue engineering.

  10. ELECTROPHORETIC SEPARATION AND COMPARATIVE ANALYSIS OF SILK GLAND PROTEINS FROM BOMBYX AND PHILOSAMIA

    Muzafar A Bhat, Punyavathi and Manjunatha H Boregowda*

    2014-07-01

    Full Text Available A comparative analysis of protein extracted from different regions of silk glands in the Bombyx mori L. and Philosamia ricini Hutt was performed employing single-dimensional-electrophoresis technique. Notably, a protein extracted directly from the lumen of the middle silk gland yielded two discrete protein bands with molecular mass of 325 and 26 kDa representing fibroin heavy (H and low (L chains than whole silk gland of B. mori. Contrastingly, such differentiation in protein separation could not be achieved due to undifferentiated regions of silk gland in P. ricini. Interestingly, copious amount of proteins with different molecular weight were noticed in the whole silk gland of B. mori and P. ricini, whose functional properties in the silk gland remain enigmatic. Thus, middle and whole silk glands of B. mori shall be an ideal source for the extraction of fibroin and sericin respectively as potent bio-materials for biotechnological and biomedical applications.

  11. A novel electrospun silk fibroin/hydroxyapatite hybrid nanofibers

    A novel electrospinning of silk fibroin/hydroxyapatite hybrid nanofibers with different composition ratios was performed with methanoic acid as a spinning solvent. The silk fibroin/hydroxyapatite hybrids containing up to 30% hydroxyapatite nanoparticles could be electrospun into the continuous fibrous structure. The electrospun silk fibroin/hydroxyapatite hybrid nanofibers showed bigger diameter and wider diameter distribution than pure silk fibroin nanofibers, and the average diameter gradually increased from 95 to 582 nm. At the same time, the secondary structure of silk fibroin/hydroxyapatite nanofibers was characterized by X-ray diffraction, Fourier transform infrared analysis, and DSC measurement. Comparing with the pure silk fibroin nanofibers, the crystal structure of silk fibroin was mainly amorphous structure in the hybrid nanofibers. X-ray diffraction results demonstrated the hydroxyapatite crystalline nature remained as evidenced from the diffraction planes (002), (211), (300), and (202) of the hydroxyapatite crystallites, which was also confirmed by Fourier transform infrared analysis. The thermal behavior of hybrid nanofibers exhibited the endothermic peak of moisture evaporation ranging from 86 to 113 °C, and the degradation peak at 286 °C appeared. The SF/HAp nanofibers mats containing 30% HAp nanoparticles showed higher breaking tenacity and extension at break for 1.1688 ± 0.0398 MPa and 6.55 ± 1.95%, respectively. Therefore, the electrospun silk fibroin/hydroxyapatite hybrid nanofibers should be provided potentially useful options for the fabrication of biomaterial scaffolds for bone tissue engineering. -- Highlights: ► The novel SF/HAp nanofibers were directly prepared by electrospinning method. ► The nanofiber diameter had significant related to the content of HAp. ► The crystal structure of silk fibroin was mainly amorphous structure in the hybrid nanofibers. ► The HAp crystals existing in the hybrid nanofibers were characterized using XRD and FTIR.

  12. A novel electrospun silk fibroin/hydroxyapatite hybrid nanofibers

    Ming, Jinfa, E-mail: jinfa.ming@gmail.com [National Engineering Laboratory for Modern Silk, Soochow University, Suzhou 215123 (China); College of Textile and Clothing Engineering, Soochow University, Suzhou 215021 (China); Zuo, Baoqi, E-mail: bqzuo@suda.edu.cn [National Engineering Laboratory for Modern Silk, Soochow University, Suzhou 215123 (China); College of Textile and Clothing Engineering, Soochow University, Suzhou 215021 (China)

    2012-11-15

    A novel electrospinning of silk fibroin/hydroxyapatite hybrid nanofibers with different composition ratios was performed with methanoic acid as a spinning solvent. The silk fibroin/hydroxyapatite hybrids containing up to 30% hydroxyapatite nanoparticles could be electrospun into the continuous fibrous structure. The electrospun silk fibroin/hydroxyapatite hybrid nanofibers showed bigger diameter and wider diameter distribution than pure silk fibroin nanofibers, and the average diameter gradually increased from 95 to 582 nm. At the same time, the secondary structure of silk fibroin/hydroxyapatite nanofibers was characterized by X-ray diffraction, Fourier transform infrared analysis, and DSC measurement. Comparing with the pure silk fibroin nanofibers, the crystal structure of silk fibroin was mainly amorphous structure in the hybrid nanofibers. X-ray diffraction results demonstrated the hydroxyapatite crystalline nature remained as evidenced from the diffraction planes (002), (211), (300), and (202) of the hydroxyapatite crystallites, which was also confirmed by Fourier transform infrared analysis. The thermal behavior of hybrid nanofibers exhibited the endothermic peak of moisture evaporation ranging from 86 to 113 Degree-Sign C, and the degradation peak at 286 Degree-Sign C appeared. The SF/HAp nanofibers mats containing 30% HAp nanoparticles showed higher breaking tenacity and extension at break for 1.1688 {+-} 0.0398 MPa and 6.55 {+-} 1.95%, respectively. Therefore, the electrospun silk fibroin/hydroxyapatite hybrid nanofibers should be provided potentially useful options for the fabrication of biomaterial scaffolds for bone tissue engineering. -- Highlights: Black-Right-Pointing-Pointer The novel SF/HAp nanofibers were directly prepared by electrospinning method. Black-Right-Pointing-Pointer The nanofiber diameter had significant related to the content of HAp. Black-Right-Pointing-Pointer The crystal structure of silk fibroin was mainly amorphous structure in the hybrid nanofibers. Black-Right-Pointing-Pointer The HAp crystals existing in the hybrid nanofibers were characterized using XRD and FTIR.

  13. Sodium ion effect on silk fibroin conformation characterized by solid-state NMR and generalized 2D NMR NMR correlation

    Ruan, Qing-Xia; Zhou, Ping

    2008-07-01

    In the present work, we investigated Na + ion effect on the silk fibroin (SF) conformation. Samples are Na +-involved regenerated silk fibroin films. 13C CP-MAS NMR demonstrates that as added [Na +] increases, partial silk fibroin conformation transit from helix-form to ?-form at certain Na + ion concentration which is much higher than that in Bombyx mori silkworm gland. The generalized two-dimensional NMR-NMR correlation analysis reveals that silk fibroin undergoes several intermediate states during its conformation transition process as [Na +] increase. The appearance order of the intermediates is followed as: helix and/or random coil ? helix-like ? ?-sheet-like ? ?-sheet, which is the same as that produced by pH decrease from 6.8 to 4.8 in the resultant regenerated silk fibroin films. The binding sites of Na + to silk fibroin might involve the carbonyl oxygen atom of certain amino acids sequence which could promote the formation of ?-sheet conformation. Since the Na +sbnd O bond is weak, the ability of Na + inducing the secondary structure transition is weaker than those of Ca 2+, Cu 2+ and even K +. It is maybe a reason why the sodium content is much lower than potassium in the silkworm gland.

  14. Impact of Protein-Metal Ion Interactions on the Crystallization of Silk Fibroin Protein

    Hu, Xiao; Lu, Qiang; Kaplan, David; Cebe, Peggy

    2009-03-01

    Proteins can easily form bonds with a variety of metal ions, which provides many unique biological functions for the protein structures, and therefore controls the overall structural transformation of proteins. We use advanced thermal analysis methods such as temperature modulated differential scanning calorimetry and quasi-isothermal TMDSC, combined with Fourier transform infrared spectroscopy, and scanning electron microscopy, to investigate the protein-metallic ion interactions in Bombyx mori silk fibroin proteins. Silk samples were mixed with different metal ions (Ca^2+, K^+, Ma^2+, Na^+, Cu^2+, Mn^2+) with different mass ratios, and compared with the physical conditions in the silkworm gland. Results show that all metallic ions can directly affect the crystallization behavior and glass transition of silk fibroin. However, different ions tend to have different structural impact, including their role as plasticizer or anti-plasticizer. Detailed studies reveal important information allowing us better to understand the natural silk spinning and crystallization process.

  15. Silk Fibroin as Edible Coating for Perishable Food Preservation.

    Marelli, B; Brenckle, M A; Kaplan, D L; Omenetto, F G

    2016-01-01

    The regeneration of structural biopolymers into micelles or nanoparticles suspended in water has enabled the design of new materials with unique and compelling properties that can serve at the interface between the biotic and the abiotic worlds. In this study, we leveraged silk fibroin quintessential properties (i.e. polymorphism, conformability and hydrophobicity) to design a water-based protein suspension that self-assembles on the surface of food upon dip coating. The water-based post-processing control of the protein polymorphism enables the modulation of the diffusion of gases through the silk fibroin thin membranes (e.g. O2 and CO2 diffusion, water vapour permeability), which is a key parameter to manage food freshness. In particular, an increased beta-sheet content corresponds to a reduction in oxygen diffusion through silk fibroin thin films. By using the dip coating of strawberries and bananas as proof of principle, we have shown that the formation of micrometre-thin silk fibroin membranes around the fruits helps the management of postharvest physiology of the fruits. Thus, silk fibroin coatings enhance fruits' shelf life at room conditions by reducing cell respiration rate and water evaporation. The water-based processing and edible nature of silk fibroin makes this approach a promising alternative for food preservation with a naturally derived material. PMID:27151492

  16. ELECTROPHORETIC SEPARATION AND COMPARATIVE ANALYSIS OF SILK GLAND PROTEINS FROM BOMBYX AND PHILOSAMIA

    Muzafar A Bhat, Punyavathi and Manjunatha H Boregowda*

    2014-01-01

    A comparative analysis of protein extracted from different regions of silk glands in the Bombyx mori L. and Philosamia ricini Hutt was performed employing single-dimensional-electrophoresis technique. Notably, a protein extracted directly from the lumen of the middle silk gland yielded two discrete protein bands with molecular mass of 325 and 26 kDa representing fibroin heavy (H) and low (L) chains than whole silk gland of B. mori. Contrastingly, such differentiation in protein separation cou...

  17. Amorphous Silk Fibroin Membranes for Separation of CO2

    Aberg, Christopher M.; Patel, Anand K.; Gil, Eun Seok; Spontak, Richard J.; Hagg, May-Britt

    2009-01-01

    Amorphous silk fibroin has shown promise as a polymeric material derivable from natural sources for making membranes for use in removing CO2 from mixed-gas streams. For most applications of silk fibroin, for purposes other than gas separation, this material is used in its highly crystalline, nearly natural form because this form has uncommonly high tensile strength. However, the crystalline phase of silk fibroin is impermeable, making it necessary to convert the material to amorphous form to obtain the high permeability needed for gas separation. Accordingly, one aspect of the present development is a process for generating amorphous silk fibroin by treating native silk fibroin in an aqueous methanol/salt solution. The resulting material remains self-standing and can be prepared as thin film suitable for permeation testing. The permeability of this material by pure CO2 has been found to be highly improved, and its mixed-gas permeability has been found to exceed the mixed-gas permeabilities of several ultrahigh-CO2-permeable synthetic polymers. Only one of the synthetic polymers poly(trimethylsilylpropyne) [PTMSP] may be more highly permeable by CO2. PTMSP becomes unstable with time, whereas amorphous silk should not, although at the time of this reporting this has not been conclusively proven.

  18. Silk Fibroin/Starch Blend Films: Preparation and Characterization

    Prasong Srihanam

    2011-01-01

    This study was aimed to prepare silk fibroin (SF) of Thai silk (Bombyx mori) and cassava starch blend films by solvent evaporation method. The SF solution was firstly prepared from the cocoons by dissolving pure silk fibroin with tertiary solvent system of CaCl2: Ethanol: H2O (1:2:8 by mol). Cassava starch solution could be prepared by boiling at high temperature. The mixture of the SF and cassava starch solution was then blended and cast on the polystyrene plates. The obtained films were inv...

  19. Nanorheology of regenerated silk fibroin solution

    A Raghu; Sharath Ananthamurthy

    2008-06-01

    We have investigated the rheological properties of regenerated silk fibroin (RSF), a viscoelastic material at micro and nano length scales, by video microscopy. We describe here the principles and technique of video microscopy as a tool in such investigations. In this work, polystyrene beads were dispersed in the matrix of RSF polymer and the positions of the embedded beads diffusing were tracked using video microscopy. An optical tweezer was used to transport and locate the bead at any desired site within the micro-volume of the sample, to facilitate the subsequent free-bead video analysis. The position information of the beads was used to obtain the time dependant mean squared displacement (MSD) of the beads in the medium and hence to calculate the dynamic moduli of the medium. We present here the results of rheological measurements of the silk polymer network in solution over a frequency range, whose upper limit is the frame capture rate of our camera at full resolution. The technique is complementary to other microrheological techniques to characterize the material, but additionally enables one to characterize local inhomogeneities in the medium, features that get averaged out in bulk characterization procedures.

  20. Microdissection of black widow spider silk-producing glands.

    Jeffery, Felicia; La Mattina, Coby; Tuton-Blasingame, Tiffany; Hsia, Yang; Gnesa, Eric; Zhao, Liang; Franz, Andreas; Vierra, Craig

    2011-01-01

    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. PMID:21248709

  1. Wet-spinning of osmotically stressed silk fibroin.

    Sohn, Sungkyun; Gido, Samuel P

    2009-08-10

    Based on the phase diagram constructed for water-silk fibroin-LiBr using the osmotic stress method, wet-spinning of osmotically stressed, regenerated Bombyx mori silk fibroin was performed, without the necessity of using expensive or toxic organic solvents. The osmotic stress was applied to prestructure the regenerated silk fibroin molecule from its original random coil state to a more oriented state, manipulating the phase of the silk solution in the phase diagram before the start of spinning. Various starting points for spinning were selected from the phase diagram to evaluate the spinning performance and also physical properties of fibers produced. Monofilament fiber with a diameter of 20 microm was produced. It was found that the fibers whose starting point in the phase diagram were around the phase boundary between silk I and silk II, at very low LiBr concentrations, showed the best spinning process stability and physical properties. This underpins the prediction that the enhanced control over structure and phase behavior using the osmotic stress method helps improve the physical properties of wet-spun regenerated silk fibroin fibers. PMID:19572633

  2. Study on improving antioxidant and antibacterial activities of silk fibroin by irradiation treatment

    The silk fibroin solutions were prepared in solvent system of CaCl2. CH3CH2OH. H2O (mole ratio = 1:2:8) followed dialysis against deionized water. The 3% silk fibroin solutions were irradiated under gamma Co-60 source with dose ranging from 0 to 50 kGy at Hanoi Irradiation Centre and bioactivities of the irradiated silk fibroin solutions were investigated with different radiation doses. The results indicated that the antioxidant and antibacterial activities of fibroin were much improved by gamma irradiation. Maximum value of DPPH radical scavenging activity was 70.4% for the solution of silk fibroin irradiated at 10 kGy. Silk fibroin solutions irradiated at doses higher than 10 kGy also exhibited rather high antibacterial activity against E. coli and S. aureus. In order to estimate the applicability of our irradiated fibroin, the silk fibroin solutions were lyophilized to obtain a pure fibroin powder, then their bio-activities were compared with those of commercial silk fibroin (Proteines De Soie/ Zijdeproteine, Bioflore, Canada). Our fibroin powder revealed higher antioxidant and antibacterial activities. The amino acid compositions of our irradiated fibroin were also higher than that of the commercial product. Thus, the irradiated silk fibroin can be used for further application in cosmetic and other related fields. (author)

  3. Preparation and characterization of blends containing silk fibroin and chitosan

    The aim of this study was to prepare and characterize blend membranes of silk fibroin and chitosan. Moreover, a conformation of fibroin to a more stable form induced by the addition of chitosan was verified. Blend membranes of fibroin/chitosan were prepared in different proportions and had their crystallinity, structural conformation and thermal stability characterized. The results of crystallographic analysis (XRD) indicated the tendency to higher structural organization caused by the addition of chitosan. FTIR showed that, mainly in a content of chitosan of only 25%, fibroin is present in a more stable form. Thermal analyzes indicate that fibroin is thermally stable and that when its proportion in the blend increases, the temperature in which the degradation is initiated also does so. (author)

  4. Silk Fibroin: a biopolymer platform for innovative pharmaceutical formulation and biomedical devices.

    Sagnella, Anna

    2015-01-01

    The protein silk fibroin (SF) from the silkworm Bombyx mori is a FDA-approved biomaterial used over centuries as sutures wire. Importantly, several evidences highlighted the potential of silk biomaterials obtained by using so-called regenerated silk fibroin (RSF) in biomedicine, tissue engineering and drug delivery. Indeed, by a water-based protocol, it is possible to obtain protein water-solution, by extraction and purification of fibroin from silk fibres. Notably, RSF can be processed in a ...

  5. Study On Improving Antioxydant And Antibacterial Activities Of Silk Fibroin By Irradiation Treatment

    Silk fibroin at dry state and the solution of 3% were irradiated by Co-60 source at dose ranges 0 - 1000 kGy and 0 - 50 kGy respectively. The results showed that irradiation treatment for fibroin solution have higher effectiveness for improvement of some bio-activities of silk fibroin compared with dry state irradiation treatment due to remarkably reducing of irradiation doses. The antioxidant activity of fibroin was significantly increase by irradiation. The maximum value of DPPH radical scavenging activity was 70.4% when fibroin solution was irradiated at dose of 10 kGy. Irradiated fibroin solution also shown antibacterial activity against tested bacteria strains (E. coli, and S. aureus). In order to estimate the applicability of our irradiated fibroin, the silk fibroin solutions were lyophilized to obtain a pure fibroin powder, then their bio-activities were compared with those of commercial silk fibroin (Proteines De Soie/ Zijdeproteine, Bioflore, Canada). Our fibroin powder revealed higher antioxidant and antibacterial activities. The amino acid compositions of our irradiated fibroin were also higher than that of the commercial product. Thus, the irradiated silk fibroin can be used for further application in cosmetic and other related fields. (author)

  6. Macro/micro porous silk fibroin scaffolds obtained via combined methodologies for articular cartilage and meniscus tissue engineering

    Yan, Leping; Oliveira, Joaquim M.; Oliveira, Ana Leite; Caridade, S. G.; Mano, J.F; Reis, R.L

    2011-01-01

    This study proved that the salt leached silk fibroin scaffolds can be developed by using more than 10 wt% aqueous silk fibroin solutions. There were some previous studies shown that this was impossible.

  7. Orientational structure formation of silk fibroin with anisotropic properties in solutions

    Key words:silk fibroin, dissolution, solution's model systems, gelation, orientational crystallization, optical polarization, longitudinal stream, α - β transition, structure formation, phase transformations, relaxation, anisotropy of swelling and desorption, thermo- and biodegradation. Subjects of the inquiry: silk fibroin is the main subject of investigation. Fibroin's solutions were obtained on the base of water and organic solvents, containing salts. Comparative investigations were carried out by using biosolution - secretion of silkworm, solutions of silk sericin, cotton cellulose, methylcellulose, polystyrene and (co) polycrylonitrile. Aim of the inquiry: the elucidation of the regularities of silk fibroin anisotropic structures formation in the direct generation of orientational ordering in solutions taking into account of influences of its the molecular structures, configuration information, α - β conformational transformations, and development jointly using polarization-optical and hydrodynamic methods to control of structure formation. And also definition of possibility fields for use biopolymers anisotropic structure formation principles. Method of inquiry: birefringence, dispersion optical rotation, circular dichroism, polarization- ultramicroscope, ultracentrifuge, viscosimetry, potentiometry, differential thermal analysis, chromatography, x-ray analysis, spectroscopy. The results achieved and their novelty: the physical regularity amorphous-crystalline fibroin dissolutions in salt-containing solvents based on chains melting, distribution and redistribution were recognized; fibroin statistical parameters, molecular-mass and conformational characteristics were established; It was shown that fibroin molecules turned into fully uncoiled and oriented state with the breakdown decay of α-spiral chain sections by I type phase transition mechanism, but in oriented state with α-spiral conservation by II type transition; the presence of longitudinal field on the boundary 'reservoir - withdrawing capillary' of gland, initiating the transition of α-spiral in β-structures as well as phase separation of fibroin and sericin in stream were discovered; the phase diagram of liquidus for secret in the framework of polymers orientation crystallization conception were suggested; the mechanism of fibroin orientational crystallization under the longitudinal flow of solutions and gels was conformed experimentally; the scientific principles of oriented-crystallized fibrillar biopolymer materials receipt were established on the base of fibroin model solutions with properties of anisotropy of moisture absorption, swelling, desorption, thermo- and biodegradation; the approach of jointly using polarization-optical and hydrodynamic methods to the investigation of fibrillar biopolymers structure formation and phase transformations in solution was developed. Practical value - the elucidated physical regularities of fibroin solution allow to formulate a new idea on fibrillar biopolymer solutions formation and to get on their late model systems for practical use; the established scientific principles of orientational structure-formation and phase transformation of fibroin will be the base for development of original methods of anisotropic biopolymer materials from solutions of the polarization-optical and developed hydrodynamic methods can be used for the investigation of structure and phase transformations of wide range of fibrillar biopolymer samples. Sphere of usage: physics of anisotropic polymer systems, macromolecular compounds solutions, gels, crystals, biopolymers materials science, optical polarization and hydrodynamics. (author)

  8. Structural study on methacrylamide-grafted Tussah silk fibroin fibres.

    Pavoni, Eleonora; Tozzi, Silvia; Tsukada, Masuhiro; Taddei, Paola

    2016-07-01

    Tussah silk fibroin fibres were modified by grafting with methacrylamide (MAA), with weight gains ranging between 2.6% and 71.4%. Raman and IR spectroscopic analyses showed that upon grafting the fibres underwent slight conformational changes towards a more unordered state, due to the covalent and hydrogen bonds interactions occurring between the polymer (polyMAA) and the amorphous domains of silk fibres. To test the stability towards alkaline hydrolysis, the untreated and MAA-grafted silk fibres (weight gain of 71.4%) were immersed in NaOH 5% at 50°C for different times; the IR and Raman spectroscopic techniques were utilized to elucidate the degradation mechanism as well as the rearrangements of the fibres induced by the treatment. Upon hydrolysis, both the untreated and grafted fibres underwent an enrichment in β-sheet conformation, due to the preferential removal of the unordered domains. As a result of the covalent interactions with silk fibroin, the polymer increased its stability towards alkaline hydrolysis, since its complete solubilization was avoided and the transformation of its CONH2 groups into COO(-) and COOH was delayed. Vibrational spectroscopy proved to be a valid technique to investigate the mechanism and the effects of the hydrolytic attack, which are both fundamental to design new-generation silk-based materials. PMID:27032490

  9. Silk fibroin based antibacterial bionanotextiles as wound dressing materials

    Çalamak, Semih [Hacettepe University, Faculty of Pharmacy, Department of Basic Pharmaceutical Sciences, 06100 Ankara (Turkey); Hacettepe University, Department of Nanotechnology and Nanomedicine, 06800 Ankara (Turkey); Erdoğdu, Ceren; Özalp, Meral [Hacettepe University, Faculty of Pharmacy, Department of Pharmaceutical Microbiology, 06100 Ankara (Turkey); Ulubayram, Kezban, E-mail: ukezban@hacettepe.edu.tr [Hacettepe University, Faculty of Pharmacy, Department of Basic Pharmaceutical Sciences, 06100 Ankara (Turkey); Hacettepe University, Department of Nanotechnology and Nanomedicine, 06800 Ankara (Turkey)

    2014-10-01

    New applications for medical biotextiles have been identified with the development of nanotechnological manufacturing technologies. Combination of nanotechnology and biotextile technology has resulted into a new field called bionanotextiles. Bionanotextiles are used in many areas which include wound dressings, bandages and tissue scaffolds. Silk fibroin (SF) from the cocoon of Bombyx mori, is one of the most favorable wound dressing materials due to its unique properties including biocompatibility, permeability, biodegradability, morphologic flexibility, and proper mechanical properties. The modification of antimicrobial properties of SFs can provide a barrier for bacterial penetration as wound dressing materials. In the present study, antibacterial polyethylenimine (PEI) (10, 20 and 30% (w/w)) was blended with SF and bionanotextiles were successfully fabricated by electrospinning. In addition, silk fibroin nanofibers were also functionalized with sulphate group in order to test whether they exhibit an antibacterial activity or not. Fibroin based bionanotextiles were characterized by scanning electron microscope (SEM), Fourier transform infrared spectroscopy (ATR-FTIR), differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA). The cytotoxicity evaluations were carried out by L929 fibroblasts with MTT assay. The indirect cytotoxicity results demonstrate that all fibroin and PEI/fibroin extracts have no cytotoxicity on L929 cancer cell line. PEI/fibroin bionanotextiles showed strong antibacterial activities against gram positive Staphylococcus aureus and gram negative Pseudomonas aeruginosa. - Highlights: • Bionanotextiles are combination of nanotechnology and biotextile technology. • Bionanotextiles have good antibacterial activity against both of S. aureus and P. aeruginosa. • Antibacterial bionanotextiles are applicable to most of the infected wounds. • No cytotoxicity was observed on L929 cell line.

  10. A pilot study of macrophage responses to silk fibroin particles.

    Cui, Xidong; Wen, Jianchuan; Zhao, Xia; Chen, Xin; Shao, Zhengzhong; Jiang, Jack J

    2013-05-01

    Silk fibroin (SF) shows promise for tissue engineering and other biomedical applications due to its excellent biocompatibility, unique biomechanical properties, and controllable biodegradability. The particulate form of SF materials may have many potential uses, including the use as a filler for tissue defects or as a controlled-release agent for drug delivery. However, many past in vivo and in vitro studies evaluating the biocompatibility and biodegradability of SF have involved bulk implants. It is essential to evaluate the inflammatory effects of SF particles before further use. In this study, two different sizes of SF particles were evaluated to assess their impact on the release of tumor necrosis factor (TNF)-?, interleukin (IL)-1?, and IL-6, in comparison with lipopolysaccharide positive control stimulation. The inflammatory processes were characterized using real-time reverse transcription polymerase chain reaction, enzyme-linked immunosorbent assay, and light microscopy evaluations. The results indicated that small silk fibroin particles and large silk fibroin particles, in culture with RAW 264.7 murine macrophage cells for 24 h, caused up-regulation of mRNA coding for TNF-?, which indicated that both size of particles have potential inflammatory effects. There was a statistically significant increase in this up-regulation under small silk fibroin stimulation. However, the immunosorbent assay suggested that there was virtually no observed release of IL-1?, IL-6, or TNF-?, relative to the control group. The results suggest that SF particles of the chosen dimensions may have good biocompatibility in culture with RAW 264.7 murine macrophages. PMID:23225634

  11. Preparation and characterization of silk fibroin/HPMC blend film

    Shetty, G. Rajesha; Kumar, R. Madhu; Rao, B. Lakshmeesha; Asha, S.; Sangappa

    2015-06-01

    In this work, the structural and mechanical stability of silk fibroin/Hydroxypropylmethyl cellulose (SF-HPMC) blend films were characterized by X-ray diffraction (XRD) and Universal Testing Machine (UTM). The results indicate that with the introduction of HPMC, the interactions between SF and HPMC results in improved crystallite size and increase in mechanical properties. The blend film obtained is more flexible compared to pure SF film.

  12. Silk fibroin gelation via non-solvent induced phase separation

    Kasoju, Naresh; Hawkins, N.; Pop-Georgievski, Ognen; Kubies, Dana; Vollrath, F.

    2016-01-01

    Roč. 4, č. 3 (2016), s. 460-473. ISSN 2047-4830 R&D Projects: GA MŠk(CZ) EE2.3.30.0029; GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:61389013 Keywords : silk fibroin * non-solvent induced phase separation * desolvation Subject RIV: CE - Biochemistry Impact factor: 3.831, year: 2014

  13. Preparation and characterization of silk fibroin as a biomaterial with potential for drug delivery

    Zhang Hao

    2012-06-01

    Full Text Available Abstract Background Degummed silk fibroin from Bombyx mori (silkworm has potential carrier capabilities for drug delivery in humans; however, the processing methods have yet to be comparatively analyzed to determine the differential effects on the silk protein properties, including crystalline structure and activity. Methods In this study, we treated degummed silk with four kinds of calcium-alcohol solutions, and performed secondary structure measurements and enzyme activity test to distinguish the differences between the regenerated fibroins and degummed silk fibroin. Results Gel electrophoresis analysis revealed that Ca(NO32-methanol, Ca(NO32-ethanol, or CaCl2-methanol treatments produced more lower molecular weights of silk fibroin than CaCl2-ethanol. X-ray diffraction and Fourier-transform infrared spectroscopy showed that CaCl2-ethanol produced a crystalline structure with more silk I (α-form, type II β-turn, while the other treatments produced more silk II (β-form, anti-parallel β-pleated sheet. Solid-State 13C cross polarization and magic angle spinning-nuclear magnetic resonance measurements suggested that regenerated fibroins from CaCl2-ethanol were nearly identical to degummed silk fibroin, while the other treatments produced fibroins with significantly different chemical shifts. Finally, enzyme activity test indicated that silk fibroins from CaCl2-ethanol had higher activity when linked to a known chemotherapeutic drug, L-asparaginase, than the fibroins from other treatments. Conclusions Collectively, these results suggest that the CaCl2-ethanol processing method produces silk fibroin with biomaterial properties that are appropriate for drug delivery.

  14. Analysis of proteome dynamics inside the silk gland lumen of Bombyx mori.

    Dong, Zhaoming; Zhao, Ping; Zhang, Yan; Song, Qianru; Zhang, Xiaolu; Guo, Pengchao; Wang, Dandan; Xia, Qingyou

    2016-01-01

    The silk gland is the only organ where silk proteins are synthesized and secreted in the silkworm, Bombyx mori. Silk proteins are stored in the lumen of the silk gland for around eight days during the fifth instar. Determining their dynamic changes is helpful for clarifying the secretion mechanism of silk proteins. Here, we identified the proteome in the silk gland lumen using liquid chromatography-tandem mass spectrometry, and demonstrated its changes during two key stages. From day 5 of the fifth instar to day 1 of wandering, the abundances of fibroins, sericins, seroins, and proteins of unknown functions increased significantly in different compartments of the silk gland lumen. As a result, these accumulated proteins constituted the major cocoon components. In contrast, the abundances of enzymes and extracellular matrix proteins decreased in the silk gland lumen, suggesting that they were not the structural constituents of silk. Twenty-five enzymes may be involved in the regulation of hormone metabolism for proper silk gland function. In addition, the metabolism of other non-proteinous components such as chitin and pigment were also discussed in this study. PMID:27102218

  15. Silk fibroin and sodium alginate blend: Miscibility and physical characteristics

    Agostini de Moraes, Mariana; Silva, Mariana Ferreira; Weska, Raquel Farias; Beppu, Marisa Masumi, E-mail: beppu@feq.unicamp.br

    2014-07-01

    Films of silk fibroin (SF) and sodium alginate (SA) blends were prepared by solution casting technique. The miscibility of SF and SA in those blends was evaluated and scanning electron microscopy (SEM) revealed that SF/SA 25/75 wt.% blends underwent microscopic phase separation, resulting in globular structures composed mainly of SF. X-ray diffraction indicated the amorphous nature of these blends, even after a treatment with ethanol that turned them insoluble in water. Thermal analyses of blends showed the peaks of degradation of pristine SF and SA shifted to intermediate temperatures. Water vapor permeability, swelling capacity and tensile strength of SF films could be enhanced by blending with SA. Cell viability remained between 90 and 100%, as indicated by in vitro cytotoxicity test. The SF/SA blend with self-assembled SF globules can be used to modulate structural and mechanical properties of the final material and may be used in designing high performance wound dressing. - Highlights: • Blend films of fibroin and alginate were prepared with microscopic phase separation; • Self-assembled globular microdomains were mainly composed by fibroin; • It was possible to obtain a film with better mechanical and physical properties; • Blend films of fibroin and alginate represent a novel material in biomaterials field.

  16. Silk Fibroin/Starch Blend Films: Preparation and Characterization

    Prasong Srihanam

    2011-01-01

    Full Text Available This study was aimed to prepare silk fibroin (SF of Thai silk (Bombyx mori and cassava starch blend films by solvent evaporation method. The SF solution was firstly prepared from the cocoons by dissolving pure silk fibroin with tertiary solvent system of CaCl2: Ethanol: H2O (1:2:8 by mol. Cassava starch solution could be prepared by boiling at high temperature. The mixture of the SF and cassava starch solution was then blended and cast on the polystyrene plates. The obtained films were investigated on their morphology, secondary structures and thermal properties by using SEM, FTIR and thermogravimetry, respectively. The results found that SF blended with starch could be formed the higher ratio of ?-sheet. With cross-section images, the blend films showed homogeneously texture. In addition, the blend film gradually increased its thermal properties when the starch blended ratio was increased. In conclusion, the SF/starch blend films could be adjusted their properties by varying the blend ratio.

  17. Characterization and Evolutionary Analyses of Silk Fibroins from Two Insect Orders: Embioptera and Lepidoptera

    Collin, Matthew Aric

    2010-01-01

    Silk production has independently evolved in several insect lineages, such as Hymenoptera (bees, ants and wasps), Siphonaptera (fleas), and Archeognatha (bristletails and silverfish). Primarily composed of proteins, silks are used for functions, such as protection, prey capture, dispersal, and reproduction. Because of the ecological significance of silks, natural selection can act on silk fibers and thus the underlying silk proteins (fibroins). Lepidoptera (moths and butterflies) and Embiopte...

  18. Acylation Modification of Antheraea pernyi Silk Fibroin Using Succinic Anhydride and Its Effects on Enzymatic Degradation Behavior

    Xiufang Li (Leah); Ceng Zhang; Lingshuang Wang; Caili Ma; Weichao Yang; Mingzhong Li

    2013-01-01

    The degradation rate of tissue engineering scaffolds should match the regeneration rate of new tissues. Controlling the degradation behavior of silk fibroin is an important subject for silk-based tissue engineering scaffolds. In this study, Antheraea pernyi silk fibroin was successfully modified with succinic anhydride and then characterized by zeta potential, ninhydrin method, and FTIR. In vitro, three-dimensional scaffolds prepared with modified silk fibroin were incubated in collagenase IA...

  19. Electrophoretic deposition of tetracycline modified silk fibroin coatings for functionalization of titanium surfaces

    Zhang, Zhen; Qu, Yinying; Li, Xiaoshuang; Zhang, Sheng; Wei, Qingsong; Shi, Yusheng; Chen, Lili

    2014-06-01

    Electrophoretic deposition has been widely used for the fabrication of functional coatings onto metal implant. A characteristic feature of this process is that positively charged materials migrate toward the cathode and can deposit on it. In this study, silk fibroin was decorated with tetracycline in aqueous solution to impart positive charge, and then deposited on negatively titanium cathode under certain electric field. The characterization of the obtained coatings indicated that the intermolecular hydrogen bonds formed between the backbone of silk fibroin and tetracycline molecular. In vitro biological tests demonstrated that osteoblast-like cells achieved acceptable cell affinity on the tetracycline cross-linked silk fibroin coatings, although greater cell viability was seen on pure silk fibroin coatings. The cationic silk fibroin coatings showed remarkable antibacterial activity against gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) bacteria. Therefore, we concluded that electrophoretic deposition was an effective and efficient technique to prepare cationic silk fibroin coatings on the titanium surface and that cationic silk fibroin coatings with acceptable biocompatibility and antibacterial property were promising candidates for further loading of functional agents.

  20. Wound healing effect of electrospun silk fibroin nanomatrix in burn-model.

    Ju, Hyung Woo; Lee, Ok Joo; Lee, Jung Min; Moon, Bo Mi; Park, Hyun Jung; Park, Ye Ri; Lee, Min Chae; Kim, Soo Hyeon; Chao, Janet Ren; Ki, Chang Seok; Park, Chan Hum

    2016-04-01

    Silk fibroin has recently become an important biomaterial for tissue engineering application. In this study, silk fibroin nanomatrix was fabricated by electrospinning and evaluated as wound dressing material in a burn rat model. The wound size reduction, histological examination, and the quantification of transforming growth factor TGF-?1 and interleukin IL-1?, 6, and 10 were measured to evaluate the healing effects. The silk fibroin nanomatrix treatment exhibited effective performance in decreasing the wound size and epithelialization. Histological finding also revealed that the deposition of collagen in the dermis was organized by covering the wound area in the silk fibroin nanomatrix treated group. The expression level of pro-inflammatory cytokine (IL-1?) was significantly reduced in the injured skin following the silk fibroin nanomatrix treatment compared to the medical gauze (control) at 7 days after burn. Also, the expression level of TGF-?1 in the wound treated with silk fibroin nanomatrix peaked 21-days post-treatment whereas expression level of TGF-?1 was highest at day 7 in the gauze treated group. In conclusion, this data demonstrates that silk fibroin nanomatrix enhances the burn wound healing, suggesting it is a good candidate for burn wound treatment. PMID:26718866

  1. Silk fibroin-based scaffolds for tissue engineering

    Li, Zi-Heng; Ji, Shi-Chen; Wang, Ya-Zhen; Shen, Xing-Can; Liang, Hong

    2013-09-01

    Silk fibroin (SF) from the Bombyx mori silkworm exhibits attractive potential applications as biomechanical materials, due to its unique mechanical and biological properties. This review outlines the structure and properties of SF, including of its biocompatibility and biodegradability. It highlights recent researches on the fabrication of various SF-based composites scaffolds that are promising for tissue engineering applications, and discusses synthetic methods of various SF-based composites scaffolds and valuable approaches for controlling cell behaviors to promote the tissue repair. The function of extracellular matrices and their interaction with cells are also reviewed here.

  2. Novel two-step method to form silk fibroin fibrous hydrogel.

    Ming, Jinfa; Li, Mengmeng; Han, Yuhui; Chen, Ying; Li, Han; Zuo, Baoqi; Pan, Fukui

    2016-02-01

    Hydrogels prepared by silk fibroin solution have been studied. However, mimicking the nanofibrous structures of extracellular matrix for fabricating biomaterials remains a challenge. Here, a novel two-step method was applied to prepare fibrous hydrogels using regenerated silk fibroin solution containing nanofibrils in a range of tens to hundreds of nanometers. When the gelation process of silk solution occurred, it showed a top-down type gel within 30min. After gelation, silk fibroin fibrous hydrogels exhibited nanofiber network morphology with ?-sheet structure. Moreover, the compressive stress and modulus of fibrous hydrogels were 31.92.6 and 2.80.8kPa, respectively, which was formed using 2.0wt.% concentration solutions. In addition, fibrous hydrogels supported BMSCs attachment and proliferation over 12days. This study provides important insight in the in vitro processing of silk fibroin into useful new materials. PMID:26652363

  3. Preparation and characterization of regenerated fiber from the aqueous solution of Bombyx mori cocoon silk fibroin

    Zhu Zhenghua [Department of Biotechnology, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588 (Japan); Department of Application Engineering, ZheJiang Vocational College of Economic and Trade, HangZhou, ZheJiang 310018 (China); Imada, Takuzo [Department of Biotechnology, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588 (Japan); Asakura, Tetsuo, E-mail: asakura@cc.tuat.ac.jp [Department of Biotechnology, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588 (Japan)

    2009-10-15

    The regenerated silk fibers with high strength and high biodegradability were prepared from the aqueous solution of Bombyx mori silk fibroin from cocoons with wet spinning method. Although the tensile strength of the regenerated silk fibroin fiber, 210 MPa is still half of the strength of native silk fiber, the diameter of the fiber is about 100 {mu}m which is suitable for monofilament of suture together with high biodegradability. The high concentration (30%, w/v) of the aqueous solution of the silk fibroin which corresponds to the high concentration in the middle silkgland of silkworm was obtained. This was performed by adjusting the pH of the aqueous solution to 10.4 which corresponds to pK{sub a} value of the OH group of Tyr residues in the silk fibroin. The mixed solvent, methanol/acetic acid (7:3 in volume ratio) was used as coagulant solvent for preparing the regenerated fiber. The structural change of silk fibroin fiber by stretching was monitored with both {sup 13}C solid state NMR and X-ray diffraction methods, indicating that the high strength of the fiber is related with the long-range orientation of the silk fibroin chain with {beta}-sheet structure.

  4. Preparation and characterization of regenerated fiber from the aqueous solution of Bombyx mori cocoon silk fibroin

    The regenerated silk fibers with high strength and high biodegradability were prepared from the aqueous solution of Bombyx mori silk fibroin from cocoons with wet spinning method. Although the tensile strength of the regenerated silk fibroin fiber, 210 MPa is still half of the strength of native silk fiber, the diameter of the fiber is about 100 ?m which is suitable for monofilament of suture together with high biodegradability. The high concentration (30%, w/v) of the aqueous solution of the silk fibroin which corresponds to the high concentration in the middle silkgland of silkworm was obtained. This was performed by adjusting the pH of the aqueous solution to 10.4 which corresponds to pKa value of the OH group of Tyr residues in the silk fibroin. The mixed solvent, methanol/acetic acid (7:3 in volume ratio) was used as coagulant solvent for preparing the regenerated fiber. The structural change of silk fibroin fiber by stretching was monitored with both 13C solid state NMR and X-ray diffraction methods, indicating that the high strength of the fiber is related with the long-range orientation of the silk fibroin chain with ?-sheet structure.

  5. Preparation and hemostatic property of low molecular weight silk fibroin.

    Lei, Caihong; Zhu, Hailin; Li, Jingjing; Feng, Xinxing; Chen, Jianyong

    2016-04-01

    Effective hemorrhage control becomes increasingly significant in today's military and civilian trauma, while the topical hemostats currently available in market still have various disadvantages. In this study, three low molecular weight silk fibroins (LMSF) were prepared through hydrolysis of silk fibroin in a ternary solvent system of CaCl2/H2O/EtOH solution at different hydrolysis temperatures. Fourier transform infrared spectroscopy analysis showed that the content of ? sheet structure in the LMSF decreased with the increase in hydrolysis temperature. The results of thromboelastographic and activated partial thromboplastin time methods showed that the LMSF hydrolyzed at 50?C can significantly strengthen the coagulation in blood and activate the intrinsic pathway of coagulation cascade. In the murine hepatic injury model, the LMSF hydrolyzed at 50?C can promote the blood clotting and decrease the blood loss and bleeding time. Based on these results, it can be suggested that the developed LMSF has the excellent hemostatic effect and may be a promising material in clinical hemostatic application. PMID:26732018

  6. Novel silk fibroin films prepared by formic acid/hydroxyapatite dissolution method

    Ming, Jinfa, E-mail: jinfa.ming@gmail.com [National Engineering Laboratory for Modern Silk, Soochow University, Suzhou 215123 (China); College of Textile and Clothing Engineering, Soochow University, Suzhou 215021 (China); Liu, Zhi; Bie, Shiyu [National Engineering Laboratory for Modern Silk, Soochow University, Suzhou 215123 (China); College of Textile and Clothing Engineering, Soochow University, Suzhou 215021 (China); Zhang, Feng [Jiangsu Province Key Laboratory of Stem Cell Research, Medical College, Soochow University, Suzhou 215006 (China); Zuo, Baoqi, E-mail: bqzuo@suda.edu.cn [National Engineering Laboratory for Modern Silk, Soochow University, Suzhou 215123 (China); College of Textile and Clothing Engineering, Soochow University, Suzhou 215021 (China)

    2014-04-01

    Bombyx mori silk fibroin from the silkworm was firstly found to be soluble in formic acid/hydroxyapatite system. The rheological behavior of silk fibroin solution was significantly influenced by HAp contents in dissolved solution. At the same time, silk fibroin nanofibers were observed in dissolved solution with 103.6 ± 20.4 nm in diameter. Moreover, the structure behavior of SF films prepared by formic acid/hydroxyapatite dissolution method was examined. The secondary structure of silk fibroin films was attributed to silk II structure (β-sheet), indicating that the hydroxyapatite contents in dissolved solution were not significantly affected by the structure of silk fibroin. The X-ray diffraction results exhibited obviously hydroxyapatite crystalline nature existing in silk fibroin films; however, when the hydroxyapatite content was 5.0 wt.% in dissolved solution, some hydroxyapatite crystals were converted to calcium hydrogen phosphate dehydrate in silk fibroin dissolution process. This result was also confirmed by Fourier transform infrared analysis and DSC measurement. In addition, silk fibroin films prepared by this dissolution method had higher breaking strength and extension at break. Based on these analyses, an understanding of novel SF dissolution method may provide an additional tool for designing and synthesizing advanced materials with more complex structures, which should be helpful in different fields, including biomaterial applications. - Highlights: • SF fibers were firstly successfully dissolved in FA/HAp solution. • The rheological behavior of SF solution was significantly influenced by HAp contents. • SF nanofibrils were observed in FA/HAp solution with 103.6 ± 20.4 nm in diameter. • SF films prepared by FA/HAp dissolution method had higher mechanical properties.

  7. Novel silk fibroin films prepared by formic acid/hydroxyapatite dissolution method

    Bombyx mori silk fibroin from the silkworm was firstly found to be soluble in formic acid/hydroxyapatite system. The rheological behavior of silk fibroin solution was significantly influenced by HAp contents in dissolved solution. At the same time, silk fibroin nanofibers were observed in dissolved solution with 103.6 20.4 nm in diameter. Moreover, the structure behavior of SF films prepared by formic acid/hydroxyapatite dissolution method was examined. The secondary structure of silk fibroin films was attributed to silk II structure (?-sheet), indicating that the hydroxyapatite contents in dissolved solution were not significantly affected by the structure of silk fibroin. The X-ray diffraction results exhibited obviously hydroxyapatite crystalline nature existing in silk fibroin films; however, when the hydroxyapatite content was 5.0 wt.% in dissolved solution, some hydroxyapatite crystals were converted to calcium hydrogen phosphate dehydrate in silk fibroin dissolution process. This result was also confirmed by Fourier transform infrared analysis and DSC measurement. In addition, silk fibroin films prepared by this dissolution method had higher breaking strength and extension at break. Based on these analyses, an understanding of novel SF dissolution method may provide an additional tool for designing and synthesizing advanced materials with more complex structures, which should be helpful in different fields, including biomaterial applications. - Highlights: SF fibers were firstly successfully dissolved in FA/HAp solution. The rheological behavior of SF solution was significantly influenced by HAp contents. SF nanofibrils were observed in FA/HAp solution with 103.6 20.4 nm in diameter. SF films prepared by FA/HAp dissolution method had higher mechanical properties

  8. Structure modifications induced in silk fibroin by enzymatic treatments. A Raman study

    Monti, Patrizia; Freddi, Giuliano; Sampaio, Sandra; Tsukada, Masuhiro; Taddei, Paola

    2005-06-01

    Raman spectroscopy was used to investigate various enzyme-catalyzed reactions onto silk fibroin, i.e. the biodegradation of Tussah ( Antheraea pernyi) silk fibroin films by a proteolytic enzyme, the oxidation of domestic ( Bombyx mori) silk fibroin by mushroom tyrosinase and the subsequent grafting of chitosan onto oxidized silk. The spectra of Tussah silk fibroin films exposed to a bacterial protease for different times demonstrated that the cleavage of sensitive peptide bonds in the amorphous glycine-rich domains resulted in the loss of various amino acid residues (Tyr, Trp, Asp, etc.). The bands attributed to the crystalline alanine-rich sequences increased in intensity, and the β-sheet molecular conformation was not affected by biodegradation. Following oxidation with mushroom tyrosinase, the tyrosine bands of Bombyx mori fibroin decreased in intensity but did not disappear. The increase of the I853/ I829 intensity ratio indicated that the Tyr residues not accessible to the enzyme were located in a strongly hydrophobic environment. Raman spectroscopy provided evidence that chitosan was effectively grafted onto oxidized silk, probably via the Schiff-base mechanism, as shown by the behavior of the imine band at about 1646 cm -1. Grafting chitosan onto silk fibroin resulted in a β-sheet→random coil conformational transition of the protein component in the bioconjugated product.

  9. Inkjet Printing of Regenerated Silk Fibroin: From Printable Forms to Printable Functions.

    Tao, Hu; Marelli, Benedetto; Yang, Miaomiao; An, Bo; Onses, M Serdar; Rogers, John A; Kaplan, David L; Omenetto, Fiorenzo G

    2015-08-01

    A formulation of regenerated silk fibroin solution that can be easily functionalized and inkjet printed on numerous surfaces is developed. As an example, the inks can be printed on laboratory gloves that change color when exposed to bacteria. PMID:26079217

  10. Electromechanical response of silk fibroin hydrogel and conductive polycarbazole/silk fibroin hydrogel composites as actuator material.

    Srisawasdi, Thanida; Petcharoen, Karat; Sirivat, Anuvat; Jamieson, Alexander M

    2015-11-01

    Pure silk fibroin (SF) hydrogel and polycarbazole/silk fibroin (SF/PCZ) hydrogels were fabricated by solvent casting technique to evaluate electromechanical responses, dielectric properties, and cantilever deflection properties as functions of electric field strength, SF concentration, glutaraldehyde concentration, and PCZ concentration in the blends. Electromechanical properties were characterized in oscillatory shear mode at electric field strengths ranging from 0 to 600V/mm and at a temperature of 27°C. For both the pristine SF and SF/PCZ hydrogels, the storage modulus response (ΔG') and the storage modulus sensitivity (ΔG'/G'0) increased dramatically with increasing electric field strength. The pristine hydrogel possessed the highest storage modulus sensitivity value of 5.87, a relatively high value when compared with other previously studied electroactive polymers. With the addition of conductive PCZ in SF hydrogel, the storage modulus sensitivity and the relative dielectric constant decreased; the conductive polymer thus provided the softening effect under electric field. In the deflection response, the dielectrophoresis force and deflection distance increased monotonically with electric field strength, where the pure SF hydrogel showed the highest deflection distance and dielectrophoresis force. PMID:26249559

  11. Study On Degradation Of Silk Fibroin By Irradiation Treatment For Cosmetic And Pharmaceutical Applications

    As a kind of protein, silk fibroin is created with silkworm Bombyx mori in products of silk. The fibroin was irradiated using Co-60 gamma source and its degradability and solubility were investigated with various radiation doses to apply in pharmacy and cosmetic. Addition to the morphological changes of irradiated fibroin fibers shows that its mechanical properties were much influenced by the irradiation. Tensile strength and elongation at break of the silk fibroin significantly decreased with increasing of radiation dose up to 1000 kGy. The tensile strength and elongation at break of the irradiated fibroin at 1000 kGy reduced to 71% and 94% respectively in compared with non-irradiated one. The solubility of silk fibroin in both calcium chloride (CaCl2/C2H5OH/H2O=1:2:8) in mole ratio and distilled water were improved by the irradiation. UV spectrometry revealed the structure of silk fibroin was also changed by irradiation. (author)

  12. Fabrication of silk fibroin film using centrifugal casting technique for corneal tissue engineering.

    Lee, Min Chae; Kim, Dong-Kyu; Lee, Ok Joo; Kim, Jung-Ho; Ju, Hyung Woo; Lee, Jung Min; Moon, Bo Mi; Park, Hyun Jung; Kim, Dong Wook; Kim, Su Hyeon; Park, Chan Hum

    2016-04-01

    Films prepared from silk fibroin have shown potential as biomaterials in tissue engineering applications for the eye. Here, we present a novel process for fabrication of silk fibroin films for corneal application. In this work, fabrication of silk fibroin films was simply achieved by centrifugal force. In contrast to the conventional dry casting method, we carried out the new process in a centrifuge with a rotating speed of 4000 rpm, where centrifugal force was imposed on an aluminum tube containing silk fibroin solution. In the present study, we also compared the surface roughness, mechanical properties, transparency, and cell proliferation between centrifugal and dry casting method. In terms of surface morphology, films fabricated by the centrifugal casting have less surface roughness than those by the dry casting. For elasticity and transparency, silk fibroin films obtained from the centrifugal casting had favorable results compared with those prepared by dry casting. Furthermore, primary human corneal keratocytes grew better in films prepared by the centrifugal casting. Therefore, our results suggest that this new fabrication process for silk fibroin films offers important potential benefits for corneal tissue regeneration. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 508-514, 2016. PMID:25939800

  13. Development and characterization of silk fibroin coated quantum dots

    Nathwani, B. B.; Needham, C.; Mathur, A. B.; Meissner, K. E.

    2008-02-01

    Recent progress in the field of semiconductor nanocrystals or Quantum Dots (QDs) has seen them find wider acceptance as a tool in biomedical research labs. As produced, high quality QDs, synthesized by high temperature organometallic synthesis, are coated with a hydrophobic ligand. Therefore, they must be further processed to be soluble in water and to be made biocompatible. To accomplish this, the QDs are generally coated with a synthetic polymer (eg. block copolymers) or the hydrophobic surface ligands exchanged with hydrophilic material (eg. thiols). Advances in this area have enabled the QDs to experience a smooth transition from being simple inorganic fluorophores to being smart sensors, which can identify specific cell marker proteins and help in diagnosis of diseases such as cancer. In order to improve the biocompatibility and utility of the QDs, we report the development of a procedure to coat QDs with silk fibroin, a fibrous crystalline protein extracted from Bombyx Mori silkworm. Following the coating process, we characterize the size, quantum yield and two-photon absorption cross section of the silk coated QDs. Additionally, the results of biocompatibility studies carried out to compare the properties of these QD-silks with conventional QDs are presented. These natural polymer coatings on QDs could enhance the intracellular delivery and enable the use of these nanocrystals as an imaging tool for studying subcellular machinery at the molecular level.

  14. Microrheological Studies of Regenerated Silk Fibroin Solution by Video Microscopy

    Raghu, A; Somashekar, R; Ananthamurthy, Sharath

    2007-01-01

    We have carried out studies on the rheological properties of regenerated silk fibroin (RSF) solution using video microscopy. The degummed silk from the Bombyx mori silkworm was used to prepare RSF solution by dissolving it in calcium nitrate tetrahydrate-methanol solvent. Measurements were carried out by tracking the position of an embedded micron-sized polystyrene bead within the RSF solution through video imaging. The time dependent mean squared displacement (MSD) of the bead in solution and hence, the complex shear modulus of this solution was calculated from the bead's position information. An optical tweezer was used to transport and locate the bead at any desired site within the micro-volume of the sample, to facilitate the subsequent free-bead video analysis. We present here the results of rheological measurements of the silk polymer network in solution over a frequency range, whose upper limit is the frame capture rate of our camera, at full resolution. By examining the distribution of MSD of beads at...

  15. Research On Degradation Of Silk Fibroin By Combination Of Electron Beam Irradiation And Hydrothermal Processing

    Silk fibers and silk proteins have been demonstrated to be useful to apply in the textile industry, biomedical, cosmetics, pharmaceuticals. In this study, the effects of electron beam (EB) irradiation combined with hydrothermal processing to the solubility of silk fibroin and generation of soluble silk protein were investigated. The solubility of unirradiated and irradiated fibroin were greater than 80 % when hydrothermal degradation was performed in the sodium hydroxide solution at appropriate concentration of 0.05 M. However, the solubility of irradiated fibroin was greater than that of unirradiated sample. The protein content increased from 0.4617 to 0.6530 mg/mg when irradiation doses increased from 0 to 200 kGy, respectively. The molecular weight of protein was determined by SDS-PAGE method. The characteristics of silk protein were confirmed by scanning electron microscope (SEM), Fourier transform infrared spectroscopy (FT-IR), thermogravimetric analysis (TGA) and X-ray diffraction (XRD). (author)

  16. Silk Fibroin Film Loaded Chlorhexidine Diacetate: Interaction and Characteristics

    N. Noi

    2009-01-01

    Full Text Available This study aimed to prepare Silk Fibroin (SF films with different weight (0.5, 1 and 1.5% for loading chlorhexidine diacetate as substrate for study their characteristics including morphology, structure and thermal properties. The morphological observation under scanning electron spectroscopy found that all of films have a smooth surface. With cross-section micrographs, the lowest SF content was smoother texture than other. The secondary structures of various films were determined by Fourier transform infrared (FTIR spectrometer. The results showed that SF film composed of β-sheet structures in different ratio depending on the SF content. The result suggested that high SF content formed crystalline by interaction between amino acids molecules in higher ratio than other. However, thermal properties of the SF films did not dramatically differ compared between low and high SF content. It was also found that CHX did not affect on the SF characteristics.

  17. Artificial crawler model for texture analysis on silk fibroin scaffolds

    Texture plays an important role in computer vision tasks. Several methods of texture analysis are available. However, these methods are not capable of extracting rich detail in images. This paper presents a novel approach to image texture classification based on the artificial crawler model. Here, we propose a new rule of movement that moves artificial crawler agents not only toward higher intensities but also toward lower ones. This strategy is able of capturing more detail because the agents explore the peaks as well as the valleys. Thus, compared with the state-of-the-art method, this approach shows an increased discriminatory power. Experiments on the most well known benchmark demonstrate the superior performance of our approach. We also tested our approach on silk fibroin scaffold analysis, and results indicate that our method is consistent and can be applied in real-world situations. (paper)

  18. Antimicrobial Silk Fibroin Hydrogel Instantaneously Induced by Cationic Surfactant

    Shan Sun

    2013-01-01

    Full Text Available Current surgical dressings and woundplast have drawbacks such as poor permeability, airtightness and biocompatibility, thus a new material is obligatory to replace the traditional ones. Due to outstanding biocompatibility of Silk Fibroin (SF and excellent antibacterial properties of Dodecyl Trimethyl Ammonium Bromide (DTAB, a cationic surfactant, the SF/DTAB hydrogel formed instantaneously is recommended. By means of SEM, FTIR and XRD, SF/DTAB hydrogel displayed clustered nanofilaments structure and similar secondary structure and morphology with SF solution mainly consisted of random coils in 1 h, then it tended to be the same as pure SF hydrogel which was composed of porous network structure and primarily contained ?-sheet structure. Excellent elasticity and antibacterial effect were verified by rheological and antibacterial measurements.

  19. Silk fibroin gelation via non-solvent induced phase separation.

    Kasoju, Naresh; Hawkins, Nicholas; Pop-Georgievski, Ognen; Kubies, Dana; Vollrath, Fritz

    2016-02-23

    Tissue engineering benefits from novel materials with precisely tunable physical, chemical and mechanical properties over a broad range. Here we report a practical approach to prepare Bombyx mori silk fibroin hydrogels using the principle of non-solvent induced phase separation (NIPS). A combination of reconstituted silk fibroin (RSF) and methanol (non-solvent), with a final concentration of 2.5% w/v and 12.5% v/v respectively, maintained at 22 °C temperature turned into a hydrogel within 10 hours. Freeze-drying of this gel gave a foam with a porosity of 88%, a water uptake capacity of 89% and a swelling index of 8.6. The gelation kinetics and the loss tangent of the gels were investigated by rheometry. The changes in the morphology of the porous foams were visualized by SEM. The changes in RSF chemical composition and the relative fraction of its secondary structural elements were analyzed by ATR-FTIR along with Fourier self-deconvolution. And, the changes in the glass transition temperature, specific heat capacity and the relative fraction of crystallinity of RSF were determined by TM-DSC. Data suggested that RSF-water-methanol behaved as a polymer-solvent-non-solvent ternary phase system, wherein the demixing of the water-methanol phases altered the thermodynamic equilibrium of RSF-water phases and resulted in the desolvation and eventual separation of the RSF phase. Systematic analysis revealed that both gelation time and the properties of hydrogels and porous foams could be controlled by the ratios of RSF and non-solvent concentration as well as by the type of non-solvent and incubation temperature. Due to the unique properties we envisage that the herein prepared NIPS induced RSF hydrogels and porous foams can possibly be used for the encapsulation of cells and/or for the controlled release of both hydrophilic and hydrophobic drugs. PMID:26730413

  20. Silk fibroin porous scaffolds for nucleus pulposus tissue engineering

    Intervertebral discs (IVDs) are structurally complex tissue that hold the vertebrae together and provide mobility to spine. The nucleus pulposus (NP) degeneration often results in degenerative IVD disease that is one of the most common causes of back and neck pain. Tissue engineered nucleus pulposus offers an alternative approach to regain the function of the degenerative IVD. The aim of this study is to determine the feasibility of porous silk fibroin (SF) scaffolds fabricated by paraffin-sphere-leaching methods with freeze-drying in the application of nucleus pulposus regeneration. The prepared scaffold possessed high porosity of 92.38 ± 5.12% and pore size of 165.00 ± 8.25 μm as well as high pore interconnectivity and appropriate mechanical properties. Rabbit NP cells were seeded and cultured on the SF scaffolds. Scanning electron microscopy, histology, biochemical assays and mechanical tests revealed that the porous scaffolds could provide an appropriate microstructure and environment to support adhesion, proliferation and infiltration of NP cells in vitro as well as the generation of extracellular matrix. The NP cell–scaffold construction could be preliminarily formed after subcutaneously implanted in a nude mice model. In conclusion, The SF porous scaffold offers a potential candidate for tissue engineered NP tissue. - Highlights: • Paraffin microsphere-leaching method is used to fabricate silk fibroin scaffold. • The scaffold has appropriate mechanical property, porosity and pore size • The scaffold supports growth and infiltration of nucleus pulposus cells. • Nucleus pulposus cells can secrete extracellular matrix in the scaffolds. • The scaffold is a potential candidate for tissue engineered nucleus pulposus

  1. Silk fibroin porous scaffolds for nucleus pulposus tissue engineering

    Zeng, Chao; Yang, Qiang [Department of Spine Surgery, Tianjin Hospital, Tianjin 300211 (China); Tianjin Medical University, Tianjin 300070 (China); Zhu, Meifeng [The Key Laboratory of Bioactive Materials, Ministry of Education, College of Life Sciences, Nankai University, Tianjin 300071 (China); Du, Lilong [Department of Spine Surgery, Tianjin Hospital, Tianjin 300211 (China); Tianjin Medical University, Tianjin 300070 (China); Zhang, Jiamin [The Key Laboratory of Bioactive Materials, Ministry of Education, College of Life Sciences, Nankai University, Tianjin 300071 (China); Ma, Xinlong [Department of Spine Surgery, Tianjin Hospital, Tianjin 300211 (China); Xu, Baoshan, E-mail: xubaoshan99@126.com [Department of Spine Surgery, Tianjin Hospital, Tianjin 300211 (China); Wang, Lianyong, E-mail: wly@nankai.edu.cn [The Key Laboratory of Bioactive Materials, Ministry of Education, College of Life Sciences, Nankai University, Tianjin 300071 (China)

    2014-04-01

    Intervertebral discs (IVDs) are structurally complex tissue that hold the vertebrae together and provide mobility to spine. The nucleus pulposus (NP) degeneration often results in degenerative IVD disease that is one of the most common causes of back and neck pain. Tissue engineered nucleus pulposus offers an alternative approach to regain the function of the degenerative IVD. The aim of this study is to determine the feasibility of porous silk fibroin (SF) scaffolds fabricated by paraffin-sphere-leaching methods with freeze-drying in the application of nucleus pulposus regeneration. The prepared scaffold possessed high porosity of 92.38 ± 5.12% and pore size of 165.00 ± 8.25 μm as well as high pore interconnectivity and appropriate mechanical properties. Rabbit NP cells were seeded and cultured on the SF scaffolds. Scanning electron microscopy, histology, biochemical assays and mechanical tests revealed that the porous scaffolds could provide an appropriate microstructure and environment to support adhesion, proliferation and infiltration of NP cells in vitro as well as the generation of extracellular matrix. The NP cell–scaffold construction could be preliminarily formed after subcutaneously implanted in a nude mice model. In conclusion, The SF porous scaffold offers a potential candidate for tissue engineered NP tissue. - Highlights: • Paraffin microsphere-leaching method is used to fabricate silk fibroin scaffold. • The scaffold has appropriate mechanical property, porosity and pore size • The scaffold supports growth and infiltration of nucleus pulposus cells. • Nucleus pulposus cells can secrete extracellular matrix in the scaffolds. • The scaffold is a potential candidate for tissue engineered nucleus pulposus.

  2. Self-assembly model, hepatocytes attachment and inflammatory response for silk fibroin/chitosan scaffolds

    Silk fibroin is an attractive natural fibrous protein for biomedical application due to its good biocompatibility and high tensile strength. Silk fibroin is apt to form a sheet-like structure during the freeze-drying process, which is not suitable for the scaffold of tissue engineering. In our former study, the adding of chitosan promoted the self-assembly of silk fibroin/chitosan (SFCS) into a three-dimensional (3D) homogeneous porous structure. In this study, a model of the self-assembly is proposed; furthermore, hepatocytes attachment and inflammatory response for the SFCS scaffold were examined. The rigid chain of chitosan may be used as a template for β-sheet formation of silk fibroin, and this may break the sheet structure of the silk fibroin scaffold and promote the formation of a 3D porous structure of the SFCS scaffold. Compared with the polylactic glycolic acid scaffold, the SFCS scaffold further facilitates the attachment of hepatocytes. To investigate the inflammatory response, SFCS scaffolds were implanted into the greater omentum of rats. From the results of implantation, we could demonstrate in vivo that the implantation of SFCS scaffolds resulted in only slight inflammation. Keeping the good histocompatibility and combining the advantages of both fibroin and chitosan, the SFCS scaffold could be a prominent candidate for soft tissue engineering, for example, in the liver.

  3. Optimization of the silk scaffold sericin removal process for retention of silk fibroin protein structure and mechanical properties

    Teh, Thomas K H; Toh, Siew-Lok; Goh, James C H, E-mail: dosgohj@nus.edu.s, E-mail: dostkh@nus.edu.s, E-mail: bietohsl@nus.edu.s [Division of Bioengineering, National University of Singapore (Singapore)

    2010-06-01

    In the process of removing sericin (degumming) from a raw silk scaffold, the fibroin structural integrity is often challenged, leading to mechanical depreciation. This study aims to identify the factors and conditions contributing to fibroin degradation during alkaline degumming and to perform an optimization study of the parameters involved to achieve preservation of fibroin structure and properties. The methodology involves degumming knitted silk scaffolds for various durations (5-90 min) and temperatures (60-100 {sup 0}C). Mechanical agitation and use of the refreshed solution during degumming are included to investigate how these factors contribute to degumming efficiency and fibroin preservation. Characterizations of silk fibroin morphology, mechanical properties and protein components are determined by scanning electron microscopy (SEM), single fiber tensile tests and gel electrophoresis (SDS-PAGE), respectively. Sericin removal is ascertained via SEM imaging and a protein fractionation method involving SDS-PAGE. The results show that fibroin fibrillation, leading to reduced mechanical integrity, is mainly caused by prolonged degumming duration. Through a series of optimization, knitted scaffolds are observed to be optimally degummed and experience negligible mechanical and structural degradation when subjected to alkaline degumming with mechanical agitation for 30 min at 100 {sup 0}C.

  4. Optimization of the silk scaffold sericin removal process for retention of silk fibroin protein structure and mechanical properties

    In the process of removing sericin (degumming) from a raw silk scaffold, the fibroin structural integrity is often challenged, leading to mechanical depreciation. This study aims to identify the factors and conditions contributing to fibroin degradation during alkaline degumming and to perform an optimization study of the parameters involved to achieve preservation of fibroin structure and properties. The methodology involves degumming knitted silk scaffolds for various durations (5-90 min) and temperatures (60-100 0C). Mechanical agitation and use of the refreshed solution during degumming are included to investigate how these factors contribute to degumming efficiency and fibroin preservation. Characterizations of silk fibroin morphology, mechanical properties and protein components are determined by scanning electron microscopy (SEM), single fiber tensile tests and gel electrophoresis (SDS-PAGE), respectively. Sericin removal is ascertained via SEM imaging and a protein fractionation method involving SDS-PAGE. The results show that fibroin fibrillation, leading to reduced mechanical integrity, is mainly caused by prolonged degumming duration. Through a series of optimization, knitted scaffolds are observed to be optimally degummed and experience negligible mechanical and structural degradation when subjected to alkaline degumming with mechanical agitation for 30 min at 100 0C.

  5. Preparation and characterization of silk fibroin as a biomaterial with potential for drug delivery

    Zhang, Hao; Li, Ling-ling; Dai, Fang-yin; Zhang, Hao-hao; Ni, Bing; Zhou, Wei; Yang, Xia; Wu, Yu-Zhang

    2012-01-01

    Background Degummed silk fibroin from Bombyx mori (silkworm) has potential carrier capabilities for drug delivery in humans; however, the processing methods have yet to be comparatively analyzed to determine the differential effects on the silk protein properties, including crystalline structure and activity. Methods In this study, we treated degummed silk with four kinds of calcium-alcohol solutions, and performed secondary structure measurements and enzyme activity test to distinguish the d...

  6. Preparation and characterization of silk fibroin as a biomaterial with potential for drug delivery

    Zhang Hao; Li Ling-ling; Dai Fang-yin; Zhang Hao-hao; Ni Bing; Zhou Wei; Yang Xia; Wu Yu-zhang

    2012-01-01

    Abstract Background Degummed silk fibroin from Bombyx mori (silkworm) has potential carrier capabilities for drug delivery in humans; however, the processing methods have yet to be comparatively analyzed to determine the differential effects on the silk protein properties, including crystalline structure and activity. Methods In this study, we treated degummed silk with four kinds of calcium-alcohol solutions, and performed secondary structure measurements and enzyme activity test to distingu...

  7. Tyrosinase-Mediated Construction of a Silk Fibroin/Elastin Nanofiber Bioscaffold.

    Hong, Yanqing; Zhu, Xueke; Wang, Ping; Fu, Haitian; Deng, Chao; Cui, Li; Wang, Qiang; Fan, Xuerong

    2016-04-01

    Elastin has characteristics of elasticity, biological activity, and mechanical stability. In the present work, tyrosinase-mediated construction of a bioscaffold with silk fibroin and elastin was carried out, aiming at developing a novel medical biomaterial. The efficiency of enzymatic oxidation of silk fibroin and the covalent reaction between fibroin and elastin were examined by spectrophotometry, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and size exclusion chromatography (SEC). The properties of composite air-dried and nanofiber scaffolds were investigated. The results reveal that elastin was successfully bonded to silk fibroins, resulting in an increase in molecular weight of fibroin proteins. ATR-FTIR spectra indicated that tyrosinase treatment impacted the conformational structure of fibroin-based membrane. The thermal behaviors and mechanical properties of the tyrosinase-treated scaffolds were also improved compared with the untreated group. NIH/3T3 cells exhibited optimum densities when grown on the nanofiber scaffold, implying that the nanofiber scaffold has enhanced biocompatibility compared to the air-dried scaffold. A biological nanofiber scaffold constructed from tyrosinase-treated fibroin and elastin could potentially be utilized in biomedical applications. PMID:26679706

  8. Collagen/silk fibroin composite scaffold incorporated with PLGA microsphere for cartilage repair.

    Wang, Jianhua; Yang, Qiu; Cheng, Niangmei; Tao, Xiaojun; Zhang, Zhihua; Sun, Xiaomin; Zhang, Qiqing

    2016-04-01

    For cartilage repair, ideal scaffolds should mimic natural extracellular matrix (ECM) exhibiting excellent characteristics, such as biocompatibility, suitable porosity, and good cell affinity. This study aimed to prepare a collagen/silk fibroin composite scaffold incorporated with poly-lactic-co-glycolic acid (PLGA) microsphere that can be applied in repairing cartilage. To obtain optimum conditions for manufacturing a composite scaffold, a scaffold composed of different collagen-to-silk fibroin ratios was evaluated by determining porosity, water absorption, loss rate in hot water, and cell proliferation. Results suggested that the optimal ratio of collagen and silk fibroin composite scaffold was 7:3. The microstructure and morphological characteristics of the obtained scaffold were also examined through scanning electron microscopy and Fourier transform infrared spectroscopy. The results of in vitro fluorescence staining of bone marrow stromal cells revealed that collagen/silk fibroin composite scaffold enhanced cell proliferation without eliciting side effects. The prepared composite scaffold incorporated with PLGA microsphere was implanted in fully thick articular cartilage defects in rabbits. Collagen/silk fibroin composite scaffold with PLGA microspheres could enhance articular cartilage regeneration and integration between the repaired cartilage and the surrounding cartilage. Therefore, this composite will be a promising material for cartilage repair and regeneration. PMID:26838900

  9. Silk Fibroin Aqueous-Based Adhesives Inspired by Mussel Adhesive Proteins.

    Burke, Kelly A; Roberts, Dane C; Kaplan, David L

    2016-01-11

    Silk fibroin from the domesticated silkworm Bombyx mori is a naturally occurring biopolymer with charged hydrophilic terminal regions that end-cap a hydrophobic core consisting of repeating sequences of glycine, alanine, and serine residues. Taking inspiration from mussels that produce proteins rich in L-3,4-dihydroxyphenylalanine (DOPA) to adhere to a variety of organic and inorganic surfaces, the silk fibroin was functionalized with catechol groups. Silk fibroin was selected for its high molecular weight, tunable mechanical and degradation properties, aqueous processability, and wide availability. The synthesis of catechol-functionalized silk fibroin polymers containing varying amounts of hydrophilic polyethylene glycol (PEG, 5000 g/mol) side chains was carried out to balance silk hydrophobicity with PEG hydrophilicity. The efficiency of the catechol functionalization reaction did not vary with PEG conjugation over the range studied, although tuning the amount of PEG conjugated was essential for aqueous solubility. Adhesive bonding and cell compatibility of the resulting materials were investigated, where it was found that incorporating as little as 6 wt % PEG prior to catechol functionalization resulted in complete aqueous solubility of the catechol conjugates and increased adhesive strength compared with silk lacking catechol functionalization. Furthermore, PEG-silk fibroin conjugates maintained their ability to form β-sheet secondary structures, which can be exploited to reduce swelling. Human mesenchymal stem cells (hMSCs) proliferated on the silks, regardless of PEG and catechol conjugation. These materials represent a protein-based approach to catechol-based adhesives, which we envision may find applicability as biodegradable adhesives and sealants. PMID:26674175

  10. Electrospinning of carboxyethyl chitosan/poly(vinyl alcohol)/silk fibroin nanoparticles for wound dressings.

    Zhou, Yingshan; Yang, Hongjun; Liu, Xin; Mao, Jun; Gu, Shaojin; Xu, Weilin

    2013-02-01

    Composite nanofibrous membranes of water-soluble N-carboxyethyl chitosan/poly(vinyl alcohol)/silk fibroin nanoparticles were successfully fabricated by electrospinning. The composite nanofibers were subjected to detailed analysis by scanning electron microscopy (SEM), X-ray diffraction (XRD) and differential scanning calorimetry (DSC). SEM results investigated that the morphology and diameter of the nanofibers were affected by silk fibroin nanoparticles content. XRD and DSC demonstrated that there was intermolecular hydrogen bonding among the molecules of carboxyethyl chitosan, silk fibroin and PVA. The crystalline microstructure of the electrospun fibers was not well developed. The indirect cytotoxicity assessments of the nanofibers were studied. The result showed the nanofibers had good biocompatibility. This novel electrospun matrix would be used as potential wound dressing for skin regeneration. PMID:23164753

  11. Controlling silk fibroin microspheres via molecular weight distribution

    Zeng, Dong-Mei; Pan, Jue-Jing; Wang, Qun; Liu, Xin-Fang; Wang, Hui [National Engineering Laboratory for Modern Silk, College for Textile and Clothing Engineering, Soochow University, Suzhou, Jiangsu 215123 (China); Zhang, Ke-Qin, E-mail: kqzhang@suda.edu.cn [National Engineering Laboratory for Modern Silk, College for Textile and Clothing Engineering, Soochow University, Suzhou, Jiangsu 215123 (China); Research Center of Cooperative Innovation for Functional Organic/Polymer Material Micro/Nanofabrication, Soochow University, Suzhou, Jiangsu 215123 (China)

    2015-05-01

    Silk fibroin (SF) microspheres were produced by salting out SF solution via the addition of potassium phosphate buffer solution (K{sub 2}HPO{sub 4}–KH{sub 2}PO{sub 4}). The morphology, size and polydispersity of SF microspheres were adjusted by changing the molecular weight (MW) distribution and concentration of SF, as well as the ionic strength and pH of the buffer solution. Changing the conditions under which the SF fiber dissolved in the Lithium Boride (LiBr) solution resulted in altering the MW distribution of SF solution. Under optimal salting-out conditions (ionic strength > 0.7 M and pH > 7) and using a smaller and narrower SF MW distribution, SF microspheres with smoother shapes and more uniform sizes were produced. Meanwhile, the size and polydispersity of the microspheres increased when the SF concentration was increased from 0.25 mg/mL to 20 mg/mL. The improved SF microspheres, obtained by altering the distribution of molecular weight, have potential in drug and gene delivery applications. - Highlights: • MW distribution was changed by applying different dissolving methods of SF fiber. • Smaller and narrower MW distribution improves the quality of SF microspheres. • Size and polydispersity of microspheres increase as SF concentration increases. • Improved SF microspheres have potential in drug and gene delivery applications.

  12. Preparation and Characterization of Nanocomposite and Nanoporous Silk Fibroin Films

    Yaowalak Srisuwan

    2008-01-01

    Full Text Available Nanocomposite and nanoporous silk fibroin (SF films were prepared by film casting of SF solution containing surfactant-free colloidal nanoparticles of methoxy poly (ethylene glycol-b-poly (D, L-lactide diblock copolymer (MPEG-b-PDLL. Self-condensation and nanophase separation of the nanoparticles from SF film matrix during film drying process gave nanopore structures. The colloidal nanoparticles were prepared in SF solution by modified-spontaneous emulsification solvent diffusion method without any surfactant. The interaction between SF and MPEG-b-PDLL in nanocomposite films was studied by Fourier transform infrared spectroscopy and thermogravimetry. The film transparency of SF nanocomposite films decreased as increasing the MPEG-b-PDLL ratio. Scanning Electron Microscopy (SEM results indicated that the nanoparticle sizes in the films were in the range of 50-200 nm with spherical shape. Nanopore structures with pore size of less than 150 nm can be observed from SEM images of the film surface and cross-section. The nanopores are interconnected throughout the nanocomposite films. The number and size of nanoparticles and nanopores increased when the MPEG-b-PDLL ratio was increased.

  13. Fabrication of silk fibroin nanoparticles for controlled drug delivery

    A novel solution-enhanced dispersion by supercritical CO2 (SEDS) was employed to prepare silk fibroin (SF) nanoparticles. The resulting SF nanoparticles exhibited a good spherical shape, a smooth surface, and a narrow particle size distribution with a mean particle diameter of about 50 nm. The results of X-ray powder diffraction, thermo gravimetry-differential scanning calorimetry, and Fourier transform infrared spectroscopy analysis of the SF nanoparticles before and after ethanol treatment indicated conformation transition of SF nanoparticles from random coil to β-sheet form and thus water insolubility. The MTS assay also suggested that the SF nanoparticles after ethanol treatment imposed no toxicity. A non-steroidal anti-inflammatory drug, indomethacin (IDMC), was chosen as the model drug and was encapsulated in SF nanoparticles by the SEDS process. The resulting IDMC–SF nanoparticles, after ethanol treatment, possessed a theoretical average drug load of 20%, an actual drug load of 2.05%, and an encapsulation efficiency of 10.23%. In vitro IDMC release from the IDMC–SF nanoparticles after ethanol treatment showed a significantly sustained release over 2 days. These studies of SF nanoparticles indicated the suitability of the SF nanoparticles prepared by the SEDS process as a biocompatible carrier to deliver drugs and also the feasibility of using the SEDS process to reach the goal of co-precipitation of drug and SF as composite nanoparticles for controlled drug delivery.

  14. Controlling silk fibroin microspheres via molecular weight distribution

    Silk fibroin (SF) microspheres were produced by salting out SF solution via the addition of potassium phosphate buffer solution (K2HPO4–KH2PO4). The morphology, size and polydispersity of SF microspheres were adjusted by changing the molecular weight (MW) distribution and concentration of SF, as well as the ionic strength and pH of the buffer solution. Changing the conditions under which the SF fiber dissolved in the Lithium Boride (LiBr) solution resulted in altering the MW distribution of SF solution. Under optimal salting-out conditions (ionic strength > 0.7 M and pH > 7) and using a smaller and narrower SF MW distribution, SF microspheres with smoother shapes and more uniform sizes were produced. Meanwhile, the size and polydispersity of the microspheres increased when the SF concentration was increased from 0.25 mg/mL to 20 mg/mL. The improved SF microspheres, obtained by altering the distribution of molecular weight, have potential in drug and gene delivery applications. - Highlights: • MW distribution was changed by applying different dissolving methods of SF fiber. • Smaller and narrower MW distribution improves the quality of SF microspheres. • Size and polydispersity of microspheres increase as SF concentration increases. • Improved SF microspheres have potential in drug and gene delivery applications

  15. Synthesis and characterization of dense membranes of silk fibroin with glycerin

    The addition of plasticizers seeks improvements in mechanical properties of dense membranes of silk fibroin with possible interactions by hydrogen bonds. The aim of the present study was to produce and characterize dense membranes of silk fibroin containing glycerin in two different concentrations. The characterization of the membranes was performed from scanning electron microscopy (SEM), mechanical traction tests, infrared spectroscopy (FTIR-ATR) and X-ray diffraction (XRD). The results indicated that the addition of glycerin allowed obtaining homogeneous and more crystalline membranes and improved their properties of elongation. (author)

  16. Silk fibroin membranes from solvent-crystallized silk fibroin/gelatin blends: Effects of blend and solvent composition

    Gil, Eun S. [Fiber and Polymer Science Program, North Carolina State University, Raleigh, NC 27695 (United States); Frankowski, David J. [Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695 (United States); Hudson, Samuel M. [Fiber and Polymer Science Program, North Carolina State University, Raleigh, NC 27695 (United States); Spontak, Richard J. [Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695 (United States) and Department of Materials Science and Engineering, North Carolina State University, Raleigh, NC 27695 (United States)]. E-mail: Rich_Spontak@ncsu.edu

    2007-04-15

    Protein membranes have been prepared by mixing gelatin (G) with Bombyx mori silk fibroin (SF) and using aqueous methanol (MeOH) to induce SF crystallization. Amorphous blends of these polymers appear quasi-homogeneous, as discerned from visual observation, electron microscopy and Fourier-transform infrared (FTIR) spectroscopy. Upon subsequent exposure to aqueous MeOH, SF undergoes a conformational change from random-coil to {beta}-sheet. This transformation occurs in pure SF, as well as in each of the G/SF blends, as discerned from FTIR spectroscopy and thermal calorimetry. The influence of MeOH-induced SF crystallization on structure and property development has been measured as functions of blend and solvent composition. By preserving a support scaffold above the G helix-to-coil transition temperature, the formation of crystalline SF networks in G/SF blends can be used to stabilize G-based hydrogels or generate SF membranes for biomaterial, pharmaceutical and gas-separation purposes. The present study not only examines the properties of G/SF blends before and after SF crystallization, but also establishes the foundation for future research into thermally-responsive G/SF bioconjugates.

  17. Development of macro/micro porous silk fibroin scaffolds with nano-sized calcium phosphate particles for bone tissue engineering

    Yan, Leping; Correia, Cristina; Correia, Joana Silva; Caridade, S. G.; Fernandes, E M; Mano, J. F.; Sousa, R A; Oliveira, Joaquim M.; de Oliveira, A. L.; Reis, R.L.

    2012-01-01

    Macro/micro porous silk scaffolds with nano-sized calcium phosphate (CaP) particles were developed for bone tissue engineering. Different amounts of nano-sized CaP particles 4, 8, 16 and 25% (CaP:silk fibroin, wt/wt) were generated into the highly concentrated aqueous silk fibroin solutions via an in-situ approach. Afterwards, the pure silk and silk/nano-CaP porous scaffolds were produced by a combination of salt-leaching/freeze-drying methods. Thermal gravimetric analysi...

  18. Novel genipin cross-linked chitosan-silk fibroin sponges for cartilage engineering strategies

    Silva, S.S; Motta, A.; Rodrigues, Mrcia T.; Pinheiro, A. F. M.; Gomes, Manuela E.; Mano, J. F.; Reis, R.L.; Migliaresi, C.

    2008-01-01

    The positive interaction of materials with tissues is an important step in regenerative medicine strategies. Hydrogels that are obtained from polysaccharides and proteins are expected to mimic the natural cartilage environment and thus provide an optimum milleu for tissue growth and regeneration. In this work, novel hydrogels composed of blends of chitosan and Bombyx mori silk fibroin were cross-linked with genipin (G) and were freeze dried to obtain chitosan/silk (CSG) sponges. C...

  19. Preparation of silk fibroinpoly(ethylene glycol) conjugate films through click chemistry

    Sampaio, Sandra; Miranda, Teresa; Santos, Jorge Gomes; Soares, Graa M. B.

    2011-01-01

    Azide silk fibroin (azido SF) and alkyne terminal poly(ethylene glycol) (PEG) 2000 (acetylene-terminal PEG 2000) were synthesized. Azido SF was reacted with acetylene-terminal PEG 2000 to produce films via a copper-mediated 1,3-cycloaddition (click chemistry) generating a triazole linkage as the networking forming reaction. Through click chemistry, novel silk-based films with various weight ratios were prepared and investigated. Fourier transform infrared, X-ray diffraction and differential...

  20. Enzymatic degradation behavior and cytocompatibility of silk fibroin-starch-chitosan conjugate membranes

    Baran, Erkan T., E-mail: erkantur@metu.edu.tr; Tuzlakoglu, Kadriye, E-mail: kadriye@dep.uminho.pt; Mano, Joao F., E-mail: jmano@dep.uminho.pt; Reis, Rui L., E-mail: rgreis@dep.uminho.pt

    2012-08-01

    The objective of this study was to investigate the influence of silk fibroin and oxidized starch conjugation on the enzymatic degradation behavior and the cytocompatability of chitosan based biomaterials. The tensile stress of conjugate membranes, which was at 50 Megapascal (MPa) for the lowest fibroin and starch composition (10 weight percent (wt.%)), was decreased significantly with the increased content of fibroin and starch. The weight loss of conjugates in {alpha}-amylase was more notable when the starch concentration was the highest at 30 wt.%. The conjugates were resistant to the degradation by protease and lysozyme except for the conjugates with the lowest starch concentration. After 10 days of cell culture, the proliferation of osteoblast-like cells (SaOS-2) was stimulated significantly by higher fibroin compositions and the DNA synthesis on the conjugate with the highest fibroin (30 wt.%) was about two times more compared to the native chitosan. The light microscopy and the image analysis results showed that the cell area and the lengths were decreased significantly with higher fibroin/chitosan ratio. The study proved that the conjugation of fibroin and starch with the chitosan based biomaterials by the use of non-toxic reductive alkylation crosslinking significantly improved the cytocompatibility and modulated the biodegradation, respectively. - Highlights: Black-Right-Pointing-Pointer Silk fibroin, starch and chitosan conjugates were prepared by reductive alkylation. Black-Right-Pointing-Pointer The enzymatic biodegradation and the cytocompatibility of conjugates were tested. Black-Right-Pointing-Pointer The conjugate with 30% starch composition was degraded by {alpha}-amylase significantly. Black-Right-Pointing-Pointer Higher starch composition in conjugates prevented protease and lysozyme degradation. Black-Right-Pointing-Pointer Fibroin incorporation effectively increased the cell proliferation of conjugates.

  1. Preparation and water absorption of cross-linked chitosan/silk fibroin blend films

    Natural polymer blend films composed of chitosan and silk fibroin were prepared by varying the ratio of chitosan to silk fibroin, with and without glutaraldehyde as a crosslinking agent. The effects of the ratio of chitosan to silk fibroin and crosslinking agent on swelling behavior of the blend films were studied. For the swelling behavior, the blend films exhibited a dramatic change in the degree of swelling when immersed in acidic solutions. The degree of swelling of the films increased as the chitosan content increased; the blend film with 80% chitosan content had the maximum degree of swelling. It appeared that crosslinking had occurred in the blend films which helped the films to retain their three dimensional structure. In addition, FTIR spectra of the films showed evidence of hydrogen bonding interaction between chitosan and silk fibroin. For the effect of salt type, the films were immersed in various types of aqueous salt solutions, viz NaCl, LiCl, CaCl2, AlCl3, and FeCl3. The films immersed in AlCl3 and FeCl3 aqueous solutions gave the maximum degree of swelling. The effects of AlCl3 and FeCl3 concentrations on swelling behavior were also investigated. It was found that the maximum degree of swelling of the films occurred at 1.0 x 10-2 M of AlCl3 and FeCl3 aqueous solutions. (author)

  2. Formation of silk fibroin nanoparticles in water-miscible organic solvent and their characterization

    When Silk fibre derived from Bombyx mori, a native biopolymer, was dissolved in highly concentrated neutral salts such as CaCl2, the regenerated liquid silk, a gradually degraded peptide mixture of silk fibroin, could be obtained. The silk fibroin nanoparticles were prepared rapidly from the liquid silk by using water-miscible protonic and polar aprotonic organic solvents. The nanoparticles are insoluble but well dispersed and stable in aqueous solution and are globular particles with a range of 35-125 nm in diameter by means of TEM, SEM, AFM and laser sizer. Over one half of the ε-amino groups exist around the protein nanoparticles by using a trinitrobenzenesulfonic acid (TNBS) method. Raman spectra shows the tyrosine residues on the surface of the globules are more exposed than those on native silk fibers. The crystalline polymorph and conformation transition of the silk nanoparticles from random-coil and α-helix form (Silk I) into anti-parallel β-sheet form (Silk II) are investigated in detail by using infrared, fluorescence and Raman spectroscopy, DSC, 13C CP-MAS NMR and electron diffraction. X-ray diffraction of the silk nanoparticles shows that the nanoparticles crystallinity is about four fifths of the native fiber. Our results indicate that the degraded peptide chains of the regenerated silk is gathered homogeneously or heterogeneously to form a looser globular structure in aqueous solution. When introduced into excessive organic solvent, the looser globules of the liquid silk are rapidly dispersed and simultaneously dehydrated internally and externally, resulting in the further chain-chain contact, arrangement of those hydrophobic domains inside the globules and final formation of crystalline silk nanoparticles with β-sheet configuration. The morphology and size of the nanoparticles are relative to the kinds, properties and even molecular structures of organic solvents, and more significantly to the looser globular substructure of the degraded silk fibroin in aqueous solution. It is possible that the silk protein nanoparticles are potentially useful in biomaterials such as cosmetics, anti-UV skincare products, industrial materials and surface improving materials, especially in enzyme/drug delivery system as vehicle

  3. Formation of silk fibroin nanoparticles in water-miscible organic solvent and their characterization

    Zhang Yuqing, E-mail: yqzhang@public1.sz.js.cn; Shen Weide; Xiang Ruli [Soochow University, Silk Biotechnol. Lab., School of Life Science (China); Zhuge Lanjian; Gao Weijian; Wang Wenbao [Soochow University, Analytical Center (China)

    2007-10-15

    When Silk fibre derived from Bombyx mori, a native biopolymer, was dissolved in highly concentrated neutral salts such as CaCl{sub 2}, the regenerated liquid silk, a gradually degraded peptide mixture of silk fibroin, could be obtained. The silk fibroin nanoparticles were prepared rapidly from the liquid silk by using water-miscible protonic and polar aprotonic organic solvents. The nanoparticles are insoluble but well dispersed and stable in aqueous solution and are globular particles with a range of 35-125 nm in diameter by means of TEM, SEM, AFM and laser sizer. Over one half of the {epsilon}-amino groups exist around the protein nanoparticles by using a trinitrobenzenesulfonic acid (TNBS) method. Raman spectra shows the tyrosine residues on the surface of the globules are more exposed than those on native silk fibers. The crystalline polymorph and conformation transition of the silk nanoparticles from random-coil and {alpha}-helix form (Silk I) into anti-parallel {beta}-sheet form (Silk II) are investigated in detail by using infrared, fluorescence and Raman spectroscopy, DSC, {sup 13}C CP-MAS NMR and electron diffraction. X-ray diffraction of the silk nanoparticles shows that the nanoparticles crystallinity is about four fifths of the native fiber. Our results indicate that the degraded peptide chains of the regenerated silk is gathered homogeneously or heterogeneously to form a looser globular structure in aqueous solution. When introduced into excessive organic solvent, the looser globules of the liquid silk are rapidly dispersed and simultaneously dehydrated internally and externally, resulting in the further chain-chain contact, arrangement of those hydrophobic domains inside the globules and final formation of crystalline silk nanoparticles with {beta}-sheet configuration. The morphology and size of the nanoparticles are relative to the kinds, properties and even molecular structures of organic solvents, and more significantly to the looser globular substructure of the degraded silk fibroin in aqueous solution. It is possible that the silk protein nanoparticles are potentially useful in biomaterials such as cosmetics, anti-UV skincare products, industrial materials and surface improving materials, especially in enzyme/drug delivery system as vehicle.

  4. A dry powder formulation from silk fibroin microspheres as a topical auto-gelling device.

    Faragò, Silvio; Lucconi, Giulia; Perteghella, Sara; Vigani, Barbara; Tripodo, Giuseppe; Sorrenti, Milena; Catenacci, Laura; Boschi, Alessandra; Faustini, Massimo; Vigo, Daniele; Chlapanidas, Theodora; Marazzi, Mario; Torre, Maria Luisa

    2016-06-01

    With the aim of establishing the formulation of a new hydrophilic auto-gelling medical device for biomedical applications, fibroin-based microspheres were prepared. The proposed microspheres were produced by a cost-effective and industrially scalable technique, such as the spray-drying. Spray-dried silk fibroin microspheres were obtained and the effects of different hydrophilic polymer on the process yield, microsphere morphology and conformation transition of fibroin were evaluated. The final auto-gelling formulations were obtained by adding calcium gluconate (as a calcium source for alginate crosslinking) to the prepared microspheres and tested by an in vitro gelling test. This study showed that the combination of fibroin with sodium alginate and poloxamer produced the most promising auto-gelling formulation for specific biomedical applications, such as the treatment of pressure ulcers. PMID:25757645

  5. AFM observation of silk fibroin on mica substrates: morphologies reflecting the secondary structures

    Yamada, Kazushi; Tsuboi, Yasuyuki; Itaya, Akira

    2003-09-01

    Bombyx mori silk fibroin was fixed on mica substrates by cast of aqueous fibroin solutions, and the microscopic morphologies of the samples were revealed by means of atomic force microscopy. By adjusting the method used to prepare the solution, we succeeded in forming quasi-2-dimensional thin films in which a network of fibroin molecules developed over the substrate. The film network consisted of fibroin in a random coil structure. The morphology of the network changed after thermal or methanol treatments, which are known to convert the secondary structure of fibroin from the random coil to the {beta}-sheet type. In both of these cases, the network morphology disappeared and characteristic island-like morphologies appeared. On the other hand, temporally evolving gelation occurred in a fibroin solution due to the formation of {beta}-sheet crystals. Such islands were also observable in a specimen prepared by the cast of the gel-containing solution. Based on these results, it was concluded that the islands consist of {beta}-sheet crystals. Of particular interest is the observation that all of the islands had a common thickness value of 1.3 nm. These morphologies are discussed in terms of the secondary structure of fibroin.

  6. Comparative study of silk fibroin porous scaffolds derived from salt/water and sucrose/hexafluoroisopropanol in cartilage formation.

    Makaya, Kumiko; Terada, Shinichi; Ohgo, Kosuke; Asakura, Tetsuo

    2009-07-01

    The purpose of this study is to create a new silk fibroin scaffold with sufficient three-dimensional morphology and porous structure for cartilage formation. We have applied sucrose particles sized around 300 to 500 microm as porogens compared to equal-sized salt particles. After the porogen was leached out with water, scaffolds were prepared with fibroin derived from sucrose/hexafluoroisopropanol (Su/H) or salt/water (Sa/W) based composites. A compression test indicated that the Sa/W fibroin was much harder than the Su/H fibroin, but a protease enzyme digested the Sa/W fibroin more quickly than Su/H fibroin. Rabbit ear chondrocytes were seeded onto the scaffolds for 4-8 week in vitro culture and histological analyses were performed. The distribution of cartilage formation in Safranin O staining was more homogenous in Su/H fibroin than that of Sa/W fibroin. The overall amount of cartilage was significantly better in the Su/H fibroin than that in the Sa/W fibroin. However, the inner structure of pore wall in the Sa/W fibroin was rough and microporous with cartilage matrix deposition, while that in the Su/H fibroin was thin and homogenous. Since mature cartilage gradually regenerates to fill the porous space, slowly degradable Su/H fibroin should be a better candidate for cartilage formation. PMID:19577196

  7. Potential of 3-D tissue constructs engineered from bovine chondrocytes / silk fibroin-chitosan for in vitro cartilage tissue engineering

    Bhardwaj, Nandana; Nguyen, Quynhhoa T; Chen, Albert C.; Kaplan, David L.; Sah, Robert L.; Kundu, Subhas C.

    2011-01-01

    The use of cell-scaffold constructs is a promising tissue engineering approach to repair cartilage defects and to study cartilaginous tissue formation. In this study, silk fibroin/chitosan blended scaffolds were fabricated and studied for cartilage tissue engineering. Silk fibroin served as a substrate for cell adhesion and proliferation while chitosan has a structure similar to that of glycosaminoglycans, and shows promise for cartilage repair. We compared the formation of cartilaginous tiss...

  8. Optical spectroscopy to investigate the structure of regenerated Bombyx mori silk fibroin in solution.

    Yang, Yuhong; Shao, Zhengzhong; Chen, Xin; Zhou, Ping

    2004-01-01

    Fluorescence and circular dichroism spectroscopy were used to monitor the conformational transition of regenerated Bombyx mori silk fibroin (RSF) in aqueous solutions under different conditions. According to the analysis of fluorescence spectra using anilinonaphthalene-8-sulfonic acid magnesium salt (ANS) as an external probe, the destruction of the hydrophobic core prior to the secondary structure change suggests that this collapse may initiate the conformational transition from random coil to beta-sheet for RSF. The temperature dependence of the structural changes of RSF, detected by both fluorescence spectroscopy and circular dichroism, shows a reversible process upon heating and recooling, with the midpoint around 45 degrees C. The results also indicate that most of the tryptophan (Trp) residues contained in silk fibroin are concentrated on the surface of the unfolded protein. However, they will change their location in the highly ordered structure (e.g., becoming more homogeneous) with the conformational transition of silk fibroin. Moreover, our studies also suggest that the presence of water plays a crucial role during the structure changes of fibroin. PMID:15132660

  9. The effect of gamma irradiation on some morphological and quantitative changes of Bombyx mori L. silk gland

    A study to determine the effect of gamma irradiation of silkworm eggs at doses of 1.00; 2.00 and 3.00 on silk gland weight, length and width is performed. It was found that gamma irradiation of eggs in the stage of embryo most intensive growth in length (B2) at doses 2.00 and 3.00 Gy stimulates increasing of silk glands weight in silkworms on the fifth day instar by 12-25 mg, as well as the silk worm width - by 7-33 ?m, which is of significant importance for the synthesis and secretion of silk proteins (fibroin and sericin). A breed specificity was also observed

  10. Potential of inherent RGD containing silk fibroin-poly (?-caprolactone) nanofibrous matrix for bone tissue engineering.

    Bhattacharjee, Promita; Kundu, Banani; Naskar, Deboki; Kim, Hae-Won; Bhattacharya, Debasis; Maiti, T K; Kundu, S C

    2016-02-01

    The current study deals with the fabrication and characterization of blended nanofibrous scaffolds of tropical tasar silk fibroin of Antheraea mylitta and poly (?-caprolactone) to act as an ideal scaffold for bone regeneration. The use of poly (?-caprolactone) in osteogenesis is well-recognized. At the same time, the osteoconductive nature of the non-mulberry tasar fibroin is also established due to its internal integrin binding peptide RGD (Arg-Gly-Asp) sequences, which enhance cellular interaction and proliferation. Considering that the materials have the required and favorable properties, the blends are formed using an equal volume ratio of fibroin (2 and 4 wt%) and poly (?-caprolactone) solution (10 wt%) to fabricate nanofibers. The nanofibers possess an average diameter of 152??18nm (2% fibroin/PCL) and 175??15nm (4% fibroin/PCL). The results of Fourier transform infrared spectroscopy substantiates the preservation of the secondary structure of the fibroin in the blends indicating the structural stability of the neo-matrix. With an increase in the fibroin percentage, the hydrophobicity and thermal stability of the matrices as measured from melting temperature Tm (using DSC) decrease, while the mechanical strength is improved. The blended nanofibrous scaffolds are biodegradable, and support the viability and proliferation of human osteoblast-like cells as observed through scanning electron and confocal microscopes. Alkaline phosphatase assay indicates the cell proliferation and the generation of the neo-bone matrix. Taken together, these findings illustrate that the silk-poly (?-caprolactone) blended nanofibrous scaffolds have an excellent prospect as scaffolding material in bone tissue engineering. PMID:26174955

  11. Preparation and characterization of regenerated Bombyx mori silk fibroin fiber with high strength

    2008-12-01

    Full Text Available Regenerated Bombyx mori silk fibers were spun from hexafluoro-iso- propanol solution of silk fibroin sponge in methanol used as a coagulant solvent and then elongated in water. The stress-strain curves of the regenerated fibers changed dramatically depending on the draw ratio and the structure was studied by 13C CP/MAS NMR and X-ray diffraction methods. The patterns of 13C CP/MAS NMR spectra of two regenerated fibers with different draw ratios (1× and 3× and native silk fiber are all β-sheet structure although the fraction of random coil/distorted β-turn decreases in the order of 1×, 3× and native fiber gradually. On the other hand, azimuthal scans of their X-ray fiber patterns changed remarkably with increasing the draw ratio. This indicates that long-range orientation of the fibroin chain changes remarkably during the drawing process, but the short-range local structure does not change significantly. Regenerated silk fiber with a draw ratio of 3× is a fiber with high strength which is comparable with that of natural silk fiber. The regenerated fiber is also more degradable than natural silk fiber in enzyme solution in vitro.

  12. Tyrosinase-catalyzed modification of Bombyx mori silk fibroin: grafting of chitosan under heterogeneous reaction conditions.

    Freddi, Giuliano; Anghileri, Anna; Sampaio, Sandra; Buchert, Johanna; Monti, Patrizia; Taddei, Paola

    2006-09-01

    The capability of mushroom tyrosinase to catalyze the oxidation of tyrosine residues of Bombyx mori silk fibroin was studied under heterogeneous reaction conditions, by using a series of silk substrates differing in surface and bulk morphology and structure, i.e. hydrated and insoluble gels, mechanically generated powder and fibre. Tyrosinase was able to oxidize 10-11% of the tyrosine residues of silk gels. The yield of the reaction was very low for the powder and undetectable for fibres. FT-Raman spectroscopy gave evidence of the oxidation reaction. New bands attributable to vibrations of oxidized tyrosine species (o-quinone) appeared, and the value of the I853/I829 intensity ratio of the tyrosine doublet changed following oxidation of tyrosine. The thermal behaviour of SF substrates was not affected by enzymatic oxidation. o-Quinones formed by tyrosinase onto gels and powder were able to undergo non-enzymatic coupling with chitosan. FT-IR and FT-Raman spectroscopy provided clear evidence of the formation of silk-chitosan bioconjugates under heterogeneous reaction conditions. Chitosan grafting caused a beta-sheet --> random coil conformational transition of silk fibroin and significant changes in the thermal behaviour. Chitosan grafting did not occur, or occurred at an undetectable level on silk fibres. The results reported in this study show the potential of the enzymatically initiated protein-polysaccharide grafting for the production of a new range of bio-based, environmentally friendly polymers. PMID:16621091

  13. Histochemical and ultrastructural evidence of lipid secretion by the silk gland of the sugarcane borer Diatraea saccharalis (Fabricius) (Lepidoptera: Crambidae).

    Victoriano, Eliane; Pinheiro, Daniela O; Gregrio, Elisa A

    2007-01-01

    The silk gland in Lepidoptera larvae is responsible for the silk production used for shelter or cocoon construction. The secretion of fibroin and sericin by the different silk gland regions are well established. There are few attempts to detect lipid components in the insect silk secretion, although the presence of such element may contribute to the resistance of the shelter to wet environment. This study characterizes the glandular region and detects the presence of lipid components in the secretion of the silk gland of Diatraea saccharalis(Fabricius). The silk gland was submitted to histochemical procedure for lipid detection or conventionally prepared for ultrastructural analyses. Lipid droplets were histochemically detected in both the apical cytoplasm of cell of the anterior region and in the lumen among the microvilli. Ultrastructural analyses of the anterior region showed lipid material, visualized as myelin-like structures within the vesicular Golgi complex and in the apical secretory globules, mixed up with the sericin; similar material was observed into the lumen, adjacent to the microvilli. Lipids were not detected in the cells neither in the lumen of the posterior region. Our results suggest that the silk produced by D. saccharalis has a minor lipid content that is secreted by the anterior region together with the sericin. PMID:18060296

  14. Effect of the sterilization method on the properties of Bombyx mori silk fibroin films

    We have compared the effects of different sterilization techniques on the properties of Bombyx mori silk fibroin thin films with the view to subsequent use for corneal tissue engineering. The transparency, tensile properties, corneal epithelial cell attachment and degradation of the films were used to evaluate the suitability of certain sterilization techniques including gamma-irradiation (in air or nitrogen), steam treatment and immersion in aqueous ethanol. The investigations showed that gamma-irradiation, performed either in air or in a nitrogen atmosphere, did not significantly alter the properties of films. The films sterilized by gamma-irradiation or by immersion in ethanol had a transparency greater than 98% and tensile properties comparable to human cornea and amniotic membrane, the materials of choice in the reconstruction of ocular surface. Although steam-sterilization produced stronger, stiffer films, they were less transparent, and cell attachment was affected by the variable topography of these films. It was concluded that gamma-irradiation should be considered to be the most suitable method for the sterilization of silk fibroin films, however, the treatment with ethanol is also an acceptable method. - Highlights: ► The effects of four methods of sterilization on the properties of silk fibroin films were investigated. ► Steam treatment leads to stiffer films but to lower transparency and variable surface topography. ► Degradation of fibroin is enhanced in the films that were gamma-irradiated. ► The effects on mechanical properties are explained through changes in both primary and secondary structure of fibroin. ► Gamma-irradiation and immersion in aqueous ethanol are suggested as preferred methods of sterilization.

  15. Effect of the sterilization method on the properties of Bombyx mori silk fibroin films

    George, Karina A. [Queensland Eye Institute, 41 Annerley Road, South Brisbane, Queensland 4101 (Australia); Institute of Health and Biomedical Innovation, Queensland University of Technology, Kelvin Grove, Queensland 4059 (Australia); Shadforth, Audra M.A. [Queensland Eye Institute, 41 Annerley Road, South Brisbane, Queensland 4101 (Australia); Chirila, Traian V., E-mail: traian.chirila@qei.org.au [Queensland Eye Institute, 41 Annerley Road, South Brisbane, Queensland 4101 (Australia); Australian Institute of Bioengineering and Nanotechnology, University of Queensland, St Lucia, 4072 (Australia); Faculty of Health Sciences, University of Queensland, Herston, Queensland 4006 (Australia); Faculty of Science and Engineering, Queensland University of Technology, Brisbane, Queensland 4001 (Australia); Laurent, Matthieu J. [Queensland Eye Institute, 41 Annerley Road, South Brisbane, Queensland 4101 (Australia); Ecole Superieure d' Ingenieurs de Luminy (ESIL), Universite de la Mediterranee Aix-Marseille II, Luminy case 925 13288, Marseille, Cedex 09 (France); Stephenson, Sally-Anne [Institute of Health and Biomedical Innovation, Queensland University of Technology, Kelvin Grove, Queensland 4059 (Australia); Faculty of Health, Queensland University of Technology, Brisbane, Queensland 4001 (Australia); Edwards, Grant A. [Australian Institute of Bioengineering and Nanotechnology, University of Queensland, St Lucia, 4072 (Australia); Madden, Peter W. [Queensland Eye Institute, 41 Annerley Road, South Brisbane, Queensland 4101 (Australia); Faculty of Health Sciences, University of Queensland, Herston, Queensland 4006 (Australia); and others

    2013-03-01

    We have compared the effects of different sterilization techniques on the properties of Bombyx mori silk fibroin thin films with the view to subsequent use for corneal tissue engineering. The transparency, tensile properties, corneal epithelial cell attachment and degradation of the films were used to evaluate the suitability of certain sterilization techniques including gamma-irradiation (in air or nitrogen), steam treatment and immersion in aqueous ethanol. The investigations showed that gamma-irradiation, performed either in air or in a nitrogen atmosphere, did not significantly alter the properties of films. The films sterilized by gamma-irradiation or by immersion in ethanol had a transparency greater than 98% and tensile properties comparable to human cornea and amniotic membrane, the materials of choice in the reconstruction of ocular surface. Although steam-sterilization produced stronger, stiffer films, they were less transparent, and cell attachment was affected by the variable topography of these films. It was concluded that gamma-irradiation should be considered to be the most suitable method for the sterilization of silk fibroin films, however, the treatment with ethanol is also an acceptable method. - Highlights: Black-Right-Pointing-Pointer The effects of four methods of sterilization on the properties of silk fibroin films were investigated. Black-Right-Pointing-Pointer Steam treatment leads to stiffer films but to lower transparency and variable surface topography. Black-Right-Pointing-Pointer Degradation of fibroin is enhanced in the films that were gamma-irradiated. Black-Right-Pointing-Pointer The effects on mechanical properties are explained through changes in both primary and secondary structure of fibroin. Black-Right-Pointing-Pointer Gamma-irradiation and immersion in aqueous ethanol are suggested as preferred methods of sterilization.

  16. The Micropillar Structure on Silk Fibroin Film Influence Intercellular Connection Mediated by Nanotubular Structures

    Renchuan You

    2014-06-01

    Full Text Available Tunneling nanotubes are important membrane channels for cell-to-cell communication. In this study, we investigated the effect of the microenvironment on nanotubular structures by preparing a three-dimensional silk fibroin micropillar structure. In previous reports, tunneling nanotubes were described as stretched membrane channels between interconnected cells at their nearest distance. They hover freely in the cell culture medium and do not contact with the substratum. Interestingly, the micropillars could provide supporting points for nanotubular connection on silk fibroin films, where nanotubular structure formed a stable anchor at contact points. Consequently, the extension direction of nanotubular structure was affected by the micropillar topography. This result suggests that the hovering tunneling nanotubes in the culture medium will come into contact with the raised roadblock on the substrates during long-distance extension. These findings imply that the surface microtopography of biomaterials have an important influence on cell communication mediated by tunneling nanotubes.

  17. Fabrication of Antibacterial Wound Dressings from Silk Fibroin and Silver Nano particles

    Full text: Patients with burn wounds that cover large body surface area are susceptible to infection which can lead to fatality. Wound dressings or skin grafts are needed to cover the wound during the regeneration of new skin tissue. The aim of this research is to fabricate antibacterial wound dressings from silk fibroin derived from the natural silk cocoon and silver nanoparticles (AgNPs) prepared by gamma irradiation. Fibroin mats composed of nonwoven fibers with diameter of 670± 11.5 nm were fabricated by electro spinning. Using gamma irradiation, the starting silver nitrate solution was reduced to colloidal AgNPs. The fibroin mats were coated with AgNPs at various AgNP concentration and then evaluated for their antibacterial property by disc diffusion test. The concentration of colloidal AgNP solution ≤ 1 mM was found to be as sufficient in inhibiting the growth of Pseudomonas aeruginosa and Staphylococcus aureus as commercial wound dressings embedded with silver ions. These results demonstrate that electro spun fibroin mats coated with AgNPs exhibite antibacterial property and can be further developed for the treatment of burn wounds

  18. Comparing the properties of Bombyx mori silk cocoons against sericin-fibroin regummed biocomposite sheets.

    Morin, Alexander; Alam, Parvez

    2016-08-01

    This paper considers the utility of sericin, a degumming waste product, in the regumming of Bombyx mori silk fibroin fibres to form sericin-fibroin biocomposites. Regummed biocomposites have a chemical character that is somewhat closer to fibroin than sericin, though sericin presence is confirmed through FT-IR spectroscopy. Using direct measurements we further find the weight fractions of sericin in the regummed biocomposites and the native cocoons differ by only 5%. Mechanically, B. mori cocoons exhibit brittle stress-strain characteristics, failing at strengths of X̅= 16.6MPa and at strains of X̅= 13%. Contrarily, aligning fibroin fibres to a unidirectional axis in the regummed biocomposites causes them to exhibit characteristics of strain hardening, which is itself a typical characteristic of silk fibre pulled in tension. Though they are half as strong (X̅= 7.2MPa), regummed biocomposites are able to absorb five times more mechanical energy (X̅= 5.6MJm(-3)) than the B. mori cocoons (X̅= 1.1MJm(-3)) and are furthermore able to elongate to more than ten times (X̅= 180%) that of the native cocoons prior to failure. Our research shows that degummed B. mori cocoons can be regummed into sheets that have potential for use as load bearing engineering biocomposites. PMID:27157746

  19. Improvement of early cell adhesion on Thai silk fibroin surface by low energy plasma.

    Amornsudthiwat, Phakdee; Mongkolnavin, Rattachat; Kanokpanont, Sorada; Panpranot, Joongjai; Wong, Chiow San; Damrongsakkul, Siriporn

    2013-11-01

    Low energy plasma has been introduced to treat the surface of Thai silk fibroin which should be enhanced for cell adhesion due to its native hydrophobic surface. Plasma surface treatment could introduce desirable hydrophilic functionalities on the surface without using any chemicals. In this work, nitrogen glow discharge plasma was generated by a low energy AC50Hz power supply system. The plasma operating conditions were optimized to reach the highest nitrogen active species by using optical emission spectroscopy. X-ray photoelectron spectroscopy (XPS) revealed that amine, hydroxyl, ether, and carboxyl groups were induced on Thai silk fibroin surface after plasma treatment. The results on Fourier transform infrared attenuated total reflection (FTIR-ATR) spectroscopy confirmed that the plasma treated effects were only on the outermost layer since there was no change in the bulk chemistry. The surface topography was insignificantly changed from the detection with atomic force microscopy (AFM). The plasma-treated effects were the improved surface wettability and cell adhesion. After a 90-s treatment, the water contact angle was at 20°, while the untreated surface was at 70°. The early cell adhesion of L929 mouse fibroblast was accelerated. L929 cells only took 3h to reach 100% cell adhesion on 90 s N2 plasma-treated surface, while there was less than 50% cell adhesion on the untreated Thai silk fibroin surface after 6h of culture. The cell adhesion results were in agreement with the cytoskeleton development. L929 F-actin was more evident on 90 s N2 plasma-treated surface than others. It could be concluded that a lower energy AC50Hz plasma system enhanced early L929 mouse fibroblast adhesion on Thai silk fibroin surface without any significant change in surface topography and bulk chemistry. PMID:23893032

  20. Preparation of Biodegradable Silk Fibroin/Alginate Blend Films for Controlled Release of Antimicrobial Drugs

    Yaowalak Srisuwan; Yodthong Baimark

    2013-01-01

    Silk fibroin (SF)/alginate blend films have been prepared for controlled release of tetracycline hydrochloride, an antimicrobial model drug. The blend films were analysed by Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), and UV-vis spectroscopy. The functional groups of the SF/alginate blends were monitored from their FTIR spectra. The homogeneity of the blend films was observed from SEM images. The dissolution and film transparency of the blend films depe...

  1. Effect of Methanol Treatment on Regenerated Silk Fibroin Microparticles Prepared by the Emulsification-Diffusion Technique

    Yodthong Baimark; Prasong Srihanam

    2009-01-01

    Silk Fibroin (SF) microparticles containing hollow structure were prepared by a water-in-oil emulsion solvent diffusion method without any surfactants. Aqueous SF solution and ethyl acetate were used as water and oil phases, respectively. Influences of SF concentration and post methanol treatment on microparticle characteristics were investigated. All microparticles contained open hollow structures. Microparticle sizes increased with the SF concentration. Conformation of SF microparticles det...

  2. Preparation and water absorption of cross-linked chitosan/silk fibroin blend films

    Suesat, Jantip; Rujiravanit, Ratana [Chulalongkorn University, The Petroleum and Petrochemical College, Bangkok (Thailand); Jamieson, Alexander M. [Case Western Reserve Univ., Department of Macromolecular Science, Cleveland (United States); Tokura, Seiichi [Kansai Univ., Faculty of Engineering, Osaka (Japan)

    2001-03-01

    Natural polymer blend films composed of chitosan and silk fibroin were prepared by varying the ratio of chitosan to silk fibroin, with and without glutaraldehyde as a crosslinking agent. The effects of the ratio of chitosan to silk fibroin and crosslinking agent on swelling behavior of the blend films were studied. For the swelling behavior, the blend films exhibited a dramatic change in the degree of swelling when immersed in acidic solutions. The degree of swelling of the films increased as the chitosan content increased; the blend film with 80% chitosan content had the maximum degree of swelling. It appeared that crosslinking had occurred in the blend films which helped the films to retain their three dimensional structure. In addition, FTIR spectra of the films showed evidence of hydrogen bonding interaction between chitosan and silk fibroin. For the effect of salt type, the films were immersed in various types of aqueous salt solutions, viz NaCl, LiCl, CaCl{sub 2}, AlCl{sub 3}, and FeCl{sub 3}. The films immersed in AlCl{sub 3} and FeCl{sub 3} aqueous solutions gave the maximum degree of swelling. The effects of AlCl{sub 3} and FeCl{sub 3} concentrations on swelling behavior were also investigated. It was found that the maximum degree of swelling of the films occurred at 1.0 x 10{sup -2} M of AlCl{sub 3} and FeCl{sub 3} aqueous solutions. (author)

  3. Effect of silk fibroin nanofibers containing silver sulfadiazine on wound healing

    Jeong L; Kim MH; Jung JY; Min BM; Park WH

    2014-01-01

    Lim Jeong,1 Min Hee Kim,1 Ju-Young Jung,2 Byung Moo Min,3 Won Ho Park1 1Department of Advanced Organic Materials and Textile System Engineering, 2College of Veterinary Medicine, Chungnam National University, Daejeon, 3Department of Oral Biochemistry, School of Dentistry, Seoul National University, Seoul, South Korea Background: One of the promising applications of silk fibroin (SF) in biomedical engineering is its use as a scaffolding material for skin regeneration. The purpose of this study...

  4. Facile fabrication of the porous three-dimensional regenerated silk fibroin scaffolds

    In the present work, we report a new facile method to fabricate porous three-dimensional regenerated silk fibroin (RSF) scaffolds through n-butanol- and freezing-induced conformation transition and phase separation. The effects of RSF concentration, freezing temperature and n-butanol addition on the microstructure, the secondary structures of silk fibroin and apparent mechanical properties of the RSF scaffolds were investigated by SEM, 13C CP-MAS NMR spectra and mechanical testing, respectively. By adjusting the RSF concentration and n-butanol addition, the pore size of the scaffold could be controlled in the range from of 10 μm to 350 μm with 84%–98% of porosity. The tensile strength of the wet scaffold reached the maximum of 755.2 ± 33.6 kPa when the concentration of RSF solution was increased to 15% w/w. Moreover, post-treatment with ethanol further induced conformation transition of RSF from random coil or helix to β-sheet. The porous scaffolds prepared by this facile and energy-saving method with good biocompatibility will have great potential for application in tissue engineering. Highlights: • A new facile and energy-saving method to fabricate porous silk fibroin scaffolds; • Freeze-drying step (a typical high energy consuming process) is unnecessary; • Morphology and mechanical properties of scaffolds were easily controlled; • Ethanol post-treatment can be used to tune the degradation behavior

  5. Facile fabrication of the porous three-dimensional regenerated silk fibroin scaffolds

    Cao, Zhengbing; Wen, Jianchuan [State Key Laboratory of Molecular Engineering of Polymers, Advanced Materials Laboratory, Department of Macromolecular Science, Fudan University, Shanghai 200433 (China); Yao, Jinrong, E-mail: yaoyaojr@fudan.edu.cn [State Key Laboratory of Molecular Engineering of Polymers, Advanced Materials Laboratory, Department of Macromolecular Science, Fudan University, Shanghai 200433 (China); Chen, Xin [State Key Laboratory of Molecular Engineering of Polymers, Advanced Materials Laboratory, Department of Macromolecular Science, Fudan University, Shanghai 200433 (China); Ni, Yusu [Otology and Skull Base Surgery Department, Eye and ENT Hospital of Fudan University, Shanghai 200031 (China); Shao, Zhengzhong [State Key Laboratory of Molecular Engineering of Polymers, Advanced Materials Laboratory, Department of Macromolecular Science, Fudan University, Shanghai 200433 (China)

    2013-08-01

    In the present work, we report a new facile method to fabricate porous three-dimensional regenerated silk fibroin (RSF) scaffolds through n-butanol- and freezing-induced conformation transition and phase separation. The effects of RSF concentration, freezing temperature and n-butanol addition on the microstructure, the secondary structures of silk fibroin and apparent mechanical properties of the RSF scaffolds were investigated by SEM, {sup 13}C CP-MAS NMR spectra and mechanical testing, respectively. By adjusting the RSF concentration and n-butanol addition, the pore size of the scaffold could be controlled in the range from of 10 μm to 350 μm with 84%–98% of porosity. The tensile strength of the wet scaffold reached the maximum of 755.2 ± 33.6 kPa when the concentration of RSF solution was increased to 15% w/w. Moreover, post-treatment with ethanol further induced conformation transition of RSF from random coil or helix to β-sheet. The porous scaffolds prepared by this facile and energy-saving method with good biocompatibility will have great potential for application in tissue engineering. Highlights: • A new facile and energy-saving method to fabricate porous silk fibroin scaffolds; • Freeze-drying step (a typical high energy consuming process) is unnecessary; • Morphology and mechanical properties of scaffolds were easily controlled; • Ethanol post-treatment can be used to tune the degradation behavior.

  6. Fabrication of Chitosan/Silk Fibroin Composite Nanofibers for Wound-dressing Applications

    Hong-sheng Wang

    2010-09-01

    Full Text Available Chitosan, a naturally occurring polysaccharide with abundant resources, has been extensively exploited for various biomedical applications, typically as wound dressings owing to its unique biocompatibility, good biodegradability and excellent antibacterial properties. In this work, composite nanofibrous membranes of chitosan (CS and silk fibroin(SF were successfully fabricated by electrospinning. The morphology of electrospun blend nanofibers was observed by scanning electron microscopy (SEM and the fiber diameters decreased with the increasing percentage of chitosan. Further, the mechanical test illustrated that the addition of silk fibroin enhanced the mechanical properties of CS/SF nanofibers. The antibacterial activities against Escherichia coli (Gram negative and Staphylococcus aureus (Gram positive were evaluated by the turbidity measurement method; and results suggest that the antibacterial effect of composite nanofibers varied on the type of bacteria. Furthermore, the biocompatibility of murine fibroblast on as-prepared nanofibrous membranes was investigated by hematoxylin and eosin (H&E staining and MTT assays in vitro, and the membranes were found to promote the cell attachment and proliferation. These results suggest that as-prepared chitosan/silk fibroin (CS/SF composite nanofibrous membranes could be a promising candidate for wound healing applications.

  7. Influence of Layer-by-Layer Polyelectrolyte Deposition and EDC/NHS Activated Heparin Immobilization onto Silk Fibroin Fabric

    M. Fazley Elahi

    2014-04-01

    Full Text Available To enhance the hemocompatibility of silk fibroin fabric as biomedical material, polyelectrolytes architectures have been assembled through the layer-by-layer (LbL technique on silk fibroin fabric (SFF. In particular, 1.5 and 2.5 bilayer of oppositely charged polyelectrolytes were assembled onto SFF using poly(allylamine hydrochloride (PAH as polycationic polymer and poly(acrylic acid (PAA as polyanionic polymer with PAH topmost. Low molecular weight heparin (LMWH activated with 1-ethyl-3-(dimethylaminopropyl carbodiimide hydrochloride (EDC and N-hydroxysuccinimide (NHS was then immobilized on its surface. Alcian Blue staining, toluidine blue assay and X-ray photoelectron spectroscopy (XPS confirmed the presence of heparin on modified SFF surfaces. The surface morphology of the modified silk fibroin fabric surfaces was characterized by scanning electron microscopy (SEM and atomic force microscopy (AFM, and obtained increased roughness. Negligible hemolytic effect and a higher concentration of free hemoglobin by a kinetic clotting time test ensured the improved biological performance of the modified fibroin fabric. Overall, the deposition of 2.5 bilayer was found effective in terms of biological and surface properties of the modified fibroin fabric compared to 1.5 bilayer self-assembly technique. Therefore, this novel approach to surface modification may demonstrate long term patency in future in vivo animal trials of small diameter silk fibroin vascular grafts.

  8. Mechanical property and biological performance of electrospun silk fibroin-polycaprolactone scaffolds with aligned fibers.

    Yuan, Han; Shi, Hongfei; Qiu, Xushen; Chen, Yixin

    2016-02-01

    The mechanical strength, biocompatibility, and sterilizability of silk fibroin allow it to be a possible candidate as a natural bone regenerate material. To improve mechanical character and reinforce the cell movement induction, silk fibroin (SF)-polycaprolactone (PCL) alloy was fabricated by electrospinning techniques with a rotating collector to form aligned fibrous scaffolds and random-oriented scaffolds. The scanning electron microscope image of the scaffold and the mechanical properties of the scaffold were investigated by tensile mechanical tests, which were compared to random-oriented scaffolds. Furthermore, mesenchymal stem cells were planted on these scaffolds to investigate the biocompatibility, elongation, and cell movement in situ. Scanning electron microscopy shows that 91% fibers on the aligned fibroin scaffold were distributed between the dominant direction 10. With an ideal support for stem cell proliferation in vitro, the aligned fibrous scaffold induces cell elongation at a length of 236.4682?m and distribution along the dominant fiber direction with a cell alignment angle at 6.574.45. Compared with random-oriented scaffolds made by artificial materials, aligned SF-PCL scaffolds could provide a moderate mesenchymal stem cell engraftment interface and speed up early stage cell movement toward the bone defect. PMID:26588014

  9. Incorporation of Human Recombinant Tropoelastin into Silk Fibroin Membranes with the View to Repairing Bruch’s Membrane

    Audra M. A. Shadforth

    2015-09-01

    Full Text Available Bombyx mori silk fibroin membranes provide a potential delivery vehicle for both cells and extracellular matrix (ECM components into diseased or injured tissues. We have previously demonstrated the feasibility of growing retinal pigment epithelial cells (RPE on fibroin membranes with the view to repairing the retina of patients afflicted with age-related macular degeneration (AMD. The goal of the present study was to investigate the feasibility of incorporating the ECM component elastin, in the form of human recombinant tropoelastin, into these same membranes. Two basic strategies were explored: (1 membranes prepared from blended solutions of fibroin and tropoelastin; and (2 layered constructs prepared from sequentially cast solutions of fibroin, tropoelastin, and fibroin. Optimal conditions for RPE attachment were achieved using a tropoelastin-fibroin blend ratio of 10 to 90 parts by weight. Retention of tropoelastin within the blend and layered constructs was confirmed by immunolabelling and Fourier-transform infrared spectroscopy (FTIR. In the layered constructs, the bulk of tropoelastin was apparently absorbed into the initially cast fibroin layer. Blend membranes displayed higher elastic modulus, percentage elongation, and tensile strength (p < 0.01 when compared to the layered constructs. RPE cell response to fibroin membranes was not affected by the presence of tropoelastin. These findings support the potential use of fibroin membranes for the co-delivery of RPE cells and tropoelastin.

  10. Factors controlling the deposition of silk fibroin nanofibrils during layer-by-layer assembly.

    de Moraes, Mariana Agostini; Crouzier, Thomas; Rubner, Michael; Beppu, Marisa Masumi

    2015-01-12

    The layer-by-layer technique has been used as a powerful method to produce multilayer thin films with tunable properties. When natural polymers are employed, complicated phenomena such as self-aggregation and fibrilogenesis can occur, making it more difficult to obtain and characterize high-quality films. The weak acid and base character of such materials provides multilayer systems that may differ from those found with synthetic polymers due to strong self-organization effects. Specifically, LbL films prepared with chitosan and silk fibroin (SF) often involve the deposition of fibroin fibrils, which can influence the assembly process, surface properties, and overall film functionality. In this case, one has the intriguing possibility of realizing multilayer thin films with aligned nanofibers. In this article, we propose a strategy to control fibroin fibril formation by adjusting the assembly partner. Aligned fibroin fibrils were formed when chitosan was used as the counterpart, whereas no fibrils were observed when poly(allylamine hydrochloride) (PAH) was used. Charge density, which is higher in PAH, apparently stabilizes SF aggregates on the nanometer scale, thereby preventing their organization into fibrils. The drying step between the deposition of each layer was also crucial for film formation, as it stabilizes the SF molecules. Preliminary cell studies with optimized multilayers indicated that cell viability of NIH-3T3 fibroblasts remained between 90 and 100% after surface seeding, showing the potential application of the films in the biomedical field, as coatings and functional surfaces. PMID:25469860

  11. The outermost surface properties of silk fibroin films reflect ethanol-treatment conditions used in biomaterial preparation.

    Terada, Dohiko; Yokoyama, Yoshiyuki; Hattori, Shinya; Kobayashi, Hisatoshi; Tamada, Yasushi

    2016-01-01

    Silk fibroin has attracted interest as a biomaterial, given its many excellent properties. Cell attachment to silk substrates is usually weaker than to standard culture dishes, and cells cultured on silk films or hydrogels typically form spheroids and micro-aggregates. However, too little is known about the higher order structures and behavior of fibroin under different conditions to explain the features of silk fibroin as a culture substrate. For instance, different biomaterial surfaces, with distinct effects on cell culture, can be achieved by varying the conditions of crystallization by alcohol immersion. Here, we show that treatment of fibroin film with 90% ethanol has a harder surface than the <80% ethanol-treated fibroin, to which individual cells prefer to attach (and then expand on the surface), rather than to aggregate. We discuss the influence of alcohol concentration on the surface properties, based on surface analysis of the films. The surface analysis involved assessment of static and dynamic contact angles, zeta potential, changes in crystallinity and microscopic morphology of electrospun fibers, and texture changes of the outermost surface at a nanometer-scale captured by a scanning probe microscope. PMID:26478294

  12. Bioactive macro/micro porous silk fibroin/Nano-sized calcium phosphate scaffolds with potential for bone tissue engineering applications

    Yan, Leping; Correia, Joana Silva; Correia, C.; Caridade, S. G.; Fernandes, E M; Sousa, R A; Mano, J. F.; Oliveira, Joaquim M.; de Oliveira, A. L.; Reis, R.L.

    2013-01-01

    Aim: The development of novel silk/nano-sized calcium phosphate (silk/nano-CaP) scaffolds with highly dispersed CaP nanoparticles in the silk fibroin (SF) matrix for bone tissue engineering. Materials & methods: Nano-CaP was incorporated in a concentrated aqueous SF solution (16 wt.%) by using an in situ synthesis method. The silk/nano-CaP scaffolds were then prepared through a combination of salt-leaching/ lyophilization approaches. Results: The CaP particles presented good affin...

  13. Mechanically-reinforced electrospun composite silk fibroin nanofibers containing hydroxyapatite nanoparticles

    Kim, Hyunryung [School of Mechanical Engineering, Yonsei University, Seoul 120-749 (Korea, Republic of); Che, Lihua; Ha, Yoon [Department of Neurosurgery, College of Medicine, Yonsei University, Seoul 120-749 (Korea, Republic of); Ryu, WonHyoung, E-mail: whryu@yonsei.ac.kr [School of Mechanical Engineering, Yonsei University, Seoul 120-749 (Korea, Republic of)

    2014-07-01

    Electrospun silk fibroin (SF) scaffolds provide large surface area, high porosity, and interconnection for cell adhesion and proliferation and they may replace collagen for many tissue engineering applications. Despite such advantages, electrospun SF scaffolds are still limited as bone tissue replacement due to their low mechanical strengths. While enhancement of mechanical strengths by incorporating inorganic ceramics into polymers has been demonstrated, electrospinning of a mixture of SF and inorganic ceramics such as hydroxyapatite is challenging and less studied due to the aggregation of ceramic particles within SF. In this study, we aimed to enhance the mechanical properties of electrospun SF scaffolds by uniformly dispersing hydroxyapatite (HAp) nanoparticles within SF nanofibers. HAp nanoaprticles were modified by γ-glycidoxypropyltrimethoxysilane (GPTMS) for uniform dispersion and enhanced interfacial bonding between HAp and SF fibers. Optimal conditions for electrospinning of SF and GPTMS-modified HAp nanoparticles were identified to achieve beadless nanofibers without any aggregation of HAp nanoparticles. The MTT and SEM analysis of the osteoblasts-cultured scaffolds confirmed the biocompatibility of the composite scaffolds. The mechanical properties of the composite scaffolds were analyzed by tensile tests for the scaffolds with varying contents of HAp within SF fibers. The mechanical testing showed the peak strengths at the HAp content of 20 wt.%. The increase of HAp content up to 20 wt.% increased the mechanical properties of the composite scaffolds, while further increase above 20 wt.% disrupted the polymer chain networks within SF nanofibers and weakened the mechanical strengths. - Highlights: • Electrospun composite silk fibroin scaffolds were mechanically-reinforced. • GPTMS enhanced hydroxyapatite distribution in silk fibroin nanofibers. • Mechanical property of composite scaffolds increased up to 20% of hydroxyapatite. • Composite scaffolds showed cell viability comparable to collagen.

  14. Expression of the hGM-CSF in the silk glands of germline of gene-targeted silkworm.

    Li, Yanmei; Cao, Guangli; Chen, Huimei; Jia, Haifang; Xue, Renyu; Gong, Chengliang

    2010-01-15

    To express human granulocyte-macrophage colony-stimulating factor (hGM-CSF) gene in the silk glands of transformation silkworm (Bombyx mori) based on gene-targeting, two fragments from fibroin heavy chain gene (fib-H) of silkworm were cloned and sequenced. One fragment contains the 1st exon and its downstream 1st intron's partial sequence; and the other fragment contains the 1st intron's partial sequence and the 2nd exon's partial sequence. Then the two fragments, as homologous arm, were inserted into pSK to generate a gene-targeted vector, pSK-HL-A3GFP-FLP-GM-CSF-FLPA-HR in which a gfp gene driven by A3 promoter and an hGM-CSF gene under the control of fibroin light chain (fib-L) promoter were included. The vector was transferred into the silkworm eggs using sperm-mediated gene transfer. After being screened for green fluorescent, the transformation silkworm was obtained, whose genome was verified by PCR and dot hybridization to confirm whether the target genes had been integrated into the silkworm genome. Furthermore, in the posterior silk glands of the G4 generation transformation silkworms, a specific band with the molecular weight of 22kDa could be detected by Western blotting with an antibody against hGM-CSF, and the expression level of the hGM-CSF estimated by ELISA was approximately 1.26ng per gram fresh posterior silk gland. PMID:20026302

  15. The application with tetramethyl pyrazine for antithrombogenicity improvement on silk fibroin surface

    Chuanxiongqin (tetramethyl pyrazine, TMPZ) is an active ingredient of the Chinese herb and was used to improve the anticoagulant activity of silk fibroin (SF). The side methyl of TMPZ was oxidized, and then linked to polyacrylic acid (PAA) via an ester bond. The prepared conjugate was further mixed with SF solutions at different ratios to make blend films. The resulting products were characterized by FTIR, UV spectrometer and X-ray photoelectron spectroscopy (XPS). The in vitro antithrombogenicity were evaluated by the activated partial thromboplastin time (APTT) and the prothrombin time (PT). It was shown that blend films had longer coagulation time than the pure SF film

  16. The application with tetramethyl pyrazine for antithrombogenicity improvement on silk fibroin surface

    Lian, Xiaojie; Wang, Song; Xu, Guoliang; Lin, Nini; Li, Qian; Zhu, Hesun

    2008-11-01

    Chuanxiongqin (tetramethyl pyrazine, TMPZ) is an active ingredient of the Chinese herb and was used to improve the anticoagulant activity of silk fibroin (SF). The side methyl of TMPZ was oxidized, and then linked to polyacrylic acid (PAA) via an ester bond. The prepared conjugate was further mixed with SF solutions at different ratios to make blend films. The resulting products were characterized by FTIR, UV spectrometer and X-ray photoelectron spectroscopy (XPS). The in vitro antithrombogenicity were evaluated by the activated partial thromboplastin time (APTT) and the prothrombin time (PT). It was shown that blend films had longer coagulation time than the pure SF film.

  17. The application with tetramethyl pyrazine for antithrombogenicity improvement on silk fibroin surface

    Lian Xiaojie [School of Material Science and Engineering, Beijing Institute of Technology, Beijing 100081 (China); Wang Song [School of Material Science and Engineering, Beijing Institute of Technology, Beijing 100081 (China)], E-mail: wangsongbit@126.com; Xu Guoliang; Lin Nini; Li Qian; Zhu Hesun [School of Material Science and Engineering, Beijing Institute of Technology, Beijing 100081 (China)

    2008-11-15

    Chuanxiongqin (tetramethyl pyrazine, TMPZ) is an active ingredient of the Chinese herb and was used to improve the anticoagulant activity of silk fibroin (SF). The side methyl of TMPZ was oxidized, and then linked to polyacrylic acid (PAA) via an ester bond. The prepared conjugate was further mixed with SF solutions at different ratios to make blend films. The resulting products were characterized by FTIR, UV spectrometer and X-ray photoelectron spectroscopy (XPS). The in vitro antithrombogenicity were evaluated by the activated partial thromboplastin time (APTT) and the prothrombin time (PT). It was shown that blend films had longer coagulation time than the pure SF film.

  18. Current research on electrospinning of silk fibroin and its blends with natural and synthetic biodegradable polymers

    Zhang, Jian-Guang; Mo, Xiu-Mei

    2013-06-01

    Silk fibroin (SF) is a kind of natural polymers with a great potential in biomedical application. Due to its good biocompatibility, biodegradability and minimal inflammatory reaction, SF is an excellent candidate for generating tissue engineering scaffolds. Electrospinning is a simple and effective method to fabricate nanofibers, which has several amazing characteristics such as very large surface area to volume ratio, flexibility in surface functionalities, and superior mechanical performance. The electrospun nanofibers from SF and its blends have been used for varied tissue engineering. This paper will give a brief review about the structure, properties and applications of SF and blend nanofibers via electrospinning.

  19. Impact of silk fibroin-based scaffold structures on human osteoblast MG63 cell attachment and proliferation

    Varkey, Aneesia; Venugopal, Elakkiya; Sugumaran, Ponjanani; Janarthanan, Gopinathan; Pillai, Mamatha M; Rajendran, Selvakumar; Bhattacharyya, Amitava

    2015-01-01

    The present study was carried out to investigate the impact of various types of silk fibroin (SF) scaffolds on human osteoblast-like cell (MG63) attachment and proliferation. SF was isolated from Bombyx mori silk worm cocoons after degumming. Protein concentration in the degummed SF solution was estimated using Bradford method. Aqueous SF solution was used to fabricate three different types of scaffolds, viz, electrospun nanofiber mat, sponge, and porous film. The structures of the prepared s...

  20. High-affinity integration of hydroxyapatite nanoparticles with chemically modified silk fibroin

    Wang Li; Li Chunzhong [East China University of Science and Technology, Key Laboratory for Ultrafine Materials of Ministry of Education, School of Materials Science and Engineering (China)], E-mail: czli@ecust.edu.cn; Senna, Mamoru [Keio University, Department of Applied Chemistry, Faculty of Science and Technology (Japan)

    2007-10-15

    Hydroxyapatite (HA)-based nanocomposites were prepared by a co-precipitation method with silk fibroin (SF) serving as organic matrix. Silk fibroin was chemically modified with an alkali solution or an enzyme attempting to improve the interface between the mineral and the organic matrix. The influences of the alkali and enzyme pretreatments on microstructure and physicochemical properties of HA-SF composite were examined and compared. The results reveal that both the two kinds of pretreatments facilitate the formation of highly ordered three-dimensional porous network throughout the composites, increase the microhardness of the composite, and promote the preferential growth of HA crystallites along c-axis. Among all the as-prepared samples, the composite containing the enzyme pretreated SF shows desirable hierarchical microstructure with higher degree of organization and more uniform pore size distribution. Due to the enzyme pretreatment, HA crystallites undergo obvious changes in morphology from rod-like to whisker-like and in crystal growth towards more apparent epitaxy along c-axis. The alkali pretreatment induces the stronger chemical interactions between HA and SF and thus to strengthen the inorganic-organic interfacial adhesion. The newly developed HA-SF composites are expected to be attractive biomedical materials for bone repair and remodeling.

  1. Bio-inspired capillary dry spinning of regenerated silk fibroin aqueous solution

    Wei Wei; Zhang Yaopeng, E-mail: zyp@dhu.edu.cn; Zhao Yingmei; Luo Jie; Shao Huili; Hu Xuechao

    2011-10-10

    To biomimic the spinning process of silkworm or spider, a capillary spinning equipment was applied to spin regenerated silk fibroin (RSF) fibers from RSF aqueous solutions in air. This equipment exhibits a wide processing window for various RSF aqueous solutions. The effects of pH, metal ions, RSF concentration and spinning parameters on the spinnability of the spinning dope and the mechanical properties of the obtained fibers were investigated. As a result, spinning dopes with a pH from 5.2 to 6.9 have good spinnability, especially for the dope with a pH of 6.0 and a Ca{sup 2+} concentration of 0.3 M. The RSF concentration of this dope ranges from 44% to 48%. Under optimized conditions of our dry spinning experiments (L/D, 133; take-up speed, 30 mm/s), the obtained as-spun fiber has a breaking strength of 46 MPa, which can be improved up to 359 MPa after a preliminary post-drawing in 80 vol.% ethanol aqueous solution. Highlights: {yields} Regenerated silk fibroin fibers were prepared by using a dry spinning method. {yields} Dope compositions affect dope spinnability. {yields} Spinning parameters affect dope spinnability and fiber properties. {yields} The breaking stress of the post-treated fiber was up to 359 MPa.

  2. Novel eatable silk fibroin gels containing salbutamol sulphate for dysphagic and geriatric patients

    Dixit Anil Satyanarayana

    2012-01-01

    Full Text Available The purpose of this research work is to prepare novel eatable gel formulations with suitable rheological characteristics, which provide a means of administering salbutamol sulphate to dysphagic and geriatric patients. Gels prepared using a natural polymer silk fibroin of different concentrations was subjected for in vitro characterization. The effect of concentration of the solution on gelation time, viscosity, and drug release was studied. FTIR and DSC spectra reveal that the drug was found compatible with silk fibroin. TGA curves showed weight loss as the temperature increased. Formulations F3, F4, F6, and F9 had thin, nectar like, honey like, and spoon thick viscosity range respectively, which is considered suitable for dysphagia patients as given by National Dysphagia Diet Task Force. Formulations showed shear thinning pseudoplastic behavior. Based on the concentration and viscosity of the polymer, formulation F9 was found to sustain the release of drug up to 90 min (99.4 ± 0.5%, whereas F3 showed release within 5 min (99.2 ± 2.0%. Mechanism of drug release was found to be anomalous transport. All formulations were found stable after 6 months when kept at refrigerated temperature (4°C - 8°C and room temperature. It can be concluded that the salbutamol sulphate gels prepared are suitable as vehicles for dysphagic patients.

  3. High-affinity integration of hydroxyapatite nanoparticles with chemically modified silk fibroin

    Hydroxyapatite (HA)-based nanocomposites were prepared by a co-precipitation method with silk fibroin (SF) serving as organic matrix. Silk fibroin was chemically modified with an alkali solution or an enzyme attempting to improve the interface between the mineral and the organic matrix. The influences of the alkali and enzyme pretreatments on microstructure and physicochemical properties of HA-SF composite were examined and compared. The results reveal that both the two kinds of pretreatments facilitate the formation of highly ordered three-dimensional porous network throughout the composites, increase the microhardness of the composite, and promote the preferential growth of HA crystallites along c-axis. Among all the as-prepared samples, the composite containing the enzyme pretreated SF shows desirable hierarchical microstructure with higher degree of organization and more uniform pore size distribution. Due to the enzyme pretreatment, HA crystallites undergo obvious changes in morphology from rod-like to whisker-like and in crystal growth towards more apparent epitaxy along c-axis. The alkali pretreatment induces the stronger chemical interactions between HA and SF and thus to strengthen the inorganic-organic interfacial adhesion. The newly developed HA-SF composites are expected to be attractive biomedical materials for bone repair and remodeling

  4. Contribution to the ultrastructural study of silk-excretion cells and autoradiographic analysis of intracellular fibroin transport in Bombyx mori L

    It is much easier to study the mechanisms involved in the synthesis and exportation of extracellular proteins in the biological material chosen is highly differentiated. The silk-excretion gland of the silkworm is ideal in this respect because during the larva period, especially at the end of the 5th and last stage, the cells at the rear (excreting tube) synthesize and export massive quantities of a single protein: fibroin. These phenomena were explored by a cytological study carried out mainly by electron microscopy and autoradiography. The results obtained are given. They relate first of all to the morphological development of the secretion tube cells from the end of the 4th larva stage to the spinning of the cocoon, and contribute new information on the cell changes during the 4th slough and the end of the 5th age. They also concern intracellular fibroin transport which is proved to take place through the Golgi apparatus, and finally the possible role of the microtubules and microfilaments in fibroin transport and secretion. On this last point the results so far constitute only, a preliminary approach which justifie no final conclusions; they merely suggest that the microfilaments of the apical region are involved in the secretion process

  5. Surface modification and properties of Bombyx mori silk fibroin films by antimicrobial peptide

    The Bombyx mori silk fibroin films (SFFs) were modified by a Cecropin B (CB) antimicrobial peptide, (NH2)-NGIVKAGPAIAVLGEAAL-CONH2, using the carbodiimide chemistry method. In order to avoid the dissolution of films during the modification procedure, the SFFs were first treated with 60% (v/v) ethanol aqueous solution, resulting a structural transition from unstable silk I to silk II. The investigation of modification conditions showed that the surface-modified SFFs had the satisfied antimicrobial activity and durability when they were activated by EDC.HCl/NHS solution followed by a treatment in CB peptide/PBS buffer (pH 6.5 or 8) solution at ambient temperature for 2 h. Moreover, the surface-modified SFFs showed the smaller contact angle due to the hydrophilic antimicrobial peptides coupled on the film surface, which is essential for the cell adhesion and proliferation. AFM results indicated that the surface roughness of SFFs was considerably increased after the modification by the peptides. The elemental composition analysis results also suggested that the peptides were tightly coupled to the surface of SFFs. This approach may provide a new option to engineer the surface-modified implanted materials preventing the biomaterial-centered infection (BCI)

  6. [The surface modification of eguus asinus augment fibroblast adhesion and proliferation on silk fibroin materials].

    Lian, Xiaojie; Wang, Song; Zhu, Hesun; Gao, Zhen

    2010-08-01

    Eguus asinus is one of the rare Chinese drugs famous for promoting blood circulation. In this experiment, it was employed to modify the silk fibroin (SF) by physical blending. Mouse embryonic fibroblasts NIH-3T3 were seeded on pure and modified SF surfaces. The morphological changes of cell on SF surfaces were characterized by optical microscope images. The cell adhesion diameter, the attachment force (F) of a single cell and cell proliferation on SF surfaces were measured by micropipette aspiration system and MTT assay. The results indicated that the fibroblasts seeded on modified SF had larger cell adhesion area, stronger initial attachment force and higher cell proliferation than did the pure SF. In conclusion, eguus asinus improved the adhesion and proliferation of fibroblasts on SF. The research promoted the further use of Chinese drug eguus asinus in tissue engineering. PMID:20842849

  7. Bio-inspired mineralization of hydroxyapatite in 3D silk fibroin hydrogel for bone tissue engineering.

    Jin, Yashi; Kundu, Banani; Cai, Yurong; Kundu, Subhas C; Yao, Juming

    2015-10-01

    To fabricate hard tissue implants with bone-like structure using a biomimetic mineralization method is drawing much more attentions in bone tissue engineering. The present work focuses in designing 3D silk fibroin hydrogel to modulate the nucleation and growth of hydroxyapatite crystals via a simple ion diffusion method. The study indicates that Ca(2+) incorporation within the hydrogel provides the nucleation sites for hydroxyapatite crystals and subsequently regulates their oriented growth. The mineralization process is regulated in a Ca(2+) concentration- and minerlization time-dependent way. Further, the compressive strength of the mineralized hydrogels is directly proportional with the mineral content in hydrogel. The orchestrated organic/inorganic composite supports well the viability and proliferation of human osteoblast cells; improved cyto-compatibility with increased mineral content. Together, the present investigation reports a simple and biomimetic process to fabricate 3D bone-like biomaterial with desired efficacy to repair bone defects. PMID:26209967

  8. Microstructural, thermal and antibacterial properties of electron beam irradiated Bombyx mori silk fibroin films

    Asha, S.; Sanjeev, Ganesh, E-mail: ganeshsanjeev@rediffmail.com [Microtron Center, Department of Studies in Physics, Mangalore University, Mangalagangotri - 574199 (India); Sangappa [Department of Studies in Physics, Mangalore University, Mangalagangotri - 574199 (India); Naik, Prashantha; Chandra, K. Sharat [Department of Biosciences, Mangalore University, Mangalagangotri - 574199 (India)

    2014-04-24

    The Bombyx mori silk fibroin (SF) films were prepared by solution casting method and the effects of electron beam on structural, thermal and antibacterial responses of the prepared films were studied. The electron irradiation for different doses was carried out using 8 MeV Microtron facility at Mangalore University. The changes in microstructural parameters and thermal stability of the films were investigated using Wide Angle X-ray Scattering (WAXS) and thermogravimetric analysis (TGA) respectively. Both microstructuralline parameters (crystallite size and lattice strain (g in %)) and thermal stability of the irradiated films have increased with radiation dosage. Agar diffusion method demonstrated the antibacterial activity of SF film which was increased after irradiation on both Gram-positive and Gram-negative species.

  9. Microstructural, thermal and antibacterial properties of electron beam irradiated Bombyx mori silk fibroin films

    The Bombyx mori silk fibroin (SF) films were prepared by solution casting method and the effects of electron beam on structural, thermal and antibacterial responses of the prepared films were studied. The electron irradiation for different doses was carried out using 8 MeV Microtron facility at Mangalore University. The changes in microstructural parameters and thermal stability of the films were investigated using Wide Angle X-ray Scattering (WAXS) and thermogravimetric analysis (TGA) respectively. Both microstructuralline parameters (crystallite size and lattice strain (g in %)) and thermal stability of the irradiated films have increased with radiation dosage. Agar diffusion method demonstrated the antibacterial activity of SF film which was increased after irradiation on both Gram-positive and Gram-negative species

  10. Facile method to prepare silk fibroin/hyaluronic acid films for vascular endothelial growth factor release.

    Zhou, Juan; Zhang, Bin; Liu, Xunwei; Shi, Lijun; Zhu, Jun; Wei, Daixu; Zhong, Jian; Sun, Gang; He, Dannong

    2016-06-01

    A facile approach was proposed to prepare silk fibroin (SF) and hyaluronic acid (HA) composite films from aqueous solution without crosslinking or any post treatment. Only by controlling the HA content and film formation temperature during the film casting, the HA/SF films with different composition were prepared. The films were then characterized by structural characteristics, thermal stability, morphology, water stability, water absorption, mechanical properties. After immersing in water for 24h, all of the films showed good structural integrity. The degradation rate of the HA/SF films in protease XIV can be controlled by changing the film formation temperature and HA content. Decreasing the temperature and adding HA resulted in the rapid release of VEGF (vascular endothelial growth factor) from the HA/SF films. Overall, the 5% HA/SF films formed at 37°C with more rapid VEGF release exhibited great potential in drug delivery, especially when the rapid vascularization was needed. PMID:27083373

  11. Effect of Methanol Treatment on Regenerated Silk Fibroin Microparticles Prepared by the Emulsification-Diffusion Technique

    Yodthong Baimark

    2009-01-01

    Full Text Available Silk Fibroin (SF microparticles containing hollow structure were prepared by a water-in-oil emulsion solvent diffusion method without any surfactants. Aqueous SF solution and ethyl acetate were used as water and oil phases, respectively. Influences of SF concentration and post methanol treatment on microparticle characteristics were investigated. All microparticles contained open hollow structures. Microparticle sizes increased with the SF concentration. Conformation of SF microparticles determined from FTIR spectra changed from predominantly random coil to β-sheet form after methanol treatment. Particle surfaces of methanol-treated SF microparticles were rougher than those of non-treated. The SF concentrations and methanol treatment did not affect open hollow structure of the SF microparticles. Major advantages of this method are the elimination of surfactants and extractable pore templates. These hollow SF microparticles are expected to be potential used as biodegradable microcarriers of cell and protein drug, because their hollow structures should permit cell attachment and enzyme immobilization.

  12. Preparation of wound dressing of polyvinyl alcohol/silk fibroin hydrogels by gamma radiation

    Poly vinylalcohol/silk fibroin (PVA/SF) hydrogels were prepared by γ-radiation. The preparation conditions such as absorbed doses and PVA/SF concentrations were investigated. When exposed to γ -radiation, PVA/SF was crosslinked to yield high water absorption materials with water content of 100 - 1000% of their dried weight depending on the preparation conditions. The crosslinked density seems to be the main factor governing the swelling of these gels. The swelling behaviors in NaCl aqueous solutions were also investigated. The swelling of PVA/SF hydrogels decreases when exposed to electrolyte solution. With an increase of absorbed dose, the gel fraction of PVA/SF increases

  13. Silk fibroin/gelatin blend films crosslinked with enzymes for biomedical applications.

    Taddei, Paola; Chiono, Valeria; Anghileri, Anna; Vozzi, Giovanni; Freddi, Giuliano; Ciardelli, Gianluca

    2013-11-01

    Microbial transglutaminase (mTG) and mushroom tyrosinase (MT) are used to crosslink B. mori silk fibroin/gelatin (SF/G) films. Crosslinked and uncrosslinked SF/G films show no phase separation. The thermal behavior and the conformational structure of SF/G films are strongly affected by blending and enzymatic treatment. Formation of high thermally stable crosslinked macromolecular species is observed, suggesting the occurrence of strong intermolecular interactions between the two polymers as confirmed by FT-Raman spectroscopy. Preliminary in vitro tests show that MT-crosslinked blends with G amounts ?40% and mTG-crosslinked SF/G 60/40 films support C2C12 cardiomyocyte adhesion and proliferation. PMID:23956190

  14. Patterning and photoluminescence of CdS nanocrystallites on silk fibroin fiber

    CdS nanocrystallites could be formed and assembled into nanoparticle strings and hexagons on natural silk fibroin fiber (SFF) through a room-temperature bio-inspired process. Herein, the biomaterial SFF served as reactive substrate, not only provides the in situ formation sites for CdS nanocrystallites, but also directs the arrangement of nanocrystalline CdS simultaneously. The photoluminescence (PL) of the resulting nanocomposites CdS/SFF is investigated extensively. The PL peaks observed from CdS nanoparticle strings are similar to those of separate CdS nanoparticles, corresponding to the band-edge emission of their individual building blocks (QD-CdS). Moreover, CdS nanoparticle hexagons perform a red-shifted and broadened emission peak.

  15. The application with protocatechualdehyde to improve anticoagulant activity and cell affinity of silk fibroin

    Wang Song [School of Material Science and Engineering, Beijing Institute of Technology, Beijing 100081 (China)], E-mail: wangsongbit@126.com; Gao Zhen; Li Erlin; Su Caoning; Zhu Hesun [School of Material Science and Engineering, Beijing Institute of Technology, Beijing 100081 (China)

    2008-11-15

    Protocatechualdehyde (PCA) is one of the effective ingredients extracted from Danshen (Radix Salviae Miltiorrhizae) and was employed to modify the silk fibroin (SF) by graft polymerization and surface adsorption. The surface composition of modified SF was characterized by attenuated total reflectance Fourier-transform infrared (ATR-FTIR), X-ray photoelectron spectroscopy (XPS), and UV spectrophotometer. The anticoagulant activity of modified SF was assessed by in vitro coagulation test and platelet adhesion measurement. The endothelial cell affinity was evaluated by a parallel plate flow chamber. The test results indicated that with the introduction of PCA into SF, the anticoagulant activity has been improved obviously. And the SF surface composition altered by PCA, but did not disturb its {beta}-sheet conformation. Moreover, the adsorbed PCA on SF surface can enhance the endothelial cell affinity.

  16. Preparation and characterization of a bacterial cellulose/silk fibroin sponge scaffold for tissue regeneration.

    Oliveira Barud, H G; Barud, Hernane da S; Cavicchioli, Maurício; do Amaral, Thais Silva; de Oliveira Junior, Osmir Batista; Santos, Diego M; Petersen, Antonio Luis de Oliveira Almeida; Celes, Fabiana; Borges, Valéria Matos; de Oliveira, Camila I; de Oliveira, Pollyanna Francielli; Furtado, Ricardo Andrade; Tavares, Denise Crispim; Ribeiro, Sidney J L

    2015-09-01

    Bacterial cellulose (BC) and silk fibroin (SF) are natural biopolymers successfully applied in tissue engineering and biomedical fields. In this work nanocomposites based on BC and SF were prepared and characterized by scanning electron microscopy (SEM), infrared spectroscopy (FT-IR), X-ray diffraction (XRD) and thermogravimetric analysis (TGA). In addition, the investigation of cytocompatibility was done by MTT, XTT and Trypan Blue dye technique. Cellular adhesion and proliferation were detected additionally. The evaluation of genotoxicity was realized by micronucleus assay. In vitro tests showed that the material is non-cytotoxic or genotoxic. SEM images revealed a greater number of cells attached at the BC/SF:50% scaffold surface than the pure BC one, suggesting that the presence of fibroin improved cell attachment. This could be related to the SF amino acid sequence that acts as cell receptors facilitating cell adhesion and growth. Consequently, BC/SF:50% scaffolds configured an excellent option in bioengineering depicting its potential for tissue regeneration and cultivation of cells on nanocomposites. PMID:26005138

  17. Effect of silk fibroin nanofibers containing silver sulfadiazine on wound healing

    Jeong L

    2014-11-01

    Full Text Available Lim Jeong,1 Min Hee Kim,1 Ju-Young Jung,2 Byung Moo Min,3 Won Ho Park1 1Department of Advanced Organic Materials and Textile System Engineering, 2College of Veterinary Medicine, Chungnam National University, Daejeon, 3Department of Oral Biochemistry, School of Dentistry, Seoul National University, Seoul, South Korea Background: One of the promising applications of silk fibroin (SF in biomedical engineering is its use as a scaffolding material for skin regeneration. The purpose of this study was to determine the wound healing effect of SF nanofibrous matrices containing silver sulfadiazine (SSD wound dressings.Methods: An SF nanofibrous matrix containing SSD was prepared by electrospinning. The cell attachment and spreading of normal human epidermal keratinocytes (NHEK and normal human epidermal fibroblasts (NHEF to SF nanofibers containing three different concentrations of SSD contents (0.1, 0.5, and 1.0 wt% were determined. In addition, a rat wound model was used in this study to determine the wound healing effect of SF nanofibers containing SSD compared with that of Acticoat, a commercially available wound dressing.Results: The number of NHEK and NHEF attached to SF nanofibers containing SSD decreased when the concentration of SSD increased. The number of attached NHEF cells was lower than that of attached NHEK cells. The SF matrix with 1.0 wt% SSD produced faster wound healing than Acticoat, although 1.0 wt% SSD inhibited the attachment of epidermal cells to SF nanofibers in vitro.Conclusion: The cytotoxic effects of SF nanofibers with SSD should be considered in the development of silver-release dressings for wound healing through its antimicrobial activity. It is challenging to design wound dressings that maximize antimicrobial activity and minimize cellular toxicity. Keywords: silk fibroin, silver sulfadiazine, nanofibrous matrix, wound healing

  18. Fabrication and characterization of silk fibroin-derived curcumin nanoparticles for cancer therapy

    Vishal Gupta

    2009-05-01

    Full Text Available Vishal Gupta1, Abraham Aseh1,3, Carmen N Ríos1, Bharat B Aggarwal2, Anshu B Mathur11Department of Plastic Surgery; 2Department of Experimental Therapeutics, The University of Texas M.D. Anderson Cancer Center, Houston, TX, USA; 3School of Pharmacy, Texas Southern University, Houston, TX, USAAbstract: Biologically derived nanoparticles (<100 nm were fabricated for local and sustained therapeutic curcumin delivery to cancer cells. Silk fibroin (SF and chitosan (CS polymers were blended noncovalently to encapsulate curcumin in various proportions of SF and CS (75:25, 50:50, and 25:75 SF:CS or pure SF at two concentrations (0.1% w/v and 10% w/v using the devised capillary-microdot technique. Curcumin-polymer conjugates were frozen, lyophilized, crystallized, suspended in phosphate-buffered saline for characterization, and tested for efficacy against breast cancer cells. All nanoparticle formulations except 0.1% w/v 50:50 SFCS were less than 100 nm in size as determined with the transmission electron microscopy. The entrapment and release of curcumin over eight days was highest for SF-derived nanoparticles as compared to all SFCS blends. The uptake and efficacy of SF-coated curcumin was significantly higher (p < 0.001 than SFCS-coated curcumin in both low and high Her2/neu expressing breast cancer cells. Interestingly, the uptake of curcumin was highest for the high Her2/neu expressing breast cancer cells when delivered with a 10% w/v SF coating as compared to other formulations. In conclusion, SF-derived curcumin nanoparticles show higher efficacy against breast cancer cells and have the potential to treat in vivo breast tumors by local, sustained, and long-term therapeutic delivery as a biodegradable system.Keywords: biodegradable, nanoparticles, curcumin, silk fibroin, breast cancer cells

  19. Study of the effects of different sterilization methods on the properties of dense and porous silk fibroin membranes

    Silk fibroin has been widely explored for many biomedical applications, due to its biocompatibility and biodegradability. Sterilization is a fundamental step in biomaterials processing, and it must not alter in a negative way the functionality of medical devices. The aim of this study was to analyze the influence of different sterilization methods in the physical and chemical characteristics of dense silk fibroin membranes. Dense fibroin membranes were sterilized by ultraviolet radiation, 70% ethanol, autoclave, ethylene oxide and gamma radiation, and were analyzed by SEM, FTIR-ATR and XRD. The results for sterilization indicated that the methods didn't cause degradation of the membranes, but the methods that used organic solvent, or increase of humidity and/or temperature (70% ethanol, autoclave and ethylene oxide) altered the molecular conformation of fibroin, increasing the proportion of β-sheet structure, what indicates an increase of crystallinity. This effect may be positive when a slower degradation of the membranes is desired, depending on the application as a bio material. (author)

  20. Impact of silk fibroin-based scaffold structures on human osteoblast MG63 cell attachment andproliferation

    Varkey A; Venugopal E; Sugumaran P; Janarthanan G; Pillai MM; Rajendran S; Bhattacharyya A,

    2015-01-01

    Aneesia Varkey,1,2 Elakkiya Venugopal,2 Ponjanani Sugumaran,2 Gopinathan Janarthanan,1 Mamatha M Pillai,2 Selvakumar Rajendran,2 Amitava Bhattacharyya1 1Advanced Textile and Polymer Research Laboratory, 2Tissue Engineering Laboratory, PSG Institute of Advanced Studies, Coimbatore, Tamil Nadu, India Abstract: The present study was carried out to investigate the impact of various types of silk fibroin (SF) scaffolds on human osteoblast-like cell (MG63) attachment and proliferation. SF was iso...

  1. Effects of silk fibroin fiber incorporation on mechanical properties, endothelial cell colonization and vascularization of PDLLA scaffolds

    Stoppato, M.; Stevens, H. Y.; E. Carletti; Migliaresi, C.; Motta, A; Guldberg, R. E.

    2013-01-01

    Attainment of functional vascularization of engineered constructs is one of the fundamental challenges of tissue engineering. However, the development of an extracellular matrix in most tissues, including bone, is dependent upon the establishment of a well developed vascular supply. In this study a poly-D,L-lactic acid (PDLLA) salt-leached sponge was modified by incorporation of silk fibroin fibers to create a multicomponent scaffold, in an effort to better support endothelial cell colonizati...

  2. Design and Characterization of a Silk-Fibroin-Based Drug Delivery Platform Using Naproxen as a Model Drug

    Tatyana Dyakonov; Chue Hue Yang; Derek Bush; Saujanya Gosangari; Shingai Majuru; Aqeel Fatmi

    2012-01-01

    The objective of this proof-of-concept study was to develop a platform for controlled drug delivery based on silk fibroin (SF) and to explore the feasibility of using SF in oral drug delivery. The SF-containing matrixes were prepared via spray-drying and film casting, and the release profile of the model drug naproxen sodium was evaluated. Attenuated total reflectance Fourier transform infrared spectroscopy (FTIR) has been used to observe conformational changes in SF- and drug-containing comp...

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

    Fan, Linpeng; Cai, Zengxiao; Zhang, Kuihua; Han, Feng; Li, Jingliang; He, Chuanglong; Mo, Xiumei; Wang, Xungai; Wang, Hongsheng

    2014-05-01

    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 ?-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. PMID:24632028

  4. Human Dental Pulp Stem Cells and Gingival Fibroblasts Seeded into Silk Fibroin Scaffolds Have the Same Ability in Attracting Vessels

    Woloszyk, Anna; Buschmann, Johanna; Waschkies, Conny; Stadlinger, Bernd; Mitsiadis, Thimios A.

    2016-01-01

    Neovascularization is one of the most important processes during tissue repair and regeneration. Current healing approaches based on the use of biomaterials combined with stem cells in critical-size bone defects fail due to the insufficient implant vascularization and integration into the host tissues. Therefore, here we studied the attraction, ingrowth, and distribution of blood vessels from the chicken embryo chorioallantoic membrane into implanted silk fibroin scaffolds seeded with either human dental pulp stem cells or human gingival fibroblasts. Perfusion capacity was evaluated by non-invasive in vivo Magnetic Resonance Imaging while the number and density of blood vessels were measured by histomorphometry. Our results demonstrate that human dental pulp stem cells and gingival fibroblasts possess equal abilities in attracting vessels within silk fibroin scaffolds. Additionally, the prolonged in vitro pre-incubation period of these two cell populations favors the homogeneous distribution of vessels within silk fibroin scaffolds, which further improves implant survival and guarantees successful healing and regeneration. PMID:27148078

  5. Attachment and growth of human bone marrow derived mesenchymal stem cells on regenerated antheraea pernyi silk fibroin films

    Luan Xiying [Institute of Medical Biotechnology, Jiangsu Province Key Laboratory of Stem Cell, Suzhou University, Suzhou 215007 (China); Wang Yong [Institute of Medical Biotechnology, Jiangsu Province Key Laboratory of Stem Cell, Suzhou University, Suzhou 215007 (China); Duan Xiang [Institute of Medical Biotechnology, Jiangsu Province Key Laboratory of Stem Cell, Suzhou University, Suzhou 215007 (China); Duan Qiaoyan [Institute of Medical Biotechnology, Jiangsu Province Key Laboratory of Stem Cell, Suzhou University, Suzhou 215007 (China); Li Mingzhong [School of Materials Engineering, Suzhou University, Suzhou 215006 (China); Lu Shenzhou [School of Materials Engineering, Suzhou University, Suzhou 215006 (China); Zhang Huanxiang [Institute of Medical Biotechnology, Jiangsu Province Key Laboratory of Stem Cell, Suzhou University, Suzhou 215007 (China); Zhang Xueguang [Institute of Medical Biotechnology, Jiangsu Province Key Laboratory of Stem Cell, Suzhou University, Suzhou 215007 (China)

    2006-12-15

    Silk fibroin of the silkworm Bombyx mori has been studied extensively, while the research on Antheraea pernyi silk fibroin (A. pernyi SF) in biomaterials is only at an early stage. In this study, the attachment, morphology, growth and phenotype of human bone marrow derived mesenchymal stem cells (hBMSCs) cultured on the regenerated A. pernyi SF films were studied in vitro. The results indicated that the attachment of hBMSCs on the regenerated A. pernyi SF films was almost the same as that on the collagen films. MTT and cell counting analyses demonstrated that the growth of hBMSCs on the regenerated A. pernyi SF films was better than that on controls. Moreover, electron scanning microscopy and fluorescence-activated cell sorting assays showed that the regenerated A. pernyi SF supported hBMSCs growth and functional maintenance compared with the controls. These data suggest that the regenerated A. pernyi SF, like Bombyx mori silk fibroin (B. mori SF) and collagen, can support hBMSCs attachment, growth and phenotypic maintenance, and has better biocompatibilities for hBMSCs in vitro culture.

  6. Spermine-modified Antheraea pernyi silk fibroin as a gene delivery carrier

    Yu Y

    2016-03-01

    Full Text Available Yanni Yu,1 Yongpei Hu,1 Xiufang Li,1 Yu Liu,1 Mingzhong Li,1 Jicheng Yang,2 Weihua Sheng2 1National Engineering Laboratory for Modern Silk, College of Textile and Clothing Engineering, Soochow University, 2Cell and Molecular Biology Institute, College of Medicine, Soochow University, Suzhou, People’s Republic of China Abstract: The development of a novel cationized polymer used as a gene delivery carrier that can conveniently and effectively transfect cells resulting in a stably expressed target gene remains a challenge. Antheraea pernyi silk fibroin (ASF is a cytocompatible and biodegradable natural polymer, and it possesses Arg–Gly–Asp sequences but a negative charge. In order to render ASF amenable to packaging plasmid DNA (pDNA, spermine was used to modify ASF to synthesize cationized ASF (CASF, which was used as a gene delivery carrier. CASF was characterized using trinitrobenzene sulfonic acid assay, the zeta potential determination, and a Fourier transform infrared analysis, and the results of these characterizations indicated that the –NH2 in spermine effectively reacts with the –COOH in the side chains of ASF. Spermine grafted to the side chains of ASF resulted in the conversion of the negative charge of ASF to a positive charge. CASF packaged pDNA and formed CASF/pDNA complexes, which exhibited spherical morphology with average particle sizes of 215–281 nm and zeta potential of approximately +3.0 mV to +3.2 mV. The results of the MTT assay, confocal laser scanning microscopy, and flow cytometry analysis in a human endothelial cell line revealed that CASF/pDNA complexes exhibited lower cytotoxicity and higher transfection efficiency compared to the pDNA complexes of polyethyleneimine. These results indicate that our synthesized CASF, a cationized polymer, is a potential gene delivery carrier with the advantages of biodegradability and low cytotoxicity. Keywords: silk fibroin, spermine, cationized polymer, gene delivery carrier

  7. Penicillin impregnation on oxygen plasma surface functionalized chitosan/Antheraea assama silk fibroin: Studies of antibacterial activity and antithrombogenic property.

    Choudhury, Arup Jyoti; Gogoi, Dolly; Kandimalla, Raghuram; Kalita, Sanjeeb; Chaudhari, Yogesh B; Khan, Mojibur R; Kotoky, Jibon; Chutia, Joyanti

    2016-03-01

    Low temperature plasma can effectively tailor the surface properties of natural polymeric biomaterials according to the need for various biomedical applications. Non-mulberry silk, Antheraea assama silk fibroin (AASF) is a natural polymer having excellent biocompatibility and mechanical strength yet unlike mulberry silk, Bombyx mori silk fibroin, has drawn less interest in biomedical research. In the quest for developing as potential biomaterial, surface functionalization of plasma induced chitosan (Cs) grafted AASF ((AASF/O2-CS)g/O2) yarn is carried out using oxygen (O2) plasma. The (AASF/O2-CS)g/O2 yarn exhibits enhanced antithrombogenic property as well as antimicrobial activity against Gram positive (Bacillus subtilis) and Gram negative (Escherichia coli) bacteria as compared to AASF yarn. Moreover, impregnation of antibiotic drug (penicillin G sodium salt, PEN) on (AASF/O2-CS)g/O2 yarn further improves the observed properties. In-vitro hemolysis assay reveals that O2 plasma treatment and subsequent impregnation of PEN do not affect the hemocompatibility of AASF yarn. The present research findings demonstrate that plasma induced grafting of Cs followed by penicillin impregnation could significantly improve the potential applicability of AASF in the field of surgical research. PMID:26706554

  8. Comparison of the in vitro and in vivo degradations of silk fibroin scaffolds from mulberry and nonmulberry silkworms

    Degradation behavior is very important in the field of silk-based biomaterials. Mulberry and nonmulberry silk fibroins are structurally and functionally distinguishable; however, no studies have examined the differences in the degradation behaviors of silk materials from various silkworm species. In this study, Ca(NO3)2 was used as a uniform solvent to obtain regenerated mulberry and nonmulberry (Antheraea pernyi and Antheraea yamamai) silk fibroin (SF) solutions, and the degradation behaviors of various SF scaffolds were examined. In vitro and in vivo results demonstrated that regenerated mulberry SF scaffolds exhibited significantly higher mass loss and free amino acid content release than did nonmulberry SF scaffolds. The differences in the primary structures and condensed structures between mulberry and nonmulberry SF contributed to the significant difference in degradation rates, in which the characteristic (–Ala–)n repeats, compact crystal structure and high α-helix and β-sheet contents make nonmulberry SF more resistant than mulberry SF to enzymatic degradation. Moreover, the Antheraea pernyi and Antheraea yamamai SFs possess similar primary structures and condensed structures, although a slight difference in degradation was observed; this difference might depend on the differences in molecular weight following the regeneration process. The results indicate that the original sources of SF significantly influence the degradation rates of SF-based materials; therefore, the original sources of SF should be fully considered for preparing tissue engineering scaffolds with matched degradation rates. (paper)

  9. Electrospun composites of PHBV, silk fibroin and nano-hydroxyapatite for bone tissue engineering

    Pa?cu, Elena I.; Stokes, Joseph; McGuinness, Garrett B., E-mail: garrett.mcguinness@dcu.ie

    2013-12-01

    Electrospinning of fibrous scaffolds containing nano-hydroxyapatite (nHAp) embedded in a matrix of functional biomacromolecules offers an attractive route to mimicking the natural bone tissue architecture. Functional fibrous substrates will support cell attachment, proliferation and differentiation, while the role of HAp is to induce cells to secrete extracellular matrix (ECM) for mineralization to form bone. Electrospinning of biomaterials composed of polyhydroxybutyrate-co-(3-hydroxyvalerate) with 2% valerate fraction (PHBV), nano-hydroxyapatite (nHAp), and Bombyx mori silk fibroin essence (SF), Mw = 90KDa, has been achieved for nHAp and SF solution concentrations of 2 (w/vol) % each and 5 (w/vol) % each. The structure and properties of the nanocomposite fibrous membranes were investigated by means of Scanning Electron Microscopy in combination with Energy Dispersive X-Ray Analysis (SEM/EDX), Fourier Transformed Infrared Spectroscopy (FT-IR), uniaxial tensile and compressive mechanical testing, degradation tests and in vitro bioactivity tests. SEM images showed smooth, uniform and continuous fibre deposition with no bead formation, and fibre diameters of between 10 and 15 ?m. EDX and FT-IR confirmed the presence of nHAp and SF. After one month in deionised water, tests showed less than 2% weight loss with the samples retaining their fibrous morphology, confirming that this material biodegrades slowly. After 28 days of immersion in Simulated Body Fluid (SBF) an apatite layer was visible on the surface of the fibres, proving their bioactivity. Preliminary in vitro biological assessment showed that after 1 and 3 days in culture, cells were attached to the fibres, retaining their morphology while presenting a flattened appearance and elongated shape on the surface of fibres. Young's modulus was found to increase from 0.7 kPa ( 0.33 kPa) for electrospun samples of PHBV only to 1.4 kPa ( 0.54 kPa) for samples with 2 (w/vol) % each of nHAp and SF. Samples prepared with 5 (w/vol) % each of nHAp and SF did not show a similar improvement. - Highlights: Electrospinning of PHBV/nHAp composites was first achieved using silk fibroin (SF). Smooth, uniform, continuous fibres with diameters between 10 and 15 ?m were deposited. Bioactivity was confirmed by surface apatite formation in Simulated Body Fluid. Tensile modulus was increased for 2% nHAp and SF relative to 0% and 5% cases. PHBV/nHAp/SF composites support initial osteoblast attachment and proliferation.

  10. Electrospun composites of PHBV, silk fibroin and nano-hydroxyapatite for bone tissue engineering

    Electrospinning of fibrous scaffolds containing nano-hydroxyapatite (nHAp) embedded in a matrix of functional biomacromolecules offers an attractive route to mimicking the natural bone tissue architecture. Functional fibrous substrates will support cell attachment, proliferation and differentiation, while the role of HAp is to induce cells to secrete extracellular matrix (ECM) for mineralization to form bone. Electrospinning of biomaterials composed of polyhydroxybutyrate-co-(3-hydroxyvalerate) with 2% valerate fraction (PHBV), nano-hydroxyapatite (nHAp), and Bombyx mori silk fibroin essence (SF), Mw = 90KDa, has been achieved for nHAp and SF solution concentrations of 2 (w/vol) % each and 5 (w/vol) % each. The structure and properties of the nanocomposite fibrous membranes were investigated by means of Scanning Electron Microscopy in combination with Energy Dispersive X-Ray Analysis (SEM/EDX), Fourier Transformed Infrared Spectroscopy (FT-IR), uniaxial tensile and compressive mechanical testing, degradation tests and in vitro bioactivity tests. SEM images showed smooth, uniform and continuous fibre deposition with no bead formation, and fibre diameters of between 10 and 15 μm. EDX and FT-IR confirmed the presence of nHAp and SF. After one month in deionised water, tests showed less than 2% weight loss with the samples retaining their fibrous morphology, confirming that this material biodegrades slowly. After 28 days of immersion in Simulated Body Fluid (SBF) an apatite layer was visible on the surface of the fibres, proving their bioactivity. Preliminary in vitro biological assessment showed that after 1 and 3 days in culture, cells were attached to the fibres, retaining their morphology while presenting a flattened appearance and elongated shape on the surface of fibres. Young's modulus was found to increase from 0.7 kPa (± 0.33 kPa) for electrospun samples of PHBV only to 1.4 kPa (± 0.54 kPa) for samples with 2 (w/vol) % each of nHAp and SF. Samples prepared with 5 (w/vol) % each of nHAp and SF did not show a similar improvement. - Highlights: • Electrospinning of PHBV/nHAp composites was first achieved using silk fibroin (SF). • Smooth, uniform, continuous fibres with diameters between 10 and 15 μm were deposited. • Bioactivity was confirmed by surface apatite formation in Simulated Body Fluid. • Tensile modulus was increased for 2% nHAp and SF relative to 0% and 5% cases. • PHBV/nHAp/SF composites support initial osteoblast attachment and proliferation

  11. Gelatin modified ultrathin silk fibroin films for enhanced proliferation of cells.

    Yang, Luyuan; Yaseen, Mohammed; Zhao, Xiubo; Coffey, Paul; Pan, Fang; Wang, Yuming; Xu, Hai; Webster, John; Lu, Jian R

    2015-04-01

    Silk fibroin (SF) films were modified with gelatin (G) to explore if such SF/G films could enhance the surface biocompatibility of silk as cell growth biomaterials. Ultrathin films were coated from aqueous SF solutions pre-mixed with different amounts of G. It was found that the SF/G blended films after methanol treatment were highly stable in physiological conditions. The incorporation of G smoothed the surface morphology of the SF/G films formed. Surface-exposed RGD sequences were successfully identified on the SF/G films through specific recognition of an integrin-mimicking peptide (bearing the sequence of CWDDGWLC). Cell culture experiments with 3T3 fibroblasts demonstrated that SF/G films with 1.2-20% (w/w) G gave clear improvement in promoting cell attachment and proliferation over pure SF films. Films containing 10-20% (w/w) of G showed cell attachment and growth even superior to the pure G films. The differences as observed from this study suggest that due to the lack of mechanical strength associated with its high solubility, G could not work alone as a cell growth scaffold. The enhanced cellular responses from the blended SF/G films must result from improvement in film stability arising from SF and in cytocompatibility arising from G. The results thus indicate the potential of the SF/G blends in tissue engineering and biomedical engineering where physical and biological properties could be manipulated via mixing either as bulk biomaterials or for coating purposes. PMID:25784671

  12. Preparation and characterization of blends containing silk fibroin and chitosan;Obtencao e caracterizacao de blendas de fibroina de seda e quitosana

    Moraes, Mariana A. de; Nogueira, Grinia M.; Weska, Raquel F.; Beppu, Marisa M., E-mail: beppu@feq.unicamp.b [Universidade Estadual de Campinas (FEQ/UNICAMP), SP (Brazil). Faculdade de Engenharia Quimica

    2009-07-01

    The aim of this study was to prepare and characterize blend membranes of silk fibroin and chitosan. Moreover, a conformation of fibroin to a more stable form induced by the addition of chitosan was verified. Blend membranes of fibroin/chitosan were prepared in different proportions and had their crystallinity, structural conformation and thermal stability characterized. The results of crystallographic analysis (XRD) indicated the tendency to higher structural organization caused by the addition of chitosan. FTIR showed that, mainly in a content of chitosan of only 25%, fibroin is present in a more stable form. Thermal analyzes indicate that fibroin is thermally stable and that when its proportion in the blend increases, the temperature in which the degradation is initiated also does so. (author)

  13. Enhanced Fibroblast Cellular Ligamentization Process to Polyethylene Terepthalate Artificial Ligament by Silk Fibroin Coating.

    Jiang, Jia; Ai, Chengchong; Zhan, Zufeng; Zhang, Peng; Wan, Fang; Chen, Jun; Hao, Wei; Wang, Yaxian; Yao, Jinrong; Shao, Zhengzhong; Chen, Tianwu; Zhou, Liang; Chen, Shiyi

    2016-04-01

    Artificial ligaments utilized in reconstruction of anterior cruciate ligament (ACL) are usually made of polyethylene terepthalate (PET) because of its good mechanical properties in vivo. However, it was found that the deficiencies in hydrophilicity and biocompatibility of PET hindered the process of ligamentization. Therefore, surface modification of the PET is deemed as a solution in resolving such problem. Silk fibroin (SF), which is characterized by good biocompatibility and low immunogenicity in clinical applications, was utilized to prepare a coating on the PET ligament (PET+SF) in this work. At first, decrease of hydrophobicity and appearance of amino groups were found on the surface of artificial PET ligament after coating with SF. Second, mouse fibroblasts were cultured on the two different kinds of ligament in order to clarify the possible effect of SF coating. It was proved that mouse fibroblasts display better adhesion and proliferation on PET+SF than PET ligament according to the results of several technical methods including SEM observation, cell adhesive force and spread area test, and mRNA analysis. Meanwhile, methylthiazolyldiphenyl-tetrazolium bromide and DNA content tests showed that biocompatibility of PET+SF is better than PET ligament. In addition, collagen deposition tests also indicated that the quantity of collagen in PET+SF is higher than PET ligament. Based on these results, it can be concluded that SF coating is suggested to be an effective approach to modify the surface of PET ligament and enhance the "ligamentization" process in vivo accordingly. PMID:26526301

  14. Elimination of large particulate units from silk fibroin PLD films by post-treatments

    After depositing silk fibroin (SF) thin films by pulsed IR-laser deposition, extraordinarily large particulate units up to several micrometers were observed. They include debris from the target and severely agglomerated protein units. Occurrence of those large particles was found to be minimum on the vertical substrate. We tried to eliminate large particulate units by two post-treatment operations, i.e. dry gaseous blow-off (GBO) and rinsing in water under simultaneous ultrasonication (WSU). Change in the surface structure by these post-treatments was observed by optical and electron microscopes with varying area from 1mm square down to 1?m square. GBO turned out to be suitable to eliminate the lightly attached particulates of 1-10?m, mostly those pulled out from the target while preserving morphological and chemical properties of smallest units underneath. WSU, on the other hand, pelt off more strongly attached surface irregularities. However, morphological change with an increase in the surface roughness in the range of 1nm was also observed after WSU. The latter might be associated with possible sonochemical effects

  15. Flexible organic light emitting diodes fabricated on biocompatible silk fibroin substrate

    Liu, Yuqiang; Xie, Yuemin; Liu, Yuan; Song, Tao; Zhang, Ke-Qin; Liao, Liangsheng; Sun, Baoquan

    2015-10-01

    Flexible and biodegradable electronics are currently under extensive investigation for biocompatible and environmentally-friendly applications. Synthetic plastic foils are widely used as substrates for flexible electronics. But typical plastic substrates such as polyethylene naphthalate (PEN) could not be degraded in a natural bio-environment. A great demand still exists for a next-generation biocompatible and biodegradable substrate for future application. For example, electronic devices can be potentially integrated into the human body. In this work, we demonstrate that the biocompatible and biodegradable natural silk fibroin (SF) films embedded with silver nanowires (AgNWs) mesh could be employed as conductive transparent substrates to fabricate flexible organic light emitting diodes (OLEDs). Compared with commercial PEN substrates coated with indium tin oxide, the AgNWs/SF composite substrates exhibit a similar sheet resistance of 12 Ω sq-1, a lower surface roughness, as well as a broader light transmission range. Flexible OLEDs based on AgNWs/SF substrates achieve a current efficiency of 19 cd A-1, demonstrating the potential of the flexible AgNWs/SF films as conductive and transparent substrates for next-generation biodegradable devices.

  16. The biocompatibility of silk fibroin and acellular collagen scaffolds for tissue engineering in the ear

    Recent experimental studies have shown the suitability of silk fibroin scaffold (SFS) and porcine-derived acellular collagen I/III scaffold (ACS) as onlay graft materials for tympanic membrane perforation repair. The aims of this study were to further characterize and evaluate the in vivo biocompatibility of SFS and ACS compared with commonly used materials such as Gelfoam and paper in a rat model. The scaffolds were implanted in subcutaneous (SC) tissue and middle ear (ME) cavity followed by histological and otoscopic evaluation for up to 26 weeks. Our results revealed that SFS and ACS were well tolerated and compatible in rat SC and ME tissues throughout the study. The tissue response adjacent to the implants evaluated by histology and otoscopy showed SFS and ACS to have a milder tissue response with minimal inflammation compared to that of paper. Gelfoam gave similar results to SFS and ACS after SC implantation, but it was found to be associated with pronounced fibrosis and osteoneogenesis after ME implantation. It is concluded that SFS and ACS both were biocompatible and could serve as potential alternative scaffolds for tissue engineering in the ear. (paper)

  17. Biocompatibility and osteoconduction of macroporous silk fibroin implants in cortical defects in sheep.

    Uebersax, Lorenz; Apfel, Tanja; Nuss, Katja M R; Vogt, Rainer; Kim, Hyoen Yoo; Meinel, Lorenz; Kaplan, David L; Auer, Joerg A; Merkle, Hans P; von Rechenberg, Brigitte

    2013-09-01

    The goal of the presented study was to compare the biocompatibility and cellular responses to porous silk fibroin (SF) scaffolds produced in a water-based (UPW) or a solvent based process (HFIP) using two different SF sources. For that reason, four different SF scaffolds were implanted (n=6) into drill hole defects in the cancellous bone of the sheep tibia and humerus. The scaffolds were evaluated histologically for biocompatibility, cell-material interaction, and cellular ingrowth. New bone formation was observed macroscopically and histologically at 8 weeks after implantation. For semiquantitative evaluation, the investigated parameters were scored and statistically analyzed (factorial ANOVA). All implants showed good biocompatibility as evident by low infiltration of inflammatory cells and the absent encapsulation of the scaffolds in connective tissue. Multinuclear foreign body giant cells (MFGCs) and macrophages were present in all parts of the scaffold at the material surface and actively degrading the SF material. Cell ingrowth and vascularization were uniform across the scaffold. However, in HFIP scaffolds, local regions of void pores were present throughout the scaffold, probably due to the low pore interconnectivity in this scaffold type in contrast to UPW scaffolds. The amount of newly formed bone was very low in both scaffold types but was more abundant in the periphery than in the center of the scaffolds and for HFIP scaffolds mainly restricted to single pores. PMID:23958322

  18. Fabrication and Intermolecular Interactions of Silk Fibroin/Hydroxybutyl Chitosan Blended Nanofibers

    Xiu-Mei Mo

    2011-03-01

    Full Text Available The native extracellular matrix (ECM is composed of a cross-linked porous network of multifibril collagens and glycosaminoglycans. Nanofibrous scaffolds of silk fibroin (SF and hydroxybutyl chitosan (HBC blends were fabricated using 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP and trifluoroacetic acid (TFA as solvents to biomimic the native ECM via electrospinning. Scanning electronic microscope (SEM showed that relatively uniform nanofibers could be obtained when 12% SF was blended with 6% HBC at the weight ratio of 50:50. Meanwhile, the average nanofibrous diameter increased when the content of HBC in SF/HBC blends was raised from 20% to 100%. Fourier transform infrared spectra (FTIR and 13C nuclear magnetic resonance (NMR showed SF and HBC molecules existed in hydrogen bonding interactions but HBC did not induce conformation of SF transforming from random coil form to β-sheet structure. X-ray diffraction (XRD confirmed the different structure of SF/HBC blended nanofibers from both SF and HBC. Thermogravimetry-Differential thermogravimetry (TG-DTG results demonstrated that the thermal stability of SF/HBC blend nanofibrous scaffolds was improved. The results indicated that the rearrangement of HBC and SF molecular chain formed a new structure due to stronger hydrogen bonding between SF and HBC. These electrospun SF/HBC blended nanofibers may provide an ideal tissue engineering scaffold and wound dressing.

  19. Conformation Transition and Thermal Properties Study of Silk Fibroin and Poly (ε-Caprolactone Blends

    M. Srisa-Ard

    2008-01-01

    Full Text Available In this study, influence of intermolecular interactions between Silk Fibroin (SF and Poly (ε-Caprolactone (PCL in homogeneous blend form on SF conformation changes and thermal properties of the blends was investigated and discussed. The SF/PCL blends were prepared by solution blending and precipitating method, respectively. Dimethylsulfoxide and isopropanol were used as a solvent and a non-solvent, respectively. The blends were characterized by Fourier Transform Infrared Spectroscopy (FTIR, Differential Scanning Calorimetry (DSC, Thermo Gravimetry (TG and Scanning Electron Microscopy (SEM. Conformation transition of SF component from β-sheet to random coil forms can be induced by blending with PCL. Melting temperature and heat of melting of the PCL decreased as increasing the SF ratio. Thermal stability of the SF can be enhanced by blending with PCL. FTIR, DSC and TG results suggested that intermolecular hydrogen bonds were formed between SF and PCL molecules in the blends. Homogeneous morphology of blends was illustrated by SEM micrographs.

  20. Fabrication and characterization of curcumin-loaded silk fibroin/P(LLA-CL) nanofibrous scaffold

    Lian, Yuan; Zhan, Jian-Chao; Zhang, Kui-Hua; Mo, Xiu-Mei

    2014-12-01

    Curcumin exhibited excellent properties including antioxidant, antiinflammatory, antiviral, antibacterial, antifungal, anticancer, and anticoagulant activities. In this study, curcumin was incorporated into silk fibroin (SF)/poly(L-lactic acid- co-e-caprolactone) (P(LLA-CL)) nanofibrous scaffolds via electrospinning, and changes brought about by raising the curcumin content were observed: SEM images showed that the average nanofibrous diameter decreased at the beginning and then increased, and the nanofibers became uniform; FTIR showed that the conformation of SF transforming from random coil form to β-sheet structure had not been induced, while SF conformation converted to β-sheet after being treated with 75% ethanol vapor; XRD results confirmed that the crystal structure of (P(LLA-CL)) had been destroyed; The mechanical test illustrated that nanofibrous scaffolds still maintained good mechanical properties. Further, curcumin-loaded nanofibrous scaffolds were evaluated for drug release, antioxidant and antimicrobial activities in vitro. The results showed that curcumin presented a sustained release behavior from nanofibrous scaffolds and maintained its free radical scavenging ability, and such scaffolds could effectively inhibit S. aureus growth (> 95%). Thus, curcumin-loaded SF/P(LLA-CL) nanofibrous scaffolds might be potential candidates for wound dressing and tissue engineering scaffolds.

  1. The cultivation of human retinal pigment epithelial cells on Bombyx mori silk fibroin.

    Shadforth, Audra M A; George, Karina A; Kwan, Anthony S; Chirila, Traian V; Harkin, Damien G

    2012-06-01

    We have presently evaluated membranes prepared from Bombyx mori silk fibroin (BMSF), for their potential use as a prosthetic Bruch's membrane and carrier substrate for human retinal pigment epithelial (RPE) cell transplantation. Porous BMSF membranes measuring 3 ?m in thickness were prepared from aqueous solutions (3% w/v) containing poly(ethylene oxide) (0.09%). The permeability coefficient for membranes was between 3 and 9 10(-5) cm/s by using Allura red or 70 kDa FITC-dextran respectively. Average pore size (sd) was 4.9 2.3 ?m and 2.9 1.5 ?m for upper and lower membrane surfaces respectively. Optimal attachment of ARPE-19 cells to BMSF membrane was achieved by pre-coating with vitronectin (1 ?g/mL). ARPE-19 cultures maintained in low serum on BMSF membranes for approximately 8 weeks, developed a cobble-stoned morphology accompanied by a cortical distribution of F-actin and ZO-1. Similar results were obtained using primary cultures of human RPE cells, but cultures took noticeably longer to establish on BMSF compared with tissue culture plastic. These findings encourage further studies of BMSF as a substrate for RPE cell transplantation. PMID:22406408

  2. Preparation of electrospun silk fibroin fiber mats as bone scaffolds: a preliminary study

    Meechaisue, Chidchanok [Department of Materials Technology, Faculty of Science, Ramkhamhaeng University, Bangkok 10240 (Thailand); Wutticharoenmongkol, Patcharaporn [Technological Center for Electrospun Fibers and The Petroleum and Petrochemical College, Chulalongkorn University, Bangkok 10330 (Thailand); Waraput, Rujira [Department of Materials Technology, Faculty of Science, Ramkhamhaeng University, Bangkok 10240 (Thailand); Huangjing, Thanapol [Department of Materials Technology, Faculty of Science, Ramkhamhaeng University, Bangkok 10240 (Thailand); Ketbumrung, Nantana [Department of Materials Technology, Faculty of Science, Ramkhamhaeng University, Bangkok 10240 (Thailand); Pavasant, Prasit [Department of Anatomy, Faculty of Dentistry, Chulalongkorn University, Bangkok 10330 (Thailand); Supaphol, Pitt [Technological Center for Electrospun Fibers and The Petroleum and Petrochemical College, Chulalongkorn University, Bangkok 10330 (Thailand)

    2007-09-15

    In the present contribution, electrospinning (e-spinning) was used to fabricate ultra-fine fibers of silk fibroin (SF) from cocoons of indigenous Thai silkworms (Nang-Lai) and Chinese/Japanese hybrid silkworms (DOAE-7). The effects of solution concentration (i.e., 10-40% (w/v) in 85% (v/v) formic acid) and applied electrostatic field strength (EFS; 10, 15 and 20 kV/10 cm) on morphology and size of the electrospun (e-spun) SF products were investigated by scanning electron microscopy. The average diameter of the resulting e-spun SF fibers was found to increase with an increase in both the solution concentration and the EFS value. Specifically, the average diameter of the e-spun SF fibers from Nang-Lai SF solutions ranged between 217 and 610 nm, while that of the fibers from DOAE-7 SF solutions ranged between 183 and 810 nm. The potential for use of the e-spun SF fiber mats as bone scaffolds was assessed with mouse osteoblast-like cells (MC3T3-E1) in which the cells appeared to adhere and proliferate well on their surface.

  3. Study of synthesis of nano-hydroxyapatite using a silk fibroin template

    Wang Jing; Yu Feng; Qu Lijie; Meng Xiangcai [Provincial Key Laboratory of Biomaterials, Jiamusi University, Jiamusi 154007 (China); Wen, G [School of Materials Science and Engineering, Harbin Institute of Technology, Heilongjiang Province 150001 (China)

    2010-08-01

    Nano-hydroxyapatite (HA) was directly synthesized on a silk fibroin (SF) template using the property of SF being soluable in a concentrated CaCl{sub 2} solution as a HA source of calcium at pH 7.4 and room temperature. The microstructure and bonding state were investigated by x-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry-thermogravimetry analysis (DSC-TG) and transmission electron microscopy (TEM). The results indicated that the HA crystals were poorly crystallized with a rod-like shape of 20-60 nm length and 10-20 nm diameter. Strong molecular interactions and chemical bonds might be present between SF and HA. There were other nucleation sites such as carbonyl (-C-O) and amine (-N-H-) groups on SF molecules besides the carboxyl (-COOH) and hydroxyl (-OH) groups previously reported. During the formation of HA, the coordination action between specific functional groups on SF and calcium ions (Ca{sup 2+}) played an important role. The crystallinity of HA was improved and had an orientation growth along (0 0 2) at the presence of SF, resulting in a structure similar to natural bone. It was concluded that SF could regulate the structure and morphology of HA effectively. (communication)

  4. Silk Fibroin/Gelatin Hybrid Films for Medical Applications: Study on Chlorhexidine Diacetate

    Baimark Yodthong

    2010-01-01

    Full Text Available This study aimed to prepare silk fibroin (SF/gelatin (G hybrid films by a solvent evaporation method for loading chlorhexidine diacetate (CHX. The SF and G solution in different ratios were mixed with CHX and placed on the 5 cm polystyrene plates before drying to obtain hybrid films. The films were determined their secondary structures and thermal properties by using Fourier transform infrared (FT-IR spectrometer and thermogravimetric analysis (TGA, respectively. The results found that all of film composed of α-helix and β-sheet structures. However, differences of the α-helix and β-sheet structures were differed according to each component. The hybrid films showed soft texture and decreased of brittle compared to SF film only when the G content increased. Thermal properties of the films indicated that decomposition temperature profiles of all films did not differ dramatically, however, combination of characteristics both SF and G were appeared in hybrid films. The releasing rate of CHX-loaded in the films was found that the CHX has released from the SF film in higher rate than hybrid and G films, respectively. It is a promising that polarity, flexibility as well as component ratio of each polymer play important role on the releasing of CHX.

  5. Effect of polyaspartic acid on hydroxyapatite deposition in silk fibroin blend films

    2010-05-01

    Full Text Available Polyaspartic acid/silk fibroin/hydroxyapatite (PASP/SF-HA composites have been synthesized by biomimetic processing. SF solution was mixed with different contents of PASP to prepare the PASP/SF blend membranes. After ethanol treatment and premineralization process, the blend membranes were immersed into 1.5 simulated body fluid (1.5 SBF for 24 h to induce apatite deposition at 37±0.5°C. Fourier transform infrared spectroscopy (FTIR and X-ray diffraction (XRD results revealed that a conformation transition of SF occurred after the addition of PASP and ethanol treatment. The FTIR and XRD results also confirmed that the main component of apatite deposition was HA. Scanning electron microscopy (SEM showed that the content of HA increased with increasing PASP concentration .Inductively Coupled Plasma-Atomic Emission Spectrometry (ICP results revealed that the Ca/P molar ratio could reach 1.45, which was close to the Ca/P ratio of apatite. It was appropriate to conclude that the increasing content of PASP had a distinct effect on HA deposition in the blend films.

  6. Morphology and thermal stability of silk fibroin/starch blended microparticles

    2010-12-01

    Full Text Available In the present study biodegradable microparticles of silk fibroin (SF/starch blends were prepared by a simple water-in-oil emulsion solvent diffusion technique. SF/starch blended solution and ethyl acetate were used as water and oil phases, respectively. The influence of SF/starch ratios on characteristics of the blended microparticles was investigated. The SF conformation of microparticle matrices from FTIR analysis was changed from random coil to β-sheet form by blending with starch. The blended microparticles had lower dissolution in water than those of SF and starch microparticles. The 1/3 (w/w SF/starch blended microparticles exhibited the lowest dissolution. The SF and starch microparticles showed irregular and deflated shapes, respectively. The blended microparticles were nearly spherical in shapes and smaller sizes. Thermal stability of the blended microparticles slightly increased with the starch blended ratio. The results suggested that SF conformational transition, thermal stability, morphology and dissolution of the blended microparticles can be adjusted by varying the blended ratio.

  7. Cartilage Tissue Engineering with Silk Fibroin Scaffolds Fabricated by Indirect Additive Manufacturing Technology

    Chih-Hao Chen

    2014-03-01

    Full Text Available Advanced tissue engineering (TE technology based on additive manufacturing (AM can fabricate scaffolds with a three-dimensional (3D environment suitable for cartilage regeneration. Specifically, AM technology may allow the incorporation of complex architectural features. The present study involves the fabrication of 3D TE scaffolds by an indirect AM approach using silk fibroin (SF. From scanning electron microscopic observations, the presence of micro-pores and interconnected channels within the scaffold could be verified, resulting in a TE scaffold with both micro- and macro-structural features. The intrinsic properties, such as the chemical structure and thermal characteristics of SF, were preserved after the indirect AM manufacturing process. In vitro cell culture within the SF scaffold using porcine articular chondrocytes showed a steady increase in cell numbers up to Day 14. The specific production (per cell basis of the cartilage-specific extracellular matrix component (collagen Type II was enhanced with culture time up to 12 weeks, indicating the re-differentiation of chondrocytes within the scaffold. Subcutaneous implantation of the scaffold-chondrocyte constructs in nude mice also confirmed the formation of ectopic cartilage by histological examination and immunostaining.

  8. Antheraea pernyi silk fibroin for targeted gene delivery of VEGF165-Ang-1 with PEI

    Vascularization is a crucial challenge in tissue engineering. One solution for this problem is to implant scaffolds that contain functional genes that promote vascularization by providing angiogenic growth factors via a gene delivery carrier. Poly(ethylenimine) (PEI) is a gene delivery carrier with high transfection efficiency but with cytotoxicity. To solve this problem, we utilized Antheraea pernyi silk fibroin (ASF), which has favorable cytocompatibility and biodegradability, RGD sequences and a negative charge, in conjunction with PEI, as the delivery vector for vascular endothelial growth factor (VEGF) 165-angiopoietin-1 (Ang-1) dual gene simultaneous expression plasmid, creating an ASF/PEI/pDNA complex. The results suggested that the zeta potential of the ASF/PEI/pDNA complex was significantly lower than that of the PEI/pDNA complex. Decreased nitrogen and increased oxygen on the surface of the complex demonstrated that the ASF had successfully combined with the surface of the PEI/pDNA. Furthermore, the complexes resisted digestion by nucleic acid enzymes and degradation by serum. L929 cells were cultured and transfected in vitro and improved cytotoxicity was found when the cells were transfected with ASF/PEI/pDNA compared with PEI/pDNA. In addition, the transfection efficiency and VEGF secretion increased. In general, this study provides a novel method for decreasing the cytotoxicity of PEI gene delivery vectors and increasing transfection efficiency of angiogenesis-related genes. (paper)

  9. Synthesis and characterization of dense membranes of silk fibroin with glycerin;Sintese e caracterizacao de membranas densas de fibroina de seda com glicerina

    Silva, Mariana F.; Moraes, Mariana A. de; Weska, Raquel F.; Nogueira, Grinia M.; Beppu, Marisa M., E-mail: beppu@feq.unicamp.b [Universidade Estadual de Campinas (FEQ/UNICAMP), SP (Brazil). Faculdade de Engenharia Quimica

    2009-07-01

    The addition of plasticizers seeks improvements in mechanical properties of dense membranes of silk fibroin with possible interactions by hydrogen bonds. The aim of the present study was to produce and characterize dense membranes of silk fibroin containing glycerin in two different concentrations. The characterization of the membranes was performed from scanning electron microscopy (SEM), mechanical traction tests, infrared spectroscopy (FTIR-ATR) and X-ray diffraction (XRD). The results indicated that the addition of glycerin allowed obtaining homogeneous and more crystalline membranes and improved their properties of elongation. (author)

  10. Impact of silk fibroin-based scaffold structures on human osteoblast MG63 cell attachment and proliferation

    Varkey A

    2015-10-01

    Full Text Available Aneesia Varkey,1,2 Elakkiya Venugopal,2 Ponjanani Sugumaran,2 Gopinathan Janarthanan,1 Mamatha M Pillai,2 Selvakumar Rajendran,2 Amitava Bhattacharyya1 1Advanced Textile and Polymer Research Laboratory, 2Tissue Engineering Laboratory, PSG Institute of Advanced Studies, Coimbatore, Tamil Nadu, India Abstract: The present study was carried out to investigate the impact of various types of silk fibroin (SF scaffolds on human osteoblast-like cell (MG63 attachment and proliferation. SF was isolated from Bombyx mori silk worm cocoons after degumming. Protein concentration in the degummed SF solution was estimated using Bradford method. Aqueous SF solution was used to fabricate three different types of scaffolds, viz, electrospun nanofiber mat, sponge, and porous film. The structures of the prepared scaffolds were characterized using optical micro­scopy and field emission scanning electron microscopy. The changes in the secondary structure of the proteins and the thermal behavior of the scaffolds were determined by Fourier transform infrared spectroscopy and thermo-gravimetric analysis, respectively. The biodegradation rate of scaffolds was determined by incubating the scaffolds in simulated body fluid for 4 weeks. MG63 cells were seeded on the scaffolds and their attachment and proliferation onto the scaffolds were studied. The MTT assay was carried out to deduce the toxicity of the developed scaffolds. All the scaffolds were found to be biocompatible. The amount of collagen produced by the osteoblast-like cells growing on different scaffolds was estimated. Keywords: silk fibroin scaffold, electrospun nanofiber, porous film, sponge, osteoblast

  11. Structural evolution of regenerated silk fibroin under shear: Combined wide- and small-angle x-ray scattering experiments using synchrotron radiation

    Rossle, Manfred [European Molecular Biology Laboratory (EMBL), France; Panine, Pierre [European Synchrotron Radiation Facility (ESRF); Urban, Volker S [ORNL; Riekel, Christine [European Synchrotron Radiation Facility (ESRF)

    2004-04-01

    The structural evolution of regenerated Bombyx mori silk fibroin during shearing with a Couette cell has been studied in situ by synchrotron radiation small- and wide-angle x-ray scattering techniques. An elongation of fibroin molecules was observed with increasing shear rate, followed by an aggregation phase. The aggregates were found to be amorphous with {beta}-conformation according to infrared spectroscopy. Scanning x-ray microdiffraction with a 5 {micro}m beam on aggregated material, which had solidified in air, showed silk II reflections and a material with equatorial reflections close to the silk I structure reflections, but with strong differences in reflection intensities. This silk I type material shows up to two low-angle peaks suggesting the presence of water molecules that might be intercalated between hydrogen-bonded sheets.

  12. Radiation degradation of silk

    Ishida, Kazushige; Kamiishi, Youichi [Textile Research Institute of Gunma, Kiryu, Gunma (Japan); Takeshita, Hidefumi; Yoshii, Fumio; Kume, Tamikazu [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    2001-03-01

    Silk fibroin powder was prepared from irradiated silk fibroin fiber by means of only physical treatment. Silk fibroin fiber irradiated with an accelerated electron beam in the dose range of 250 - 1000 kGy was pulverized by using a ball mill. Unirradiated silk fibroin fiber was not pulverized at all. But the more irradiation was increased, the more the conversion efficiency from fiber to powder was increased. The conversion efficiency of silk fibroin fiber irradiated 1000 kGy in oxygen was 94%. Silk fibroin powder shows remarkable solubility, which dissolved 57% into water of ambient temperature. It is a very interesting phenomenon that silk fibroin which did not treat with chemicals gets solubility only being pulverized. In order to study mechanism of solubilization of silk fibroin powder, amino acid component of soluble part of silk fibroin powder was analyzed. The more irradiation dose up, the more glycine or alanine degraded, but degradation fraction reached bounds about 50%. Other amino acids were degraded only 20% even at the maximum. To consider crystal construction of silk fibroin, it is suggested that irradiation on silk fibroin fiber selectively degrades glycine and alanine in amorphous region, which makes it possible to pulverize and to dissolve silk fibroin powder. (author)

  13. Electrospun silk fibroin/poly(lactide-co-ε-caprolactone nanofibrous scaffolds for bone regeneration

    Wang Z

    2016-04-01

    Full Text Available Zi Wang,1,* Ming Lin,1,* Qing Xie,1 Hao Sun,1 Yazhuo Huang,1 DanDan Zhang,1 Zhang Yu,1 Xiaoping Bi,1 Junzhao Chen,1 Jing Wang,2 Wodong Shi,1 Ping Gu,1 Xianqun Fan1 1Department of Ophthalmology, Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, 2Biomaterials and Tissue Engineering Laboratory, College of Chemistry & Chemical Engineering and Biotechnology, Donghua University, Shanghai, People’s Republic of China *These authors contributed equally to this work Background: Tissue engineering has become a promising therapeutic approach for bone regeneration. Nanofibrous scaffolds have attracted great interest mainly due to their structural similarity to natural extracellular matrix (ECM. Poly(lactide-co-ε-caprolactone (PLCL has been successfully used in bone regeneration, but PLCL polymers are inert and lack natural cell recognition sites, and the surface of PLCL scaffold is hydrophobic. Silk fibroin (SF is a kind of natural polymer with inherent bioactivity, and supports mesenchymal stem cell attachment, osteogenesis, and ECM deposition. Therefore, we fabricated hybrid nanofibrous scaffolds by adding different weight ratios of SF to PLCL in order to find a scaffold with improved properties for bone regeneration.Methods: Hybrid nanofibrous scaffolds were fabricated by blending different weight ratios of SF with PLCL. Human adipose-derived stem cells (hADSCs were seeded on SF/PLCL nanofibrous scaffolds of various ratios for a systematic evaluation of cell adhesion, proliferation, cytotoxicity, and osteogenic differentiation; the efficacy of the composite of hADSCs and scaffolds in repairing critical-sized calvarial defects in rats was investigated.Results: The SF/PLCL (50/50 scaffold exhibited favorable tensile strength, surface roughness, and hydrophilicity, which facilitated cell adhesion and proliferation. Moreover, the SF/PLCL (50/50 scaffold promoted the osteogenic differentiation of hADSCs by elevating the expression levels of osteogenic marker genes such as BSP, Ocn, Col1A1, and OPN and enhanced ECM mineralization. In vivo assays showed that SF/PLCL (50/50 scaffold improved the repair of the critical-sized calvarial defect in rats, resulting in increased bone volume, higher trabecular number, enhanced bone mineral density, and increased new bone areas, compared with the pure PLCL scaffold.Conclusion: The SF/PLCL (50/50 nanofibrous scaffold facilitated hADSC proliferation and osteogenic differentiation in vitro and further promoted new bone formation in vivo, suggesting that the SF/PLCL (50/50 nanofibrous scaffold holds great potential in bone tissue regeneration. Keywords: human adipose-derived mesenchymal stem cells, silk fibrin, poly(lactide-co-ε-caprolactone, proliferation, osteogenesis

  14. A novel electrospinning approach to fabricate high strength aqueous silk fibroin nanofibers.

    Singh, B N; Panda, N N; Pramanik, K

    2016-06-01

    The present paper describes a rapid method of producing concentrated aqueous regenerated Bombyx mori silk fibroin (RSF) solution by applying mild shearing under forced dehumidified air and generation of electrospun SF nanofibers from concentrated solution with high mechanical strength using free liquid surface electrospinning machine. The shear induced concentrating mechanism favoured the electrospinning process by enhancing the viscosity (>2.43Pas as onset for electrospinning) and decreasing the surface tension of the solution (40.1-37.7mN/m). Shearing reduced the β-turns and random coil molecular conformation and thereby, intensified the β-sheet content from 16.9% to 34% which is the minimum content needed to commence RSF nanofibers formation. Subsequently, electrospun nanofibrous mats were produced from different batches of concentrated SF solutions (15-21wt%). Among the concentrated RSF, 17wt% RSF solution was the most favourable concentration producing electrospun nanofibrous mat having lowest average fiber diameters of 183±55nm and good tensile strength. The mechanical strength of the nanofibrous sheet was further improved by cross-linking with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide (EDC+NHS) which might be due to enhancement of β-sheet content. These nanofibers exhibited 17.57±1.13MPa ultimate tensile strength, 12.48±1.46% tensile strain at break and 37.7% increase in root mean square surface roughness which is favourable feature for cell adhesion and neo-tissue formation. PMID:26905467

  15. Immobilization of acetylcholinesterase via biocompatible interface of silk fibroin for detection of organophosphate and carbamate pesticides

    Xue Rui [College of Environmental and Energy Engineering, Beijing University of Technology, Beijing 100124 (China); Kang Tianfang, E-mail: kangtf@yahoo.cn [College of Environmental and Energy Engineering, Beijing University of Technology, Beijing 100124 (China); Lu Liping; Cheng Shuiyuan [College of Environmental and Energy Engineering, Beijing University of Technology, Beijing 100124 (China)

    2012-06-01

    An amperometric biosensor for the detection of organophosphate and carbamate pesticides was developed based on the immobilization of acetylcholinesterase (AChE) on regenerated silk fibroin (SF) matrix by non-covalent adsorption. SF and AChE were coated sequentially on the surface of the glassy carbon electrode (GCE) which was modified with multiwall carbon nanotube (MWNTs). The obtained biosensor was denoted as AChE-SF/MWNTs/GCE. The atomic force microscopy images showed that the SF matrix provided a more homogeneous interface for the AChE immobilization. The aggregation of immobilizing AChE was therefore avoided. The cyclic voltammogram of thiocholine at this biosensor exhibited a well defined oxidation peak at 0.667 V (vs. SCE). The inhibition rate of methyl parathion to the immobilized AChE was proportional to the logarithm of the concentration of methyl parathion over the range of the concentration of methyl parathion from 3.5 Multiplication-Sign 10{sup -6} to 2.0 Multiplication-Sign 10{sup -3} M with a detection limit of 5.0 Multiplication-Sign 10{sup -7} M. Similarly, the linearly response range of carbaryl was from 1.0 Multiplication-Sign 10{sup -7} to 3.0 Multiplication-Sign 10{sup -5} M with a detection limit of 6.0 Multiplication-Sign 10{sup -8} M. The experimental results indicate that AChE not only can be immobilized steadily on the SF matrix, but also the bioactivity of immobilizing AChE can be preserved effectively.

  16. Small diameter electrospun silk fibroin vascular grafts: Mechanical properties, in vitro biodegradability, and in vivo biocompatibility.

    Catto, Valentina; Farè, Silvia; Cattaneo, Irene; Figliuzzi, Marina; Alessandrino, Antonio; Freddi, Giuliano; Remuzzi, Andrea; Tanzi, Maria Cristina

    2015-09-01

    To overcome the drawbacks of autologous grafts currently used in clinical practice, vascular tissue engineering represents an alternative approach for the replacement of small diameter blood vessels. In the present work, the production and characterization of small diameter tubular matrices (inner diameter (ID)=4.5 and 1.5 mm), obtained by electrospinning (ES) of Bombyx mori silk fibroin (SF), have been considered. ES-SF tubular scaffolds with ID=1.5 mm are original, and can be used as vascular grafts in pediatrics or in hand microsurgery. Axial and circumferential tensile tests on ES-SF tubes showed appropriate properties for the specific application. The burst pressure and the compliance of ES-SF tubes were estimated using the Laplace's law. Specifically, the estimated burst pressure was higher than the physiological pressures and the estimated compliance was similar or higher than that of native rat aorta and Goretex® prosthesis. Enzymatic in vitro degradation tests demonstrated a decrease of order and crystallinity of the SF outer surface as a consequence of the enzyme activity. The in vitro cytocompatibility of the ES-SF tubes was confirmed by the adhesion and growth of primary porcine smooth muscle cells. The in vivo subcutaneous implant into the rat dorsal tissue indicated that ES-SF matrices caused a mild host reaction. Thus, the results of this investigation, in which comprehensive morphological and mechanical aspects, in vitro degradation and in vitro and in vivo biocompatibility were considered, indicate the potential suitability of these ES-SF tubular matrices as scaffolds for the regeneration of small diameter blood vessels. PMID:26046273

  17. In vitro biocompatibility evaluation of silk-fibroin/polyurethane membrane with cultivation of HUVECs

    Zhou, Mei; Wang, Wei-Ci; Liao, Yong-Gui; Liu, Wen-Qi; Yu, Miao; Ouyang, Chen-Xi

    2014-03-01

    In order to investigate the in vitro biocompatibility of a novel polyurethane (PU) membrane modified by incorporation of superfine silk-fibroin powder (SFP), which was prepared for small-diameter vascular grafts, with the cultivation of human umbilical vein endothelial cells (HUVECs), PU and SFP were mixed with the ratios of 9:1, 7:3, 5:5, 3:7 (PU:SFP) to make four composite materials. Unmodified PU and polytetrafluoroethylene (PTFE) were added as control groups. CCK-8 assay was used to evaluate the cytotoxicity of these biomaterials. Data were processed using SPSS, and P < 0.05 was considered to be statistically significant. Adherence and spreading of HUVECs on the surface of specimens was observed using direct contact cultivation. The toxicity ratings of the novel composites were grade 0-1, which is in the acceptable range. In all the experimental groups except control, SFP/PU with ratio of 1:9 had the least cytotoxicity property, and more content of SFP in the composite showed no improvement of the biocompatibility. HUVECs strongly attached to and grew on the surface of the biomaterials, and proliferated rapidly. The proliferation ability increased with increased proportion of SFP; however the cell quantity on the surface of the materials decreased when the proportion of SFP was equal to or larger than that of PU in the composite. It is concluded that this novel material has excellent cellular affinity with no cytotoxicity to HUVECs. Adding SFP gives PU better biocompatibility, while further research on optimum blend ratios is still needed.

  18. Spermine-modified Antheraea pernyi silk fibroin as a gene delivery carrier.

    Yu, Yanni; Hu, Yongpei; Li, Xiufang; Liu, Yu; Li, Mingzhong; Yang, Jicheng; Sheng, Weihua

    2016-01-01

    The development of a novel cationized polymer used as a gene delivery carrier that can conveniently and effectively transfect cells resulting in a stably expressed target gene remains a challenge. Antheraea pernyi silk fibroin (ASF) is a cytocompatible and biodegradable natural polymer, and it possesses Arg-Gly-Asp sequences but a negative charge. In order to render ASF amenable to packaging plasmid DNA (pDNA), spermine was used to modify ASF to synthesize cationized ASF (CASF), which was used as a gene delivery carrier. CASF was characterized using trinitrobenzene sulfonic acid assay, the zeta potential determination, and a Fourier transform infrared analysis, and the results of these characterizations indicated that the -NH2 in spermine effectively reacts with the -COOH in the side chains of ASF. Spermine grafted to the side chains of ASF resulted in the conversion of the negative charge of ASF to a positive charge. CASF packaged pDNA and formed CASF/pDNA complexes, which exhibited spherical morphology with average particle sizes of 215-281 nm and zeta potential of approximately +3.0 mV to +3.2 mV. The results of the MTT assay, confocal laser scanning microscopy, and flow cytometry analysis in a human endothelial cell line revealed that CASF/pDNA complexes exhibited lower cytotoxicity and higher transfection efficiency compared to the pDNA complexes of polyethyleneimine. These results indicate that our synthesized CASF, a cationized polymer, is a potential gene delivery carrier with the advantages of biodegradability and low cytotoxicity. PMID:27042056

  19. Relationship between rheology and electro-spinning performance of regenerated silk fibroin prepared using different degumming methods

    Kim, Hyun Ju; Um, In Chul

    2014-05-01

    Electro-spun silk fibroin (SF) has been studied for biomedical applications because of its good biocompatibility, cyto-compatibility, and simple fabrication method. SF is obtained by a degumming process and the degumming method can affect the degree of molecular degradation of SF during the degumming process. In the present study, the effect of the degumming method on the rheology and electro-spinning performance of a silk solution was examined. In addition, the relationship between the rheology and electrospinnability was investigated. Regardless of the degumming method, all silk formic acid solutions exhibited almost Newtonian fluid behavior. The order of the viscosity of the silk solution was as follows: HTHP method > acid method > soap/soda method. An analysis of the correlation between the viscosity and electrospun morphology showed that the viscosity played a key role in determining the electro-spun morphology, and the critical viscosity for good fiber formation without beads in electro-spinning exists between 0.13 and 0.20 Pa·s. The viscosity also determines the maximum electro-spinning rate of the SF formic acid solution. The morphology and diameter of the electro-spun fiber were almost unaffected by the electro-spinning rate of the SF solution.

  20. Assessment of freestanding membranes prepared from Antheraea pernyi silk fibroin as a potential vehicle for corneal epithelial cell transplantation

    Freestanding membranes created from Bombyx mori silk fibroin (BMSF) offer a potential vehicle for corneal cell transplantation since they are transparent and support the growth of human corneal epithelial (HCE) cells. Fibroin derived from the wild silkworm Antheraea pernyi (APSF) might provide a superior material by virtue of containing putative cell-attachment sites that are absent from BMSF. Thus we have investigated the feasibility of producing transparent, freestanding membranes from APSF and have analysed the behaviour of HCE cells on this material. No significant differences in cell numbers or phenotype were observed in short term HCE cell cultures established on either fibroin. Production of transparent freestanding APSF membranes, however, proved to be problematic as cast solutions of APSF were more prone to becoming opaque, displayed significantly lower permeability and were more brittle than BMSF-membranes. Cultures of HCE cells established on either membrane developed a normal stratified morphology with cytokeratin pair 3/12 being immuno-localized to the superficial layers. We conclude that while it is feasible to produce transparent freestanding membranes from APSF, the technical difficulties associated with this biomaterial, along with an absence of enhanced cell growth, currently favour the continued development of BMSF as a preferred vehicle for corneal cell transplantation. Nevertheless, it remains possible that refinement of techniques for processing APSF might yet lead to improvements in the handling properties and performance of this material. (paper)

  1. Towards functional 3D-stacked electrospun composite scaffolds of PHBV, silk fibroin and nanohydroxyapatite: Mechanical properties and surface osteogenic differentiation.

    Paşcu, Elena I; Cahill, Paul A; Stokes, Joseph; McGuinness, Garrett B

    2016-04-01

    Bone tissue engineering scaffolds have two challenging functional tasks to fulfil: to encourage cell proliferation, differentiation and matrix synthesis and to provide suitable mechanical stability upon implantation. Composites of biopolymers and bioceramics combine the advantages of both types of materials, resulting in better processability and enhanced mechanical and biological properties through matrix reinforcement. In the present study, novel thick bone composite scaffolds were successfully fabricated using electrospun flat sheets of polyhydroxybutyrate-polyhydroxyvalerate/nanohydroxyapatite/silk fibroin essence (2% nanohydroxyapatite - 2% silk fibroin essence and 5% nanohydroxyapatite - 5% silk fibroin essence, respectively). Their potential asin vitrobone regeneration scaffolds was evaluated using mouse calvarian osteoblast cells (MC3T3-E1), in terms of morphology (scanning electron microscope), cell attachment, cell proliferation, Col type I, osteopontin and bone alkaline phosphatase activity (Quantitative Real Time Polymerase Chain Reaction [qRT-PCR], enzyme-linked immunosorbent assay, immunocytochemistry). Electrospun polyhydroxybutyrate-polyhydroxyvalerate scaffolds were used as reference constructs. The results showed that the compressive and tensile mechanical properties of the scaffolds are dependent on the change in their composition, and the treatment these underwent. Furthermore, methanol-treated and autoclaved (MA) P2 (2% nanohydroxyapatite, 2% silk fibroin essence) samples appeared to exhibit more promising tensile properties. Additionally, the compressive tests results confirmed that the methanol pre-treatment and the autoclaving step lead to an increase in the P2 secant modulus when compared to the non-methanol-treated ones, P2 and P5 (5% nanohydroxyapatite, 5% silk fibroin essence), respectively.Both formulations of polyhydroxybutyrate-polyhydroxyvalerate/nanohydroxyapatite/silk fibroin essence composite promoted greater cell adhesion and proliferation than the corresponding polyhydroxybutyrate-polyhydroxyvalerate control ones. Cells seeded on the composite fibrous scaffolds were extensively expanded and elongated on the fibre surface after one day in culture, whereas those seeded on the polyhydroxybutyrate-polyhydroxyvalerate scaffolds were not completely elongated. In addition, cells grown on P2 and P5 scaffolds had higher alkaline phosphatase activity when compared to those containing no nanohydroxyapatite/silk fibroin essence. PMID:26767394

  2. Optimization and evaluation of silk fibroin-chitosan freeze-dried porous scaffolds for cartilage tissue engineering application.

    Vishwanath, Varshini; Pramanik, Krishna; Biswas, Amit

    2016-05-01

    Silk fibroin/chitosan blend has been reported to be an attractive biomaterial that provides a 3D porous structure with controllable pore size and mechanical property suitable for tissue engineering applications. However, there is no systematic study for optimizing the ratio of silk fibroin (SF) and chitosan (CS) which seems to influence the scaffold property to a great extent. The present research, therefore, investigates the effect of blend ratio of SF and CS on scaffold property and establishes the optimum value of blend ratio. Among the various blends, the scaffolds with blend ratio of SF/CS (80:20) were found to be superior. The scaffold possesses pore size in the range 71-210 μm and porosity of 82.2 ± 1.3%. The compressive strength of the scaffold was measured as 190 ± 0.2 kPa. The cell supportive property of the scaffold in terms of cell attachment, cell viability, and proliferation was confirmed by cell culture study using mesenchymal stem cells derived from umbilical cord blood. Furthermore, the assessment of glycosaminoglycan secretion on the scaffolds indicates its potentiality toward cartilage tissue regeneration. PMID:26830046

  3. Silk Fibroin/Nano-CaP Bilayered scaffolds for osteochondral tissue engineering

    Yan, Leping; Oliveira, Joaquim M.; de Oliveira, A. L.; Reis, R.L.

    2014-01-01

    In this study, bilayered silk and silk/nano-CaP scaffolds were developed for osteochondral (OC) tissue engineering. Aqueous silk solution (16 wt.%) was used for preparation of the cartilage-like layer and, for generation of the silk/nano-CaP suspension and the bottom layer (CaP/Silk: 16 wt.%). The scaffolds were formed by using salt-leaching/lyophilization approach. The scanning electron microscopy revealed that the both layers presented porous structure and integrated well. Mi...

  4. Silk Fibroin/Chitosan Blend Films Loaded Methylene Blue as a Model for Polar Molecular Releasing: Comparison between Thai Silk Varieties

    Srihanam Prasong

    2011-01-01

    Full Text Available The aims of this study were to prepare Silk Fibroin (SF/Chitosan (CS blend films loaded Methylene Blue (MB and characterize their related properties. The SF was obtained from various Thai silk varieties locally called Nang Lai, Mo and Kaki. The blend films were prepared by mixing the SF, CS and MB solutions before pouring on polystyrene plates. They were then taken to an oven at 40C for 3 days to obtain the films. In addition, each native SF silk variety and CS films were also prepared as control. SEM micrographs showed that native Nang Lai film appeared the bead like particles while other film types were smooth. Comparison between the blend films, Mo variety film has smooth surfaces while Nang Lai and Kaki films composed of flat particles covered their surfaces. FTIR results indicated that all of native SF has similar absorption bands at amide regions which coexisted of a-helix and -sheet structures. Moreover, SF blended with CS showed strong bands at amide II, exhibit -sheet structure. The blend film of Mo variety rapidly decomposed whereas Nang Lai was the lowest. In vitro releasing study indicated that MB released more quickly at the initial 30 min which Mo variety was the slowest.

  5. Incorporation of Exogenous RGD Peptide and Inter-Species Blending as Strategies for Enhancing Human Corneal Limbal Epithelial Cell Growth on Bombyx mori Silk Fibroin Membranes

    Traian V. Chirila

    2013-05-01

    Full Text Available While fibroin isolated from the cocoons of domesticated silkworm Bombyx mori supports growth of human corneal limbal epithelial (HLE cells, the mechanism of cell attachment remains unclear. In the present study we sought to enhance the attachment of HLE cells to membranes of Bombyx mori silk fibroin (BMSF through surface functionalization with an arginine-glycine-aspartic acid (RGD-containing peptide. Moreover, we have examined the response of HLE cells to BMSF when blended with the fibroin produced by a wild silkworm, Antheraea pernyi, which is known to contain RGD sequences within its primary structure. A procedure to isolate A. pernyi silk fibroin (APSF from the cocoons was established, and blends of the two fibroins were prepared at five different BMSF/APSF ratios. In another experiment, BMSF surface was modified by binding chemically the GRGDSPC peptide using a water-soluble carbodiimide. Primary HLE were grown in the absence of serum on membranes made of BMSF, APSF, and their blends, as well as on RGD-modified BMSF. There was no statistically significant enhancing effect on the cell attachment due to the RGD presence. This suggests that the adhesion through RGD ligands may have a complex mechanism, and the investigated strategies are of limited value unless the factors contributing to this mechanism become better known.

  6. In vitro and in vivo characterization of a silk fibroin-coated polyester vascular prosthesis.

    Huang, Fuhua; Sun, Lizhong; Zheng, Jun

    2008-12-01

    Silk fibroin (SF) is well known to be biocompatible, degradable, and nontoxic. In this study, SF was impregnated into a porous polyester graft (InterVascular external velour, InterVascular, Inc., La Ciotat, France), 8 mm in diameter. The SF-impregnated graft was investigated in vitro and in vivo to evaluate its potential for use as a new vascular graft impervious to blood, while retaining high porosity for tissue ingrowth and biological healing. For in vitro investigation, the water permeability, coating weight, morphology, and mechanical properties of the SF-impregnated grafts were compared with collagen-coated grafts (InterGard grafts, InterVascular, Inc.). The water permeability of the controls (1388 +/- 30.5 mL/cm(2)/min at 120 mm Hg) was reduced >99% by SF impregnation, rendering the graft impervious to blood. The coating weight of the collagen was 117 +/- 22 mg/g of graft, producing a slightly lower value than the InterGard prosthesis (302 +/- 23 mg/g). For the in vivo experiment, six SF-sealed vascular grafts were implanted in the abdominal aorta of dogs for scheduled periods ranging from 4 h to 6 months. Commercial collagen-impregnated grafts (InterGard) and untreated external velour grafts (InterVascular) were also implanted for scheduled periods ranging from 1 to 6 months for comparison. Gross observation of the explanted grafts and histological examination of the representative sections were conducted for two types of grafts using a light microscope after hematoxylin-eosin staining. These SF-impregnated grafts showed less foreign body and inflammation reactions, and the SF layer was almost completely absorbed. The average of the values in each period for the SF grafts was 48% neointima at 1 month, 85% at 3 months, and 97% at 6 months, whereas those of the InterGard prostheses was 34, 46, and 90%, respectively. This study demonstrated that the use of a biodegradable SF as biological sealant can be a feasible approach to prepare impervious textile arterial prostheses. The SF-impregnated graft showed less thrombogenesis and induced host cell migration along the matrix without foreign body or inflammatory reactions. Moreover, it appears to facilitate the development of endothelial-like cells. PMID:19133021

  7. Electrospun silk fibroin/poly(lactide-co-ε-caprolactone) nanofibrous scaffolds for bone regeneration

    Wang, Zi; Lin, Ming; Xie, Qing; Sun, Hao; Huang, Yazhuo; Zhang, DanDan; Yu, Zhang; Bi, Xiaoping; Chen, Junzhao; Wang, Jing; Shi, Wodong; Gu, Ping; Fan, Xianqun

    2016-01-01

    Background Tissue engineering has become a promising therapeutic approach for bone regeneration. Nanofibrous scaffolds have attracted great interest mainly due to their structural similarity to natural extracellular matrix (ECM). Poly(lactide-co-ε-caprolactone) (PLCL) has been successfully used in bone regeneration, but PLCL polymers are inert and lack natural cell recognition sites, and the surface of PLCL scaffold is hydrophobic. Silk fibroin (SF) is a kind of natural polymer with inherent bioactivity, and supports mesenchymal stem cell attachment, osteogenesis, and ECM deposition. Therefore, we fabricated hybrid nanofibrous scaffolds by adding different weight ratios of SF to PLCL in order to find a scaffold with improved properties for bone regeneration. Methods Hybrid nanofibrous scaffolds were fabricated by blending different weight ratios of SF with PLCL. Human adipose-derived stem cells (hADSCs) were seeded on SF/PLCL nanofibrous scaffolds of various ratios for a systematic evaluation of cell adhesion, proliferation, cytotoxicity, and osteogenic differentiation; the efficacy of the composite of hADSCs and scaffolds in repairing critical-sized calvarial defects in rats was investigated. Results The SF/PLCL (50/50) scaffold exhibited favorable tensile strength, surface roughness, and hydrophilicity, which facilitated cell adhesion and proliferation. Moreover, the SF/PLCL (50/50) scaffold promoted the osteogenic differentiation of hADSCs by elevating the expression levels of osteogenic marker genes such as BSP, Ocn, Col1A1, and OPN and enhanced ECM mineralization. In vivo assays showed that SF/PLCL (50/50) scaffold improved the repair of the critical-sized calvarial defect in rats, resulting in increased bone volume, higher trabecular number, enhanced bone mineral density, and increased new bone areas, compared with the pure PLCL scaffold. Conclusion The SF/PLCL (50/50) nanofibrous scaffold facilitated hADSC proliferation and osteogenic differentiation in vitro and further promoted new bone formation in vivo, suggesting that the SF/PLCL (50/50) nanofibrous scaffold holds great potential in bone tissue regeneration.

  8. Electrospinning-aligned and random polydioxanone-polycaprolactone-silk fibroin-blended scaffolds: geometry for a vascular matrix

    Extracellular matrices are arranged with a specific geometry based on tissue type and mechanical stimulus. For blood vessels in the body, preferential alignment of fibers is in the direction of repetitive force. Electrospinning is a controllable process which can result in fiber alignment and randomization depending on the parameters utilized. In this study, arterial grafts composed of polycaprolactone (PCL), polydioxanone (PDO) and silk fibroin in blends of 100:0 and 50:50 for both PCL:silk and PDO:silk were investigated to determine if fibers could be controllably aligned using a mandrel rotational speed ranging from 500 to 8000 revolutions per minute (RPM). Results revealed that large- and small-diameter mandrels produced different degrees of fiber alignment based on a fast Fourier transform of scanning electron microscope images. Uniaxial tensile testing further demonstrated scaffold anisotropy through changes in peak stress, modulus and strain at break at mandrel rotational speeds of 500 and 8000 RPM, causing peak stress and modulus for PCL to increase 5- and 4.5-fold, respectively, as rotational speed increased. Additional mechanical testing was performed on grafts using dynamic compliance, burst strength and longitudinal strength displaying that grafts electrospun at higher rotational rates produced stiffer conduits which had lower compliance and higher burst strength compared to the lower mandrel rotational rate. Scaffold properties were found to depend on several parameters in the electrospinning process: mandrel rotational rate, polymer type, and mandrel size. Vascular scaffold design under anisotropic conditions provided interesting insights and warrants further investigation.

  9. Electrospinning-aligned and random polydioxanone-polycaprolactone-silk fibroin-blended scaffolds: geometry for a vascular matrix

    McClure, M J; Sell, S A; Bowlin, G L [Department of Biomedical Engineering, Virginia Commonwealth University, Richmond, VA 23284 (United States); Ayres, C E; Simpson, D G, E-mail: glbowlin@vcu.ed [Department of Anatomy and Neurobiology, Virginia Commonwealth University, Richmond, VA 23298 (United States)

    2009-10-15

    Extracellular matrices are arranged with a specific geometry based on tissue type and mechanical stimulus. For blood vessels in the body, preferential alignment of fibers is in the direction of repetitive force. Electrospinning is a controllable process which can result in fiber alignment and randomization depending on the parameters utilized. In this study, arterial grafts composed of polycaprolactone (PCL), polydioxanone (PDO) and silk fibroin in blends of 100:0 and 50:50 for both PCL:silk and PDO:silk were investigated to determine if fibers could be controllably aligned using a mandrel rotational speed ranging from 500 to 8000 revolutions per minute (RPM). Results revealed that large- and small-diameter mandrels produced different degrees of fiber alignment based on a fast Fourier transform of scanning electron microscope images. Uniaxial tensile testing further demonstrated scaffold anisotropy through changes in peak stress, modulus and strain at break at mandrel rotational speeds of 500 and 8000 RPM, causing peak stress and modulus for PCL to increase 5- and 4.5-fold, respectively, as rotational speed increased. Additional mechanical testing was performed on grafts using dynamic compliance, burst strength and longitudinal strength displaying that grafts electrospun at higher rotational rates produced stiffer conduits which had lower compliance and higher burst strength compared to the lower mandrel rotational rate. Scaffold properties were found to depend on several parameters in the electrospinning process: mandrel rotational rate, polymer type, and mandrel size. Vascular scaffold design under anisotropic conditions provided interesting insights and warrants further investigation.

  10. Construction of engineering adipose-like tissue in vivo utilizing human insulin gene-modified umbilical cord mesenchymal stromal cells with silk fibroin 3D scaffolds.

    Li, Shi-Long; Liu, Yi; Hui, Ling

    2015-12-01

    We evaluated the use of a combination of human insulin gene-modified umbilical cord mesenchymal stromal cells (hUMSCs) with silk fibroin 3D scaffolds for adipose tissue engineering. In this study hUMSCs were isolated and cultured. HUMSCs infected with Ade-insulin-EGFP were seeded in fibroin 3D scaffolds with uniform 50-60 µm pore size. Silk fibroin scaffolds with untransfected hUMSCs were used as control. They were cultured for 4 days in adipogenic medium and transplanted under the dorsal skins of female Wistar rats after the hUMSCs had been labelled with chloromethylbenzamido-1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (CM-Dil). Macroscopical impression, fluorescence observation, histology and SEM were used for assessment after transplantation at 8 and 12 weeks. Macroscopically, newly formed adipose tissue was observed in the experimental group and control group after 8 and 12 weeks. Fluorescence observation supported that the formed adipose tissue originated from seeded hUMSCs rather than from possible infiltrating perivascular tissue. Oil red O staining of newly formed tissue showed that there was substantially more tissue regeneration in the experimental group than in the control group. SEM showed that experimental group cells had more fat-like cells, whose volume was larger than that of the control group, and degradation of the silk fibroin scaffold was greater under SEM observation. This study provides significant evidence that hUMSCs transfected by adenovirus vector have good compatibility with silk fibroin scaffold, and adenoviral transfection of the human insulin gene can be used for the construction of tissue-engineered adipose. PMID:23509085

  11. Semi-interpenetrating polymer networks composed of silk fibroin and poly(ethylene glycol) for wound dressing

    Semi-interpenetrating polymer networks (SIPNs) composed of silk fibroin (SF) and poly(ethylene glycol) (PEG) were prepared by photopolymerization of a PEG macromer in the presence of SF to improve the mechanical properties of SF sponge as wound dressing. The morphological structure of the SF/PEG SIPNs was observed to be composed of an interconnected microporous surface and a cross-sectional area. SF/PEG SIPNs showed non-cytotoxicity evaluated by a cell proliferation method using L929 fibroblasts. Wound contraction treated with SF/PEG SIPNs sponges was faster than that of Vaseline gauze as a control. Histological observation confirmed that the deposition of collagen in the dermis was organized by covering the wound area with SF/PEG SIPNs. The above results indicated that SF/PEG SIPNs could be used as wound dressing

  12. Semi-interpenetrating polymer networks composed of silk fibroin and poly(ethylene glycol) for wound dressing

    Kweon, HaeYong; Yeo, Joo-hong; Lee, Kwang-gill [Applied Sericulture and Apiculture Division, National Institute of Agricultural Science and Technology, Suwon 441100 (Korea, Republic of); Lee, Hyun Chul; Na, Hee Sam [Department of Microbiology and Research Institute of Medical Sciences, Chonnam National University Medical School, Kwangju (Korea, Republic of); Won, Young Ho [Department of Dermatology, Chonnam National University Medical School, Kwangju (Korea, Republic of); Cho, Chong Su [School of Agricultural Biotechnology, Seoul National University, Seoul 151742 (Korea, Republic of)], E-mail: chocs@plaza.snu.ac.kr

    2008-09-01

    Semi-interpenetrating polymer networks (SIPNs) composed of silk fibroin (SF) and poly(ethylene glycol) (PEG) were prepared by photopolymerization of a PEG macromer in the presence of SF to improve the mechanical properties of SF sponge as wound dressing. The morphological structure of the SF/PEG SIPNs was observed to be composed of an interconnected microporous surface and a cross-sectional area. SF/PEG SIPNs showed non-cytotoxicity evaluated by a cell proliferation method using L929 fibroblasts. Wound contraction treated with SF/PEG SIPNs sponges was faster than that of Vaseline gauze as a control. Histological observation confirmed that the deposition of collagen in the dermis was organized by covering the wound area with SF/PEG SIPNs. The above results indicated that SF/PEG SIPNs could be used as wound dressing.

  13. Influence of Philosamia ricini Silk Fibroin Components on Morphology, Secondary Structure and Thermal Properties of Chitosan Biopolymer Film

    S. Prasong

    2009-01-01

    Full Text Available This study aimed to prepare Eri (Philosamia ricini Silk Fibroin (SF/chitosan (CS blend films by a solvent evaporation method and to compare the blend films with both native SF and CS films. Influence of SF ratios on the morphology, secondary structure and thermal decomposition of the CS blend films were investigated. The native SF and CS films were uniform and homogeneous without phase separation. For the blend films, the uniform can be found less than 60% of SF composition. All of SF/CS blend films showed both SF and CS characteristics. FT-IR results showed that the blend films composed of both random coil and ?-sheet with predominant of ?-sheet form. Interaction of intermolecular between SF and CS have occurred which were measured by thermogravimetric thermograms. Increasing of SF contents was leading to the increase of ?-sheet structures which were enhanced the thermal stability of the CS blend films.

  14. High performance pentacene organic field-effect transistors consisting of biocompatible PMMA/silk fibroin bilayer dielectric

    Pentacene organic field-effect transistors (OFETs) based on single- or double-layer biocompatible dielectrics of poly(methyl methacrylate) (PMMA) and/or silk fibroin (SF) are fabricated. Compared with those devices based on single PMMA or SF dielectric or SF/PMMA bilayer dielectric, the OFETs with biocompatible PMMA/SF bilayer dielectric exhibit optimal performance with a high field-effect mobility of 0.21 cm2/Vs and a current on/off ratio of 1.5×104. By investigating the surface morphology of the pentacene active layer through atom force microscopy and analyzing the electrical properties, the performance enhancement is mainly attributed to the crystallization improvement of the pentacene and the smaller interface trap density at the dielectric/organic interface. Meanwhile, a low contact resistance also indicates that a good electrode/organic contact is formed, thereby assisting the performance improvement of the OFET

  15. Composite biocompatible hydroxyapatite-silk fibroin coatings for medical implants obtained by Matrix Assisted Pulsed Laser Evaporation

    The aim of this study was to obtain biomimetic inorganic-organic thin films as coatings for metallic medical implants. These contain hydroxyapatite, the inorganic component of the bony tissues, and a natural biopolymer - silk fibroin - added in view to induce the surface functionalization. Hydroxyapatite (HA), silk fibroin (FIB) and composite HA-FIB films were obtained by Matrix Assisted Pulsed Laser Evaporation (MAPLE) in order to compare their physical and biological performances as coatings on metallic prostheses. We used an excimer laser source (KrF*, λ = 248 nm, τ = 25 ns) operated at 10 Hz repetition rate. Coatings were deposited on quartz, Si and Ti substrates and then subjected to physical (FTIR, XRD, AFM, SEM) analyses, correlated with the results of the cytocompatibility in vitro tests. The hybrid films were synthesized from frozen targets of aqueous suspensions with 3:2 or 3:4 weight ratio of HA:FIB. An appropriate stoichiometric and functional transfer was obtained for 0.4-0.5 J/cm2 laser fluence. FTIR spectra of FIB and HA-FIB films exhibited distinctive absorption maxima, in specific positions of FIB random coil form: 1540 cm-1 amide II, 1654 cm-1 amide I, 1243 cm-1 amide III, while the peak from 1027 cm-1 appeared only for HA and composite films. Osteosarcoma SaOs2 cells cultured 72 h on FIB and HA-FIB films showed increased viability, good spreading and normal cell morphology. The well-elongated, flattened cells are a sign of an appropriate interaction with the MAPLE FIB and composite HA-FIB coatings.

  16. Composite biocompatible hydroxyapatite-silk fibroin coatings for medical implants obtained by Matrix Assisted Pulsed Laser Evaporation

    Miroiu, F.M., E-mail: marimona.miroiu@inflpr.ro [National Institute for Lasers, Plasma, and Radiation Physics, 409 Atomistilor Street, RO-77125, MG-36, Magurele-Ilfov (Romania); Socol, G.; Visan, A.; Stefan, N.; Craciun, D.; Craciun, V.; Dorcioman, G.; Mihailescu, I.N. [National Institute for Lasers, Plasma, and Radiation Physics, 409 Atomistilor Street, RO-77125, MG-36, Magurele-Ilfov (Romania); Sima, L.E.; Petrescu, S.M. [Institute of Biochemistry, Romanian Academy, 296 Splaiul Independentei, 060031 Bucharest (Romania); Andronie, A.; Stamatin, I. [3Nano-SAE Alternative Energy Sources-University of Bucharest, Faculty of Physics, 409 Atomistilor Street, RO-77125, Magurele-Ilfov (Romania); Moga, S.; Ducu, C. [University of Pitesti, Targul din Vale Str, no. 1, 110040 Pitesti (Romania)

    2010-05-25

    The aim of this study was to obtain biomimetic inorganic-organic thin films as coatings for metallic medical implants. These contain hydroxyapatite, the inorganic component of the bony tissues, and a natural biopolymer - silk fibroin - added in view to induce the surface functionalization. Hydroxyapatite (HA), silk fibroin (FIB) and composite HA-FIB films were obtained by Matrix Assisted Pulsed Laser Evaporation (MAPLE) in order to compare their physical and biological performances as coatings on metallic prostheses. We used an excimer laser source (KrF*, {lambda} = 248 nm, {tau} = 25 ns) operated at 10 Hz repetition rate. Coatings were deposited on quartz, Si and Ti substrates and then subjected to physical (FTIR, XRD, AFM, SEM) analyses, correlated with the results of the cytocompatibility in vitro tests. The hybrid films were synthesized from frozen targets of aqueous suspensions with 3:2 or 3:4 weight ratio of HA:FIB. An appropriate stoichiometric and functional transfer was obtained for 0.4-0.5 J/cm{sup 2} laser fluence. FTIR spectra of FIB and HA-FIB films exhibited distinctive absorption maxima, in specific positions of FIB random coil form: 1540 cm{sup -1} amide II, 1654 cm{sup -1} amide I, 1243 cm{sup -1} amide III, while the peak from 1027 cm{sup -1} appeared only for HA and composite films. Osteosarcoma SaOs2 cells cultured 72 h on FIB and HA-FIB films showed increased viability, good spreading and normal cell morphology. The well-elongated, flattened cells are a sign of an appropriate interaction with the MAPLE FIB and composite HA-FIB coatings.

  17. Proteome identification of the silkworm middle silk gland.

    Li, Jian-Ying; Ye, Lu-Peng; Che, Jia-Qian; Song, Jia; You, Zheng-Ying; Wang, Shao-Hua; Zhong, Bo-Xiong

    2016-03-01

    To investigate the functional differentiation among the anterior (A), middle (M), and posterior (P) regions of silkworm middle silk gland (MSG), their proteomes were characterized by shotgun LC-MS/MS analysis with a LTQ-Orbitrap mass spectrometer. To get better proteome identification and quantification, triplicate replicates of mass spectrometry analysis were performed for each sample. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium (Vizcaíno et al., 2014) [1] via the PRIDE partner repository (Vizcaino, 2013) [2] with the dataset identifier PXD003371. The peptide identifications that were further processed by PeptideProphet program in Trans-Proteomic Pipeline (TPP) after database search with Mascot software were also available in .XML format files. Data presented here are related to a research article published in Journal of Proteomics by Li et al. (2015) [3]. PMID:26937469

  18. Silk fibroins modify the atmospheric low temperature plasma-treated poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) film for the application of cardiovascular tissue engineering

    Huaxiao Yang; Min Sun; Ping Zhou; Luanfeng Pan; Chungen Wu

    2010-01-01

    Tissue engineered scaffold is one of the hopeful therapies for the patients with organ or tissue damages. The key element for a tissue engineered scaffold material is high biocompatibility. Herein the poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) film was irradiated by the low temperature atmospheric plasma and then coated by the silk fibroins (SF). After plasma treatment, the surface of PHBHHx film became rougher and more hydrophilic than that of original film. The experiment of PH...

  19. Impact of silk fibroin-based scaffold structures on human osteoblast MG63 cell attachment and proliferation.

    Varkey, Aneesia; Venugopal, Elakkiya; Sugumaran, Ponjanani; Janarthanan, Gopinathan; Pillai, Mamatha M; Rajendran, Selvakumar; Bhattacharyya, Amitava

    2015-01-01

    The present study was carried out to investigate the impact of various types of silk fibroin (SF) scaffolds on human osteoblast-like cell (MG63) attachment and proliferation. SF was isolated from Bombyx mori silk worm cocoons after degumming. Protein concentration in the degummed SF solution was estimated using Bradford method. Aqueous SF solution was used to fabricate three different types of scaffolds, viz, electrospun nanofiber mat, sponge, and porous film. The structures of the prepared scaffolds were characterized using optical microscopy and field emission scanning electron microscopy. The changes in the secondary structure of the proteins and the thermal behavior of the scaffolds were determined by Fourier transform infrared spectroscopy and thermo-gravimetric analysis, respectively. The biodegradation rate of scaffolds was determined by incubating the scaffolds in simulated body fluid for 4 weeks. MG63 cells were seeded on the scaffolds and their attachment and proliferation onto the scaffolds were studied. The MTT assay was carried out to deduce the toxicity of the developed scaffolds. All the scaffolds were found to be biocompatible. The amount of collagen produced by the osteoblast-like cells growing on different scaffolds was estimated. PMID:26491306

  20. Synthesis and cellular compatibility of Co-doped ZnO particles in silk-fibroin peptides.

    Zou, Yuanwen; Huang, Zhongbing; Wang, Yan; Liao, Xiaoming; Yin, Guangfu; Gu, Jianwen

    2013-02-01

    Co-doped ZnO particles were successfully prepared via a facile biomineralization process in the template of silk-fibroin (SF) peptides at room temperature, and SF peptides were coated onto the surface of particles. The ratio of Zn/Co in reactive solution could substantially influence the morphology of as-prepared particles, and the rough spherical particles including some nanoparticles of 50 nm diameters could be obtained at 4:1 ratio of Zn/Co. The saturation magnetization of SF-coated Co-ZnO particles was 8.63 emu/g, much larger than that of Co-ZnO without SF. L929 cell test revealed that the Co-doped ZnO particles had a good cellular compatibility at the concentration of less than 0.25 mg/mL due to silk-peptide coating, indicating that the prepared Co-doped ZnO particles have a potential for the biomedical applications. PMID:23010108

  1. Surface modification of silk fibroin fibers with poly(methyl methacrylate) and poly(tributylsilyl methacrylate) via RAFT polymerization for marine antifouling applications

    In this study, silk fibroin surface containing hydroxyl and aminogroups was firstly modified using a polymerizable coupling agent 3-(trimethoxysilyl) propyl methacrylate (MPS), in order to induce vinyl groups onto the fiber surface. The reversible addition–fragmentation chain transfer (RAFT)-mediated polymerization of methyl methacrylate (MMA) and tributylsilyl methacrylate (TBSiMA) through the immobilized vinyl bond on the silk fibroin surface in the presence of 2-cyanoprop-2-yl dithiobenzoate (CPDB) as chain-transfer agent and 2,2′-azobis(isobutyronitrile) (AIBN) as initiator was conducted in toluene solution at 70 °C for 24 h. The structure and properties of the modified fiber were characterized by Fourier Transform Infrared Spectroscopy, 13C, 29Si Nuclear Magnetic Resonance (NMR) spectroscopy, thermogravimetric analysis (TGA), scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS), confirming the presence of the coupling molecule and the methacrylate groups onto the silk fibroin fiber surface. Molecular weight distributions were assessed by triple detection size exclusion chromatography (TD-SEC) in order to verify the livingness of the polymerization. - Highlights: • SF surface containing hydroxyl and amino groups was firstly modified with MPS. • RAFT polymerizations of MMA and TBSiMA were studied. • TD-SEC was used to verify the livingness of the RAFT polymerization. • The grafted polymer chains enhance the thermal stability of the SF fibers. • The grafted fibers could be potentially promising candidates as antifouling agents

  2. Bio-hybrid silk fibroin/calcium phosphate/PLGA nanocomposite scaffold to control the delivery of vascular endothelial growth factor

    Farokhi, Mehdi, E-mail: mehdi13294@yahoo.com [Department of Tissue Engineering, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Mottaghitalab, Fatemeh, E-mail: fatemeh.motaghi@gmail.com [Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University (TMU), Tehran (Iran, Islamic Republic of); Shokrgozar, Mohammad Ali, E-mail: mashokrgozar@pasteur.ac.ir [National Cell Bank of Iran, Pasteur Institute of Iran, Tehran (Iran, Islamic Republic of); Ai, Jafar, E-mail: jafar_ai@tums.ac.ir [Department of Tissue Engineering, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Hadjati, Jamshid; Azami, Mahmoud [Department of Tissue Engineering, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of)

    2014-02-01

    This study investigated the efficacy of bio-hybrid silk fibroin/Calcium phosphate/PLGA nanocomposite scaffold as vascular endothelial growth factor (VEGF) delivery system. The scaffold was fabricated using freeze-drying and electrospinning. Here, we highlight the structural changes of the scaffold using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy and differential scanning calorimetry (DSC). The uniform dispersion of calcium phosohate (CaP) powder within silk fibroin (SF) solution was also confirmed using Zeta potential analysis. Moreover, good biocompatibility of osteoblast cells next to the scaffold was approved by cell adhesion, proliferation and alkaline phosphatase production. The release profile of VEGF during 28 days has established the efficacy of the scaffold as a sustained delivery system. The bioactivity of the released VEGF was maintained about 83%. The histology analysis has shown that the new bone tissue formation happened in the defected site after 10 weeks of implantation. Generally, our data showed that the fabricated scaffold could be considered as an effective scaffold for bone tissue engineering applications. - Highlights: • Silk fibroin/calcium phosphate/PLGA scaffold was successfully fabricated using freeze-drying and electrospinning. • The scaffold could control the release of VEGF during 28 days. • The bioactivity of electrospun VEGF was above 80%. • VEGF loaded scaffold could induce bone regeneration after 10 weeks in rabbit.

  3. Bio-hybrid silk fibroin/calcium phosphate/PLGA nanocomposite scaffold to control the delivery of vascular endothelial growth factor

    This study investigated the efficacy of bio-hybrid silk fibroin/Calcium phosphate/PLGA nanocomposite scaffold as vascular endothelial growth factor (VEGF) delivery system. The scaffold was fabricated using freeze-drying and electrospinning. Here, we highlight the structural changes of the scaffold using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy and differential scanning calorimetry (DSC). The uniform dispersion of calcium phosohate (CaP) powder within silk fibroin (SF) solution was also confirmed using Zeta potential analysis. Moreover, good biocompatibility of osteoblast cells next to the scaffold was approved by cell adhesion, proliferation and alkaline phosphatase production. The release profile of VEGF during 28 days has established the efficacy of the scaffold as a sustained delivery system. The bioactivity of the released VEGF was maintained about 83%. The histology analysis has shown that the new bone tissue formation happened in the defected site after 10 weeks of implantation. Generally, our data showed that the fabricated scaffold could be considered as an effective scaffold for bone tissue engineering applications. - Highlights: • Silk fibroin/calcium phosphate/PLGA scaffold was successfully fabricated using freeze-drying and electrospinning. • The scaffold could control the release of VEGF during 28 days. • The bioactivity of electrospun VEGF was above 80%. • VEGF loaded scaffold could induce bone regeneration after 10 weeks in rabbit

  4. Enhanced bone formation in electrospun poly(l-lactic-co-glycolic acid)-tussah silk fibroin ultrafine nanofiber scaffolds incorporated with graphene oxide.

    Shao, Weili; He, Jianxin; Sang, Feng; Wang, Qian; Chen, Li; Cui, Shizhong; Ding, Bin

    2016-05-01

    To engineer bone tissue, it is necessary to provide a biocompatible, mechanically robust scaffold. In this study, we fabricated an ultrafine nanofiber scaffold by electrospinning a blend of poly(l-lactic-co-glycolic acid), tussah silk fibroin, and graphene oxide (GO) and characterized its morphology, biocompatibility, mechanical properties, and biological activity. The data indicate that incorporation of 10wt.% tussah silk and 1wt.% graphene oxide into poly(l-lactic-co-glycolic acid) nanofibers significantly decreased the fiber diameter from 280 to 130nm. Furthermore, tussah silk and graphene oxide boosted the Young's modulus and tensile strength by nearly 4-fold and 3-fold, respectively, and significantly enhanced adhesion, proliferation in mouse mesenchymal stem cells and functionally promoted biomineralization-relevant alkaline phosphatase (ALP) and mineral deposition. The results indicate that composite nanofibers could be excellent and versatile scaffolds for bone tissue engineering. PMID:26952489

  5. An optimized sericin-1 expression system for mass-producing recombinant proteins in the middle silk glands of transgenic silkworms.

    Wang, Feng; Xu, Hanfu; Yuan, Lin; Ma, Sanyuan; Wang, Yuancheng; Duan, Xiaoli; Duan, Jianping; Xiang, Zhonghuai; Xia, Qingyou

    2013-10-01

    The middle silk gland (MSG) of silkworm is thought to be a potential host for mass-producing valuable recombinant proteins. Transgenic MSG expression systems based on the usage of promoter of sericin1 gene (sericin-1 expression system) have been established to produce various recombinant proteins in MSG. However, further modifying the activity of the sericin-1 expression system to yield higher amounts of recombinant proteins is still necessary. In this study, we provide an alternative modification strategy to construct an efficient sericin-1 expression system by using the hr3 enhancer (hr3 CQ) from a Chongqing strain of the Bombyx mori nuclear polyhedrosis virus (BmNPV) and the 3'UTRs of the fibroin heavy chain (Fib-HPA), the fibroin light chain (Fib-LPA), and Sericin1 (Ser1PA) genes. We first analyzed the effects of these DNA elements on expression of luciferase, and found that the combination of hr3 CQ and Ser1PA was most effective to increase the activity of luciferase. Then, hr3 CQ and Ser1PA were used to modify the sericin1 expression system. Transgenic silkworms bearing these modified sericin1 expression vectors were generated by a piggyBac transposon mediated genetic transformation method. Our results showed that mRNA level of DsRed reporter gene in transgenic silkworms containing hr3 CQ and Ser1PA significantly increased by 9 fold to approximately 83% of that of endogenous sericin1. As the results of that, the production of recombinant RFP increased by 16 fold to 9.5% (w/w) of cocoon shell weight. We conclude that this modified sericin-1 expression system is efficient and will contribute to the MSG as host to mass produce valuable recombinant proteins. PMID:23435751

  6. Spider Transcriptomes Identify Ancient Large-Scale Gene Duplication Event Potentially Important in Silk Gland Evolution.

    Clarke, Thomas H; Garb, Jessica E; Hayashi, Cheryl Y; Arensburger, Peter; Ayoub, Nadia A

    2015-07-01

    The evolution of specialized tissues with novel functions, such as the silk synthesizing glands in spiders, is likely an influential driver of adaptive success. Large-scale gene duplication events and subsequent paralog divergence are thought to be required for generating evolutionary novelty. Such an event has been proposed for spiders, but not tested. We de novo assembled transcriptomes from three cobweb weaving spider species. Based on phylogenetic analyses of gene families with representatives from each of the three species, we found numerous duplication events indicative of a whole genome or segmental duplication. We estimated the age of the gene duplications relative to several speciation events within spiders and arachnids and found that the duplications likely occurred after the divergence of scorpions (order Scorpionida) and spiders (order Araneae), but before the divergence of the spider suborders Mygalomorphae and Araneomorphae, near the evolutionary origin of spider silk glands. Transcripts that are expressed exclusively or primarily within black widow silk glands are more likely to have a paralog descended from the ancient duplication event and have elevated amino acid replacement rates compared with other transcripts. Thus, an ancient large-scale gene duplication event within the spider lineage was likely an important source of molecular novelty during the evolution of silk gland-specific expression. This duplication event may have provided genetic material for subsequent silk gland diversification in the true spiders (Araneomorphae). PMID:26058392

  7. Silk formation mechanisms in the larval salivary glands of Apis mellifera (Hymenoptera: Apidae)

    Elaine C M Silva-Zacarin; Regina L M Silva De Moraes; S R Taboga

    2003-12-01

    The mechanism of silk formation in Apis mellifera salivary glands, during the 5th instar, was studied. Larval salivary glands were dissected and prepared for light and polarized light microscopy, as well as for scanning and transmission electron microscopy. The results showed that silk formation starts at the middle of the 5th instar and finishes at the end of the same instar. This process begins in the distal secretory portion of the gland, going towards the proximal secretory portion; and from the periphery to the center of the gland lumen. The silk proteins are released from the secretory cells as a homogeneous substance that polymerizes in the lumen to form compact birefringent tactoids. Secondly, the water absorption from the lumen secretion, carried out by secretory and duct cells, promotes aggregation of the tactoids that form a spiral-shape filament with a zigzag pattern. This pattern is also the results of the silk compression in the gland lumen and represents a high concentration of macromolecularly well-oriented silk proteins.

  8. Silk fibroin film from golden-yellow Bombyx mori is a biocomposite that contains lutein and promotes axonal growth of primary neurons.

    Pistone, Assunta; Sagnella, Anna; Chieco, Camilla; Bertazza, Gianpaolo; Varchi, Greta; Formaggio, Francesco; Posati, Tamara; Saracino, Emanuela; Caprini, Marco; Bonetti, Simone; Toffanin, Stefano; Di Virgilio, Nicola; Muccini, Michele; Rossi, Federica; Ruani, Giampiero; Zamboni, Roberto; Benfenati, Valentina

    2016-05-01

    The use of doped silk fibroin (SF) films and substrates from Bombyx mori cocoons for green nanotechnology and biomedical applications has been recently highlighted. Cocoons from coloured strains of B. mori, such as Golden-Yellow, contain high levels of pigments that could have a huge potential for the fabrication of SF based biomaterials targeted to photonics, optoelectronics and neuroregenerative medicine. However, the features of extracted and regenerated SF from cocoons of B. mori Golden-Yellow strain have never been reported. Here we provide a chemophysical characterization of regenerated silk fibroin (RSF) fibers, solution, and films obtained from cocoons of a Golden-Yellow strain of B. mori, by SEM, (1) H-NMR, HPLC, FT-IR, Raman and UV-Vis spectroscopy. We found that the extracted solution and films from B. mori Golden-Yellow fibroin displayed typical Raman spectroscopic and optical features of carotenoids. HPLC-analyses revealed that lutein was the carotenoid contained in the fiber and RSF biopolymer from yellow cocoons. Notably, primary neurons cultured on yellow SF displayed a threefold higher neurite length than those grown of white SF films. The results we report pave the way to expand the potential use of yellow SF in the field of neuroregenerative medicine and provide green chemistry approaches in biomedicine. © 2016 Wiley Periodicals, Inc. Biopolymers 105: 287-299, 2016. PMID:26756916

  9. Antimicrobial and antioxidant surface modification toward a new silk-fibroin (SF)-L-Cysteine material for skin disease management

    Nogueira, Frederico; Granadeiro, Luíza; Mouro, Claudia; Gouveia, Isabel C.

    2016-02-01

    A novel dressing material - silk fibroin fabric (SF)-L-Cysteine (L-Cys) - is here developed to be used as standard treatment for atopic dermatitis (AD), which combines comfort, thermic, and tensile strength properties of silk materials with antioxidant and antimicrobial effects of L-Cys. A careful understanding about the linking strategies is needed in order not to compromise the bioavailability of L-Cys and deplenish its bioactivity. Durability was also addressed through washing cycles and compared with hospital requirements, according to international Standard EN ISO 105-C06:2010. The present research also analyze the interactions between Staphylococcus aureus and SF-L-Cys under simulating conditions of AD and demonstrated the effectiveness of a double covalent grafting, with the importance of SF tyrosine (Tyr) covalent linkage with L-Cys (SF-g-L-Cys/Tyr-g-L-Cys) even after several washing cycles, twenty five, whereas for a disposable application a single covalent mechanism of grafting L-Cys proved to be sufficient (SF-g-L-Cys). Results showed effective antimicrobial activities exhibiting higher inhibition ratios of 98.65% for SF-g-L-Cys after 5 washing cycles, whereas 97.55% for SF-g-L-Cys/Tyr-g-L-Cys after 25 washing cycles, both at pH 9.5 grafting strategy. Furthermore, it is also reported a non-protumoral effect of L-Cys. A new advance is herein achieved at the world of medical antimicrobial textiles tailored to address wound moisture environment and exudate self-cleaning, which may open novel applications as complementary therapy for AD disease.

  10. Development of novel electrospun nanofibrous scaffold from P. ricini and A. mylitta silk fibroin blend with improved surface and biological properties

    Biomaterials that stimulate cell attachment and proliferation without any surface modification (e.g. RGD coating) provide potent and cost effective scaffold for regenerative medicine. This study assessed the physico-chemical properties and cell supportive potential of a silk fibroin blend scaffold derived from eri (Philosamia ricini) and tasar (Antheraea mylitta) silk (ET) respectively by electrospinning process. The scanning electron microscopy and transmission electron microscopy study found that the fiber diameters are in 200 to 800 nm range with flat morphology. The porosity of ET scaffold is found to be 79 ± 5% with majority of pore diameter between 2.5 to 5 nm. Similarly, Bombyx mori (BM) silk fibroin and gelatin nanofibrous scaffolds were prepared and taken as control. The ultimate tensile strength of the ET and BM scaffold are found to be 1.83 ± 0.13 MPa and 1.47 ± 0.10 MPa respectively. The measured contact angle (a measure of hydrophilicity) for ET (54.7° ± 1.8°) is found to be lower than BM (62° ± 2.3°). The ability to deposit apatite over ET is comparable to that of BM nanofibers. All the scaffolds were seeded with cord blood derived mesenchymal stem cells (hMSCs) and cultured for 14 days in vitro. The immunofluorescence study reveals enhanced cell attachment with higher metabolic activity for MSCs grown over ET than BM and gelatin. The ET scaffold also demonstrated expression of higher amount cell adhesion molecules (CD29/CD44) and higher proliferation rate than BM and gelatin as confirmed by MTT assay, DNA content estimation assay, flow cytometry study and SEM study. Overall, it may be concluded that ET scaffold may have potential in developing bone tissue grafts for clinical applications in the future. - Highlights: • We have fabricated eri–tasar blended electrospun silk fibroin nanofiber with superior surface property. • The hydrophilicity is higher than the silk fibroin nanofiber derived from Bombyx mori (BM). • The nanofibrous scaffold showed improved cell adhesion and proliferation property. • The expression of CD44 and CD29 over the MSC grown over this is substantially higher than BM. • The metabolic activity of MSC grown over this scaffold is greater than those from BM

  11. Development of novel electrospun nanofibrous scaffold from P. ricini and A. mylitta silk fibroin blend with improved surface and biological properties

    Panda, N.; Bissoyi, A.; Pramanik, K.; Biswas, A., E-mail: amitb79@gmail.com

    2015-03-01

    Biomaterials that stimulate cell attachment and proliferation without any surface modification (e.g. RGD coating) provide potent and cost effective scaffold for regenerative medicine. This study assessed the physico-chemical properties and cell supportive potential of a silk fibroin blend scaffold derived from eri (Philosamia ricini) and tasar (Antheraea mylitta) silk (ET) respectively by electrospinning process. The scanning electron microscopy and transmission electron microscopy study found that the fiber diameters are in 200 to 800 nm range with flat morphology. The porosity of ET scaffold is found to be 79 ± 5% with majority of pore diameter between 2.5 to 5 nm. Similarly, Bombyx mori (BM) silk fibroin and gelatin nanofibrous scaffolds were prepared and taken as control. The ultimate tensile strength of the ET and BM scaffold are found to be 1.83 ± 0.13 MPa and 1.47 ± 0.10 MPa respectively. The measured contact angle (a measure of hydrophilicity) for ET (54.7° ± 1.8°) is found to be lower than BM (62° ± 2.3°). The ability to deposit apatite over ET is comparable to that of BM nanofibers. All the scaffolds were seeded with cord blood derived mesenchymal stem cells (hMSCs) and cultured for 14 days in vitro. The immunofluorescence study reveals enhanced cell attachment with higher metabolic activity for MSCs grown over ET than BM and gelatin. The ET scaffold also demonstrated expression of higher amount cell adhesion molecules (CD29/CD44) and higher proliferation rate than BM and gelatin as confirmed by MTT assay, DNA content estimation assay, flow cytometry study and SEM study. Overall, it may be concluded that ET scaffold may have potential in developing bone tissue grafts for clinical applications in the future. - Highlights: • We have fabricated eri–tasar blended electrospun silk fibroin nanofiber with superior surface property. • The hydrophilicity is higher than the silk fibroin nanofiber derived from Bombyx mori (BM). • The nanofibrous scaffold showed improved cell adhesion and proliferation property. • The expression of CD44 and CD29 over the MSC grown over this is substantially higher than BM. • The metabolic activity of MSC grown over this scaffold is greater than those from BM.

  12. The ESR signals in silk fibroin and wool keratin under both the effect of UV-irradiation and without any external effects and the formation of free radicals.

    Mamedov, Sh V; Aktas, B; Cantrk, M; Aksakal, B; Alekperov, V; Blbl, F; Yilgin, R; Aslanov, R B

    2002-08-01

    ESR studies have been done on natural and UV-irradiated silk fibroins and wool keratins at the temperature range of -196 degrees C to 20 C. The intensities of ESR signals obtained from the irradiated samples at -196 C remarkably increase with respect to those of natural samples. While the signals mainly consist of triplet peaks at -196 C. a doublet arises around the room temperatures. For the first time, at room temperature without any external effect the complicated ESR spectra of fibrous proteins (wool keratin and silk fibroin) whose components are as follows have been observed: (1) (for white wool keratin) a central doublet with deltaHm = 1.1 mT and g = 2.0075; deltaHm = 5mT and g = 2.1911; (2) a wide peak with deltaHm approximately 66 mT and g approximately 2.1575; (3) the 'sulfur' peak given in the literature with deltaHm = 2.2 mT and g = 2.0218; (4) the signal with deltaHm = 0.6 mT and g = 2.0065, and for silk fibroin, (a) a very wide signal with deltaHm approximately 70 mT and g approximately 2.084; (b) a very sharp signal with deltaHm approximately 1.1 mT and g approximately 2.01; and (c) relatively narrower signal with deltaHm approximately 5 mT and g approximately 2.336. It has been shown by recombination kinetic method that 30-50% of the free radicals formed by UV-irradiation do not undergo recombination up to 220 degrees C and 15 degrees C for silk libroin and wool keratin, respectively, even they keep their concentration constant for long period of time (weeks, months, even longer). In this article, considering above-mentioned results, the mechanism of signals observed in natural wool keratin and silk fibroin without any external effects is examined. We can briefly explain the role of the subject of the article, by considering fibrous proteins and some applications of the reactions by free radical occurring in these proteins tinder the effects of different factors in medicine and biology and the important role of oxidation and the other kinds of degradations on these processes. as well as the significant applications of ESR investigations on comprehending the processes by free radical. PMID:12099283

  13. Composite biodegradable biopolymer coatings of silk fibroin - Poly(3-hydroxybutyric-acid-co-3-hydroxyvaleric-acid) for biomedical applications

    Miroiu, Floralice Marimona; Stefan, Nicolaie; Visan, Anita Ioana; Nita, Cristina; Luculescu, Catalin Romeo; Rasoga, Oana; Socol, Marcela; Zgura, Irina; Cristescu, Rodica; Craciun, Doina; Socol, Gabriel

    2015-11-01

    Composite silk fibroin-poly(3-hydroxybutyric-acid-co-3-hydroxyvaleric-acid) (SF-PHBV) biodegradable coatings were grown by Matrix Assisted Pulsed Laser Evaporation on titanium substrates. Their physico-chemical properties and particularly the degradation behavior in simulated body fluid at 37 °C were studied as first step of applicability in local controlled release for tissue regeneration applications. SF and PHBV, natural biopolymers with excellent biocompatibility, but different biodegradability and tensile strength properties, were combined in a composite to improve their properties as coatings for biomedical uses. FTIR analyses showed the stoichiometric transfer from targets to coatings by the presence in the spectra of the main absorption maxima characteristic of both polymers. XRD investigations confirmed the FTIR results showing differences in crystallization behavior with respect to the SF and PHBV content. Contact angle values obtained through wettability measurements indicated the MAPLE deposited coatings were highly hydrophilic; surfaces turning hydrophobic with the increase of the PHBV component. Degradation assays proved that higher PHBV contents resulted in enhanced resistance and a slower degradation rate of composite coatings in SBF. Distinct drug-release schemes could be obtained by adjusting the SF:PHBV ratio to controllably tuning the coatings degradation rate, from rapid-release formulas, where SF predominates, to prolonged sustained ones, for larger PHBV content.

  14. Fabrication and structure analysis of poly(lactide-co-glycolic acid)/silk fibroin hybrid scaffold for wound dressing applications.

    Shahverdi, Sheida; Hajimiri, Mirhamed; Esfandiari, Mohammad Amin; Larijani, Bagher; Atyabi, Fatemeh; Rajabiani, Afsaneh; Dehpour, Ahmad Reza; Gharehaghaji, Ali Akbar; Dinarvand, Rassoul

    2014-10-01

    Silk fibroin (SF) and poly(lactide-co-glycolic acid) (PLGA) have been proved to be invaluable polymers in the field wound healing. This study aims at optimizing the electrospinning process of those polymers to make a hybrid membrane as a chronic wounds dressing. After characterizing the scaffolds, PLGA/SF (2:1), and PLGA scaffolds were selected for further study according to their superior tensile mechanical properties. The attachment and proliferation of mouse fibroblasts (L929) on scaffolds were measured using colorimetric assay and scanning electron microscopy. Furthermore, to evaluate the wound healing effect of the scaffolds in comparison with gauze and Comfeel() dressings, an excision wound model was conducted on diabetic rats. On the postoperative days of 3, 6, 9, 12, and 15, residual wound area was calculated using macroscopic data. In vitro results showed that the attachment and proliferation of L929 were significantly increased on PLGA/SF (2:1) hybrid scaffold. Animal study and histopathological evaluation outcomes confirmed the in vitro results as well. On day 15, the residual wound area in PLGA/SF (2:1) hybrid membrane group was significantly smaller than PLGA and control groups. This promising scaffold has the potential to be used for the upcoming development of wound dressings with or without biological drugs. PMID:25051110

  15. The Effect of Temperatures and Incubation Times on Some Properties of Silk Fibroin/Chitosan Blend Films

    Prasong Srihanam

    2011-01-01

    Full Text Available The objective of this work was to study the effect of temperatures and incubation times on Silk Fibroin (SF/Chitosan (CS blend films properties. The films were firstly prepared by mixing the SF and CS solution in homogeneously blended, then cast on the polystyrene culture plates before taking into the oven at 40C for 3 days. The obtained SF/CS blend films were treated at 90 and 120C, each for 30, 60 and 90 min, respectively. All of films were characterized for their morphology and secondary structures by using SEM and FTIR, respectively. The results showed that SF/CS films have homogeneous texture without phase separation. However, they also appeared some particles dispersed and embedded on their surfaces. The surfaces of the films slightly increased smooth texture when increased both temperatures and incubation times. The results were also noted that the characteristics were varied by those different temperatures. FTIR spectra indicated that the absorption bands at amide regions (I, II and III of the SF were similar profile. Moreover, treatment the blended films with temperatures resulted to changed the secondary structures of the films. This means the ratio of -sheet were increased. It can be expected that water evaporation by increasing temperature resulted to make the molecules of SF and CS close up and help to form H-bonds between them.

  16. The anticoagulant ability of ferulic acid and its applications for improving the blood compatibility of silk fibroin

    Wang Song; Gao Zhen; Chen Xiaomeng; Lian Xiaojie; Zhu Hesun [School of Material Science and Engineering, Beijing Institute of Technology, Beijing 100081 (China); Zheng Jun; Sun Lizhong [Department of Cardiac Surgery, Cardiovascular Institute and Fu Wai Hospital, CAMS and PUMC, Beijing 100037 (China)], E-mail: wangsongbit@hotmail.com

    2008-12-15

    The hemocompatibility of silk fibroin (SF) was improved with ferulic acid (FA) by graft polymerization. Ferulic acid is an active ingredient of many Chinese herbal medicines, such as Chuanxiong (Rhizoma ligustici wallichii), Danggui (Angelica sinensis) and Awei (Asafoetida giantfennel), which have been used to treat cardiovascular diseases by Chinese physicians for thousands of years. The inhibitory functions of FA on blood coagulation and erythrocyte agglutination were first characterized by a Lee-White test tube method and a micropipette technique, respectively. Then, FA was immobilized on SF by graft polymerization and the surface composition of modified SF was characterized by attenuated total reflectance Fourier-transform infrared (ATR-FTIR), x-ray photoelectron spectroscopy (XPS) and optical microscopy. The anticoagulant activity of modified SF was assessed, respectively, by in vitro clotting time measurements on a photo-optical clot detection instrument and with the Lee-White test tube method. The test results indicated that in comparison to untreated SF, the anticoagulant activity of modified SF has been improved significantly. Moreover, the SF surface composition is altered by FA but its {beta}-sheet conformation is not disturbed.

  17. The anticoagulant ability of ferulic acid and its applications for improving the blood compatibility of silk fibroin

    The hemocompatibility of silk fibroin (SF) was improved with ferulic acid (FA) by graft polymerization. Ferulic acid is an active ingredient of many Chinese herbal medicines, such as Chuanxiong (Rhizoma ligustici wallichii), Danggui (Angelica sinensis) and Awei (Asafoetida giantfennel), which have been used to treat cardiovascular diseases by Chinese physicians for thousands of years. The inhibitory functions of FA on blood coagulation and erythrocyte agglutination were first characterized by a Lee-White test tube method and a micropipette technique, respectively. Then, FA was immobilized on SF by graft polymerization and the surface composition of modified SF was characterized by attenuated total reflectance Fourier-transform infrared (ATR-FTIR), x-ray photoelectron spectroscopy (XPS) and optical microscopy. The anticoagulant activity of modified SF was assessed, respectively, by in vitro clotting time measurements on a photo-optical clot detection instrument and with the Lee-White test tube method. The test results indicated that in comparison to untreated SF, the anticoagulant activity of modified SF has been improved significantly. Moreover, the SF surface composition is altered by FA but its β-sheet conformation is not disturbed.

  18. Nano-scaled hydroxyapatite/silk fibroin sheets support osteogenic differentiation of rat bone marrow mesenchymal cells

    Tanaka, Toshimitsu [Research Institute for Cell Engineering (RICE), National Institute of Advanced Industrial Science and Technology (AIST) 3-11-46 Nakoji, Amagasaki, Hyogo 661-0974 (Japan); First Department of Oral and Maxillofacial Surgery, Tsurumi University School of Dental Medicine, 2-1-3 Tsurumi, Tsurumi-ku, Yokohama, Kanagawa 230-0063 (Japan); Hirose, Motohiro [Research Institute for Cell Engineering (RICE), National Institute of Advanced Industrial Science and Technology (AIST), 3-11-46 Nakoji, Amagasaki, Hyogo 661-0974 (Japan)]. E-mail: motohiro-hirose@aist.go.jp; Kotobuki, Noriko [Research Institute for Cell Engineering (RICE), National Institute of Advanced Industrial Science and Technology (AIST), 3-11-46 Nakoji, Amagasaki, Hyogo 661-0974 (Japan); Ohgushi, Hajime [Research Institute for Cell Engineering (RICE), National Institute of Advanced Industrial Science and Technology (AIST), 3-11-46 Nakoji, Amagasaki, Hyogo 661-0974 (Japan); Furuzono, Tsutomu [Department of Biomedical Engineering, National Cardiovascular Center Research Institute, 5-7-1 Fujishiro-dai, Suita, Osaka 565-8565 (Japan); Innovation Plaza Osaka, Japan Science and Technology Agency (JST), 3-1-10 Techno-stage, Izumi, Osaka 594-1144 (Japan); Sato, Junichi [First Department of Oral and Maxillofacial Surgery, Tsurumi University School of Dental Medicine, 2-1-3 Tsurumi, Tsurumi-ku, Yokohama, Kanagawa 230-0063 (Japan)

    2007-05-16

    A novel biomaterial that was composed of nano-scaled sintered hydroxyapatite (HAp) and silk fibroin (SF) was fabricated. We cultured rat marrow mesenchymal cells (MMCs) on this biomaterial (nano-HAp/SF sheet), on bare SF sheets, and on tissue culture polystyrene (TCPS) dishes as controls, then evaluated cell adhesion, proliferation, and differentiation of the MMCs. After 1 h of culture, a large number of viable cells were observed on the nano-HAp/SF sheets in comparison to the controls. In addition, after 3 h of culture, the morphology of the cells on the nano-HAp/SF sheets was quite different from that on the SF sheets. MMCs extrude their cytoplasmic processes to nano-HAp particles and are well attached to the sheets. After 14 days of culture, under osteogenic conditions, the alkaline phosphatase (ALP) activity and bone-specific osteocalcin secretion of the cells on nano-HAp/SF sheets were higher than were those on the controls. These results indicated that the surface of the nano-HAp/SF sheets is covered with appropriate HAp crystal for MMC adhesion/proliferation and that the sheets effectively support the osteogenic differentiation of MMCs. Therefore, the nano-HAp/SF sheet is an effective biomaterial that is applicable in bone reconstruction surgery.

  19. Silk fibroin/gelatin–chondroitin sulfate–hyaluronic acid effectively enhances in vitro chondrogenesis of bone marrow mesenchymal stem cells

    Tissue engineering is becoming promising for cartilage repair due to the limited self-repair capacity of cartilage tissue. We previously fabricated and characterized a three-dimensional silk fibroin/gelatin–chondroitin sulfate–hyaluronic acid (SF–GCH) scaffold and showed that it could promote proliferation of human bone marrow mesenchymal stem cells (BM-MSCs). This study aimed to evaluate its biological performance as a new biomimetic material for chondrogenic induction of BM-MSCs in comparison to an SF scaffold and conventional pellet culture. We found that the SF–GCH scaffold significantly enhanced the proliferation and chondrogenic differentiation of BM-MSCs compared to the SF scaffold and pellet culture in which the production of sulfated glycoaminoglycan was increased in concordance with the up-regulation of chondrogenic-specific gene markers. Our findings indicate the significant role of SF–GCH by providing a supportive structure and the mimetic cartilage environment for chondrogenesis which enables cartilage regeneration. Thus, our fabricated SF–GCH scaffold may serve as a potential biomimetic material for cartilage tissue engineering. - Highlights: • SF–GCH scaffold enhances proliferation and chondrogenic differentiation of BM-MSCs. • SF–GCH acts as a supportive and biomimetic material for BM-MSC chondrogenesis. • SF–GCH is a potential biomimetic scaffold suitable for cartilage tissue engineering

  20. Influence of the protocol used for fibroin extraction on the mechanical properties and fiber sizes of electrospun silk mats.

    Aznar-Cervantes, Salvador D; Vicente-Cervantes, Daniel; Meseguer-Olmo, Luis; Cenis, José L; Lozano-Pérez, A Abel

    2013-05-01

    Silk fibroin (SF) was regenerated using three of the most common protocols described in the bibliography for the dissolution of raw SF (LiBr 9.3M, CaCl2 50 wt.% or CaCl2:EtOH:H2O 1:2:8 in molar ratio). The integrity of regenerated SF in aqueous solution was analyzed by SDS-PAGE and different profiles of degradation were observed depending on the protocol used. This fact was found to affect also the aqueous solubility of the freeze dried protein. These different SFs were used to produce electrospun mats using SF solutions of SF 17 wt.% in 1,1,1,1',1',1'-hexafluoro-2-propanol (HFIP) and significant differences in fiber sizes, elongation and ultimate strength values were found. This work provides a global overview of the manner that different methods of SF extraction can affect the properties of electrospun SF-mats and consequently it should be considered depending on the use they are going to be made for. PMID:23498216

  1. Fabrication and characterization of layered chitosan/silk fibroin/nano-hydroxyapatite scaffolds with designed composition and mechanical properties.

    Zhou, Ting; Wu, Jingjing; Liu, Jiaoyan; Luo, Ying; Wan, Ying

    2015-08-01

    Chitosan/nano-hydroxyapatite (HA) composites were first prepared and then used together with chitosan and silk fibroin (SF) to produce a type of four-layer porous scaffold that is potentially applicable for articular cartilage repair. The bottom layer of the scaffold was built with the chitosan/HA composite and the other three layers of the scaffold were fabricated using chitosan/SF composites in which the content of the chitosan and SF was altered in a mutually reversed trend. The so-produced chitosan/SF/HA scaffolds were further crosslinked using tripolyphosphate to achieve enhanced mechanical properties. Interconnected porous microstructures throughout the scaffolds were constructed using a temperature gradient processing technique, and the resultant scaffolds were endowed with graded pore-sizes and porosities as well as porous interface zones between contiguous layers without visual clefts. The compressive modulus and stress at 10% strain of the scaffolds in wet state showed a gradient-changed trend which partially mimics the compressive mechanical properties of an articular cartilage matrix. Cell culture on some chitosan/SF/HA scaffolds for a period of time of up to 14 d showed that the scaffolds were able to well support the growth and infiltration of cells, suggesting that the presently developed chitosan/SF/HA scaffolds have promising potential for articular cartilage repair. PMID:26225911

  2. Tissue performance of bladder following stretched electrospun silk fibroin matrix and bladder acellular matrix implantation in a rabbit model.

    Huang, Jian-Wen; Xu, Yue-Min; Li, Zhao-Bo; Murphy, Sean V; Zhao, Weixin; Liu, Qiang-Qiang; Zhu, Wei-Dong; Fu, Qiang; Zhang, Yao-Peng; Song, Lu-Jie

    2016-01-01

    The goal of this study was to investigate the tissue performance of bladder following stretched electrospun silk fibroin matrix (SESFM) implantation compared with bladder acellular matrix (BAM). We compared SESFM with BAM based on porosity and pore size. Scaffolds were separately transplanted into opposite walls of the bladder of 30 rabbits after stripping the bladder mucosa and smooth muscle (1.5 2.0 cm(2) ). Gross anatomical observation, histological analysis and muscle contractility studies were performed at 2, 4, and 8 weeks post-op. SESFM has higher porosity and larger pore size compared with BAM (p??0.05). However, SESFM promoted a higher degree of smooth muscle and vessel regeneration compared to BAM (p?muscle strips supported by SESFM displayed higher contractile responses to carbachol, KCl, and phenylephrine compared with BAM. At 8 weeks, both matrices elicited similar mild acute and chronic inflammatory reactions. Our results demonstrated that SESFM has greater ability to promote bladder tissue regeneration with structural and functional properties compared to BAM, and with similar biocompatibility. 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 9-16, 2016. PMID:26148477

  3. Nano-scaled hydroxyapatite/silk fibroin sheets support osteogenic differentiation of rat bone marrow mesenchymal cells

    A novel biomaterial that was composed of nano-scaled sintered hydroxyapatite (HAp) and silk fibroin (SF) was fabricated. We cultured rat marrow mesenchymal cells (MMCs) on this biomaterial (nano-HAp/SF sheet), on bare SF sheets, and on tissue culture polystyrene (TCPS) dishes as controls, then evaluated cell adhesion, proliferation, and differentiation of the MMCs. After 1 h of culture, a large number of viable cells were observed on the nano-HAp/SF sheets in comparison to the controls. In addition, after 3 h of culture, the morphology of the cells on the nano-HAp/SF sheets was quite different from that on the SF sheets. MMCs extrude their cytoplasmic processes to nano-HAp particles and are well attached to the sheets. After 14 days of culture, under osteogenic conditions, the alkaline phosphatase (ALP) activity and bone-specific osteocalcin secretion of the cells on nano-HAp/SF sheets were higher than were those on the controls. These results indicated that the surface of the nano-HAp/SF sheets is covered with appropriate HAp crystal for MMC adhesion/proliferation and that the sheets effectively support the osteogenic differentiation of MMCs. Therefore, the nano-HAp/SF sheet is an effective biomaterial that is applicable in bone reconstruction surgery

  4. Silk fibroin/gelatin–chondroitin sulfate–hyaluronic acid effectively enhances in vitro chondrogenesis of bone marrow mesenchymal stem cells

    Sawatjui, Nopporn [Biomedical Sciences, Graduate School, Khon Kaen University, Khon Kaen 40002 (Thailand); Centre for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen 40002 (Thailand); Damrongrungruang, Teerasak [Department of Oral Diagnosis, Faculty of Dentistry, Khon Kaen University, Khon Kaen 40002 (Thailand); Leeanansaksiri, Wilairat [Stem Cell Therapy and Transplantation Research Group, Suranaree University of Technology, Nakhon Ratchasima 30000 (Thailand); School of Microbiology, Suranaree University of Technology, Nakhon Ratchasima 30000 (Thailand); Jearanaikoon, Patcharee [Centre for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen 40002 (Thailand); Hongeng, Suradej [Department of Pediatrics, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok 10400 (Thailand); Limpaiboon, Temduang, E-mail: temduang@kku.ac.th [Centre for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen 40002 (Thailand)

    2015-07-01

    Tissue engineering is becoming promising for cartilage repair due to the limited self-repair capacity of cartilage tissue. We previously fabricated and characterized a three-dimensional silk fibroin/gelatin–chondroitin sulfate–hyaluronic acid (SF–GCH) scaffold and showed that it could promote proliferation of human bone marrow mesenchymal stem cells (BM-MSCs). This study aimed to evaluate its biological performance as a new biomimetic material for chondrogenic induction of BM-MSCs in comparison to an SF scaffold and conventional pellet culture. We found that the SF–GCH scaffold significantly enhanced the proliferation and chondrogenic differentiation of BM-MSCs compared to the SF scaffold and pellet culture in which the production of sulfated glycoaminoglycan was increased in concordance with the up-regulation of chondrogenic-specific gene markers. Our findings indicate the significant role of SF–GCH by providing a supportive structure and the mimetic cartilage environment for chondrogenesis which enables cartilage regeneration. Thus, our fabricated SF–GCH scaffold may serve as a potential biomimetic material for cartilage tissue engineering. - Highlights: • SF–GCH scaffold enhances proliferation and chondrogenic differentiation of BM-MSCs. • SF–GCH acts as a supportive and biomimetic material for BM-MSC chondrogenesis. • SF–GCH is a potential biomimetic scaffold suitable for cartilage tissue engineering.

  5. Mimicked cartilage scaffolds of silk fibroin/hyaluronic acid with stem cells for osteoarthritis surgery: Morphological, mechanical, and physical clues.

    Jaipaew, Jirayut; Wangkulangkul, Piyanun; Meesane, Jirut; Raungrut, Pritsana; Puttawibul, Puttisak

    2016-07-01

    Osteoarthritis is a critical disease that comes from degeneration of cartilage tissue. In severe cases surgery is generally required. Tissue engineering using scaffolds with stem cell transplantation is an attractive approach and a challenge for orthopedic surgery. For sample preparation, silk fibroin (SF)/hyaluronic acid (HA) scaffolds in different ratios of SF/HA (w/w) (i.e., 100:0, 90:10, 80:20, and 70:30) were formed by freeze-drying. The morphological, mechanical, and physical clues were considered in this research. The morphological structure of the scaffolds was observed by scanning electron microscope. The mechanical and physical properties of the scaffolds were analyzed by compressive and swelling ratio testing, respectively. For the cell experiments, scaffolds were seeded and cultured with human umbilical cord-derived mesenchymal stem cells (HUMSCs). The cultured scaffolds were tested for cell viability, histochemistry, immunohistochemistry, and gene expression. The SF with HA scaffolds showed regular porous structures. Those scaffolds had a soft and elastic characteristic with a high swelling ratio and water uptake. The SF/HA scaffolds showed a spheroid structure of the cells in the porous structure particularly in the SF80 and SF70 scaffolds. Cells could express Col2a, Agg, and Sox9 which are markers for chondrogenesis. It could be deduced that SF/HA scaffolds showed significant clues for suitability in cartilage tissue engineering and in surgery for osteoarthritis. PMID:27127042

  6. Slip knots and unfastening topologies enhance toughness without reducing strength of silk fibroin fibres.

    Berardo, Alice; Pantano, Maria F; Pugno, Nicola M

    2016-02-01

    The combination of high strength and high toughness is a desirable feature that structural materials should display. However, while in the past, engineers had to compromise on either strength or toughness depending on the requested application, nowadays, new toughening strategies are available to provide strong materials with high toughness. In this paper, we focus on one of such strategy, which requires no chemical treatment, but the implementation of slip knots with optimized shape and size in the involved material, which is silkworm silk in this case. In particular, a variety of slip knot topologies with different unfastening mechanisms are investigated, including even complex knots usually used in the textile industry, and their efficiency in enhancing toughness of silk fibres is discussed. PMID:26855750

  7. Spermine-modified Antheraea pernyi silk fibroin as a gene delivery carrier

    Yu Y.; Hu Y.; Li X.; Liu Y.; Li M.; Yang J.; Sheng W.

    2016-01-01

    Yanni Yu,1 Yongpei Hu,1 Xiufang Li,1 Yu Liu,1 Mingzhong Li,1 Jicheng Yang,2 Weihua Sheng2 1National Engineering Laboratory for Modern Silk, College of Textile and Clothing Engineering, Soochow University, 2Cell and Molecular Biology Institute, College of Medicine, Soochow University, Suzhou, People’s Republic of China Abstract: The development of a novel cationized polymer used as a gene delivery carrier that can conveniently and effectively transfect cells resulting in a stably ...

  8. ToF-SIMS Characterization of Silk Fibroin and Polypyrrole Composite Actuators

    Bradshaw, Nathan P.; Severt, Sean Y.; Wang, Zhaoying; Fengel, Carly V.; Larson, Jesse D.; Zhu, Zihua; Murphy, Amanda; Leger, Janelle M.

    2015-08-31

    Biocompatible materials capable of controlled actuation under biologically relevant conditions are in high demand for use in a number of biomedical applications. Recently, we demonstrated that a composite material composed of silk biopolymer and the conducting polymer poly(pyrrole) can bend under an applied voltage using a simple bilayer device. Here we present further characterization of these bilayer actuators using time of flight secondary ion mass spectrometry, and provide clarification on the mechanism of actuation and factors affecting device performance and stability. We will discuss the results of this study in the context of strategies for optimization of device performance.

  9. Bioactivity of porous biphasic calcium phosphate enhanced by recombinant human bone morphogenetic protein 2/silk fibroin microsphere.

    Chen, Liang; Gu, Yong; Feng, Yu; Zhu, Xue-Song; Wang, Chun-Zeng; Liu, Hai-Long; Niu, Hai-Yun; Zhang, Chi; Yang, Hui-Lin

    2014-07-01

    To prepare a bioactive bone substitute, which integrates biphasic calcium phosphate (BCP) and rhBMP-2/silk fibroin (SF) microsphere, and to evaluate its characteristics. Hydroxyapatite and ?-tricalcium phosphate were integrated with a ratio of 6040%. RhBMP-2/SF (0.5 ?g/1 mg) microsphere was prepared, and its rhBMP-2-release kinetics was assed. After joining pore-forming agent (Sodium chloride, NaCl), porous BCP/rhBMP-2/SF were manufactured, and its characteristics and bioactivity in vitro were evaluated. Mean diameter of rhBMP-2/SF microsphere was 398.7 99.86 nm, with a loading rate of 4.53 0.08%. RhBMP-2 was released in a dual-phase pattern, of which fast-release (nearly half of protein released) focused on the initial 3 days, and slow-release sustained more than 28 days. With the increase in concentration of NaCl, greater was porosity and pore size, but smaller mechanical strength of BCP/rhBMP-2/SF. Material with 150% (w/v) NaCl had an optimal performance, with a porosity of 78.83%, pore size of 293.25 42.77?m and mechanical strength of 31.03 MPa. Proliferation of human placenta-derived mesenchymal stem cells (hPMSCs) on leaching extract medium was similar to the normal medium (P = 0.89), which was better than that on control group (P = 0.03). Activity of alkaline phosphatase on BCP/rhBMP-2/SF surface was higher than on pure BCP at each time point except at 1 day (P agent has excellent porosity, pore size and mechanical strength. The biomaterial induces proliferation and differentiation hPMSCs effectively. PMID:24659100

  10. Silk fibroin/sodium alginate composite nano-fibrous scaffold prepared through thermally induced phase-separation (TIPS) method for biomedical applications

    To mimic the natural fibrous structure of the tissue extracellular matrix, a nano-fibrous silk fibroin (SF)/sodium alginate (SA) composite scaffold was fabricated by a thermally-induced phase-separation method. The effects of SF/SA ratio on the structure and the porosity of the composite scaffolds were examined. Scanning electron microscopy and porosity results showed that the 5SF/1SA and 3SF/1SA scaffolds possessed an excellent nano-fibrous structure and a porosity of more than 90%. Fourier transform infrared, X-ray diffraction, and differential scanning calorimetry results indicated the physical interaction between SF and SA molecules and their good compatibility in the 5SF/1SA and 3SF/1SA scaffolds, whereas they showed less compatibility in the 1SF/1SA scaffold. Cell culture results showed that MG-63 cells can attach and grow well on the surface of the SF/SA scaffolds. The nano-fibrous SF/SA scaffold can be potentially used in tissue engineering. - Highlights: • We fabricate a nano-fibrous silk fibroin (SF)/sodium alginate (SA) composite scaffold. • The scaffold was prepared through a thermally induced phase separation method. • SF molecules are physically interacted with SA molecules. • Good molecular compatibility can be found in 5SF/1SA and 3SF/1SA scaffolds. • The nano-fibrous SF/SA scaffold is biocompatible

  11. Silk fibroin/sodium alginate composite nano-fibrous scaffold prepared through thermally induced phase-separation (TIPS) method for biomedical applications

    Zhang, Haiping, E-mail: zhp9810_a@163.com; Liu, Xiaotian, E-mail: xtianliu@126.com; Yang, Mingying, E-mail: yangm@zju.edu.cn; Zhu, Liangjun, E-mail: ljzhu@zju.edu.cn

    2015-10-01

    To mimic the natural fibrous structure of the tissue extracellular matrix, a nano-fibrous silk fibroin (SF)/sodium alginate (SA) composite scaffold was fabricated by a thermally-induced phase-separation method. The effects of SF/SA ratio on the structure and the porosity of the composite scaffolds were examined. Scanning electron microscopy and porosity results showed that the 5SF/1SA and 3SF/1SA scaffolds possessed an excellent nano-fibrous structure and a porosity of more than 90%. Fourier transform infrared, X-ray diffraction, and differential scanning calorimetry results indicated the physical interaction between SF and SA molecules and their good compatibility in the 5SF/1SA and 3SF/1SA scaffolds, whereas they showed less compatibility in the 1SF/1SA scaffold. Cell culture results showed that MG-63 cells can attach and grow well on the surface of the SF/SA scaffolds. The nano-fibrous SF/SA scaffold can be potentially used in tissue engineering. - Highlights: • We fabricate a nano-fibrous silk fibroin (SF)/sodium alginate (SA) composite scaffold. • The scaffold was prepared through a thermally induced phase separation method. • SF molecules are physically interacted with SA molecules. • Good molecular compatibility can be found in 5SF/1SA and 3SF/1SA scaffolds. • The nano-fibrous SF/SA scaffold is biocompatible.

  12. DNA Synthesis in the Giant Nuclei of Insects - Control Machinery and Structures Observed in the Silk-Producing Gland of Bombyx Mori

    The existence in many insect organs of giant nuclei without visible chromosomes raises the question of possible homologies between the chromatin structures of these nuclei and those of polytene nuclei or common euploid cells. Studies have been made of the nuclei in the silk-producing gland of Bombyx mori. The DNA synthesis is cyclic. During the third stage there are three successive synthesis cycles, which appear to be relatively autonomous in the individual nuclei. For more than 24 hours after moulting, however, synthesis is greatly reduced; moulting factors thus cause synchronization of all the nuclei. This leads to the conclusion that the triggering of a synthesis cycle is controlled by general factors external to the cell. At the end of larval development, DNA synthesis is suspended at the moment when large-scale secretion of silk begins. Evaluation of the pool of endogenic precursors of DNA shows that it is considerably reduced at the end of the DNA synthesis period. The hypothesis proposed is that large-scale synthesis of fibroin requires polarization of the metabolism, hence the depletion of the nucleotide pool and the end of DNA synthesis. DNA synthesis within a single nucleus is to some extent asynchronic. In particular, a well-defined, delayed-synthesis structure visible only in the female seems to be a possible homologue of a sex chromosome. Other asynchronisms are also apparent, though less clearly. Functional studies thus allow the supposition that in the giant nucleus replication units retain an individuality comparable to that of a polytene chromosome. These observations together lead to the conclusion that a nucleus in the silk-producing gland has physiological and structural characteristics similar to those of a polytene nucleus, differing from it essentially in the lesser degree of condensation of its structures. (author)

  13. New application of silk protein

    Gunma prefecture is famous for sericulture and silk textile industry district in Japan. In Gunma prefecture, some kinds of new generation silk as high performance and high quality silk were developed. These silk are used not only for the new textile materials but also for new industrial materials. New application of silk protein, fibroin and sericin, is considered. (author)

  14. New application of silk protein

    Kamiishi, Youichi [Textile Research Institute of Gunma, Kiryu, Gunma (Japan)

    2000-03-01

    Gunma prefecture is famous for sericulture and silk textile industry district in Japan. In Gunma prefecture, some kinds of new generation silk as high performance and high quality silk were developed. These silk are used not only for the new textile materials but also for new industrial materials. New application of silk protein, fibroin and sericin, is considered. (author)

  15. Development of novel electrospun nanofibrous scaffold from P. Ricini And A. Mylitta silk fibroin blend with improved surface and biological properties.

    Panda, N; Bissoyi, A; Pramanik, K; Biswas, A

    2015-03-01

    Biomaterials that stimulate cell attachment and proliferation without any surface modification (e.g. RGD coating) provide potent and cost effective scaffold for regenerative medicine. This study assessed the physico-chemical properties and cell supportive potential of a silk fibroin blend scaffold derived from eri (Philosamia ricini) and tasar (Antheraea mylitta) silk (ET) respectively by electrospinning process. The scanning electron microscopy and transmission electron microscopy study found that the fiber diameters are in 200 to 800nm range with flat morphology. The porosity of ET scaffold is found to be 79±5% with majority of pore diameter between 2.5 to 5nm. Similarly, Bombyx mori (BM) silk fibroin and gelatin nanofibrous scaffolds were prepared and taken as control. The ultimate tensile strength of the ET and BM scaffold are found to be 1.83±0.13MPa and 1.47±0.10MPa respectively. The measured contact angle (a measure of hydrophilicity) for ET (54.7°±1.8°) is found to be lower than BM (62°±2.3°). The ability to deposit apatite over ET is comparable to that of BM nanofibers. All the scaffolds were seeded with cord blood derived mesenchymal stem cells (hMSCs) and cultured for 14days in vitro. The immunofluorescence study reveals enhanced cell attachment with higher metabolic activity for MSCs grown over ET than BM and gelatin. The ET scaffold also demonstrated expression of higher amount cell adhesion molecules (CD29/CD44) and higher proliferation rate than BM and gelatin as confirmed by MTT assay, DNA content estimation assay, flow cytometry study and SEM study. Overall, it may be concluded that ET scaffold may have potential in developing bone tissue grafts for clinical applications in the future. PMID:25579953

  16. Bioengineered neo-corneal endothelium using collagen type-I coated silk fibroin film.

    Kim, Eun Young; Tripathy, Nirmalya; Cho, Sun Ah; Joo, Choun-Ki; Lee, Dongwon; Khang, Gilson

    2015-12-01

    Corneal transplantation, a common surgical protocol for visual acuity improvement, is limited owing to shortage of high quality donor corneas and/or lack of accurate replication of structural and biochemical composition of native cornea in a scaffold. Construction of neo-corneas utilizing novel, biocompatible and biodegradable scaffold/film source, could possibly address such formidable challenges. Herein, we designed optically transparent, micro-structurally stable silk films surface-coated with collagen type-I (Col-I/SF) as an alternative scaffold source for bioengineering of neo-cornea. Morphological, structural characteristics and in vitro biological studies were performed using primary rabbit corneal endothelial cells (rCEnCs) as models. The Col-I/SF films demonstrated higher Ra (nm) values compared to the bare SF surfaces. In vitro biological studies showed a significant increment in initial cell attachment and proliferation of cultured rCEnCs on the Col-I/SF films with well-maintained characteristic polygonal shape of rCEnCs. Although any remarkable changes regarding the morphology, expression of ZO-1 and Na(+)/K(+)-ATPase were absent, however the cells were found to be capable of well-expressing their functional proteins which regulates functions of corneal endothelium. Collectively, these results strongly suggest Col-I/SF film for future corneal transplantation therapy. PMID:26433646

  17. Genome editing of BmFib-H gene provides an empty Bombyx mori silk gland for a highly efficient bioreactor.

    Ma, Sanyuan; Shi, Run; Wang, Xiaogang; Liu, Yuanyuan; Chang, Jiasong; Gao, Jie; Lu, Wei; Zhang, Jianduo; Zhao, Ping; Xia, Qingyou

    2014-01-01

    Evolution has produced some remarkable creatures, of which silk gland is a fascinating organ that exists in a variety of insects and almost half of the 34,000 spider species. The impressive ability to secrete huge amount of pure silk protein, and to store proteins at an extremely high concentration (up to 25%) make the silk gland of Bombyx mori hold great promise to be a cost-effective platform for production of recombinant proteins. However, the extremely low production yields of the numerous reported expression systems greatly hindered the exploration and application of silk gland bioreactors. Using customized zinc finger nucleases (ZFN), we successfully performed genome editing of Bmfib-H gene, which encodes the largest and most abundant silk protein, in B. mori with efficiency higher than any previously reported. The resulted Bmfib-H knocked-out B. mori showed a smaller and empty silk gland, abnormally developed posterior silk gland cells, an extremely thin cocoon that contain only sericin proteins, and a slightly heavier pupae. We also showed that removal of endogenous Bmfib-H protein could significantly increase the expression level of exogenous protein. Furthermore, we demonstrated that the bioreactor is suitable for large scale production of protein-based materials. PMID:25359576

  18. In vitro and in vivo studies on the cytotoxicity of irradiated silk fibroin against mouse melanoma tumor cell

    Byun, Eui-Baek [Team for Radiation Food Science and Biotechnology, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup 580-185 (Korea, Republic of); Division of Bioresources and Biosciences, Faculty of Agriculture, Graduate school of Kyushu University, 6-10-1 Hakozaki, Fukuoka 812-8581 (Japan); Sung, Nak-Yun [Team for Radiation Food Science and Biotechnology, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup 580-185 (Korea, Republic of); Kwon, Sun-Kyu [Team for Radiation Food Science and Biotechnology, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup 580-185 (Korea, Republic of); Graduate school of Food and Biotechnology, Korea University, Jochiwon 339-800 (Korea, Republic of); Song, Beom-Seok; Kim, Jae-Hun; Choi, Jong-il [Team for Radiation Food Science and Biotechnology, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup 580-185 (Korea, Republic of); Hwang, Han-Joon [Graduate school of Food and Biotechnology, Korea University, Jochiwon 339-800 (Korea, Republic of); Byun, Myung-Woo [Team for Radiation Food Science and Biotechnology, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup 580-185 (Korea, Republic of); Lee, Ju-Woon [Team for Radiation Food Science and Biotechnology, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup 580-185 (Korea, Republic of)], E-mail: sjwlee@kaeri.re.kr

    2009-07-15

    The physicochemical properties of proteins can be altered by irradiation. But, it is rarely that the researches on the functional properties of irradiated proteins have been reported. Fibroin is a fibrous protein derived from silkworm Bombyx mori and has been suggested as a biomaterial for biomedical application. Therefore, fibroin was selected as a model protein and was examined with the irradiation effects on the cytotoxicity of fibroin on tumor cell. The cytotoxicity of fibroin against mouse melanoma cell (B16BL6) showed a significant increase dependent upon the increase of irradiation dose. And also, the splenocyte proliferation activities of fibroin were increased by gamma irradiation. In addition, the oral administration of irradiated fibroin significantly increased the inhibition rate of tumor growth in tumor-bearing mouse model. The reason might be due to the change of protein structure by gamma irradiation and is being studied. From these result, it could be concluded that the irradiated fibroin might be a potential candidate as a valuable product in food and medical industry.

  19. In vitro and in vivo studies on the cytotoxicity of irradiated silk fibroin against mouse melanoma tumor cell

    Byun, Eui-Baek; Sung, Nak-Yun; Kwon, Sun-Kyu; Song, Beom-Seok; Kim, Jae-Hun; Choi, Jong-il; Hwang, Han-Joon; Byun, Myung-Woo; Lee, Ju-Woon

    2009-07-01

    The physicochemical properties of proteins can be altered by irradiation. But, it is rarely that the researches on the functional properties of irradiated proteins have been reported. Fibroin is a fibrous protein derived from silkworm Bombyx mori and has been suggested as a biomaterial for biomedical application. Therefore, fibroin was selected as a model protein and was examined with the irradiation effects on the cytotoxicity of fibroin on tumor cell. The cytotoxicity of fibroin against mouse melanoma cell (B16BL6) showed a significant increase dependent upon the increase of irradiation dose. And also, the splenocyte proliferation activities of fibroin were increased by gamma irradiation. In addition, the oral administration of irradiated fibroin significantly increased the inhibition rate of tumor growth in tumor-bearing mouse model. The reason might be due to the change of protein structure by gamma irradiation and is being studied. From these result, it could be concluded that the irradiated fibroin might be a potential candidate as a valuable product in food and medical industry.

  20. Physico-chemical properties and efficacy of silk fibroin fabric coated with different waxes as wound dressing.

    Kanokpanont, Sorada; Damrongsakkul, Siriporn; Ratanavaraporn, Juthamas; Aramwit, Pornanong

    2013-04-01

    Silk fibroin (SF) has been widely used as a wound dressing material due to its suitable physical and biological characteristics. In this study, a non-adhesive wound dressing which applies to cover the wound surface as an absorbent pad that would absorb wound fluid while accelerate wound healing was developed. The modification of SF fabrics by wax coating was purposed to prepare the non-adhesive wound dressing that is required in order to minimize pain and risk of repeated injury. SF woven fabrics were coated with different types of waxes including shellac wax, beeswax, or carnauba wax. Physical and mechanical properties of the wax-coated SF fabrics were characterized. It was clearly observed that all waxes could be successfully coated on the SF fabrics, possibly due to the hydrophobic interactions between hydrophobic domains of SF and waxes. The wax coating improved tensile modulus and percentage of elongation of the SF fabrics due to the denser structure and the thicker fibers coated. The in vitro degradation study demonstrated that all wax-coated SF fabrics remained up to 90% of their original weights after 7 weeks of incubation in lysozyme solution under physiological conditions. The wax coating did not affect the degradation behavior of the SF fabrics. A peel test of the wax-coated SF fabrics was carried out in the partial- and full-thickness wounds of porcine skin in comparison to that of the commercial wound dressing. Any wax-coated SF fabrics were less adhesive than the control, as confirmed by less number of cells attached and less adhesive force. This might be that the wax-coated SF fabrics showed the hydrophobic property, allowing the loosely adherence to the hydrophilic wound surface. In addition, the in vivo biocompatibility test of the wax-coated SF fabrics was performed in Sprague-Dawley rats with subcutaneous model. The irritation scores indicated that the carnauba wax-coated SF fabric was not irritant while the shellac wax or beeswax-coated SF fabrics were slightly irritant, comparing with the commercial wound dressing. Therefore, SF fabrics coated with waxes, particularly carnauba wax, would be promising choices of non-adhesive wound dressing. PMID:23313451

  1. Reducing blood glucose level in TIDM mice by orally administering the silk glands of transgenic hIGF-I silkworms.

    Cong, Liang; Cao, Guangli; Renyu, Xue; Zhonghua, Pan; Xiaojian, Zheng; Zhou, Wenlin; Gong, Chengliang

    2011-07-15

    To realize the secretory expression of human insulin-like growth factor-I (hIGF-I) in the posterior silk glands (PSGs) of transgenic silkworms, the piggyBac transposon vector pigA3GFP-fibHS-hIGF-i.e.-neo containing a neomycin-resistance gene (neo), green fluorescent protein gene (gfp) and human insulin-like growth factor I (hIGF-I) gene controlled by the Bombyxmori fibroin heavy chain gene (fib-H) promoter with its downstream signal peptide sequence, and a helper plasmid containing the piggyBac transposase sequence under the control of the B. mori actin 3 gene (A3) promoter were transferred into silkworm eggs by sperm-mediated gene transfer. Transformed silkworms were obtained after being screened for green fluorescence and by the antibiotic G418. In the PSGs of the transformed silkworms, a specific band representing hIGF-I could be detected by Western blotting, and the content of the hIGF-I estimated by ELISA was approximately 1.84 ?g/gram of cocoon and 19.18 ?g/gram of freeze-dried PSG powder. To further estimate the biological activity of the expressed hIGF-I, streptozotocin-induced TIDM mice were orally administered with the PSG powder of the transgenic silkworms, the results showed the blood glucose levels of mice were significantly reduced, suggesting that the the PSGs powder of transgenic hIGF-I silkworms could possibly be used as a perorally administered medicine. PMID:21672515

  2. Purification and some properties of G-factor from the silk gland of silkworm

    Purification of G-factor from the silk gland of silkworms and some of its properties have been studied in comparison with the G-factor from E. coli and rat liver transferase II. In the experiment, GTP-?-32P was prepared from GDP and 32Pi by photophosphorylation with spinach chloroplasts, and purified by Dowex-1 chromatography. The purified G-factor was homogeneous on disc gel electrophoresis. Molecular weight of 80,000 was estimated from a calibrated Sephadex G-200 column. The purified G-factor exhibited uncoupled ribosome-dependent GTPase activity, and was inhibited by fusidic acid, SH inhibitor, and diphtheria toxin plus NAD. The G-factor from E. coli and that from the silk gland were not interchangeable in GTPase reaction. (Yamanoto, Y.)

  3. Removal of copper ions from aqueous solution by adsorption onto novel polyelectrolyte film-coated nanofibrous silk fibroin non-wovens

    Zhou, Weitao, E-mail: weitao_zhou@yahoo.com [Key Laboratory of Functional Textiles, The Education Department of Henan Province, Zhongyuan University of Technology, Zhengzhou 450007 (China); Huang, Haitao [School of Textile, Henan Institute of Engineering, Zhengzhou 451191 (China); Du, Shan [Australian Future Fibers Research and Innovation Centre for Frontier Materials, Deakin University, Geelong, VIC 3217 (Australia); Huo, Yingdong; He, Jianxin [Key Laboratory of Functional Textiles, The Education Department of Henan Province, Zhongyuan University of Technology, Zhengzhou 450007 (China); Cui, Shizhong, E-mail: snowballer@163.com [Key Laboratory of Functional Textiles, The Education Department of Henan Province, Zhongyuan University of Technology, Zhengzhou 450007 (China)

    2015-08-01

    Graphical abstract: - Highlights: • Polyethylenimine coated silk fibroin nanofibrous nonwovens were fabricated. • The characteristics such as the fiber shape and porous structure were well maintained. • The structure and adsorption properties were studied. The adsorption property for copper ions is good. - Abstract: In this approach, polyelectrolyte film-coated nanofibrous silk fibroin (SF) nonwovens were prepared from the alternate deposition of positively charged polyethylenimine (PEI) and negatively charged SF using electrostatic layer-by-layer (LBL) self-assembled technology. The composite membranes were characterized by scanning electron microscopy (SEM) and Fourier transform infrared (FTIR) spectrometer. The SF-PEI multilayer-assembled nanofibers (less than five layers) were fine and uniform with the fiber diameter from 400 nm to 600 nm, and had very large surface area and high porosity (more than 70%). The amino groups of PEI were proved to be deposited onto SF nonwovens, which granted the coated nonwovens with potential applicability for copper ions adsorption. The PEI films coated SF substrate showed much higher copper ions adsorption capacity than that of ethanol treated SF nanofibers. Adding the number of PEI coated could enhance the Cu{sup 2+} adsorption capacity significantly. The maximum milligrams per gram of copper ions adsorbed reached 59.7 mg/g when the SF substrate was coated with 5 bilayers of SF-PEI. However, the copper ions adsorption capacity had no obvious change as the number of PEI continued to increase. These results suggest potential for PEL film-coated nanofibrous nonwovens as a new adsorbent for metal ions.

  4. Silk fibroin immobilization on poly(ethylene terephthalate) films: Comparison of two surface modification methods and their effect on mesenchymal stem cells culture

    Silk fibroin (SF) has played a curial role for the surface modification of conventional materials to improve the biocompatibility, and SF modified poly(ethylene terephthalate) (PET) materials have potential applications on tissue engineering such as artificial ligament, artificial vessel, artificial heart valve sewing cuffs dacron and surgical mesh engineering. In this work, SF was immobilized onto PET film via two different methods: 1) plasma pretreatment followed by SF dip coating (PET-SF) and 2) plasma-induce acrylic acid graft polymerization and subsequent covalent immobilization of SF on PET film (PET-PAA-SF). It could be found that plasma treatment provided higher surface roughness which was suitable for further SF dip coating, while grafted poly(acrylic acid) (PAA) promised the covalent bonding between SF and PAA. ATR-FTIR adsorption band at 3284 cm−1, 1623 cm−1 and 1520 cm−1 suggested the successful introduction of SF onto PET surface, while the amount of immobilized SF of PET-SF was higher than PET-PAA-SF according to XPS investigation (0.29 vs 0.23 for N/C ratio). Surface modified PET film was used as substrate for mesenchymal stem cells (MSCs) culture, the cells on PET-SF surface exhibited optimum density compared to PET-PAA-SF according to CCK-8 assays, which indicated that plasma pretreatment followed by SF dip coating was a simple and effective way to prepare biocompatible PET surface. Highlights: ► Silk fibroins were immobilized onto PET films with or without the linker of PAA. ► Various techniques were performed to characterize the modified surfaces ► Plasma treatment followed by SF dip coating introduced more SF onto PET films. ► Compare to PET-PAA-SF, PET-SF has better biocompatibility base on MSCs culture

  5. Silk fibroin immobilization on poly(ethylene terephthalate) films: Comparison of two surface modification methods and their effect on mesenchymal stem cells culture

    Liang, Meini; Yao, Jinrong; Chen, Xin; Huang, Lei; Shao, Zhengzhong, E-mail: zzshao@fudan.edu.cn

    2013-04-01

    Silk fibroin (SF) has played a curial role for the surface modification of conventional materials to improve the biocompatibility, and SF modified poly(ethylene terephthalate) (PET) materials have potential applications on tissue engineering such as artificial ligament, artificial vessel, artificial heart valve sewing cuffs dacron and surgical mesh engineering. In this work, SF was immobilized onto PET film via two different methods: 1) plasma pretreatment followed by SF dip coating (PET-SF) and 2) plasma-induce acrylic acid graft polymerization and subsequent covalent immobilization of SF on PET film (PET-PAA-SF). It could be found that plasma treatment provided higher surface roughness which was suitable for further SF dip coating, while grafted poly(acrylic acid) (PAA) promised the covalent bonding between SF and PAA. ATR-FTIR adsorption band at 3284 cm{sup ?1}, 1623 cm{sup ?1} and 1520 cm{sup ?1} suggested the successful introduction of SF onto PET surface, while the amount of immobilized SF of PET-SF was higher than PET-PAA-SF according to XPS investigation (0.29 vs 0.23 for N/C ratio). Surface modified PET film was used as substrate for mesenchymal stem cells (MSCs) culture, the cells on PET-SF surface exhibited optimum density compared to PET-PAA-SF according to CCK-8 assays, which indicated that plasma pretreatment followed by SF dip coating was a simple and effective way to prepare biocompatible PET surface. Highlights: ? Silk fibroins were immobilized onto PET films with or without the linker of PAA. ? Various techniques were performed to characterize the modified surfaces ? Plasma treatment followed by SF dip coating introduced more SF onto PET films. ? Compare to PET-PAA-SF, PET-SF has better biocompatibility base on MSCs culture.

  6. Silk fibroin nanoparticles constitute avector for controlled release of resveratrol inanexperimental model of inflammatory boweldisease in rats

    Lozano-Prez AA

    2014-09-01

    Full Text Available Antonio Abel Lozano-Prez,1 Alba Rodriguez-Nogales,2 Vctor Ortiz-Cullera,1 Francesca Algieri,2 Jos Garrido-Mesa,2 Pedro Zorrilla,2 M Elena Rodriguez-Cabezas,2 Natividad Garrido-Mesa,2 M Pilar Utrilla,2 Laura De Matteis,3 Jess Martnez delaFuente,3 Jos Luis Cenis,1 Julio Glvez2 1Instituto Murciano de Investigacin y Desarrollo Agrario y Alimentario, Murcia, Spain; 2Centro de Investigaciones Biomdicas en Red Enfermedades Hepticas y Digestivas, Department of Pharmacology, ibs Granada, Center for Biomedical Research, University of Granada, Granada, Spain; 3Instituto de Nanociencia de Aragn, Universidad de Zaragoza, Zaragoza, Spain Purpose: We aimed to evaluate the intestinal anti-inflammatory properties of silk fibroin nanoparticles, around 100nm in size, when loaded with the stilbene compound resveratrol, in an experimental model of rat colitis. Methods: Nanoparticles were loaded with resveratrol by adsorption. The biological effects of the resveratrol-loaded nanoparticles were tested both in vitro, in a cell culture of RAW 264.7cells (mouse macrophages, and in vivo, in the trinitrobenzenesulfonic acid model of rat colitis, when administered intracolonically.Results: The resveratrol liberation in 1 phosphate-buffered saline (PBS; pH 7.4 was characterized by fast liberation, reaching the solubility limit in 3hours, which was maintained over a period of 80hours. The in vitro assays revealed immunomodulatory properties exerted by these resveratrol-loaded nanoparticles since they promoted macrophage activity in basal conditions and inhibited this activity when stimulated with lipopolysaccharide. The in vivo experiments showed that after evaluation of the macroscopic symptoms, inflammatory markers, and intestinal barrier function, the fibroin nanoparticles loaded with resveratrol had a better effect than the single treatments, being similar to that produced by the glucocorticoid dexamethasone. Conclusion: Silk fibroin nanoparticles constitute an attractive strategy for the controlled release of resveratrol, showing immunomodulatory properties and intestinal anti-inflammatory effects. Keywords: immunomodulatory, cytokines, TNBS rat colitis, RAW 264.7 macrophage cells, antioxidant

  7. Enhancement of osseointegration of polyethylene terephthalate artificial ligament by coating of silk fibroin and depositing of hydroxyapatite

    Jiang J

    2014-09-01

    Full Text Available Jia Jiang,1,2 Fang Wan,1 Jianjun Yang,1 Wei Hao,3 Yaxian Wang,3 Jinrong Yao,3 Zhengzhong Shao,3 Peng Zhang,1 Jun Chen,1 Liang Zhou,4 Shiyi Chen11Fudan University Sports Medicine Center and Department of Sports Medicine and Arthroscopy Surgery, Huashan Hospital, 2State Key Laboratory of Molecular Engineering of Polymers, 3Laboratory of Advanced Materials, National Key Laboratory of Molecular Engineering of Polymers, Department of Macromolecular Science, Fudan University, Shanghai, People’s Republic of China; 4Department of Forest Products, Anhui Agricultural University, Hefei Anhui Province, People’s Republic of ChinaBackground: Application of artificial ligament in anterior cruciate ligament reconstruction is one of the research focuses of sports medicine but the biological tendon–bone healing still remains a problem. The preliminary study of hydroxyapatite (HAP coating on the polyethylene terephthalate (PET surface could effectively induce the osteoblast differentiation, but the tendon–bone healing was still not stable. As a green synthesis process, the biomimetic mineralization can simulate the natural bone growth in vitro and in vivo.Methods: HAP crystals were grown under the guide of silk fibroin (SF PET surface by biomimetic route. Several techniques including scanning electron microscopy, attenuated total reflectance Fourier transform infrared spectroscopy, X-ray diffraction, and energy-dispersive X-ray spectroscopy were utilized for proving the introduction of both SF and HAP. The viability and osseointegration of bone marrow stromal cells on the surface of three kinds of ligament, including PET group (non-coating group, PET+SF group (SF-coating group, and PET+SF+HAP group (combined HAP- and SF-coating group, were analyzed by CCK-8 assays and alkaline phosphatase (ALP detection. Seventy-two mature male New Zealand rabbits were randomly divided into three groups. Among them, 36 rabbits were sacrificed for mechanical testing, and histological examination for the others. Results: The SF and SF+HAP were successfully coated on the surface of PET fiber. The CCK-8 assay showed that the cell proliferation on PET+SF+HAP group was better than the other two groups from 24 to 120 hours. After 14 days of culture, the cells in the PET+SF+HAP group delivered higher levels of ALP than the other two groups. After 3 days of culture, the expression level of integrin β1 in the PET+SF+HAP group and PET+SF group were higher than in the PET group. The mean load to failure and the stiffness value of the PET+SF+HAP group were both higher than the other two groups. Hematoxylin and eosin staining showed that new bone tissue formation was only found in the PET+SF+HAP group 8 weeks postoperatively. Masson staining showed that in the PET+SF+HAP group 8 weeks postoperatively, the PET fibers were almost completely encircled by collagen. Histomorphometric analysis showed that the width of the graft–bone interface in the PET+SF+HAP group was narrower than that in the other two groups 4 and 8 weeks postoperatively. The mRNA level of BMP-7 in the PET+SF+HAP groups was significantly higher than those in the other two groups 4 and 8 weeks postoperatively.Conclusion: The study showed that the combined SF and HAP coating by biomimetic route on the surface of PET artificial ligament could induce graft osseointegration in the bone tunnel, providing theoretical and experimental foundation for manufacturing novel artificial ligaments meeting the clinical needs.Keywords: biomineralization, tendon–bone healing, ligament reconstruction

  8. Study of the effects of different sterilization methods on the properties of dense and porous silk fibroin membranes;Estudo dos efeitos de diferentes metodos de esterilizacao nas propriedades de membranas densas de fibroina de seda

    Weska, Raquel F.; Moraes, Mariana A. de; Beppu, Marisa M., E-mail: raquelweska@terra.com.b [Universidade Estadual de Campinas (UNICAMP), SP (Brazil). Fac. de Engenharia Quimica

    2009-07-01

    Silk fibroin has been widely explored for many biomedical applications, due to its biocompatibility and biodegradability. Sterilization is a fundamental step in biomaterials processing, and it must not alter in a negative way the functionality of medical devices. The aim of this study was to analyze the influence of different sterilization methods in the physical and chemical characteristics of dense silk fibroin membranes. Dense fibroin membranes were sterilized by ultraviolet radiation, 70% ethanol, autoclave, ethylene oxide and gamma radiation, and were analyzed by SEM, FTIR-ATR and XRD. The results for sterilization indicated that the methods didn't cause degradation of the membranes, but the methods that used organic solvent, or increase of humidity and/or temperature (70% ethanol, autoclave and ethylene oxide) altered the molecular conformation of fibroin, increasing the proportion of beta-sheet structure, what indicates an increase of crystallinity. This effect may be positive when a slower degradation of the membranes is desired, depending on the application as a bio material. (author)

  9. Differential binding of the Bombyx silk gland-specific factor SGFB to its target DNA sequence drives posterior-cell-restricted expression.

    Horard, B; Julien, E; Nony, P; Garel, A; Couble, P

    1997-01-01

    The gene encoding the silk protein P25 in Bombyx mori is expressed in the posterior silk gland (PSG) cells and repressed in the middle silk gland (MSG) cells. To identify the factors involved in this transcription-dependent spatial restriction, we examined the P25 chromatin in PSG and MSG nuclei by DNase I-aided ligation-mediated PCR and analyzed the expression of various P25-lacZ constructs in biolistically treated silk glands. P25 promoter activation depends on two cis-acting elements. One ...

  10. Designing silk fibroin-based matrices with ionic liquids for tissue engineering strategies using human adipose stem cells

    Silva, S.S; Popa, Elena Geta; Gomes, Manuela E.; Oliveira, Mariana B.; Mano, J.F; Reis, R.L

    2012-01-01

    Silk (SF) is an attractive biomaterial to be used in tissue engineering applications because of its excellent mechanical properties and biocompatibility [1, 2]. In this work, the cellular response of silk hydrogels produced through dissolution of this protein in ionic liquids (ILs) was investigated. For that, degummed fibers obtained from the cocoons of mulberry silkworm (Bombyx mori) were dissolved in an IL and the solution was gelified in ethanol, followed by IL removal from gels using soxh...

  11. Response of human mesenchymal stem cells to intrafibrillar nanohydroxyapatite content and extrafibrillar nanohydroxyapatite in biomimetic chitosan/silk fibroin/nanohydroxyapatite nanofibrous membrane scaffolds

    Lai GJ

    2015-01-01

    Full Text Available Guo-Jyun Lai,1,* KT Shalumon,1,* Jyh-Ping Chen1,2 1Department of Chemical and Materials Engineering, 2Research Center for Industry of Human Ecology, Chang Gung University of Science and Technology, Taoyuan, Taiwan, Republic of China *These authors contributed equally to this work Abstract: Incorporation of nanohydroxyapatite (nHAP within a chitosan (CS/silk fibroin (SF nanofibrous membrane scaffold (NMS may provide a favorable microenvironment that more closely mimics the natural bone tissue physiology and facilitates enhanced osteogensis of the implanted cell population. In this study, we prepared pristine CS/SF NMS, composite CS/SF/nHAP NMS containing intrafibrillar nHAP by in situ blending of 10% or 30% nHAP before the electrospinning step, and composite CS/SF/nHAP NMS containing extrafibrillar nHAP by depositing 30% nHAP through alternative soaking surface mineralization. We investigated the effect of the incorporation of HAP nanoparticles on the physicochemical properties of pristine and composite NMS. We confirmed the presence of ~30 nm nHAP in the composite nanofibrous membranes by thermogravimetry analysis (TGA, X-ray diffraction (XRD, and scanning electron microscopy (SEM, either embedded in or exposed on the nanofiber. Nonetheless, the alternative soaking surface mineralization method drastically influenced the mechanical properties of the NMS with 88% and 94% drop in Young’s modulus and ultimate maximum stress. Using in vitro cell culture experiments, we investigated the effects of nHAP content and location on proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs. The proliferation of hMSCs showed no significant difference among pristine and composite NMS. However, the extent of osteogenic differentiation of hMSCs was found to be positively correlated with the content of nHAP in the NMS, while its location within the nanofiber played a less significant role. In vivo experiments were carried out with hMSCs seeded in CS/SF/30%nHAP NMS prepared by in situ blending and subcutaneous implantation in nude mice. Micro-computed tomography images as well as histological and immunohistochemical analysis of the retrieved hMSCs/NMS construct 1 and 2 months postimplantation indicated that NMS had the potential for bone regeneration and can be suggested as a promising scaffold for bone tissue engineering. Keywords: chitosan, silk fibroin, hydroxyapatite, electrospinning, nanofibers, human bone marrow stem cells, bone tissue engineering

  12. Critical electrolyte concentration of silk gland chromatin of the sugarcane borer Diatraea saccharalis, induced using agrochemicals.

    Santos, S A; Fermino, F; Moreira, B M T; Araujo, K F; Falco, J R P; Ruvolo-Takasusuki, M C C

    2014-01-01

    The sugarcane borer Diatraea saccharalis is widely known as the main pest of sugarcane crop, causing increased damage to the entire fields. Measures to control this pest involve the use of chemicals and biological control with Cotesia flavipes wasps. In this study, we evaluated the insecticides fipronil (Frontline; 0.0025%), malathion (Malatol Bio Carb; 0.4%), cipermetrina (Galgotrin; 10%), and neem oil (Natuneem; 100%) and the herbicide nicosulfuron (Sanson 40 SC; 100%) in the posterior region silk glands of 3rd- and 5th-instar D. saccharalis by studying the variation in the critical electrolyte concentration (CEC). Observations of 3rd-instar larvae indicated that malathion, cipermetrina, and neem oil induced increased chromatin condensation that may consequently disable genes. Tests with fipronil showed no alteration in chromatin condensation. With the use of nicosulfuron, there was chromatin and probable gene decompaction. In the 5th-instar larvae, the larval CEC values indicated that malathion and neem oil induced increased chromatin condensation. The CEC values for 5th-instar larvae using cipermetrina, fipronil, and nicosulfuron indicated chromatin unpacking. These observations led us to conclude that the quantity of the pesticide does not affect the mortality of these pests, can change the conformation of complexes of DNA, RNA, and protein from the posterior region of silk gland cells of D. saccharalis, activating or repressing the expression of genes related to the defense mechanism of the insect and contributing to the selection and survival of resistant individuals. PMID:25299111

  13. DNA Synthesis during Endomitosis Is Stimulated by Insulin via the PI3K/Akt and TOR Signaling Pathways in the Silk Gland Cells of Bombyx mori

    Yaofeng Li

    2015-03-01

    Full Text Available Silk gland cells undergo multiple endomitotic cell cycles during silkworm larval ontogeny. Our previous study demonstrated that feeding is required for continued endomitosis in the silk gland cells of silkworm larvae. Furthermore, the insulin signaling pathway is closely related to nutritional signals. To investigate whether the insulin signaling pathway is involved in endomitosis in silk gland cells, in this study, we initially analyzed the effects of bovine insulin on DNA synthesis in endomitotic silk gland cells using 5-bromo-2'-deoxyuridine (BrdU labeling technology, and found that bovine insulin can stimulate DNA synthesis. Insulin signal transduction is mainly mediated via phosphoinositide 3-kinase (PI3K/Akt, the target of rapamycin (TOR and the extracellular signal-regulated kinase (ERK pathways in vertebrates. We ascertained that these three pathways are involved in DNA synthesis in endomitotic silk gland cells using specific inhibitors against each pathway. Moreover, we investigated whether these three pathways are involved in insulin-stimulated DNA synthesis in endomitotic silk gland cells, and found that the PI3K/Akt and TOR pathways, but not the ERK pathway, are involved in this process. These results provide an important theoretical foundation for the further investigations of the mechanism underlying efficient endomitosis in silk gland cells.

  14. Quantitative analyses of the effect of silk fibroin/nano-hydroxyapatite composites on osteogenic differentiation of MG-63 human osteosarcoma cells.

    Lin, Linxue; Hao, Runsong; Xiong, Wei; Zhong, Jian

    2015-05-01

    Silk fibroin (SF)/nano-hydroxyapatite (n-HA) composites are potential biomaterials for bone defect repair. Up to now, the biological evaluation studies of SF/n-HA composites have primarily concentrated on their biocompatibility at cell level such as cell viability and proliferation and tissue level such as material absorption and new bone formation. In this work, SF/n-HA composites were fabricated using a simplified coprecipitation methods and were deposited onto Ti alloy substrates. Then the cell adhesion ability of SF/n-HA composites was observed by SEM and cell proliferation ability of SF/n-HA composites was determined by MTT assay. The ALP activity, BGP contents, and Col I contents of MG-63 human osteosarcoma cells on SF/n-HA composites were quantitatively analyzed. HA nanocrystals were used as controls. These experiments showed that SF/n-HA composites had better cell adhesion and osteogenic differentiation abilities than n-HA materials. This work provides quantitative data to analyze the effect of SF/n-HA composites on cell osteogenic differentiation. PMID:25454062

  15. Amperometric biosensor based on tyrosinase immobilized onto multiwalled carbon nanotubes-cobalt phthalocyanine-silk fibroin film and its application to determine bisphenol A

    An amperometric bisphenol A (BPA) biosensor was fabricated by immobilizing tyrosinase on multiwalled carbon nanotubes (MWNTs)-cobalt phthalocyanine (CoPc)-silk fibroin (SF) composite modified glassy carbon electrode (GCE). In MWNTs-CoPc-SF composite film, SF provided a biocompatible microenvironment for the tyrosinase to retain its bioactivity, MWNTs possessed excellent inherent conductivity to enhance the electron transfer rate and CoPc showed good electrocatalytic activity to electrooxidation of BPA. The cyclic voltammogram of BPA at this biosensor exhibited a well defined anodic peak at 0.625 V. Compared with bare GCE, the oxidation signal of BPA significantly increased; therefore, this oxidation signal was used to determine BPA. The effect factors were optimized and the electrochemical parameters were calculated. The possible oxidation mechanism was also discussed. Under optimum conditions, the oxidation current was proportional to BPA concentration in the range from 5.0 x 10-8 to 3.0 x 10-6 M with correlation coefficient of 0.9979 and detection limit of 3.0 x 10-8 M (S/N = 3). The proposed method was successfully applied to determine BPA in plastic products and the recovery was in the range from 95.36% to 104.39%.

  16. Amperometric biosensor based on tyrosinase immobilized onto multiwalled carbon nanotubes-cobalt phthalocyanine-silk fibroin film and its application to determine bisphenol A

    Yin Huanshun [College of Chemistry and Material Science, Shandong Agricultural University, Taian 271018, Shandong (China); College of Resources and Environment, Shandong Agricultural University, Taian 271018, Shandong (China); Zhou Yunlei; Xu Jing [College of Chemistry and Material Science, Shandong Agricultural University, Taian 271018, Shandong (China); Ai Shiyun, E-mail: ashy@sdau.edu.cn [College of Chemistry and Material Science, Shandong Agricultural University, Taian 271018, Shandong (China); Cui Lin [College of Chemistry and Material Science, Shandong Agricultural University, Taian 271018, Shandong (China); Zhu Lusheng, E-mail: lushzhu@sdau.edu.cn [College of Resources and Environment, Shandong Agricultural University, Taian 271018, Shandong (China)

    2010-02-05

    An amperometric bisphenol A (BPA) biosensor was fabricated by immobilizing tyrosinase on multiwalled carbon nanotubes (MWNTs)-cobalt phthalocyanine (CoPc)-silk fibroin (SF) composite modified glassy carbon electrode (GCE). In MWNTs-CoPc-SF composite film, SF provided a biocompatible microenvironment for the tyrosinase to retain its bioactivity, MWNTs possessed excellent inherent conductivity to enhance the electron transfer rate and CoPc showed good electrocatalytic activity to electrooxidation of BPA. The cyclic voltammogram of BPA at this biosensor exhibited a well defined anodic peak at 0.625 V. Compared with bare GCE, the oxidation signal of BPA significantly increased; therefore, this oxidation signal was used to determine BPA. The effect factors were optimized and the electrochemical parameters were calculated. The possible oxidation mechanism was also discussed. Under optimum conditions, the oxidation current was proportional to BPA concentration in the range from 5.0 x 10{sup -8} to 3.0 x 10{sup -6} M with correlation coefficient of 0.9979 and detection limit of 3.0 x 10{sup -8} M (S/N = 3). The proposed method was successfully applied to determine BPA in plastic products and the recovery was in the range from 95.36% to 104.39%.

  17. A Silk Fibroin/Collagen Nerve Scaffold Seeded with a Co-Culture of Schwann Cells and Adipose-Derived Stem Cells for Sciatic Nerve Regeneration

    Li, Ruixin; Li, Dong; Feng, Shiqing

    2016-01-01

    As a promising alternative to autologous nerve grafts, tissue-engineered nerve grafts have been extensively studied as a way to bridge peripheral nerve defects and guide nerve regeneration. The main difference between autogenous nerve grafts and tissue-engineered nerve grafts is the regenerative microenvironment formed by the grafts. If an appropriate regenerative microenvironment is provided, the repair of a peripheral nerve is feasible. In this study, to mimic the bodys natural regenerative microenvironment closely, we co-cultured Schwann cells (SCs) and adipose-derived stem cells (ADSCs) as seed cells and introduced them into a silk fibroin (SF)/collagen scaffold to construct a tissue-engineered nerve conduit (TENC). Twelve weeks after the three different grafts (plain SF/collagen scaffold, TENC, and autograft) were transplanted to bridge 1-cm long sciatic nerve defects in rats, a series of electrophysiological examinations and morphological analyses were performed to evaluate the effect of the tissue-engineered nerve grafts on peripheral nerve regeneration. The regenerative outcomes showed that the effect of treatment with TENCs was similar to that with autologous nerve grafts but superior to that with plain SF/collagen scaffolds. Meanwhile, no experimental animals had inflammation around the grafts. Based on this evidence, our findings suggest that the TENC we developed could improve the regenerative microenvironment and accelerate nerve regeneration compared to plain SF/collagen and may serve as a promising strategy for peripheral nerve repair. PMID:26799619

  18. Surface modification of strontium-doped porous bioactive ceramic scaffolds via poly(DOPA) coating and immobilizing silk fibroin for excellent angiogenic and osteogenic properties.

    Wang, Xu; Gu, Zhipeng; Jiang, Bo; Li, Li; Yu, Xixun

    2016-04-22

    For bioceramic scaffolds employed in clinical applications, excellent bioactivity and tenacity were of great importance. Modifying inorganic SCPP scaffolds with biological macromolecules could obviously improve its bioactivity and eliminate its palpable brittleness. However, it was hard to execute directly due to extremely bad interfacial compatibility between them. In this research, dopamine (DOPA) was introduced onto strontium-doped calcium polyphosphate (SCPP) scaffolds, subsequently the preliminary material was successfully further modified by silk fibroin (SF). SCPP/D/SF possessed suitable biomechanical properties, ability to stimulate angiogenic factor secretion and excellent biocompatibility. Biomechanical examination demonstrated that SCPP/D/SF scaffolds yielded better compressive strength because of improved interfacial compatibility. MTT assay and CLSM observation showed that SCPP/D/SF scaffolds had good cytocompatibility and presented better inducing-cell-migration potential than pure SCPP scaffolds. Meanwhile, its ability to stimulate angiogenic factor secretion was measured through the ELISA assay and immunohistological analysis in vitro and in vivo respectively. The results revealed, superior to SCPP, SCPP/D/SF could effectively promote VEGF and bFGF expression, possibly leading to enhancing angiogenesis and osteogenesis. In a word, SCPP/D/SF could serve as a potential bone tissue engineering scaffold for comparable biomechanical properties and excellent bioactivity. It provided a novel idea for modification of inorganic materials to prepare promising bone tissue engineering scaffolds with the ability to accelerate bone regeneration and vascularization. PMID:26870855

  19. Electron beam induced water-soluble silk fibroin nanoparticles as a natural antioxidant and reducing agent for a green synthesis of gold nanocolloid

    Wongkrongsak, Soraya; Tangthong, Theeranan; Pasanphan, Wanvimol

    2016-01-01

    The research proposes a novel water-soluble silk fibroin nanoparticles (WSSF-NPs) created by electron beam irradiation. In this report, we demonstrate the effects of electron beam irradiation doses ranging from 1 to 30 kGy on the molecular weight (MW), nanostructure formation, antioxidant activity and reducing power of the WSSF-NPs. Electron beam-induced degradation of SF causing MW reduction from 250 to 37 kDa. Chemical characteristic functions of SF still remained after exposing to electron beam. The WSSF-NPs with the MW of 37 kDa exhibited spherical morphology with a nanoscaled size of 40 nm. Antioxidant activities and reducing powers were investigated using 2,2-diphenyl-1-picrylhryl free radical (DPPH•) scavenging activity and ferric reducing antioxidant power (FRAP) assays, respectively. The WSSF-NPs showed greater antioxidant activity and reducing power than non-irradiated SF. By increasing their antioxidant and reducing power efficiencies, WSSF-NPs potentially created gold nanocolloid. WSSF-NPs produced by electron beam irradiation would be a great merit for the uses as a natural antioxidant additive and a green reducing agent in biomedical, cosmetic and food applications.

  20. Change in silk protein by radiation

    Ishida, Kazushige; Kamiishi, Youichi [Textile Research Institute of Gunma, Kiryu, Gunma (Japan); Takeshita, Hidefumi; Yoshii, Fumio; Kume, Tamikazu [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    2002-03-01

    Silk fibroin fiber irradiated with an accelerated electron beam in the dose range of 250 - 1,000 kGy was pulverized by using a ball mill. As irradiation dose increased, the conversion efficiency from fiber to powder increased, which reached 94% at 1,000 kGy. Silk fibroin powder obtained by this method dissolved 57% into water of ambient temperature. It is a very interesting phenomenon that silk fibroin obtains solubility without chemical treatment. In order to study mechanism of solubilization of silk fibroin powder, amino acid component of soluble part of silk powder was performed. The more irradiation dose up, the more recovery fraction of glycine or alanine decreased, which is, however, reached the minimum about 50%. To consider this result with crystal structure of silk fibroin, it is suggested that irradiation on silk fibroin fiber selectively degrades glycine and alanine in amorphous region, which makes it possible to pulverize and to dissolve for silk fibroin powder. Molecular weight of soluble part was also measured, but it had no serious concern with irradiation dose. Particle size distribution of silk fibroin powder was measured in order to study reduction of irradiation dose needed for pulverization. This measurement exhibited the possibility that lengthening of pulverization time reduces of irradiation dose. In addition, structure of particle was inferred from result of this measurement. (author)

  1. MicroRNA of the fifth-instar posterior silk gland of silkworm identified by Solexa sequencing

    Jisheng Li

    2014-12-01

    Full Text Available No special studies have been focused on the microRNA (miRNA in the fifth-instar posterior silk gland of Bombyx mori. Here, using next-generation sequencing, we acquired 93.2 million processed reads from 10 small RNA libraries. In this paper, we tried to thoroughly describe how our dataset generated from deep sequencing which was recently published in BMC genomics. Results showed that our findings are largely enriched silkworm miRNA depository and may benefit us to reveal the miRNA functions in the process of silk production.

  2. Silk Nanospheres and Microspheres from Silk/PVA Blend Films for Drug Delivery

    WANG, XIAOQIN; Yucel, Tuna; Lu, Qiang; Hu, Xiao; Kaplan, David L.

    2009-01-01

    Silk fibroin protein-based micro- and nanospheres provide new options for drug delivery due to their biocompatibility, biodegradability and their tunable drug loading and release properties. In the present study, we report a new aqueous-based preparation method for silk spheres with controllable sphere size and shape. The preparation was based on phase separation between silk fibroin and polyvinyl alcohol (PVA) at a weight ratio of 1/1 and 1/4. Water-insoluble silk spheres were easily obtaine...

  3. Evaluating degradation of silk's fibroin by attenuated total reflectance infrared spectroscopy: Case study of ancient banners from Polish collections

    Koperska, M. A.; ?ojewski, T.; ?ojewska, J.

    2015-01-01

    In this study a part of research where artificially aged model samples were used as a guideline to the mechanism of degradation is presented. In previous work Bombyx Mori silk samples were exposed to various environments such as different oxygen, water vapour and volatile organic products content, all at the temperature of 150 C [11]. Based on those results gathered with by Attenuated Total Reflectance/Fourier Transform Infrared Spectroscopy (ATR-FTIR) the degradation estimators were proposed and classified as follows: Primary functional groups estimators EAmideI/II - intensity ratios of Amide I Cdbnd O stretching vibration to Amide II Nsbnd H in-plane bending and Csbnd N stretching vibrations A1620/A1514. ECOOH - band 1318 cm-1 integral to band integral of CH3 bending vibration band located at 1442 cm-1P1318/P1442. Secondary conformational estimators EcCdbndO2 - intensity ratios within Amide I Cdbnd O stretching vibration of parallel ?-sheet to antiparallel ?-sheet A1620/A1699. In this work estimators were verified against estimators calculated from spectra of silk samples from 8 museum objects: 3 from 19th, 2 from 18th, 1 from 17th and 2 from 16th century including 3 banners from the storage resources of the Wawel Royal Castle in Cracow, Poland.

  4. An Australian webspinner species makes the finest known insect silk fibers

    Okada, Shoko; Weisman, Sarah; Trueman, Holly E.; Mudie, Stephen T.; Haritos, Victoria S.; Sutherland, Tara D. (CSIRO/MSE); (CSIRO)

    2009-01-15

    Aposthonia gurneyi, an Australian webspinner species, is a primitive insect that constructs and lives in a silken tunnel which screens it from the attentions of predators. The insect spins silk threads from many tiny spines on its forelegs to weave a filmy sheet. We found that the webspinner silk fibers have a mean diameter of only 65 nm, an order of magnitude smaller than any previously reported insect silk. The purpose of such fine silk may be to reduce the metabolic cost of building the extensive tunnels. At the molecular level, the A. gurneyi silk has a predominantly beta-sheet protein structure. The most abundant clone in a cDNA library produced from the webspinner silk glands encoded a protein with extensive glycine-serine repeat regions. The GSGSGS repeat motif of the A. gurneyi silk protein is similar to the well-known GAGAGS repeat motif found in the heavy fibroin of silkworm silk, which also has beta-sheet structure. As the webspinner silk gene is unrelated to the silk gene of the phylogenetically distant silkworm, this is a striking example of convergent evolution.

  5. Recent progress in development of transgenic silkworms overexpressing recombinant human proteins with therapeutic potential in silk glands.

    Itoh, Kohji; Kobayashi, Isao; Nishioka, So-Ichiro; Sezutsu, Hideki; Machii, Hiroaki; Tamura, Toshiki

    2016-01-01

    Since 2000, transgenic silkworms have been developed to produce recombinant proteins with therapeutic potential for future clinical use, including antibody preparations. Lysosomal storage diseases (LSDs) are inherited metabolic disorders caused by mutations of lysosomal enzymes associated with excessive accumulation of natural substrates and neurovisceral symptoms. Over the past few years, enzyme replacement therapy (ERT) with human lysosomal enzymes produced by genetically engineered mammalian cell lines has been used clinically to treat several patients with an LSD involving multi-organ symptoms. ERT is based on the incorporation of recombinant glycoenzymes by their binding to glycan receptors on the surface of target cells and their subsequent delivery to lysosomes. However, ERT has several disadvantages, including difficulty mass producing human enzymes, dangers of pathogen contamination, and high costs. Recently, the current authors have succeeded in producing transgenic silkworms overexpressing human lysosomal enzymes in the silk glands and the authors have purified catalytically active enzymes from the middle silk glands. Silk gland-derived human enzymes carrying high-mannose and pauci-mannose N-glycans were endocytosed by monocytes via the mannose receptor pathway and were then delivered to lysosomes. Conjugates with cell-penetrating peptides were also taken up by cultured fibroblasts derived from patients with enzyme deficiencies to restore intracellular catalytic activity and reduce the excessive accumulation of substrates in patient fibroblasts. Transgenic silkworms overexpressing human lysosomal enzymes in the silk glands could serve as future bioresources that provide safe therapeutic enzymes for the treatment of LSDs. Combining recent developments in transglycosylation technology with microbial endoglycosidases will promote the development of therapeutic glycoproteins as bio-medicines. PMID:26971553

  6. Response of human mesenchymal stem cells to intrafibrillar nanohydroxyapatite content and extrafibrillar nanohydroxyapatite in biomimetic chitosan/silk fibroin/nanohydroxyapatite nanofibrous membrane scaffolds

    Lai, Guo-Jyun; Shalumon, KT; Chen, Jyh-Ping

    2015-01-01

    Incorporation of nanohydroxyapatite (nHAP) within a chitosan (CS)/silk fibroin (SF) nanofibrous membrane scaffold (NMS) may provide a favorable microenvironment that more closely mimics the natural bone tissue physiology and facilitates enhanced osteogensis of the implanted cell population. In this study, we prepared pristine CS/SF NMS, composite CS/SF/nHAP NMS containing intrafibrillar nHAP by in situ blending of 10% or 30% nHAP before the electrospinning step, and composite CS/SF/nHAP NMS containing extrafibrillar nHAP by depositing 30% nHAP through alternative soaking surface mineralization. We investigated the effect of the incorporation of HAP nanoparticles on the physicochemical properties of pristine and composite NMS. We confirmed the presence of ~30 nm nHAP in the composite nanofibrous membranes by thermogravimetry analysis (TGA), X-ray diffraction (XRD), and scanning electron microscopy (SEM), either embedded in or exposed on the nanofiber. Nonetheless, the alternative soaking surface mineralization method drastically influenced the mechanical properties of the NMS with 88% and 94% drop in Youngs modulus and ultimate maximum stress. Using in vitro cell culture experiments, we investigated the effects of nHAP content and location on proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs). The proliferation of hMSCs showed no significant difference among pristine and composite NMS. However, the extent of osteogenic differentiation of hMSCs was found to be positively correlated with the content of nHAP in the NMS, while its location within the nanofiber played a less significant role. In vivo experiments were carried out with hMSCs seeded in CS/SF/30%nHAP NMS prepared by in situ blending and subcutaneous implantation in nude mice. Micro-computed tomography images as well as histological and immunohistochemical analysis of the retrieved hMSCs/NMS construct 1 and 2 months postimplantation indicated that NMS had the potential for bone regeneration and can be suggested as a promising scaffold for bone tissue engineering. PMID:25609962

  7. Production of an active feline interferon in the cocoon of transgenic silkworms using the fibroin H-chain expression system

    We constructed the fibroin H-chain expression system to produce recombinant proteins in the cocoon of transgenic silkworms. Feline interferon (FeIFN) was used for production and to assess the quality of the product. Two types of FeIFN fusion protein, each with N- and C-terminal sequences of the fibroin H-chain, were designed to be secreted into the lumen of the posterior silk glands. The expression of the FeIFN/H-chain fusion gene was regulated by the fibroin H-chain promoter domain. The transgenic silkworms introduced these constructs with the piggyBac transposon-derived vector, which produced the normal sized cocoons containing each FeIFN/H-chain fusion protein. Although the native-protein produced by transgenic silkworms have almost no antiviral activity, the proteins after the treatment with PreScission protease to eliminate fibroin H-chain derived N- and C-terminal sequences from the products, had very high antiviral activity. This H-chain expression system, using transgenic silkworms, could be an alternative method to produce an active recombinant protein and silk-based biomaterials

  8. Analyses of the Molecular Mechanisms Associated with Silk Production in Silkworm by iTRAQ-Based Proteomics and RNA-Sequencing-Based Transcriptomics.

    Wang, Shaohua; You, Zhengying; Feng, Mao; Che, Jiaqian; Zhang, Yuyu; Qian, Qiujie; Komatsu, Setsuko; Zhong, Boxiong

    2016-01-01

    Silkworm is used as a model organism to analyze two standard complex traits, which are high and low silk yields. To understand the molecular mechanisms of silk production, the posterior silk glands aged to the third day of the fifth instar were analyzed from the ZB strain with low silk production and from the control strain Lan10. Using isobaric tags for relative and absolute quantification (iTRAQ) quantitative shotgun proteomics and RNA-sequencing-based transcriptomics, 139 proteins and 630 transcripts were identified as novel in the ZB strain compared with the Lan10 strain, indicating that these results significantly expand the coverage of proteins and transcripts of the posterior silk glands in the silkworm. Of the 89 differently changed proteins, 23 were increased, and 66 were decreased. Of the 788 transcripts, 779 were upregulated, and 9 were downregulated. These results confirm that decreased energy utilization/protein translation and enhanced protein degradation are the key factors in lower silk production. Moreover, this study provides novel insight into the molecular changes that may result in lower silk production, namely, a combination of impaired transcription activity, missed protein folding/transport, and lowered yields of the main components of fibroin, along with weakened growth/development of the posterior silk gland. PMID:26626507

  9. Presence of phosphorus in Nephila clavipes dragline silk.

    Michal, C A; Simmons, A H; Chew, B G; Zax, D B; Jelinski, L W

    1996-01-01

    Solid-state 31P-NMR of Nephila clavipes dragline silk indicates the presence of phosphorus in at least two chemically distinct environments. Amino acid analyses of acid-hydrolyzed silk confirm the presence of phosphotyrosine as one of the phosphorus-containing components. The unusual chemical shift (18.9 ppm downfield from 85% H3PO4), proton chemical shift, and acid lability of a second component suggest that it is part of a strained five-membered cyclic phosphate that might be found on a beta-D-ribose. The five-membered cyclic phosphate is not removed from the silk fibers by exhaustive aqueous extraction. It is absent in nascent silk fibroin from the glands, suggesting that its formation is part of the fiber processing that occurs in the ducts leading to the spinnerets. High-resolution NMR spectra of silk dissolved in propionic acid/12 N HCl (50:50 v/v) show five phosphorus sites assigned to phosphorylated tyrosine residues, phosphorylated serine residues, inorganic phosphate, and two hydrolysis products of the cyclic phosphate compound. The observed posttranslational phosphorylation may be important in the processing and modulation of the physical properties of dragline silk. PMID:8770226

  10. Effects of Titanium Dioxide Nanoparticles on the Synthesis of Fibroin in Silkworm (Bombyx mori).

    Ni, Min; Li, FanChi; Tian, JiangHai; Hu, JingSheng; Zhang, Hua; Xu, KaiZun; Wang, BinBin; Li, YangYang; Shen, WeiDe; Li, Bing

    2015-08-01

    Silkworm (Bombyx mori) is an economically important insect, and its silk production capacity largely depends on its ability to synthesize fibroin. While breeding of B. mori varieties has been a key strategy to improve silk production, little improvement of B. mori silk production has been achieved to date. As a result, the development of sericulture economy has not progressed well, pointing to the need of new ways for improvement of B. mori silk production. Titanium dioxide nanoparticles (TiO2 NPs), a food additive widely used for livestock, have been shown to promote animal growth and increase the protein synthesis in animals. However, no studies on effect of TiO2 NPs on fibroin synthesis in B. mori have been available. In this study, the differential expression profiles of genes and proteins in the silk gland of B. mori fed without or with TiO2 NPs (5 μg ml(-1)) were analyzed and compared using digital gene expression (DGE), reverse transcription quantitative polymerase chain reaction (RT-qPCR), semi-qPCR, and Western blot analysis. The effects of TiO2 NPs feeding on the activity of proteases in the midgut and the synthesis and transportation of amino acids in hemolymph were also investigated. DGE analyses showed that among a total of 4,741 genes detected, 306 genes were differentially expressed after the TiO2 NPs feeding, of which 137 genes were upregulated whereas 169 genes were downregulated. 106 genes were shown to be involved in fibroin synthesis, of which 97 genes, including those encoding cuticular protein glycine-rich 10, serine protease inhibitor 28, aspartate aminotransferase, lysyl-tRNA synthetase, and splicing factor arginine/serine-rich 6, and silk gland factor-1 (SGF-1), were upregulated with the maximum induction of 8.52-folds, whereas nine genes, including those encoding aspartylglucosaminidase, the cathepsin L in Tribolium castaneum, and similar to SPRY domain-containing SOCS box protein 3, were downregulated with the maximum reduction of 8.11-folds. Transcription levels of nine genes were further verified by RT-qPCR, and the results were consistent with those with DGE. Transcription and expression levels of fibroin light chain (Fib-L) gene were increased after TiO2 NPs feeding, indicating that TiO2 NPs improves fibroin synthesis. Compared with that of control, the mean protease activity was increased by 56.67% in the B. mori fed with TiO2 NPs, and the transport of four key amino acids used for fibroin synthesis in hemolymph was also increased. These findings indicated that TiO2 NPs feeding can improve the absorption and utilization of amino acids from the feed and could be a new way to increase the fibroin synthesis in B. mori. PMID:25876086

  11. Molecular mechanisms of silk gland damage caused by phoxim exposure and protection of phoxim-induced damage by cerium chloride in Bombyx mori.

    Li, Bing; Sun, Qingqing; Yu, Xiaohong; Xie, Yi; Hong, Jie; Zhao, Xiaoyang; Sang, Xuezi; Shen, Weide; Hong, Fashui

    2015-09-01

    It is known that exposure to organophosphorus pesticides (OP) including phoxim can produce oxidative stress, neurotoxicity, and greatly attenuate cocooning rate in the silkworm, Bombyx mori. Cerium treatment has been demonstrated to relieve phoxim-induced toxicity in B. mori; however, very little is known about the molecular mechanisms of silk gland injury due to OP exposure and protection of gland damage due to cerium pretreatment. The aim of this study was to evaluate silk gland damage and its molecular mechanisms in phoxim-induced silkworm toxicity and the protective mechanisms of cerium following exposure to phoxim. The results showed that phoxim exposure resulted in severe gland damage, reductions in protein synthesis and the cocooning rate of silkworms. Cerium (Ce) attenuated gland damage caused by phoxim, promoted protein synthesis, increased the antioxidant capacity of the gland and increased the cocooning rate of B. mori. Furthermore, digital gene expression data suggested that phoxim exposure led to significant up-regulation of 714 genes and down-regulation of 120 genes. Of these genes, 122 were related to protein metabolism, specifically, the down-regulated Ser2, Ser3, Fib-L, P25, and CYP450. Ce pretreatment resulted in up-regulation of 162 genes, and down-regulation of 141 genes, importantly, Ser2, Ser3, Fib-L, P25, and CYP333B8 were up-regulated. Treatment with CeCl3 + phoxim resulted in higher levels of Fib-L, P25, Ser2, Ser3, CAT, TPx, and CYP333B8 expression in the silk gland of silkworms. These findings indicated that Ce increased cocooning rate via the promotion of silk protein synthesis-related gene expression in the gland under phoxim-induced toxicity. These findings may expand the application of rare earths in sericulture. PMID:24616058

  12. Fabrication and Biocompatibility of Electrospun Silk Biocomposites

    Ick-Soo Kim

    2011-10-01

    Full Text Available Silk fibroin has attracted great interest in tissue engineering because of its outstanding biocompatibility, biodegradability and minimal inflammatory reaction. In this study, two kinds of biocomposites based on regenerated silk fibroin are fabricated by electrospinning and post-treatment processes, respectively. Firstly, regenerated silk fibroin/tetramethoxysilane (TMOS hybrid nanofibers with high hydrophilicity are prepared, which is superior for fibroblast attachment. The electrospinning process causes adjacent fibers to ‘weld’ at contact points, which can be proved by scanning electron microscope (SEM. The water contact angle of silk/tetramethoxysilane (TMOS composites shows a sharper decrease than pure regenerated silk fibroin nanofiber, which has a great effect on the early stage of cell attachment behavior. Secondly, a novel tissue engineering scaffold material based on electrospun silk fibroin/nano-hydroxyapatite (nHA biocomposites is prepared by means of an effective calcium and phosphate (Ca–P alternate soaking method. nHA is successfully produced on regenerated silk fibroin nanofiber within several min without any pre-treatments. The osteoblastic activities of this novel nanofibrous biocomposites are also investigated by employing osteoblastic-like MC3T3-E1 cell line. The cell functionality such as alkaline phosphatase (ALP activity is ameliorated on mineralized silk nanofibers. All these results indicate that this silk/nHA biocomposite scaffold material may be a promising biomaterial for bone tissue engineering.

  13. Expression of hIGF-I in the silk glands of transgenic silkworms and in transformed silkworm cells.

    Zhao, Yue; Li, Xi; Cao, GuangLi; Xue, RenYu; Gong, ChengLiang

    2009-12-01

    To express human insulin-like growth factor-I (hIGF-I) in transformed Bombyx mori cultured cells and silk glands, the transgenic vector pigA3GFP-hIGF-ie-neo was constructed with a neomycin resistance gene driven by the baculovirus ie-1 promoter, and with the hIGF-I gene under the control of the silkworm sericin promoter Ser-1. The stably transformed BmN cells expressing hIGF-I were selected by using the antibiotic G418 at a final concentration of 700-800 microg/mL after the BmN cells were transfected with the piggyBac vector and the helper plasmid. The specific band of hIGF-I was detected in the transformed cells by Western blot. The expression level of hIGF-I, determined by ELISA, was about 7800 pg in 5x10(5) cells. Analysis of the chromosomal insertion sites by inverse PCR showed that exogenous DNA could be inserted into the cell genome randomly or at TTAA target sequence specifically for piggyBac element transposition. The transgenic vector pigA3GFP-hIGF-ie-neo was transferred into the eggs using sperm-mediated gene transfer. Finally, two transgenic silkworms were obtained after screening for the neo and gfp genes and verified by PCR and dot hybridization. The expression level of hIGF-I determined by ELISA was about 2440 pg/g of silk gland of the transgenic silkworms of the G1 generation. PMID:20016970

  14. Radiation degradation of silk protein

    Pewlong, W.; Sudatis, B. [Office of Atomic Energy for Peace, Bangkok (Thailand); Takeshita, Hidefumi; Yoshii, Fumio; Kume, Tamikazu [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    2000-03-01

    Silk fibroin fiber from the domesticated silkworm Bombyx mori was irradiated using an electron beam accelerator to investigate the application of the radiation degradation technique as a means to solubilize fibroin. The irradiation caused a significant degradation of the fiber. The tensile strength of fibroin fiber irradiated up to 2500 kGy decreased rapidly with increasing dose. The presence of oxygen in the irradiation atmosphere enhanced degradation of the tensile strength. The solubilization of irradiated fibroin fiber was evaluated using the following three kinds of solutions: a calcium chloride solution(CaCl{sub 2}/C{sub 2}H{sub 5}OH/H{sub 2}O=1:2:8 in mole ratio), a hydrochloric acid (0.5 N) and a distilled water. Dissolution of fibroin fiber into these solutions was significantly enhanced by irradiation. Especially, an appreciable amount of water soluble proteins was extracted by a distilled water. (author)

  15. Radiation degradation of silk protein

    Wachiraporn Pewlong; Boonya Sudatis [Office of Atomic Energy for Peace, Bangkok (Thailand); Takeshita, Hidefumi; Yoshii, Fumio; Kume, Tamikazu [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    2000-09-01

    Silk fibroin fiber from the domesticated silkworm Bombyx mori was irradiated in the dose range up to 2500 kGy using an electron beam accelerator to apply the radiation degradation technique as a means to solubilize fibroin. The tensile strength of irradiated fibroin fiber decreased with increasing dose and the presence of oxygen in the irradiation atmosphere enhanced the degradation. The solubilization of irradiated fibroin fiber was evaluated using the following three kinds of solutions: calcium chloride solution (CaCl{sub 2}/C{sub 2}H{sub 5}OH/H{sub 2}O = 1 : 2 : 8 in mole ratio), hydrochloric acid (0.5N) and distilled water. Dissolution of fibroin fiber into these solutions was significantly enhanced by irradiation. Especially, an appreciable amount of water-soluble protein was extracted by distilled water. (author)

  16. Radiation degradation of silk protein

    Silk fibroin fiber from the domesticated silkworm Bombyx mori was irradiated using an electron beam accelerator to investigate the application of the radiation degradation technique as a means to solubilize fibroin. The irradiation caused a significant degradation of the fiber. The tensile strength of fibroin fiber irradiated up to 2500 kGy decreased rapidly with increasing dose. The presence of oxygen in the irradiation atmosphere enhanced degradation of the tensile strength. The solubilization of irradiated fibroin fiber was evaluated using the following three kinds of solutions: a calcium chloride solution(CaCl2/C2H5OH/H2O=1:2:8 in mole ratio), a hydrochloric acid (0.5 N) and a distilled water. Dissolution of fibroin fiber into these solutions was significantly enhanced by irradiation. Especially, an appreciable amount of water soluble proteins was extracted by a distilled water. (author)

  17. Different properties of electrospun fibrous scaffolds of separated heavy-chain and light-chain fibroins of Bombyx mori.

    Wadbua, Paweena; Promdonkoy, Boonhiang; Maensiri, Santi; Siri, Sineenat

    2010-06-01

    This study is the first to report on the fabrication and properties of electrospun scaffolds derived from separated light-chain fibroin and heavy-chain fibroin, two major proteins of silk fibroin. Among seven different extraction conditions, which were commonly used to extract fibroin from cocoons of Bombyx mori, only Ajisawa's reagent and 9 M lithium thiocyanate could extract both heavy-chain fibroin and light-chain fibroin, while the other conditions could yield only the light-chain fibroin. Mixed fibroin, light-chain fibroin, and heavy-chain fibroin were fabricated using electrospinning methods. Average diameters of the fibers were 658+/-208, 517+/-162, and 518+/-171 nm, respectively and their sizes after treatment with 50% methanol for 60 min were slightly increased to 747+/-244, 556+/-164 and 521+/-201 nm, respectively. FTIR spectra showed similar predominant beta-sheet conformation of mixed fibroin and heavy-chain fibroin scaffolds after treated with methanol, whereas the predominant structure of light-chain fibroin was random coil conformation. Although, scaffolds derived from mixed fibroin and heavy-chain fibroin showed similar properties, the light-chain fibroin scaffold clearly exhibited different properties, including more hydrophilic character, water uptake ability, degradation rate, and cell adhesion capability. PMID:20338193

  18. Ultrastructure of the silk glands in three adult females of sphecid wasps of the genus Microstigmus (Hymenoptera: Pemphredoninae) Ultraestructura de las glndulas de seda en hembras adultas de tres especies de avispas del gnero Microstigmus (Hymenoptera: Sphecidae: Pemphredoninae)

    JOS EDUARDO SERRO

    2005-01-01

    The ultrastructure of silk glands in adult females of sphecid wasps is described. Individual glands are scattered in the more posterior portion of the metasoma being each gland formed by an enlarged secretory unity with one conducting canal. Each secretory unit is a gland cell containing a large lumen and numerous secretory granules, which are delimited by membrane and filled with middle electron dense content of homogeneous aspect, which are exocytosed into the extra cellular lumen. Cell cyt...

  19. COMPARATIVE ANALYSIS OF THREE-DIMENSIONAL NANOSTRUCTURE OF POROUS BIOCOMPATIBLE SCAFFOLDS MADE OF RECOMBINANT SPIDROIN AND SILK FIBROIN FOR REGENERATIVE MEDICINE

    O. I. Agapova

    2015-05-01

    Full Text Available Aim. To perform a comparison of three-dimensional nanostructure of porous biocompatible scaffolds made of fibroin Bombix mori and recombinant spidroin rS1/9. Materials and methods. Three-dimensional porous scaffolds were produced by salt leaching technique. The comparison of biological characteristics of the scaffolds shows that adhesion and proliferation of mouse fibroblasts in vitro on these two types of scaffolds do not differ significantly. Comparative experiments in vivo show that regeneration of bone tissue of rats is faster with implantation of recombinant spidroin scaffolds. Three-dimensional nanostructure of scaffolds and interconnectivity of nanopores were studied with scanning probe nanotomography (SPNT to explain higher regenerative activity of spidroin-based scaffolds. Results. Significant differences were detected in the integral density and volume of pores: the integral density of nanopores detected on 2D AFM images is 46 μm–2    and calculated volume porosity is 24% in rS1/9-based scaffolds; in fibroin-based three-dimensional structures density of nanopores and calculated volume porosity were 2.4 μm–2  and 0.5%, respectively. Three-dimensional reconstruction system of nanopores and clusters of interconnected nanopores in rS1/9-based scaffolds showed that volume fraction of pores interconnected in percolation clusters is 35.3% of the total pore volume or 8.4% of the total scaffold volume. Conclusion. Scanning probe nanotomography method allows obtaining unique information about topology of micro – and nanopore systems of artificial biostructures. High regenerative activity of rS1/9-based scaffolds can be explained by higher nanoporosity of the scaffolds.

  20. In vivo analysis of fibroin heavy chain signal peptide of silkworm Bombyx mori using recombinant baculovirus as vector

    In order to investigate the functional signal peptide of silkworm fibroin heavy chain (FibH) and the effect of N- and C-terminal parts of FibH on the secretion of FibH in vivo, N- and C-terminal segments of fibh gene were fused with enhanced green fluorescent protein (EGFP) gene. The fused gene was then introduced into silkworm larvae and expressed in silk gland using recombinant AcMNPV (Autographa californica multiple nuclear polyhedrosis virus) as vector. The fluorescence of EGFP was observed with fluorescence microscope. FibH-EGFP fusion proteins extracted from silk gland were analyzed by Western blot. Results showed that the two alpha helices within N-terminal 163 amino acid residues and the C-terminal 61 amino acid residues were not necessary for cleavage of signal peptide and secretion of the fusion protein into silk gland. Then the C-terminal 61 amino acid residues were substituted with a His-tag in the fusion protein to facilitate the purification. N-terminal sequencing of the purified protein showed that the signal cleavage site is between position 21 and 22 amino acid residues

  1. Isolation and processing of silk proteins for biomedical applications.

    Kundu, Banani; Kurland, Nicholas E; Yadavalli, Vamsi K; Kundu, Subhas C

    2014-09-01

    Silk proteins of silkworms are chiefly composed of core fibroin protein and glycoprotein sericin that glues fibroin. Unique mechanical properties, cyto-compatibility and controllable biodegradability facilitate the use of fibroin in biomedical applications. Sericin serves as additive in cosmetic and food industries, as mitotic factor in cell culture media, anti-cancerous drug, anticoagulant and as biocompatible coating. For all these uses; aqueous solutions of silk proteins are preferred. Therefore, an accurate understanding of extraction procedure of silk proteins from their sources is critical. A number of protocols exist, amongst which it is required to settle a precise and easy one with desired yield and least down-stream processing. Here, we report extraction of proteins employing methods mentioned in literature using cocoons of mulberry and nonmulberry silks. This study reveals sodium carbonate salt-boiling system is the most efficient sericin extraction procedure for all silk variants. Lithium bromide is observed as the effective fibroin dissolution system for mulberry silk cocoons; whereas heterogeneous species-dependent result is obtained in case of nonmulberry species. We further show the effect of common post processing on nanoscale morphology of mulberry silk fibroin films. This knowledge eases the adoption and fabrication of silk biomaterials in devices and therapeutic delivery systems. PMID:24971560

  2. Production of scFv-Conjugated Affinity Silk Powder by Transgenic Silkworm Technology

    Sato, Mitsuru; Kojima, Katsura; Sakuma, Chisato; Murakami, Maria; Aratani, Eriko; Takenouchi, Takato; Tamada, Yasushi; Kitani, Hiroshi

    2012-01-01

    Bombyx mori (silkworm) silk proteins are being utilized as unique biomaterials for medical applications. Chemical modification or post-conjugation of bioactive ligands expand the applicability of silk proteins; however, the processes are elaborate and costly. In this study, we used transgenic silkworm technology to develop single-chain variable fragment (scFv)-conjugated silk fibroin. The cocoons of the transgenic silkworm contain fibroin L-chain linked with scFv as a fusion protein. After di...

  3. Production of fine powder from silk by radiation

    Takeshita, Hidefumi; Yoshii, Fumio; Kume, Tamikazu [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment; Ishida, Kazushige; Kamiishi, Youichi [Textile Research Inst. of Gunma, Kiryu, Gunma (Japan)

    2000-09-01

    To produce silk fine powder, silk fibroin fibers were irradiated with an accelerated electron beam. Though unirradiated silk fibers were not pulverized at all, irradiated silk fibers were able to be crushed only by physical means using a ball mill. In the dose range of 250- 1000 kGy, as the dose to silk fibroin increased, the conversion rate from fiber to powder was increased. The electron microscope observation showed that the particle size of silk powder was less than 10 micrometers, and that those particles composed aggregate. Although silk fibroin fiber was not soluble in water, silk powders from fiber irradiated 500 kGy dissolved more than 50 weight %. Amino acid analysis of soluble fraction in silk powder showed that Gly content is lower than in normal silk and other amino acids contents except for Ala are 1.5 times as large as in normal silk. From these results, it is reasonable to suppose that irradiation and pulverization decomposed Gly and Ala. (author)

  4. Sequential steps of macroautophagy and chaperone-mediated autophagy are involved in the irreversible process of posterior silk gland histolysis during metamorphosis of Bombyx mori.

    Shiba, Hajime; Yabu, Takeshi; Sudayama, Makoto; Mano, Nobuhiro; Arai, Naoto; Nakanishi, Teruyuki; Hosono, Kuniaki

    2016-04-15

    To elucidate the degradation process of the posterior silk gland during metamorphosis of the silkworm ITALIC! Bombyx mori, tissues collected on the 6th day after entering the 5th instar (V6), prior to spinning (PS), during spinning (SP) and after cocoon formation (CO) were used to analyze macroautophagy, chaperone-mediated autophagy (CMA) and the adenosine triphosphate (ATP)-dependent ubiquitin proteasome. Immediately after entering metamorphosis stage PS, the levels of ATP and phosphorylated p70S6 kinase protein decreased spontaneously and continued to decline at SP, followed by a notable restoration at CO. In contrast, phosphorylated AMP-activated protein kinase α (AMPKα) showed increases at SP and CO. Most of the Atg8 protein was converted to form II at all stages. The levels of ubiquitinated proteins were high at SP and CO, and low at PS. The proteasome activity was high at V6 and PS but low at SP and CO. In the isolated lysosome fractions, levels of Hsc70/Hsp70 protein began to increase at PS and continued to rise at SP and CO. The lysosomal cathepsin B/L activity showed a dramatic increase at CO. Our results clearly demonstrate that macroautophagy occurs before entering the metamorphosis stage and strongly suggest that the CMA pathway may play an important role in the histolysis of the posterior silk gland during metamorphosis. PMID:26944491

  5. Aqueous multiphoton lithography with multifunctional silk-centred bio-resists

    Sun, Yun-Lu; Li, Qi; Sun, Si-Ming; Huang, Jing-Chun; Zheng, Bo-Yuan; Chen, Qi-Dai; Shao, Zheng-Zhong; Sun, Hong-Bo

    2015-10-01

    Silk and silk fibroin, the biomaterial from nature, nowadays are being widely utilized in many cutting-edge micro/nanodevices/systems via advanced micro/nanofabrication techniques. Herein, for the first time to our knowledge, we report aqueous multiphoton lithography of diversiform-regenerated-silk-fibroin-centric inks using noncontact and maskless femtosecond laser direct writing (FsLDW). Initially, silk fibroin was FsLDW-crosslinked into arbitrary two/three-dimensional micro/nanostructures with good elastic properties merely using proper photosensitizers. More interestingly, silk/metal composite micro/nanodevices with multidimension-controllable metal content can be FsLDW-customized through laser-induced simultaneous fibroin oxidation/crosslinking and metal photoreduction using the simplest silk/Ag+ or silk/[AuCl4]- aqueous resists. Noticeably, during FsLDW, fibroin functions as biological reductant and matrix, while metal ions act as the oxidant. A FsLDW-fabricated prototyping silk/Ag microelectrode exhibited 104-Ω-1 m-1-scale adjustable electric conductivity. This work not only provides a powerful development to silk micro/nanoprocessing techniques but also creates a novel way to fabricate multifunctional metal/biomacromolecule complex micro/nanodevices for applications such as micro/nanoscale mechanical and electrical bioengineering and biosystems.

  6. Bilayered silk/silk-nanoCaP scaffolds for osteochondral tissue engineering : in vitro and In vivo assessment of biological performance

    Yan, Leping; Correia, Joana Silva; Oliveira, Mariana B.; Vilela, Carlos; Pereira, H.; Sousa, R A; Mano, J. F.; de Oliveira, A. L.; Oliveira, Joaquim M.; Reis, R.L.

    2015-01-01

    Novel porous bilayered scaffolds, fully integrating a silk fibroin (SF) layer and a silk-nano calcium phosphate (Silk-NanoCaP) layer for osteochondral defect (OCD) regeneration were developed. Homogeneous porosity distribution was achieved in the scaffolds, with calcium phosphate phase only retained in the Silk-NanoCaP layer. The scaffold presented compressive moduli of 0.4 MPa in wet state. Rabbit bone marrow mesenchymal stromal cells (RBMSCs) were cultured on the scaffolds, and good adhesio...

  7. Production of fine powder from silk by radiation

    Takeshita, Hidefumi; Yoshii, Fumio; Kume, Tamikazu [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment; Ishida, Kazunari; Kamiishi, Youichi [Textile Research Institute of Gunma, Kiryu, Gunma (Japan)

    2000-03-01

    Silk fine power was prepared directly from silk fiber irradiated with an accelerated electron beam(EB). Irradiated silk fiber was well pulverized only by physical crushing using ball mill without any chemical pretreatment. Raw and degummed silk fibers were irradiated at ambient temperature in the dose range of 250-1000 kGy. Although unirradiated silk fibers were not pulverized at all, irradiated fibers were easily pulverized and showed higher conversion from fiber to powder for higher doses. The presence of oxygen in the irradiation atmosphere enhanced pulverization of silk fiber. Raw silk fibers were less pulverized compared to degummed ones. The electron microscope observation showed that the minimum particle size of silk powder obtained from fiber irradiated by 1000 kGy in oxygen was less than 10 microns. It was found that fibroin powder obtained in this work dissolved remarkably into cold water, thought unirradiated fibroin fiber had little solubility even in hot water. A typical soluble fraction was about 60% for fibroin powder obtained from fiber irradiated by 1000 kGy in oxygen. (author)

  8. Bio-inspired fabrication of fibroin cryogels from the muga silkworm Antheraea assamensis for liver tissue engineering

    Conventional scaffold fabrication techniques result in narrow pore architectures causing a limited interconnectivity and use of porogens, which affects the bio- or cyto-compatibility. To ameliorate this, cryogels are immensely explored due to their macro-porous nature, ease in fabrication, using ice crystals as porogens, the shape property, easy reproducibility and cost-effective fabrication technique. Cryogels in the present study are prepared from nonmulberry Indian muga silk gland protein fibroin of Antheraea assamensis using two different fabrication temperatures (−20 and −80 °C). Anionic surfactant sodium dodecyl sulfate is used to solubilize fibroin, which in turn facilitates gelation by accelerating the ß-sheet formation. Ethanol is employed to stabilize the 3D network and induces bimodal porosity. The gels thus formed demonstrate increased ß-sheet content (FTIR) and a considerable effect of pre-freezing temperatures on 3D micro-architectures. The cryogels are capable of absorbing large amounts of water and withstanding mechanical compression without structure deformation. Further, cell impregnated cryogels well support the viability of human hepatocarcinoma cells (live/dead assay). The formation of cellular aggregates (confocal laser and scanning electron microscope), derivation in metabolic activity and proliferation rate are obtained in constructs fabricated at different temperatures. In summary, the present work reveals promising insights in the development of a biomimetic functional template for biomedical therapeutics and liver tissue engineering. (paper)

  9. Bio-inspired fabrication of fibroin cryogels from the muga silkworm Antheraea assamensis for liver tissue engineering.

    Kundu, Banani; Kundu, S C

    2013-10-01

    Conventional scaffold fabrication techniques result in narrow pore architectures causing a limited interconnectivity and use of porogens, which affects the bio- or cyto-compatibility. To ameliorate this, cryogels are immensely explored due to their macro-porous nature, ease in fabrication, using ice crystals as porogens, the shape property, easy reproducibility and cost-effective fabrication technique. Cryogels in the present study are prepared from nonmulberry Indian muga silk gland protein fibroin of Antheraea assamensis using two different fabrication temperatures (-20 and -80 °C). Anionic surfactant sodium dodecyl sulfate is used to solubilize fibroin, which in turn facilitates gelation by accelerating the ß-sheet formation. Ethanol is employed to stabilize the 3D network and induces bimodal porosity. The gels thus formed demonstrate increased ß-sheet content (FTIR) and a considerable effect of pre-freezing temperatures on 3D micro-architectures. The cryogels are capable of absorbing large amounts of water and withstanding mechanical compression without structure deformation. Further, cell impregnated cryogels well support the viability of human hepatocarcinoma cells (live/dead assay). The formation of cellular aggregates (confocal laser and scanning electron microscope), derivation in metabolic activity and proliferation rate are obtained in constructs fabricated at different temperatures. In summary, the present work reveals promising insights in the development of a biomimetic functional template for biomedical therapeutics and liver tissue engineering. PMID:24002731

  10. Fluid Mechanical Properties of Silkworm Fibroin Solutions

    Matsumoto, Akira

    2005-11-01

    The aqueous solution behavior of silk fibroin is of interest due to the assembly and processing of this protein related to the spinning of protein fibers that exhibit remarkable mechanical properties. To gain insight into the origins of this functional feature, it is desired to determine how the protein behaves under a range of solution conditions. Pure fibroin at different concentrations in water was studied for surface tension, as a measure of surfactancy. In addition, shear induced changes on these solutions in terms of structure and morphology was also determined. Fibroin solutions exhibited shear rate-sensitive viscosity changes and precipitated at a critical shear rate where a dramatic increase of 75-150% of the initial value was observed along with a decrease in viscosity. In surface tension measurements, critical micelle concentrations were in the range of 3-4% w/v. The influence of additional factors, such as sericin protein, divalent and monovalent cations, and pH on the solution behavior in relation to structural and morphological features will also be described.

  11. Solubilization of silk protein by radiation

    Sudatis, Boonya; Pongpat, Suchada [Office of Atomic Energy of Peace, Bangkok (Thailand)

    2002-03-01

    Gamma irradiated silk fibroin at doses of 0, 5, 10, 20, 40, 60, 80, 100, 125, 250, 500, 750 and 1000 kGy were soaked in water for 1 hr. Silk fibroin solubilized percentage was investigated from lost weight of sample (dried at 105{sup 0}C), they were 0, 0, 0.7, 0, 0.11, 0.11, 0, 0.73, 0.77, 4.38, 8.32, 10.22 and 18.52 respectively. It showed that at the higher dose up to 250 kGy had direct effect to solubility, and increased with increasing dose. In addition, silk sericin dissolved 77.76, 82.22, 83.55, 84.31, 86.04, 86.67 and 87.37% after gamma irradiation at the doses of 0, 50, 100, 200, 500, 750 and 1000 kGy respectively. It presents that radiation can cause silk protein, fibroin and sericin dissolve because of their degradation. (author)

  12. Functional silk: colored and luminescent.

    Tansil, Natalia C; Koh, Leng Duei; Han, Ming-Yong

    2012-03-15

    Silkworm silk is among the most widely used natural fibers for textile and biomedical applications due to its extraordinary mechanical properties and superior biocompatibility. A number of physical and chemical processes have also been developed to reconstruct silk into various forms or to artificially produce silk-like materials. In addition to the direct use and the delicate replication of silk's natural structure and properties, there is a growing interest to introduce more new functionalities into silk while maintaining its advantageous intrinsic properties. In this review we assess various methods and their merits to produce functional silk, specifically those with color and luminescence, through post-processing steps as well as biological approaches. There is a highlight on intrinsically colored and luminescent silk produced directly from silkworms for a wide range of applications, and a discussion on the suitable molecular properties for being incorporated effectively into silk while it is being produced in the silk gland. With these understanding, a new generation of silk containing various functional materials (e.g., drugs, antibiotics and stimuli-sensitive dyes) would be produced for novel applications such as cancer therapy with controlled release feature, wound dressing with monitoring/sensing feature, tissue engineering scaffolds with antibacterial, anticoagulant or anti-inflammatory feature, and many others. PMID:22302383

  13. Preparation and characterization of silk/silica hybrid biomaterials by sol-gel crosslinking process

    Hou Aiqin, E-mail: aiqinhou@dhu.edu.c [National Engineering Research Center for Dyeing and Finishing of Textiles, Donghua University, 3H, 2999 North Renmin Road, Songjiang, Shanghai 201620 (China); Chen Huawei [National Engineering Research Center for Dyeing and Finishing of Textiles, Donghua University, 3H, 2999 North Renmin Road, Songjiang, Shanghai 201620 (China)

    2010-03-15

    The silk/silica hybrid biomaterials are synthesized by sol-gel crosslinking process. The chemical and morphological structures of silk/silica hybrids are investigated with micro-FT-IR spectra, X-ray diffraction, SEM, AFM, and DSC. The results show that the crosslinking reactions among inorganic nano-particles, fibroin and 2,4,6-tri[(2-epihydrin-3-bimethyl-ammonium)propyl]-1,3,5-triazine chloride (Tri-EBAC) take place during sol-gel process. The silk/silica hybrids form new molecular structures containing not only organic fibroin but also inorganic nano-silica particles. The inorganic particles are bounded to the fibroin through covalent bonds. The silk/silica hybrids can form excellent film with very even nanometer particles. The thermal properties of organic/inorganic hybrid are improved.

  14. Preparation and characterization of silk/silica hybrid biomaterials by sol-gel crosslinking process

    The silk/silica hybrid biomaterials are synthesized by sol-gel crosslinking process. The chemical and morphological structures of silk/silica hybrids are investigated with micro-FT-IR spectra, X-ray diffraction, SEM, AFM, and DSC. The results show that the crosslinking reactions among inorganic nano-particles, fibroin and 2,4,6-tri[(2-epihydrin-3-bimethyl-ammonium)propyl]-1,3,5-triazine chloride (Tri-EBAC) take place during sol-gel process. The silk/silica hybrids form new molecular structures containing not only organic fibroin but also inorganic nano-silica particles. The inorganic particles are bounded to the fibroin through covalent bonds. The silk/silica hybrids can form excellent film with very even nanometer particles. The thermal properties of organic/inorganic hybrid are improved.

  15. The Performance of Silk Scaffolds in a Rat Model of Augmentation Cystoplasty

    Seth, Abhishek; Chung, Yeun Goo; Gil, Eun Seok; Tu, Duong; Franck, Debra; Di Vizio, Dolores; Adam, Rosalyn M.; Kaplan, David L.; Estrada, Carlos R.; Mauney, Joshua R.

    2013-01-01

    The diverse processing plasticity of silk-based biomaterials offers a versatile platform for understanding the impact of structural and mechanical matrix properties on bladder regenerative processes. Three distinct groups of 3-D matrices were fabricated from aqueous solutions of Bombyx mori silk fibroin either by a gel spinning technique (GS1 and GS2 groups) or a solvent-casting/salt-leaching method in combination with silk film casting (FF group). SEM analyses revealed that GS1 matrices cons...

  16. Differential scanning fluorimetry illuminates silk feedstock stability and processability

    Dicko, C.; Kasoju, Naresh; Hawkins, N.; Vollrath, F.

    2016-01-01

    Roč. 12, č. 1 (2016), s. 255-262. ISSN 1744-683X R&D Projects: GA MŠk(CZ) EE2.3.30.0029 Institutional support: RVO:61389013 Keywords : differential scanning fluorimetry * biomaterials * silk fibroin Subject RIV: CE - Biochemistry Impact factor: 4.029, year: 2014

  17. Silk constructs for delivery of muskuloskeletal therapeutics

    Meinel, Lorenz; Kaplan, David L.

    2012-01-01

    Silk fibroin (SF) is a biopolymer with distinguishing features from many other bio- as well as synthetic polymers. From a biomechanical and drug delivery perspective, SF combines remarkable versatility for scaffolding (solid implants, hydrogels, threads, solutions), with advanced mechanical properties and good stabilization and controlled delivery of entrapped protein and small molecule drugs, respectively. It is this combination of mechanical and pharmaceutical features which render SF so ex...

  18. Silk film biomaterials for ocular surface repair

    Lawrence, Brian David

    Current biomaterial approaches for repairing the cornea's ocular surface upon injury are partially effective due to inherent material limitations. As a result there is a need to expand the biomaterial options available for use in the eye, which in turn will help to expand new clinical innovations and technology development. The studies illustrated here are a collection of work to further characterize silk film biomaterials for use on the ocular surface. Silk films were produced from regenerated fibroin protein solution derived from the Bombyx mori silkworm cocoon. Methods of silk film processing and production were developed to produce consistent biomaterials for in vitro and in vivo evaluation. A wide range of experiments was undertaken that spanned from in vitro silk film material characterization to in vivo evaluation. It was found that a variety of silk film properties could be controlled through a water-annealing process. Silk films were then generated that could be use in vitro to produce stratified corneal epithelial cell sheets comparable to tissue grown on the clinical standard substrate of amniotic membrane. This understanding was translated to produce a silk film design that enhanced corneal healing in vivo on a rabbit injury model. Further work produced silk films with varying surface topographies that were used as a simplified analog to the corneal basement membrane surface in vitro. These studies demonstrated that silk film surface topography is capable of directing corneal epithelial cell attachment, growth, and migration response. Most notably epithelial tissue development was controllably directed by the presence of the silk surface topography through increasing cell sheet migration efficiency at the individual cellular level. Taken together, the presented findings represent a comprehensive characterization of silk film biomaterials for use in ocular surface reconstruction, and indicate their utility as a potential material choice in the development of innovative procedures and technologies for corneal repair.

  19. Mechanical behavior comparison of spider and silkworm silks using molecular dynamics at atomic scale.

    Lee, Myeongsang; Kwon, Junpyo; Na, Sungsoo

    2016-02-01

    Spider and silkworm silk proteins have received much attention owing to their inherent structural stability, biodegradability, and biocompatibility. These silk protein materials have various mechanical characteristics such as elastic modulus, ultimate strength and fracture toughness. While the considerable mechanical characteristics of the core crystalline regions of spider silk proteins at the atomistic scale have been investigated through several experimental techniques and computational studies, there is a lack of comparison between spider and silkworm fibroins in the atomistic scale. In this study, we investigated the differences between the mechanical characteristics of spider and silkworm fibroin structures by applying molecular dynamics and steered molecular dynamics. We found that serine amino acids in silkworm fibroins not only increased the number of hydrogen bonds, but also altered their structural characteristics and mechanical properties. PMID:26806791

  20. SPIDER SILK

    PORAV Viorica

    2014-05-01

    Full Text Available The strengthness and toughness of spider fiber and its multifunctional nature is only surpassed in some cases by synthetic high performance fibers. In the world of natural fibers, spider silk has been long time recognized as a wonder fiber for its unique combination of high strength and rupture elongation. Scientists in civil military engineering reveal that the power of biological material (spider silk lies in the geometric configuration of structural protein, and the small cluster of week hydrogen bonds that works together to resist force and dissipate energy. Each spider and each type of silk has a set of mechanical properties optimized for their biological function. Most silks, in particular deagline silk, have exceptional mechanical properties. They exhibit a unique combination of high tensile strength and extensibility (ductility. This enables a silk fiber to absorb a lot of energy before breaking (toughness, the area under a stress- strain curve. A frequent mistake made in the mainstream media is to confuse strength and toughness when comparing silk to other materials. As shown below in detail, weight for weight, silk is stronger than steel, but not as strong as Kevlar. Silk is,however, tougher than both.This paper inform about overview on the today trend in the world of spider silk.

  1. An axial distribution of seeding, proliferation, and osteogenic differentiation of MC3T3-E1 cells and rat bone marrow-derived mesenchymal stem cells across a 3D Thai silk fibroin/gelatin/hydroxyapatite scaffold in a perfusion bioreactor.

    Sinlapabodin, Salita; Amornsudthiwat, Phakdee; Damrongsakkul, Siriporn; Kanokpanont, Sorada

    2016-01-01

    In cell culture, a perfusion bioreactor provides effective transportation of nutrients, oxygen, and waste removal to and from the core of the scaffold. In addition, it provides mechanical stimuli for enhancing osteogenic differentiation. In this study, we used an axial distribution of cell numbers, alkaline phosphatase (ALP) enzyme activity, and calcium content across 4 cross-sections of 10mm thick scaffold, made of Thai silk fibroin (SF)/gelatin (G)/hydroxyapatite (HA), as a tool to evaluate the suitable perfusion flow rate. These evaluations cover all cellular developmental phases starting from seeding, to proliferation, and later osteogenic differentiation. Mouse pre-osteoblastic MC3T3-E1 cell lines were used as a cell model during seeding and proliferation. The bioreactor seeded scaffold provided more uniform cell distribution across the scaffold compared to centrifugal and agitation seeding, while the overall number of adhered cells from bioreactor seeding was slightly lower than agitation seeding. The dynamic culture using 1 ml/min perfusion flow rate (initial shear stress of 0.1 dyn/cm(2)) enabled statistically higher MC3T3-E1 proliferation, ALP activity, and calcium deposition than those observed in the static-culturing condition. However, the perfusion flow rate of 1 ml/min seemed not to be enough for enhancing ALP expression across all sections of the scaffold. Rat bone marrow derived stromal cells (rMSC) were used in the detachment test and osteogenic differentiation. It was found that perfusion flow rate of 5 ml/min caused statistically higher cell detachment than that of 1 and 3 ml/min. The perfusion flow rate of 3 ml/min gave the highest rMSC osteogenic differentiation on a SF/G/HA scaffold than other flow rates, as observed from the significantly highest number of ALP enzyme activity and the calcium content without any significant cell growth. In addition, all of these parameters were evenly distributed across all scaffold sections. PMID:26478392

  2. Expression of the hIGF-I gene driven by the Fhx/P25 promoter in the silk glands of germline silkworm and transformed BmN cells.

    Li, Yanmei; Cao, Guangli; Wang, Yang; Xue, Renyu; Zhou, Wenlin; Gong, Chengliang

    2011-03-01

    The expression of the human insulin-like growth factor (hIGF-I) gene driven by the Fhx/P25 promoter in the silk glands of transgenic silkworms (Bombyx mori) and in transformed silkworm cells, was achieved using BmN cells transfected with a piggyBac vector, pigA3GFP-Fhx/P25-hIGF-ie-neo containing a neomycin-resistance gene (neo), a green fluorescent protein gene (gfp), an hIGF-I gene, and a helper plasmid containing the piggyBac transposase sequence under the control of the B. mori actin 3 (A3) promoter. We selected stably transformed BmN cells expressing hIGF-I using the antibiotic G418. The expression level of hIGF-I was about 450 pg in 3 10(6) cells, determined by ELISA. The piggyBac vector was transferred into the silkworm eggs using sperm-mediated gene transfer. The expression level of hIGF-I per gram fresh posterior silk glands of G4 transgenic silkworms was approx. 150 ng. PMID:21072564

  3. Metal nanoparticles triggered persistent negative photoconductivity in silk protein hydrogels

    Gogurla, Narendar; Sinha, Arun K.; Naskar, Deboki; Kundu, Subhas C.; Ray, Samit K.

    2016-03-01

    Silk protein is a natural biopolymer with intriguing properties, which are attractive for next generation bio-integrated electronic and photonic devices. Here, we demonstrate the negative photoconductive response of Bombyx mori silk protein fibroin hydrogels, triggered by Au nanoparticles. The room temperature electrical conductivity of Au-silk hydrogels is found to be enhanced with the incorporation of Au nanoparticles over the control sample, due to the increased charge transporting networks within the hydrogel. Au-silk lateral photoconductor devices show a unique negative photoconductive response under an illumination of 325 nm, with excitation energy higher than the characteristic metal plasmon resonance band. The enhanced photoconductance yield in the hydrogels over the silk protein is attributed to the photo-oxidation of amino groups in the β-pleated sheets of the silk around the Au nanoparticles followed by the breaking of charge transport networks. The Au-silk nanocomposite does not show any photoresponse under visible illumination because of the localization of excited charges in Au nanoparticles. The negative photoconductive response of hybrid Au-silk under UV illumination may pave the way towards the utilization of silk for future bio-photonic devices using metal nanoparticle platforms.

  4. Hierarchical charge distribution controls self-assembly process of silk in vitro

    Zhang, Yi; Zhang, Cencen; Liu, Lijie; Kaplan, David L.; Zhu, Hesun; Lu, Qiang

    2015-12-01

    Silk materials with different nanostructures have been developed without the understanding of the inherent transformation mechanism. Here we attempt to reveal the conversion road of the various nanostructures and determine the critical regulating factors. The regulating conversion processes influenced by a hierarchical charge distribution were investigated, showing different transformations between molecules, nanoparticles and nanofibers. Various repulsion and compressive forces existed among silk fibroin molecules and aggregates due to the exterior and interior distribution of charge, which further controlled their aggregating and deaggregating behaviors and finally formed nanofibers with different sizes. Synergistic action derived from molecular mobility and concentrations could also tune the assembly process and final nanostructures. It is suggested that the complicated silk fibroin assembly processes comply a same rule based on charge distribution, offering a promising way to develop silk-based materials with designed nanostructures.

  5. Particulate Sol Route Hydroxyapatite Thin Film-Silk Protein Interface Interactions

    Özcan, Selçuk; ÇİFTÇİOĞLU, Muhsin

    2010-01-01

    Hydroxyapatite (HAp) thin film coatings were prepared on bioinert glass slides by a particulate sol method and the effects of intermediate silk fibroin and silk sericin coatings on the HAp film formation and surface topography were examined. The films prepared with smaller crushed particle sols had a higher agglomeration tendency during the drying consolidation step of the thin film formation, and contained agglomerates larger in number and size, which was demonstrated experimentally and in a...

  6. Controlled Release from Multilayer Silk Biomaterial Coatings to Modulate Vascular Cell Responses

    Wang, Xianyan; Zhang, Xiaohui; Castellot, John; Herman, Ira; Iafrati, Mark; Kaplan, David L.

    2007-01-01

    A multilayered silk fibroin protein coating system was employed as a drug carrier and delivery system to evaluate vascular cell responses to heparin, paclitaxel, and clopidogrel. The results demonstrated that the silk coating system was an effective system for drug-eluting coatings, such as for stent applications, based on its useful micromechanical properties and biological outcomes. Cell attachment and viability studies with human aortic endothelial cells (HAECs) and human coronary artery s...

  7. Characterization on Eri (Philosamia ricini) Fibroin Film: Morphology, Structure and Thermal Properties

    Y. Srisuwan; N. Narkkong; P. Srihanam

    2009-01-01

    This study aims to prepare Eri Silk Fibroin (SF) solution and construct of its film. The Eri cocoons were firstly removed of sericin and then dissolved by 6 M Ca(NO3)2. The SF solution was dialyzed before measurement of silk percentage by weight. The solvate protein was then taken to prepare of SF film using 0.5 and 1% weight. The 10 mL of SF solution was cast on the polystyrene plates, left in room temperature for 2 days. The obtained SF films were then observed under Scanning Electron Micro...

  8. Enzymatic degradation behavior and cytocompatibility of silk fibroin–starch–chitosan conjugate membranes

    The objective of this study was to investigate the influence of silk fibroin and oxidized starch conjugation on the enzymatic degradation behavior and the cytocompatability of chitosan based biomaterials. The tensile stress of conjugate membranes, which was at 50 Megapascal (MPa) for the lowest fibroin and starch composition (10 weight percent (wt.%)), was decreased significantly with the increased content of fibroin and starch. The weight loss of conjugates in α-amylase was more notable when the starch concentration was the highest at 30 wt.%. The conjugates were resistant to the degradation by protease and lysozyme except for the conjugates with the lowest starch concentration. After 10 days of cell culture, the proliferation of osteoblast-like cells (SaOS-2) was stimulated significantly by higher fibroin compositions and the DNA synthesis on the conjugate with the highest fibroin (30 wt.%) was about two times more compared to the native chitosan. The light microscopy and the image analysis results showed that the cell area and the lengths were decreased significantly with higher fibroin/chitosan ratio. The study proved that the conjugation of fibroin and starch with the chitosan based biomaterials by the use of non-toxic reductive alkylation crosslinking significantly improved the cytocompatibility and modulated the biodegradation, respectively. - Highlights: ► Silk fibroin, starch and chitosan conjugates were prepared by reductive alkylation. ► The enzymatic biodegradation and the cytocompatibility of conjugates were tested. ► The conjugate with 30% starch composition was degraded by α-amylase significantly. ► Higher starch composition in conjugates prevented protease and lysozyme degradation. ► Fibroin incorporation effectively increased the cell proliferation of conjugates.

  9. TOWARDS A ROUTINE METHODOLOGY FOR ASSESSING THE CONDITION OF HISTORIC SILK

    Jeongjin Kim

    2009-01-01

    Full Text Available To ensure continued access and long-term preser vation itis essential to understand the condition of an artefact, andthe behaviour of its component materials, in order to setlimits on display and handling. In support of curators andconservators of collections with historic textiles, we aredeveloping rapid and routine technology, involving nearinfrared spectroscopy coupled with multivariate analysis,which, when applied non-invasively on-site, will allow theestimation of the state of deterioration of silk fabrics.Pursuing this aim, we have now carried out further analyticalstudies on silks artificially aged under different regimesfor up to 20 days: dry thermal, high heat and humidity, andsunlight equivalent irradiation. The tensile strengths, yellownessindices, apparent molecular weights of fibroin, andnear infrared spectral sorbed moisture parameters for theaged silks were determined. The mechanical performanceof silk diminished exponentially over each ageing timecourse. For the various silks, the increases in yellownessindices and decreases in fibroin molecular weights andmoisture contents showed similar kinetics to the mechanicalchanges. While each parameter was correlated with thetensile strength, silks exposed to the different acceleratedageing factors exhibited diverse correlations. It is concludedthat, when formulating a general model for characterisingthe condition of silk using chemometrics, the referenceset should include examples of silks which have beenexposed to the full variety of ageing factors.

  10. The expression of GFP under the control of fibroin promotor in primary ovarian cells of Antheraea pernyi

    Wenli Li; Liji Jin; Pengcheng Bu; Lijia An

    2003-12-01

    The fibroin promoter can stably express foreign gene in lepidopteran cells. Total RNA was extracted from the gland of silkworm, Antheraea pernyi and the transcription initiation site of fibroin gene of A. pernyi was identified by RNA ligase mediated rapid amplification of cDNA ends (RLM-RACE). The expression vector (pGFP-N2/Fib) was constructed by use of replacing the CMV promoter with the fibroin promoter. The results of visual screening under a fluorescent inverted microscope and Western blot analysis indicated that the GFP gene was expressed in the primary cells of ovary origins from A. pernyi.

  11. SPIDER SILK

    PORAV Viorica

    2014-01-01

    The strengthness and toughness of spider fiber and its multifunctional nature is only surpassed in some cases by synthetic high performance fibers. In the world of natural fibers, spider silk has been long time recognized as a wonder fiber for its unique combination of high strength and rupture elongation. Scientists in civil military engineering reveal that the power of biological material (spider silk) lies in the geometric configuration of structural protein, and the small cluster of we...

  12. Silk microgels formed by proteolytic enzyme activity.

    Samal, Sangram K; Dash, Mamoni; Chiellini, Federica; Kaplan, David L; Chiellini, Emo

    2013-09-01

    The proteolytic enzyme α-chymotrypsin selectively cleaves the amorphous regions of silk fibroin protein (SFP) and allows the crystalline regions to self-assemble into silk microgels (SMGs) at physiological temperature. These microgels consist of lamellar crystals in the micrometer scale, in contrast to the nanometer-scaled crystals in native silkworm fibers. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and zeta potential results demonstrated that α-chymotrypsin utilized only the non-amorphous domains or segments of the heavy chain of SFP to form negatively charged SMGs. The SMGs were characterized in terms of size, charge, structure, morphology, crystallinity, swelling kinetics, water content and thermal properties. The results suggest that the present technique of preparing SMGs by α-chymotrypsin is simple and efficient, and that the prepared SMGs have useful features for studies related to biomaterial and pharmaceutical needs. This process is also an easy way to obtain the amorphous peptide chains for further study. PMID:23756227

  13. In vitro phosphorylation as tool for modification of silk and keratin fibrous materials.

    Volkov, Vadim; Cavaco-Paulo, Artur

    2016-05-01

    An overview is given of the recent work on in vitro enzymatic phosphorylation of silk fibroin and human hair keratin. Opposing to many chemical "conventional" approaches, enzymatic phosphorylation is in fact a mild reaction and the treatment falls within "green chemistry" approach. Silk and keratin are not phosphorylated in vivo, but in vitro. This enzyme-driven modification is a major technological breakthrough. Harsh chemical chemicals are avoided, and mild conditions make enzymatic phosphorylation a real "green chemistry" approach. The current communication presents a novel approach stating that enzyme phosphorylation may be used as a tool to modify the surface charge of biocompatible materials such as keratin and silk. PMID:27075736

  14. Processing and characterization of silk sericin from Bombyx mori and its application in biomaterials and biomedicines.

    Cao, Ting-Ting; Zhang, Yu-Qing

    2016-04-01

    Bombyx mori silk is composed of 60-80% fibroin, 15-35% sericin and 1-5% non-sericin component including wax, pigments, sugars and other impurities. For two decades, the protein-based silk fibroin was extensively used in the research and development of medical biomaterials and biomedicines. Sericin is frequently ignored and abandoned as a byproduct or waste in the processing of traditional silk fabrics, silk floss or modern silk biomaterials. However, similar to fibroin, sericin is not only a highly useful biological material, but also a lot of biological activity. Moreover, the non-sericin component present with sericin in the cocoon shell also has a strong biological activity. In this review, the extraction and recovery methods of sericin and the non-sericin component from the cocoon layer are reported, and their composition, properties and biological activity are described to produce a comprehensive report on biomedical materials and biological drugs. In addition, related problems or concerns present in the research and development of sericin are discussed, and a potential application of sericin in sustainable development is also presented. PMID:26838924

  15. Novel and simple route to fabricate fully biocompatible plasmonic mushroom arrays adhered on silk biopolymer

    Park, Joonhan; Choi, Yunkyoung; Lee, Myungjae; Jeon, Heonsu; Kim, Sunghwan

    2014-12-01

    A fully biocompatible plasmonic quasi-3D nanostructure is demonstrated by a simple and reliable fabrication method using strong adhesion between gold and silk fibroin. The quasi-3D nature gives rise to complex photonic responses in reflectance that are prospectively useful in bio/chemical sensing applications. Laser interference lithography is utilized to fabricate large-area plasmonic nanostructures.A fully biocompatible plasmonic quasi-3D nanostructure is demonstrated by a simple and reliable fabrication method using strong adhesion between gold and silk fibroin. The quasi-3D nature gives rise to complex photonic responses in reflectance that are prospectively useful in bio/chemical sensing applications. Laser interference lithography is utilized to fabricate large-area plasmonic nanostructures. Electronic supplementary information (ESI) available: The incident angle dependence of reflectance spectra and the atomic force microscopy image of the Au nanoparticle array on a silk film after 1 hour of ultrasonication. See DOI: 10.1039/c4nr05172f

  16. High level expression and facile purification of recombinant silk-elastin-like polymers in auto induction shake flask cultures

    Machado, Raul; Azevedo-Silva, João; Correia, Cristina; Collins, Tony; Arias, Francisco Javier; Rodríguez-Cabello, Jose Carlos; Casal, Margarida

    2013-01-01

    Silk-elastin-like polymers (SELPs) are protein-based polymers composed of repetitive amino acid sequence motifs found in silk fibroin (GAGAGS) and mammalian elastin (VPGVG). These polymers are of much interest, both from a fundamental and applied point of view, finding potential application in biomedicine, nanotechnology and as materials. The successful employment of such polymers in such diverse fields, however, requires the ready availability of a variety of different forms with novel enhan...

  17. Effect of Sericin on Mechanical Behavior of Composite Material Reinforced by Silk Woven Fabric

    Kimura, Teruo; Ino, Haruhiro; Hanada, Koji; Katori, Sigetaka

    Recent, attention has been given to shift from glass fibers and carbon fibers to natural fibers for FRP composites for the goal of protecting the environment. This paper concerned with the application of silk fabric for composite materials. Polypropylene (PP) was used for the matrix material and the silk fabric composites were molded using a compression molding method. Especially, the effect of sericin on mechanical behaviors of composite materials was discussed. Good adhesion between silk and PP was obtained by removing the sericin existing around the fibroin. The tensile modulus of composite decreased with decreasing the sericin because of the flexibility of silk fibers without sericin. In particular, the higher Izod impact value was obtained for the composites containing the silk fibers without sericin.

  18. Production of fibroin nanopowder through electrospraying

    Fibroin is a biomaterial and in the powder form, has found applications such as food and cosmetic additive as well as drug delivery. Various methods have been employed to produce fibroin powder with different particle size range. In this study, a novel and original application of electrospraying technique capable of producing fibroin nanopowder is presented. Our technique is based on electrospraying of dilute fibroin solution in formic acid. Moreover, the effect of variables in electrospraying, namely, concentration of fibroin solution, voltage, feed rate, and needle–collector distance, on average particle size of fibroin nanopowder has been studied. The result of this study showed that electrospraying is capable of producing fibroin nanopowder with average particle size as low as 80 nm. In fact in comparison to other methods reported in the literature, electrospraying alongside with the precipitation method produce fibroin nanopowder with the lowest particle size. However, nanopowder obtained through electrospraying technique enjoys a more uniform spherical shape and size. As far as the variables are concerned, it was ascertained that lower concentrations, lower feed rates and longer needle–collector distances lead to a decrease in the average particle size of fibroin nanopowder. Increasing voltage up to 20 kV decreases the particle size; but with higher voltages the average particle size increases. FT-IR and XRD studies showed that the fibroin nanopowder has a β-sheets structure, similar to fibroin filaments but with a lower crystallinity index.

  19. Production of fibroin nanopowder through electrospraying

    Gholami, A.; Tavanai, H., E-mail: tavanai@cc.iut.ac.ir; Moradi, A. R. [Isfahan University of Technology, Department of Textile Engineering, Center of Excellence for Environmental Nanotechnology (Iran, Islamic Republic of)

    2011-05-15

    Fibroin is a biomaterial and in the powder form, has found applications such as food and cosmetic additive as well as drug delivery. Various methods have been employed to produce fibroin powder with different particle size range. In this study, a novel and original application of electrospraying technique capable of producing fibroin nanopowder is presented. Our technique is based on electrospraying of dilute fibroin solution in formic acid. Moreover, the effect of variables in electrospraying, namely, concentration of fibroin solution, voltage, feed rate, and needle-collector distance, on average particle size of fibroin nanopowder has been studied. The result of this study showed that electrospraying is capable of producing fibroin nanopowder with average particle size as low as 80 nm. In fact in comparison to other methods reported in the literature, electrospraying alongside with the precipitation method produce fibroin nanopowder with the lowest particle size. However, nanopowder obtained through electrospraying technique enjoys a more uniform spherical shape and size. As far as the variables are concerned, it was ascertained that lower concentrations, lower feed rates and longer needle-collector distances lead to a decrease in the average particle size of fibroin nanopowder. Increasing voltage up to 20 kV decreases the particle size; but with higher voltages the average particle size increases. FT-IR and XRD studies showed that the fibroin nanopowder has a {beta}-sheets structure, similar to fibroin filaments but with a lower crystallinity index.

  20. Silkomics: Insight into the Silk Spinning Process of Spiders.

    Dos Santos-Pinto, José Roberto Aparecido; Garcia, Ana Maria Caviquioli; Arcuri, Helen Andrade; Esteves, Franciele Grego; Salles, Heliana Clara; Lubec, Gert; Palma, Mario Sergio

    2016-04-01

    The proteins from the silk-producing glands were identified using both a bottom-up gel-based proteomic approach as well as from a shotgun proteomic approach. Additionally, the relationship between the functions of identified proteins and the spinning process was studied. A total of 125 proteins were identified in the major ampullate, 101 in the flagelliform, 77 in the aggregate, 75 in the tubuliform, 68 in the minor ampullate, and 23 in aciniform glands. On the basis of the functional classification using Gene Ontology, these proteins were organized into seven different groups according to their general function: (i) web silk proteins-spidroins, (ii) proteins related to the folding/conformation of spidroins, (iii) proteins that protect silk proteins from oxidative stress, (iv) proteins involved in fibrillar preservation of silks in the web, (v) proteins related to ion transport into and out of the glands during silk fiber spinning, (vi) proteins involved in prey capture and pre-digestion, and (vii) housekeeping proteins from all of the glands. Thus, a general mechanism of action for the identified proteins in the silk-producing glands from the Nephila clavipes spider was proposed; the current results also indicate that the webs play an active role in prey capture. PMID:26923066

  1. Radiation processing of silk protein (Bilateral research cooperation OAEP and JAERI. December 1998 - December 2002)

    Thailand's production of silk, about 1,200 ton per year, also gives about 10% of silk waste which is expected to be recycled into new material (non-textile application) and to avoid environmental pollution. For this purpose, cooperative program 'radiation processing of silk protein' was conducted between OAEP (Thailand) and JAERI. Among the results already obtained are: radiation degradation of silk protein (fibroin) with gamma rays at 160 kGy, production of fine silk milled powder (<90 microns) by electron beam irradiation at 250-1000 kGy (dry method) using electron accelerator (1 MeV, 1 mA), use of antioxidant effect of silk protein on lipid peroxidation and antibacterial activity of irradiated silk protein powder, and wound dressing hydrogel mixed with silk protein and use of antibacterial activity of cross-linked silk protein/PVA hydrogel. Other topics of interest are gamma irradiation of anionic natural polymer solution for use as latex protein scavenger and gamma radiation degradation of chitosan for use as plant growth promoter and fungicide. (S. Ohno)

  2. Role of pH and charge on silk protein assembly in insects and spiders

    Foo, C. Wong Po; Bini, E.; Hensman, J.; Knight, D. P.; Lewis, R. V.; Kaplan, D. L.

    2006-02-01

    Silk fibers possess impressive mechanical properties, dependant, in part, on the crystalline ?-sheets silk II conformation. The transition to silk II from soluble silk I-like conformation in silk glands, is thought to originate in the spinning ducts immediately before the silk is drawn down into a fiber. However the assembly process of these silk molecules into fibers, whether in silkworms or spiders, is not well understood. Extensional flow, protein concentration, pH and metal ion concentrations are thought to be most important in in vivo silk processing and in affecting structural conformations. We look at how parameters such as pH, [Ca2+], [K+], and [Cu2+], and water content, interact with the domain structure of silk proteins towards the successful storage and processing of these concentrated hydrophobic silk proteins. Our recent domain mapping studies of all known silk proteins, and 2D Raman spectroscopy, NMR, and DLS studies performed on sections of silkworm gland, suggest that low pH and gradual water removal promote intermolecular over intramolecular hydrogen bonding. This discussion helps to provide the necessary ground rules towards the design of silk protein analogues with specific hydrophobicity and charge profiles to optimize expression, solubility and assembly with implications in structural biology and material science.

  3. Physical characterization of functionalized spider silk: electronic and sensing properties

    Eden Steven, Jin Gyu Park, Anant Paravastu, Elsa Branco Lopes, James S Brooks, Ongi Englander, Theo Siegrist, Papatya Kaner and Rufina G Alamo

    2011-01-01

    Full Text Available This work explores functional, fundamental and applied aspects of naturally harvested spider silk fibers. Natural silk is a protein polymer where different amino acids control the physical properties of fibroin bundles, producing, for example, combinations of β-sheet (crystalline and amorphous (helical structural regions. This complexity presents opportunities for functional modification to obtain new types of material properties. Electrical conductivity is the starting point of this investigation, where the insulating nature of neat silk under ambient conditions is described first. Modification of the conductivity by humidity, exposure to polar solvents, iodine doping, pyrolization and deposition of a thin metallic film are explored next. The conductivity increases exponentially with relative humidity and/or solvent, whereas only an incremental increase occurs after iodine doping. In contrast, iodine doping, optimal at 70 °C, has a strong effect on the morphology of silk bundles (increasing their size, on the process of pyrolization (suppressing mass loss rates and on the resulting carbonized fiber structure (that becomes more robust against bending and strain. The effects of iodine doping and other functional parameters (vacuum and thin film coating motivated an investigation with magic angle spinning nuclear magnetic resonance (MAS-NMR to monitor doping-induced changes in the amino acid-protein backbone signature. MAS-NMR revealed a moderate effect of iodine on the helical and β-sheet structures, and a lesser effect of gold sputtering. The effects of iodine doping were further probed by Fourier transform infrared (FTIR spectroscopy, revealing a partial transformation of β-sheet-to-amorphous constituency. A model is proposed, based on the findings from the MAS-NMR and FTIR, which involves iodine-induced changes in the silk fibroin bundle environment that can account for the altered physical properties. Finally, proof-of-concept applications of functionalized spider silk are presented for thermoelectric (Seebeck effects and incandescence in iodine-doped pyrolized silk fibers, and metallic conductivity and flexibility of micron-sized gold-sputtered silk fibers. In the latter case, we demonstrate the application of gold-sputtered neat spider silk to make four-terminal, flexible, ohmic contacts to organic superconductor samples.

  4. Physical characterization of functionalized spider silk: electronic and sensing properties

    This work explores functional, fundamental and applied aspects of naturally harvested spider silk fibers. Natural silk is a protein polymer where different amino acids control the physical properties of fibroin bundles, producing, for example, combinations of ?-sheet (crystalline) and amorphous (helical) structural regions. This complexity presents opportunities for functional modification to obtain new types of material properties. Electrical conductivity is the starting point of this investigation, where the insulating nature of neat silk under ambient conditions is described first. Modification of the conductivity by humidity, exposure to polar solvents, iodine doping, pyrolization and deposition of a thin metallic film are explored next. The conductivity increases exponentially with relative humidity and/or solvent, whereas only an incremental increase occurs after iodine doping. In contrast, iodine doping, optimal at 70 deg. C, has a strong effect on the morphology of silk bundles (increasing their size), on the process of pyrolization (suppressing mass loss rates) and on the resulting carbonized fiber structure (that becomes more robust against bending and strain). The effects of iodine doping and other functional parameters (vacuum and thin film coating) motivated an investigation with magic angle spinning nuclear magnetic resonance (MAS-NMR) to monitor doping-induced changes in the amino acid-protein backbone signature. MAS-NMR revealed a moderate effect of iodine on the helical and ?-sheet structures, and a lesser effect of gold sputtering. The effects of iodine doping were further probed by Fourier transform infrared (FTIR) spectroscopy, revealing a partial transformation of ?-sheet-to-amorphous constituency. A model is proposed, based on the findings from the MAS-NMR and FTIR, which involves iodine-induced changes in the silk fibroin bundle environment that can account for the altered physical properties. Finally, proof-of-concept applications of functionalized spider silk are presented for thermoelectric (Seebeck) effects and incandescence in iodine-doped pyrolized silk fibers, and metallic conductivity and flexibility of micron-sized gold-sputtered silk fibers. In the latter case, we demonstrate the application of gold-sputtered neat spider silk to make four-terminal, flexible, ohmic contacts to organic superconductor samples.

  5. Physical characterization of functionalized spider silk: electronic and sensing properties

    Steven, Eden; Brooks, James S [Department of Physics and National High Magnetic Field Laboratory, Florida State University, 1800 East Paul Dirac, Tallahassee, FL 32310 (United States); Park, Jin Gyu [FAMU-FSU Department of Industrial and Manufacturing Engineering, High-Performance Materials Institute, Florida State University, 2005 Levy Ave., Tallahassee, FL 32310 (United States); Paravastu, Anant; Siegrist, Theo; Kaner, Papatya; Alamo, Rufina G [FAMU-FSU Department of Chemical and Biomedical Engineering and National High Magnetic Field Laboratory, Florida State University, 1800 East Paul Dirac, Tallahassee, FL 32310 (United States); Branco Lopes, Elsa [Departamento de Quimica, Instituto Tecnologico e Nuclear/CFMC-UL, P-2686-953 Sacavem (Portugal); Englander, Ongi, E-mail: esteven@magnet.fsu.edu [FAMU-FSU Department of Mechanical Engineering and National High Magnetic Field Laboratory, Florida State University, 1800 East Paul Dirac, Tallahassee, Florida 32310 (United States)

    2011-10-15

    This work explores functional, fundamental and applied aspects of naturally harvested spider silk fibers. Natural silk is a protein polymer where different amino acids control the physical properties of fibroin bundles, producing, for example, combinations of {beta}-sheet (crystalline) and amorphous (helical) structural regions. This complexity presents opportunities for functional modification to obtain new types of material properties. Electrical conductivity is the starting point of this investigation, where the insulating nature of neat silk under ambient conditions is described first. Modification of the conductivity by humidity, exposure to polar solvents, iodine doping, pyrolization and deposition of a thin metallic film are explored next. The conductivity increases exponentially with relative humidity and/or solvent, whereas only an incremental increase occurs after iodine doping. In contrast, iodine doping, optimal at 70 deg. C, has a strong effect on the morphology of silk bundles (increasing their size), on the process of pyrolization (suppressing mass loss rates) and on the resulting carbonized fiber structure (that becomes more robust against bending and strain). The effects of iodine doping and other functional parameters (vacuum and thin film coating) motivated an investigation with magic angle spinning nuclear magnetic resonance (MAS-NMR) to monitor doping-induced changes in the amino acid-protein backbone signature. MAS-NMR revealed a moderate effect of iodine on the helical and {beta}-sheet structures, and a lesser effect of gold sputtering. The effects of iodine doping were further probed by Fourier transform infrared (FTIR) spectroscopy, revealing a partial transformation of {beta}-sheet-to-amorphous constituency. A model is proposed, based on the findings from the MAS-NMR and FTIR, which involves iodine-induced changes in the silk fibroin bundle environment that can account for the altered physical properties. Finally, proof-of-concept applications of functionalized spider silk are presented for thermoelectric (Seebeck) effects and incandescence in iodine-doped pyrolized silk fibers, and metallic conductivity and flexibility of micron-sized gold-sputtered silk fibers. In the latter case, we demonstrate the application of gold-sputtered neat spider silk to make four-terminal, flexible, ohmic contacts to organic superconductor samples.

  6. Silks as scaffolds for skin reconstruction.

    Reimers, Kerstin; Liebsch, Christina; Radtke, Christine; Kuhbier, Jörn W; Vogt, Peter M

    2015-11-01

    In this short review, we describe the use of high molecular weight proteins produced in the glands of several arthropods-commonly called silks-for the purpose to enhance human skin wound healing. To this end an extensive literature search has been performed, the publications have been categorized concerning silk preparation and application and summarized accordingly: Scaffolds to promote wound healing were prepared by processing the silks in different ways including solubilization of the protein fibers followed by casting or electrospinning. The silk scaffolds were additionally modified by coating or blending with the intention of further functionalization. In several approaches, the scaffolds were also vitalized with skin cells or stem cells. In vitro and in vivo models were implied to test for safety and efficiency. We conclude that silk scaffolds are characterized by an advantageous biocompatibility as well as an impressive versatility rendering them ideally suited for application in wounds. Nevertheless, further investigation is needed to exploit the full capacity of silk in different wound models and to achieve clinical transfer in time. PMID:25995140

  7. Silk-tropoelastin protein films for nerve guidance.

    White, James D; Wang, Siran; Weiss, Anthony S; Kaplan, David L

    2015-03-01

    Peripheral nerve regeneration may be enhanced through the use of biodegradable thin film biomaterials as highly tuned inner nerve conduit liners. Dorsal root ganglion neuron and Schwann cell responses were studied on protein films comprising silk fibroin blended with recombinant human tropoelastin protein. Tropoelastin significantly improved neurite extension and enhanced Schwann cell process length and cell area, while the silk provided a robust biomaterial template. Silk-tropoelastin blends afforded a 2.4-fold increase in neurite extension, when compared to silk films coated with poly-d-lysine. When patterned by drying on grooved polydimethylsiloxane (3.5 μm groove width, 0.5 μm groove depth), these protein blends induced both neurite and Schwann cell process alignment. Neurons were functional as assessed using patch-clamping, and displayed action potentials similar to those cultured on poly(lysine)-coated glass. Taken together, silk-tropoelastin films offer useful biomaterial interfacial platforms for nerve cell control, which can be considered for neurite guidance, disease models for neuropathies and surgical peripheral nerve repairs. PMID:25481743

  8. Effect of degumming time on silkworm silk fibre for biodegradable polymer composites

    Ho, Mei-po; Wang, Hao; Lau, Kin-tak

    2012-02-01

    Recently, many studies have been conducted on exploitation of natural materials for modern product development and bioengineering applications. Apart from plant-based materials (such as sisal, hemp, jute, bamboo and palm fibre), animal-based fibre is a kind of sustainable natural materials for making novel composites. Silkworm silk fibre extracted from cocoon has been well recognized as a promising material for bio-medical engineering applications because of its superior mechanical and bioresorbable properties. However, when producing silk fibre reinforced biodegradable/bioresorbable polymer composites, hydrophilic sericin has been found to cause poor interfacial bonding with most polymers and thus, it results in affecting the resultant properties of the composites. Besides, sericin layers on fibroin surface may also cause an adverse effect towards biocompatibility and hypersensitivity to silk for implant applications. Therefore, a proper pre-treatment should be done for sericin removal. Degumming is a surface modification process which allows a wide control of the silk fibre's properties, making the silk fibre possible to be used for the development and production of novel bio-composites with unique/specific mechanical and biodegradable properties. In this paper, a cleaner and environmentally friendly surface modification technique for tussah silk in polymer based composites is proposed. The effectiveness of different degumming parameters including degumming time and temperature on tussah silk is discussed through the analyses of their mechanical and morphological properties. Based on results obtained, it was found that the mechanical properties of tussah silk are affected by the degumming time due to the change of the fibre structure and fibroin alignment.

  9. Sex differences in cooperative silk-spinning by weaver ant larvae

    Wilson, Edward O.; Hölldobler, Bert

    1980-01-01

    Final-instar larvae of weaver ants (Oecophylla longinoda) use their silk for nest construction rather than for cocoon spinning; this commitment represents a shift from entirely selfish to entirely cooperative behavior that occurred after the emergence of the phylogenetically advanced subfamily Formicinae. Male larvae were found to have smaller silk glands and to contribute substantially less silk to nest construction. The hypothesis of kin selection appears to be the most plausible and parsim...

  10. Inner ear delivery of dexamethasone using injectable silk-polyethylene glycol (PEG) hydrogel.

    Yu, Dehong; Sun, Changling; Zheng, Zhaozhu; Wang, Xueling; Chen, Dongye; Wu, Hao; Wang, Xiaoqin; Shi, Fuxin

    2016-04-30

    Minimally invasive delivery and sustained release of therapeutics to the inner ear are of importance to the medical treatment of inner ear disease. In this study, the injectable silk fibroin-polyethylene glycol (Silk-PEG) hydrogel was investigated as a drug delivery carrier to deliver poorly soluble micronized dexamethasone (mDEX) to the inner ear of guinea pigs. Encapsulation of mDEX with a loading up to 5% (w/v) did not significantly change the silk gelation time, and mDEX were evenly distributed in the PEG-Silk hydrogel as visualized by SEM. The loading of mDEX in Silk-PEG hydrogel largely influenced in vitro drug release kinetics. The optimized Silk-PEG-mDEX hydrogel (2.5% w/v loading, in situ-forming,10μl) was administered directly onto the round window membrane of guinea pigs. The DEX concentration in perilymph maintained above 100ng/ml for at least 10 days for the Silk-PEG formulation while less than 12h for the control sample of free mDEX. Minimal systemic exposure was achieved with low DEX concentrations (<0.2μg/ml) in cerebrospinal fluid (CSF) and plasma in the first 2h after the local application of the Silk-PEG-mDEX hydrogel. A transient hearing threshold shift was found but then resolved after 14days as revealed by auditory brainstem response (ABR), showing minimal inflammatory responses on the round window membrane and scala taympani. The Silk-PEG hydrogel completely degraded in 21 days. Thus, the injectable PEG-Silk hydrogel is an effective and safe vehicle for inner ear delivery and sustained release of glucocorticoid. PMID:26972377

  11. Study on spinning process of silkworm by using synchrotron radiation X-ray diffraction

    The spinning process of a fifth instar Bombyx mori silkworm was studied in situ by synchrotron radiation wide angle X-ray diffraction (SR-WAXD). Silk glands dissected out from the larvae, and degummed cocoon silk were also examined by SR-WAXD. The effects of drying and drawing on the development of ordered structure of silk fibroin in the silk gland were investigated. The results show that ordered structures related to silk I and silk II are formed in the spinning dope near the spinneret of the silkworm head. During drying process of the silk gland, dehydration results in the formation of ordered structure of silk fibroin. Its conformation transforms to anti-parallel ?-sheet which tends to construct perfect lamellar crystalline structure of silk II during drying. In addition, drawing is another important factor to induce the formation of ordered structure of silk fibroin and resultant fiber. (authors)

  12. Reproducing Natural Spider Silks' Copolymer Behavior in Synthetic Silk Mimics

    An, Bo; Jenkins, Janelle E; Sampath, Sujatha; Holland, Gregory P; Hinman, Mike; Yarger, Jeffery L; Lewis, Randolph [Wyoming; (Sandia); (Utah SU); (AZU)

    2012-10-30

    Dragline silk from orb-weaving spiders is a copolymer of two large proteins, major ampullate spidroin 1 (MaSp1) and 2 (MaSp2). The ratio of these proteins is known to have a large variation across different species of orb-weaving spiders. NMR results from gland material of two different species of spiders, N. clavipes and A. aurantia, indicates that MaSp1 proteins are more easily formed into β-sheet nanostructures, while MaSp2 proteins form random coil and helical structures. To test if this behavior of natural silk proteins could be reproduced by recombinantly produced spider silk mimic protein, recombinant MaSp1/MaSp2 mixed fibers as well as chimeric silk fibers from MaSp1 and MaSp2 sequences in a single protein were produced based on the variable ratio and conserved motifs of MaSp1 and MaSp2 in native silk fiber. Mechanical properties, solid-state NMR, and XRD results of tested synthetic fibers indicate the differing roles of MaSp1 and MaSp2 in the fiber and verify the importance of postspin stretching treatment in helping the fiber to form the proper spatial structure.

  13. Natural Silk as a Photonics Component: a Study on Its Light Guiding and Nonlinear Optical Properties

    Kujala, Sami; Mannila, Anna; Karvonen, Lasse; Kieu, Khanh; Sun, Zhipei

    2016-03-01

    Silk fibers are expected to become a pathway to biocompatible and bioresorbable waveguides, which could be used to deliver localized optical power for various applications, e.g., optical therapy or imaging inside living tissue. Here, for the first time, the linear and nonlinear optical properties of natural silk fibers have been studied. The waveguiding properties of silk fibroin of largely unprocessed Bombyx mori silkworm silk are assessed using two complementary methods, and found to be on the average 2.8 dB mm-1. The waveguide losses of degummed silk are to a large extent due to scattering from debris on fiber surface and helical twisting of the fiber. Nonlinear optical microscopy reveals both configurational defects such as torsional twisting, and strong symmetry breaking at the center of the fiber, which provides potential for various nonlinear applications. Our results show that nonregenerated B. mori silk can be used for delivering optical power over short distances, when the waveguide needs to be biocompatible and bioresorbable, such as embedding the waveguide inside living tissue.

  14. Natural Silk as a Photonics Component: a Study on Its Light Guiding and Nonlinear Optical Properties

    Kujala, Sami; Mannila, Anna; Karvonen, Lasse; Kieu, Khanh; Sun, Zhipei

    2016-01-01

    Silk fibers are expected to become a pathway to biocompatible and bioresorbable waveguides, which could be used to deliver localized optical power for various applications, e.g., optical therapy or imaging inside living tissue. Here, for the first time, the linear and nonlinear optical properties of natural silk fibers have been studied. The waveguiding properties of silk fibroin of largely unprocessed Bombyx mori silkworm silk are assessed using two complementary methods, and found to be on the average 2.8 dB mm−1. The waveguide losses of degummed silk are to a large extent due to scattering from debris on fiber surface and helical twisting of the fiber. Nonlinear optical microscopy reveals both configurational defects such as torsional twisting, and strong symmetry breaking at the center of the fiber, which provides potential for various nonlinear applications. Our results show that nonregenerated B. mori silk can be used for delivering optical power over short distances, when the waveguide needs to be biocompatible and bioresorbable, such as embedding the waveguide inside living tissue. PMID:26926272

  15. Characterization and mechanical performance study of silk/PVA cryogels: towards nucleus pulposus tissue engineering

    Poly (vinyl) alcohol (PVA) cryogels are reported in the literature for application in nucleus pulposus (NP) replacement strategies. However, these studies are mainly limited to acellular approaches—in part due to the high hydrophilicity of PVA gels that renders cellular adhesion difficult. Silk is a versatile biomaterial with excellent biocompatibility. We hypothesize that the incorporation of silk with PVA will (i) improve the cell-hosting abilities of PVA cryogels and (ii) allow better tailoring of physical properties of the composite cryogels for an NP tissue engineering purpose. 5% (wt/vol) PVA is blended with 5% silk fibroin (wt/vol) to investigate the effect of silk : PVA ratios on the cryogels’ physical properties. Results show that the addition of silk results in composite cryogels that are able to swell to more than 10 times its original dry weight and rehydrate to at least 70% of its original wet weight. Adding at least 20% silk significantly improves surface hydrophobicity and is correlated with an improvement in cell-hosting abilities. Cell-seeded cryogels also display an increment in compressive modulus and hoop stress values. In all, adding silk to PVA creates cryogels that can be potentially used as NP replacements. (paper)

  16. Water-insoluble Silk Films with Silk I Structure

    Lu, Q.; Hu, X; Wang, X; Kluge, J; Lu, S; Cebe, P; Kaplan, D

    2010-01-01

    Water-insoluble regenerated silk materials are normally produced by increasing the {beta}-sheet content (silk II). In the present study water-insoluble silk films were prepared by controlling the very slow drying of Bombyx mori silk solutions, resulting in the formation of stable films with a predominant silk I instead of silk II structure. Wide angle X-ray scattering indicated that the silk films stabilized by slow drying were mainly composed of silk I rather than silk II, while water- and methanol-annealed silk films had a higher silk II content. The silk films prepared by slow drying had a globule-like structure at the core surrounded by nano-filaments. The core region was composed of silk I and silk II, surrounded by hydrophilic nano-filaments containing random turns and {alpha}-helix secondary structures. The insoluble silk films prepared by slow drying had unique thermal, mechanical and degradative properties. Differential scanning calorimetry results revealed that silk I crystals had stable thermal properties up to 250 C, without crystallization above the T{sub g}, but degraded at lower temperatures than silk II structure. Compared with water- and methanol-annealed films the films prepared by slow drying had better mechanical ductility and were more rapidly enzymatically degraded, reflecting the differences in secondary structure achieved via differences in post processing of the cast silk films. Importantly, the silk I structure, a key intermediate secondary structure for the formation of mechanically robust natural silk fibers, was successfully generated by the present approach of very slow drying, mimicking the natural process. The results also point to a new mode of generating new types of silk biomaterials with enhanced mechanical properties and increased degradation rates, while maintaining water insolubility, along with a low {beta}-sheet content.

  17. Silk Batik using Cochineal Dye

    The history of silk, including sericulture (the production of raw silk, which requires the raising of silkworms on their natural diet, mulberry leaves) and silk manufacturing, is rich and extensive. It encompasses several famous “silk roads” (trade routes), various cultures and technologies, ideas,...

  18. Towards Silk Fiber Optics: Refractive Index Characterization, Fiber Spinning, and Spinneret Analysis

    Spitzberg, Joshua David

    Of the many biologically derived materials, whose historical record of use by humans underscores an ex-vivo utility, silk is interesting for it's contemporary repurposing from textile to biocompatible substrate. And while even within this category silk is one of several materials studied for novel repurposing, it has the unique character of being evolutionarily developed specifically for fiber spinning in vivo. The work discussed here is inspired by taking what nature has given, to explore the in vitro spinning of silk towards biocompatible fiber optics applications. A common formulation of silk used in biomedical studies for re-forming it into the various structures begins with the silkworm cocoon, which is degummed and dissolved into an aqueous solution of its miscible protein, fibroin, and post-treated to fabricate solid structures. In the first aim, the optical refractive index (RI) of various post-treatment methods is discussed towards determining RI design techniques. The methods considered in this work for re-forming a solid fiber from the reconstituted silk fibroin (RSF) solution borrow from the industrial techniques of gel spinning, and dry-spinning. In the second aim, methods are applied to RSF and quality of the spun fibers discussed. A feature common to spinning techniques is passing the (silk) material through a spinneret of specific shape. In the third aim, fluid flow through a simplified native silkworm spinneret is modeled towards bio-inspired lessons in design. In chapter 1 the history, reconstitution, are discussed towards understanding the fabrication of several optical device examples. Chapter 2 then prefaces the experiments and measurements in fiber optics by reviewing electromagnetic theory of waveguide function, and loss factors, to be considered in actual device fabrication. Chapter 3 presents results and discussion for the first aim, understanding design principles for the refractive index of RSF. From this point, industrial fiber-spinning approaches are reviewed from a theoretical and methodological perspective in chapter 4. Thus, chapter 5 presents results for the second aim, efforts to apply these techniques using RSF. Chapter 6 discusses the third aim, understanding the design of the silkworm spinneret by an idealized model of natural and reconstituted silk fibroin flow. While the ultimate goal of a structurally and optically smooth and uniform fiber remains elusive, this work serves as a guide for future efforts.

  19. Microscopic structural analysis of fractured silk fibers from Bombyx mori and Samia cynthia ricini using 13C CP/MAS NMR with a 1 mm microcoil MAS NMR probehead

    Yamauchi, Kazuo

    2010-07-01

    Conformational changes have been studied in silk fibers from the domestic silkworm Bombyx mori and a wild silkworm Samia cynthia ricini as a result of fractured by stretching. About 300 samples consisting of only the fractured regions of [1-13C]Ala or [1-13C]Gly labeled silk fibers were collected and observed by 13C CP/MAS NMR spectra. The total amount of these fractured fibers is only about 1 mg and therefore we used a home-built 1 mm microcoil MAS NMR probehead. A very small increase in the fraction of random coil was noted for the alanine regions of both silk fibroins and for the glycine region of B. mori silk fibroin. However, there is no difference in the spectra before and after fractured for the glycine region of S. c. ricini silk fibroin. Thus, the influence of fracture occurs exclusively at the Ala region for S. c. ricini. The relationship between sequence, fracture and structure is discussed. © 2010 Elsevier Inc. All rights reserved.

  20. Tuning assembly and enzymatic degradation of silk/poly(N-vinylcaprolactam) multilayers via molecular weight and hydrophobicity.

    Espinosa-Dzib, Alejandra; Chen, Jun; Zavgorodnya, Oleksandra; Kozlovskaya, Veronika; Liang, Xing; Kharlampieva, Eugenia

    2015-07-01

    We report on enzymatically degradable nanothin coatings obtained by layer-by-layer (LbL) assembly of silk fibroin with poly(N-vinylcaprolactam) (PVCL) via hydrogen bonding and hydrophobic interactions. We found that both silk β-sheet content, controlled through dipping and spin-assisted LbL, and PVCL molecular weight regulate film thickness, microstructure, pH-stability, and biodegradability with a nanoscale precision. Thickness of (silk/PVCL) films increased with increase in PVCL molecular weight and decrease in deposition pH. The impact of assembly pH on film growth was more dramatic for dipped films. These systems show a significant rise in thickness with increase in PVCL molecular weight at pH films exhibited a greater stability at elevated pH and against enzymatic degradation as compared to their dipped counterparts. For both film types, the pH and enzymatic stability was improved with increasing PVCL length and β-sheet content, indicating enhanced hydrophobic and hydrogen-bonded interactions between PVCL and silk. Finally, we fabricated spherical and cubical (silk/PVCL) LbL capsules of regulated permeability and enzymatic degradation. Our approach gives a unique opportunity to tune thickness, morphology, structure, and biodegradability rate of silk films and capsules by varying silk secondary structure and PVCL length. Accounting for all-aqueous fabrication and the biocompatibility of both polymers these biodegradable materials provide novel platforms for delivery systems and medical devices. PMID:26041120

  1. Preparation, characterization and biological test of 3D-scaffolds based on chitosan, fibroin and hydroxyapatite for bone tissue engineering

    This work describes the preparation and characterization of porous 3D-scaffolds based on chitosan (CHI), chitosan/silk fibroin (CHI/SF) and chitosan/silk fibroin/hydroxyapatite (CHI/SF/HA) by freeze drying. The biomaterials were characterized by X-ray diffraction, attenuated total reflection Fourier transform infrared spectroscopy, thermogravimetric analysis, differential scanning calorimetry, scanning electron microscopy and energy dispersive spectroscopy. In addition, studies of porosity, pore size, contact angle and biological response of SaOs-2osteoblastic cells were performed. The CHI scaffolds have a porosity of 94.2 ± 0.9%, which is statistically higher than the one presented by CHI/SF/HA scaffolds, 89.7 ± 2.6%. Although all scaffolds were able to promote adhesion, growth and maintenance of osteogenic differentiation of SaOs-2 cells, the new 3D-scaffold based on CHI/SF/HA showed a significantly higher cell growth at 7 days and 21 days and the level of alkaline phosphatase at 14 and 21 days was statistically superior compared to other tested materials. - Highlights: • Preparation of 3D-scaffolds based on CHI, with or without addition of SF and HA. • Scaffolds exhibited interconnected porous structure (pore size superior to 50 μm). • The tripolyphosphate did not induce any significant cytotoxic response. • The CHI/SF/HA composite showed a higher cell growth and ALP activity

  2. Preparation, characterization and biological test of 3D-scaffolds based on chitosan, fibroin and hydroxyapatite for bone tissue engineering

    Lima, Paulo Autran Leite; Resende, Cristiane Xavier [Departamento de Ciências de Materiais, Universidade Federal de Sergipe, Av. Marechal Rondon, s/n. Jardim Rosa Elze, São Cristóvão, Sergipe CEP 49000-100 (Brazil); Dulce de Almeida Soares, Glória [Departamento de Ciências de Materiais, Universidade Federal do Rio de Janeiro, Av. Brigadeiro Trompowisk, s/n. Ilha do Fundão, Rio de Janeiro, Rio de Janeiro CEP 21900-000 (Brazil); Anselme, Karine [Institut de Science des Matériaux de Mulhouse (IS2M), CNRS LRC7228, 15, Jean Starcky Street, BP 2488, 68054 Mulhouse cedex (France); Almeida, Luís Eduardo, E-mail: lealmeida2009@gmail.com [Departamento de Ciências de Materiais, Universidade Federal de Sergipe, Av. Marechal Rondon, s/n. Jardim Rosa Elze, São Cristóvão, Sergipe CEP 49000-100 (Brazil)

    2013-08-01

    This work describes the preparation and characterization of porous 3D-scaffolds based on chitosan (CHI), chitosan/silk fibroin (CHI/SF) and chitosan/silk fibroin/hydroxyapatite (CHI/SF/HA) by freeze drying. The biomaterials were characterized by X-ray diffraction, attenuated total reflection Fourier transform infrared spectroscopy, thermogravimetric analysis, differential scanning calorimetry, scanning electron microscopy and energy dispersive spectroscopy. In addition, studies of porosity, pore size, contact angle and biological response of SaOs-2osteoblastic cells were performed. The CHI scaffolds have a porosity of 94.2 ± 0.9%, which is statistically higher than the one presented by CHI/SF/HA scaffolds, 89.7 ± 2.6%. Although all scaffolds were able to promote adhesion, growth and maintenance of osteogenic differentiation of SaOs-2 cells, the new 3D-scaffold based on CHI/SF/HA showed a significantly higher cell growth at 7 days and 21 days and the level of alkaline phosphatase at 14 and 21 days was statistically superior compared to other tested materials. - Highlights: • Preparation of 3D-scaffolds based on CHI, with or without addition of SF and HA. • Scaffolds exhibited interconnected porous structure (pore size superior to 50 μm). • The tripolyphosphate did not induce any significant cytotoxic response. • The CHI/SF/HA composite showed a higher cell growth and ALP activity.

  3. [Engineered spider silk: the intelligent biomaterial of the future. Part II].

    Kaźmierska, Katarzyna; Florczak, Anna; Piekoś, Konrad; Mackiewicz, Andrzej; Dams-Kozłowska, Hanna

    2011-01-01

    The development and progress in engineered spider silk manufacturing has enabled its practical application. Recombinant spider silk can assemble in several morphological forms such as films, hydrogels, fibers, scaffolds, microcapsules, and micro- and nanospheres. The in vitro fiber formation takes place by mimicking the natural spinning process in the spider spinning gland: in the presence of phosphate ions and dragging forces. Films are obtained by evaporation of solvent from the silk solution, while the result of evaporation of the solvent in the presence of porogens is a silk scaffold. Hydrogels are formed by spontaneous polymerization of silk particles in solutions at low pH. The silk film assembled at the interface of two immiscible phases forms microcapsules. The smallest of the described forms--silk spheres--are obtained by salting out the silk protein solution after addition of the phosphate ions. Common properties of the silk biomaterials are biocompatibility and biodegradability, which make them suitable for a number of applications in medicine and pharmacy. Moreover, the strategy of hybrid proteins which provides the desired function to biomaterial will further expand their potential use. PMID:21734323

  4. Ultrastructure of the Lyonet's glands in larvae of Diatraea saccharalis Fabricius (Lepidoptera: Pyralidae)

    Eliane Victoriano; GREGRIO, ELISA A.

    2004-01-01

    The Lyonet's gland is found in Lepidoptera larvae, close to the excretory duct of the silk gland. The role played by this gland is still uncertain. This work aims to describe the ultrastructure of the Lyonet's gland in Diatraea saccharalis larvae, offering suggestions regarding its possible function. The insects were reared under laboratory-controlled conditions. The glands were conventionally prepared for transmission (TEM) and scanning (SEM) electron microscopy. SEM showed that Lyonet's gla...

  5. Ultrastructure of the silk glands in three adult females of sphecid wasps of the genus Microstigmus (Hymenoptera: Pemphredoninae Ultraestructura de las glándulas de seda en hembras adultas de tres especies de avispas del género Microstigmus (Hymenoptera: Sphecidae: Pemphredoninae

    JOSÉ EDUARDO SERRÃO

    2005-03-01

    Full Text Available The ultrastructure of silk glands in adult females of sphecid wasps is described. Individual glands are scattered in the more posterior portion of the metasoma being each gland formed by an enlarged secretory unity with one conducting canal. Each secretory unit is a gland cell containing a large lumen and numerous secretory granules, which are delimited by membrane and filled with middle electron dense content of homogeneous aspect, which are exocytosed into the extra cellular lumen. Cell cytoplasm is filled with rough endoplasmic reticulum, polyribosomes and Golgi apparatuses profiles. The basal plasma membrane has many short infoldings and is lined by a thin basement membrane. The apical membrane is invaginated to form the boundary of the lumen, containing scattered short microvilli. The cells are mononucleated and the nucleus is pleomorphic containing disperse chromatin. The extracellular lumen is separated of the receiving canal by a convoluted sponge-like wall forming many villi, which in their tip are lined by a thin single layered cuticle. In the basal region of the villi a cuticular covering is lacking. In the conducting canal secretion acquire two different electron densities. A thin cuticle lines the lumen of conducting canal and the cell cytoplasm is scarce and the organelles are represented for few rough endoplasmic reticulum and polyribosomes. These results are discussed in relation to function and origin of silk gland in adult SphecidaeSe describe la ultraestructura de las glándulas de la seda en tres hembras adultas de avispas Sphecidae del género Microstigmus. Las glándulas individualmente se dispersan en la porción más posterior del metasoma, y cada una de ellas está formada por una unidad secretora grande y un canal que conduce la secreción. Cada célula de la unidad secretora contiene un gran lumen y numerosos gránulos secretores, los que están delimitados por una membrana y repletos de un contenido denso a los electrones de aspecto homogéneo, el que es eliminado hacia el lumen. El citoplasma de la célula está repleto de perfiles del retículo endoplasmático granular, de polirribosomas y de aparatos de Golgi. La membrana plasmática basal presenta muchos repliegues cortos hacia el interior y está revestida por una membrana basal fina. La membrana plasmática apical está invaginada para formar el límite del lumen extracelular, lleva microvellosidades cortas y dispersas. Las células son mononucleadas y el núcleo es de forma variada y contiene cromatina dispersa. El lumen está separado del canal de recepción por una pared replegada semejante a una esponja, con muchas vellosidades, las que en su extremidad están delimitadas por una cutícula fina. En la región basal, las vellosidades carecen de cubierta cuticular. El canal que conduce la secreción adquiere dos densidades de electrones diferentes. Una cutícula fina reviste el lumen del canal que conduce la secreción, siendo escaso el citoplasma de la célula y conteniendo poco retículo endoplasmático granular y polirribosomas. Estos resultados se discuten en relación a la función y al origen de la glándula de seda en los adultos de Sphecidae

  6. Silk and PEG as means to stiffen a parylene probe for insertion in the brain: toward a double time-scale tool for local drug delivery

    Lecomte, A.; Castagnola, V.; Descamps, E.; Dahan, L.; Blatché, M. C.; Dinis, T. M.; Leclerc, E.; Egles, C.; Bergaud, C.

    2015-12-01

    The use of soft materials as substrate for neural probes aims at achieving better compliance with the surrounding neurons while maintaining minimal rejection. Many strategies have emerged to enable such probes to penetrate the cortex, among which the use of resorbable polymers. We performed several tests involving two resorbable polymers considered most promising: polyethylene glycol (PEG) and silk fibroin (SF) from Bombyx Mori silkworms. Our coating method provides a repeatable, uniform structure optimized for a stress-reduced insertion of a parylene-C neural probe. Standard compression tests as well as in vitro and in vivo insertion assessments show that both SF and PEG-coated probes are stiff enough to avoid the buckling effect during insertion in the cortex. However, with a buckling force of 300 mN and a mechanical holding in vitro of tens of minutes, we assess silk fibroin to be more reliable for practical handling. In vivo first try-outs in mouse brain showed neither buckling issues of the probe nor undesired alteration of the signal recording. Moreover, we evidenced two distinct time scales in the bioresorption of our polymer coatings: silk fibroin degrades itself in a matter of weeks and PEG dissolves itself within seconds in the presence of water. We then present a hybrid PEG and SF coating that could be used as a drug delivery system with different time scales to reduce both the acute and the chronic body reaction.

  7. Untangling spider silk evolution with spidroin terminal domains

    Garb Jessica E

    2010-08-01

    Full Text Available Abstract Background Spidroins are a unique family of large, structural proteins that make up the bulk of spider silk fibers. Due to the highly variable nature of their repetitive sequences, spidroin evolutionary relationships have principally been determined from their non-repetitive carboxy (C-terminal domains, though they offer limited character data. The few known spidroin amino (N-terminal domains have been difficult to obtain, but potentially contain critical phylogenetic information for reconstructing the diversification of spider silks. Here we used silk gland expression data (ESTs from highly divergent species to evaluate the functional significance and phylogenetic utility of spidroin N-terminal domains. Results We report 11 additional spidroin N-termini found by sequencing ~1,900 silk gland cDNAs from nine spider species that shared a common ancestor > 240 million years ago. In contrast to their hyper-variable repetitive regions, spidroin N-terminal domains have retained striking similarities in sequence identity, predicted secondary structure, and hydrophobicity. Through separate and combined phylogenetic analyses of N-terminal domains and their corresponding C-termini, we find that combined analysis produces the most resolved trees and that N-termini contribute more support and less conflict than the C-termini. These analyses show that paralogs largely group by silk gland type, except for the major ampullate spidroins. Moreover, spidroin structural motifs associated with superior tensile strength arose early in the history of this gene family, whereas a motif conferring greater extensibility convergently evolved in two distantly related paralogs. Conclusions A non-repetitive N-terminal domain appears to be a universal attribute of spidroin proteins, likely retained from the origin of spider silk production. Since this time, spidroin N-termini have maintained several features, consistent with this domain playing a key role in silk assembly. Phylogenetic analyses of the conserved N- and C-terminal domains illustrate dramatic radiation of the spidroin gene family, involving extensive duplications, shifts in expression patterns and extreme diversification of repetitive structural sequences that endow spider silks with an unparalleled range of mechanical properties.

  8. Oxygen-reduction activity of silk-derived carbons

    Iwazaki, Tomoya [Department of Fine Materials Engineering, Faculty of Textile Science and Technology, Shinshu University, 3-15-1 Tokida, Ueda, Nagano 386-8567 (Japan); Shinano Kenshi Co., Ltd., 1078 Kamimaruko, Ueda, Nagano 386-0498 (Japan); Yang, Hongsheng; Obinata, Ryoujin; Sugimoto, Wataru; Takasu, Yoshio [Department of Fine Materials Engineering, Faculty of Textile Science and Technology, Shinshu University, 3-15-1 Tokida, Ueda, Nagano 386-8567 (Japan)

    2010-09-15

    Carbonized silk fibroin (CS), which is free of metallic elements, showed high catalytic activity for oxygen-reduction reaction (ORR). The catalytic activity of CS for ORR was greatly enhanced by steam activation forming silk-derived activated carbon (CS-AC). The surface morphology, surface area, pore structure and remaining nitrogen species of the CSs were compared with those of the CS-ACs. The open-circuit potential and the power density of a polymer electrolyte fuel cell using a CS900-AC, which was heat-treated at 900 C prior to the steam activation, and a platinum/C (C: carbon black) anode under pure oxygen and hydrogen gases, respectively, both at 0.2 MPa, were 0.92 V and 142 mW cm{sup -2} at 80 C. The ORR on the activated carbon, CS900-AC, proceeded with a 3.5-electron reaction at 0.6 V (vs. RHE); however, this was improved to a 3.9-electron reaction with the addition of zirconium oxide at 20 wt% to CS900-AC. (author)

  9. Biotechnologies based on silk

    Sehnal, František

    Heidelberg : Springer, 2011 - (Vilcinskas, A.), s. 211-224 ISBN 978-90-481-9640-1 R&D Projects: GA ČR GAP502/10/2382 Institutional research plan: CEZ:AV0Z50070508 Keywords : silk Subject RIV: ED - Physiology

  10. Influence of different surface modification treatments on silk biotextiles for tissue engineering applications.

    Ribeiro, Viviana P; Almeida, Lília R; Martins, Ana R; Pashkuleva, Iva; Marques, Alexandra P; Ribeiro, Ana S; Silva, Carla J; Bonifácio, Graça; Sousa, Rui A; Reis, Rui L; Oliveira, Ana L

    2016-04-01

    Biotextile structures from silk fibroin have demonstrated to be particularly interesting for tissue engineering (TE) applications due to their high mechanical strength, interconnectivity, porosity, and ability to degrade under physiological conditions. In this work, we described several surface treatments of knitted silk fibroin (SF) scaffolds, namely sodium hydroxide (NaOH) solution, ultraviolet radiation exposure in an ozone atmosphere (UV/O3 ) and oxygen (O2 ) plasma treatment followed by acrylic acid (AAc), vinyl phosphonic acid (VPA), and vinyl sulfonic acid (VSA) immersion. The effect of these treatments on the mechanical properties of the textile constructs was evaluated by tensile tests in dry and hydrated states. Surface properties such as morphology, topography, wettability and elemental composition were also affected by the applied treatments. The in vitro biological behavior of L929 fibroblasts revealed that cells were able to adhere and spread both on the untreated and surface-modified textile constructs. The applied treatments had different effects on the scaffolds' surface properties, confirming that these modifications can be considered as useful techniques to modulate the surface of biomaterials according to the targeted application. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 496-507, 2016. PMID:25939722

  11. Comparative Transcriptome Analysis Reveals Different Silk Yields of Two Silkworm Strains

    Li, Juan; Qin, Sheng; Yu, Huanjun; Zhang, Jing; Liu, Na; Yu, Ye; Hou, Chengxiang; Li, Muwang

    2016-01-01

    Cocoon and silk yields are the most important characteristics of sericulture. However, few studies have examined the genes that modulate these features. Further studies of these genes will be useful for improving the products of sericulture. JingSong (JS) and Lan10 (L10) are two strains having significantly different cocoon and silk yields. In the current study, RNA-Seq and quantitative polymerase chain reaction (qPCR) were performed on both strains in order to determine divergence of the silk gland, which controls silk biosynthesis in silkworms. Compared with L10, JS had 1375 differentially expressed genes (DEGs; 738 up-regulated genes and 673 down-regulated genes). Nine enriched gene ontology (GO) terms were identified by GO enrichment analysis based on these DEGs. KEGG enrichment analysis results showed that the DEGs were enriched in three pathways, which were mainly associated with the processing and biosynthesis of proteins. The representative genes in the enrichment pathways and ten significant DEGs were further verified by qPCR, the results of which were consistent with the RNA-Seq data. Our study has revealed differences in silk glands between the two silkworm strains and provides a perspective for understanding the molecular mechanisms determining silk yield. PMID:27159277

  12. Structure, composition and mechanical properties of the silk fibres of the egg case of the Joro spider, Nephila clavata (Araneae, Nephilidae)

    Ping Jiang; Cong Guo; Taiyong Lv; Yonghong Xiao; Xinjun Liao; Bing Zhou

    2011-12-01

    The silk egg case and orb web of spiders are elaborate structures that are assembled from a number of components. We analysed the structure, the amino acid and fibre compositions, and the tensile properties of the silk fibres of the egg case of Nephila clavata. SEM shows that the outer and inner covers of the egg case consist of thick, medium and thin silk fibres. The silk fibres of the outer cover of the egg case are probably produced by the major and minor ampullate glands. The silk fibres of the inner cover of the egg case from cylindrical glands appears to be distinct from the silk fibres of the major ampullate glands based on their micro-morphology, mole percent amino acid composition and types, and tensile behaviour and properties. Collectively, our investigations show that N. clavata uses silk fibres from relatively few glands in varying combinations to achieve different physical and chemical properties (e.g., color, diameter, morphology and amino acid composition) and functional and mechanical properties in the different layers of the egg case.

  13. Silk to steel

    Adkins, Monty

    2005-01-01

    Details: Silk to Steel is a multi-channel (5.1 and 7.1) composition. It was a Finalist at CIMESP 2005, Brazil, and received an Honourable Mention at the Bourges International Electroacoustic Music Competition 2005. There have been 17 further performances, including VI CIMESP 2005, Goethe-Institut, São Paulo; Elektrophonie, Dijon, France; Sounds Electric’05, Aula Maxima, Maynooth, Ireland; Fylkingen Institute, Stockholm; Concordia University, Montreal; VI International Festival of ...

  14. Silk Road Traject

    Hamsea, Christian Lucian (Hrsg.)

    2009-01-01

    The catalogue Silk Road Traject presents the art projects of the artist and lecturer of the Chair of Pedagogics at the Erlangen-Nuremberg University Christian Lucian Hamsea during his stay in Tbilisi as guest professor at the State Academy of Art: "Tagging My Soul Underground" with stencils and graffiti paintings and “Lost space”, carried out in one of the Tbilisi abandoned factory territory in the style of “trash art”.

  15. Nutritional value of silk powder from irradiated silk waste

    Bunjob, Malee; Lakshanasomya, Niphaporn [Department of Medical Sciences, Nonthaburi (Thailand); Meesilpa, Prateep [Department of Agriculture, Bangkok (Thailand); Sudatis, Boonya [Office of Atomic Energy for Peace, Bangkok (Thailand)

    2002-03-01

    Silk waste from Thai reeling factory was developed into purified silk protein. Cleanliness of silk filaments by boiling in water was firstly needed. After air drying, they were irradiated with Gamma ray using Cobalt-60 at doses of 750 and 1,000 kGy, then the irradiated silk filaments were dissolved in calcium chloride-ethanol-water solution. The next steps would be dialysis followed by freezed dried process to obtain irradiated silk powder. Two samples of 750 and 1,000 kGy irradiated silk powder were analyzed for nutritional components compared with non irradiated silk filaments. It was found that, the fat content in two irradiated samples was distinctive lower than the non irradiated one, however the protein content was nearly the same in three samples. In addition the moisture content in two irradiated samples was distinctive higher than the non irradiated one. These results show that irradiation technique is useful for development of valuable silk protein as biomaterial. (author)

  16. 21 CFR 184.1262 - Corn silk and corn silk extract.

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Corn silk and corn silk extract. 184.1262 Section... SAFE Listing of Specific Substances Affirmed as GRAS 184.1262 Corn silk and corn silk extract. (a) Corn silk is the fresh styles and stigmas of Zea mays L. collected when the corn is in milk....

  17. Molecular spring: from spider silk to silkworm silk

    Wu, Xiang; Du, Ning; Xu, Gang-Qin; Li, Bao-Wen

    2009-01-01

    In this letter, we adopt a new approach combining theoretical modeling with silk stretching measurements to explore the mystery of the structures between silkworm and spider silks, leading to the differences in mechanical response against stretching. Hereby the typical stress-strain profiles are reproduced by implementing the newly discovered and verified "$\\beta$-sheet splitting" mechanism, which primarily varies the secondary structure of protein macromolecules; our modeling and simulation results show good accordance with the experimental measurements. Hence, it can be concluded that the post-yielding mechanical behaviors of both kinds of silks are resulted from the splitting of crystallines while the high extensibility of spider dragline is attributed to the tiny $\\beta$-sheets solely existed in spider silk fibrils. This research reveals for the first time the structural factors leading to the significant difference between spider and silkworm silks in mechanical response to the stretching force. Addition...

  18. Secondary Structure Transition and Critical Stress for a Model of Spider Silk Assembly.

    Giesa, Tristan; Perry, Carole C; Buehler, Markus J

    2016-02-01

    Spiders spin their silk from an aqueous solution to a solid fiber in ambient conditions. However, to date, the assembly mechanism in the spider silk gland has not been satisfactorily explained. In this paper, we use molecular dynamics simulations to model Nephila clavipes MaSp1 dragline silk formation under shear flow and determine the secondary structure transitions leading to the experimentally observed fiber structures. While no experiments are performed on the silk fiber itself, insights from this polypeptide model can be transferred to the fiber scale. The novelty of this study lies in the calculation of the shear stress (300-700 MPa) required for fiber formation and identification of the amino acid residues involved in the transition. This is the first time that the shear stress has been quantified in connection with a secondary structure transition. By study of molecules containing varying numbers of contiguous MaSp1 repeats, we determine that the smallest molecule size giving rise to a "silk-like" structure contains six polyalanine repeats. Through a probability analysis of the secondary structure, we identify specific amino acids that transition from ?-helix to ?-sheet. In addition to portions of the polyalanine section, these amino acids include glycine, leucine, and glutamine. The stability of ?-sheet structures appears to arise from a close proximity in space of helices in the initial spidroin state. Our results are in agreement with the forces exerted by spiders in the silking process and the experimentally determined global secondary structure of spidroin and pulled MaSp1 silk. Our study emphasizes the role of shear in the assembly process of silk and can guide the design of microfluidic devices that attempt to mimic the natural spinning process and predict molecular requirements for the next generation of silk-based functional materials. PMID:26669270

  19. Tailoring the properties and functions of phosphate/silk/Ag/chitosan scaffolds

    Two novel silk composites of phosphatic phases with nanosilver/chitosan having enhanced biocompatibility were achieved. Hydroxyapatite and octa calcium phosphates were synthesized in situ within silk fibroin/chitosan/nanosilver composites recently studied. Thermo-gravimetric analysis (TGA) and Differential Scanning Calorimetry (DSC) verified their thermal behavior. The structural aspects were characterized applying X-ray diffraction analysis (XRD), transmission electron microscopy (TEM) and field emission scanning electron microscope (FESEM) with EDAX. Additionally X-ray Photoelectron Spectroscopy (XPS) and Fourier Transform Infrared spectroscopy (FTIR) were applied. Mercury porosimeter was used to verify the pore size distribution. The in vitro degradation was followed in D-MEM for 48 h in a cumulative manner for five successive periods. Biochemical analyses of Ca, P and total protein using relevant chemical kits and atomic absorption for silver were performed. ANOVA statistics was carried out. Phosphatic crystalline phases along with the presence of silk, chitosan and nano-silver were developed. The diameters of hydroxyapatite and octa calcium phosphate particles were ~ 8–17 nm and 15–22 nm respectively. Comparatively higher degradation of Octa composite possessing higher porosity proved in turn more osteoinduction with in situ apatitic development. - Highlights: • A bottom–up approach controlled the achieved in situ configurations. • The calculated (CI)XDR and (CI)FTIR for both phases are highly conformable. • Post-immersion trimodal pore system was achieved in OCP composite. • Silver nanoparticle inclusion expected to enhance composite bactericidal activity

  20. Tailoring the properties and functions of phosphate/silk/Ag/chitosan scaffolds.

    Abdel-Fattah, Wafa I; Sallam, Abdel Sattar M; Diab, A M; Ali, Ghareib W

    2015-09-01

    Two novel silk composites of phosphatic phases with nanosilver/chitosan having enhanced biocompatibility were achieved. Hydroxyapatite and octa calcium phosphates were synthesized in situ within silk fibroin/chitosan/nanosilver composites recently studied. Thermo-gravimetric analysis (TGA) and Differential Scanning Calorimetry (DSC) verified their thermal behavior. The structural aspects were characterized applying X-ray diffraction analysis (XRD), transmission electron microscopy (TEM) and field emission scanning electron microscope (FESEM) with EDAX. Additionally X-ray Photoelectron Spectroscopy (XPS) and Fourier Transform Infrared spectroscopy (FTIR) were applied. Mercury porosimeter was used to verify the pore size distribution. The in vitro degradation was followed in D-MEM for 48 h in a cumulative manner for five successive periods. Biochemical analyses of Ca, P and total protein using relevant chemical kits and atomic absorption for silver were performed. ANOVA statistics was carried out. Phosphatic crystalline phases along with the presence of silk, chitosan and nano-silver were developed. The diameters of hydroxyapatite and octa calcium phosphate particles were ~8-17 nm and 15-22 nm respectively. Comparatively higher degradation of Octa composite possessing higher porosity proved in turn more osteoinduction with in situ apatitic development. PMID:26046279

  1. Tailoring the properties and functions of phosphate/silk/Ag/chitosan scaffolds

    Abdel-Fattah, Wafa I., E-mail: nrcfifi@yahoo.com [National Research Centre, Biomaterials Department, Elbehoos St. Dokki, Giza (Egypt); Sallam, Abdel Sattar M., E-mail: ab_m_sallam@yahoo.com [Faculty of Science, Physics Department, Ain Shams University, Cairo (Egypt); Diab, A.M., E-mail: amdiabdr55@yahoo.com [High Institute of Engineering, Physics Department, El-Shorouk Academy, Cairo (Egypt); Ali, Ghareib W., E-mail: wafaa_ghareeeb@yahoo.com [National Research Centre, Biomaterials Department, Elbehoos St. Dokki, Giza (Egypt)

    2015-09-01

    Two novel silk composites of phosphatic phases with nanosilver/chitosan having enhanced biocompatibility were achieved. Hydroxyapatite and octa calcium phosphates were synthesized in situ within silk fibroin/chitosan/nanosilver composites recently studied. Thermo-gravimetric analysis (TGA) and Differential Scanning Calorimetry (DSC) verified their thermal behavior. The structural aspects were characterized applying X-ray diffraction analysis (XRD), transmission electron microscopy (TEM) and field emission scanning electron microscope (FESEM) with EDAX. Additionally X-ray Photoelectron Spectroscopy (XPS) and Fourier Transform Infrared spectroscopy (FTIR) were applied. Mercury porosimeter was used to verify the pore size distribution. The in vitro degradation was followed in D-MEM for 48 h in a cumulative manner for five successive periods. Biochemical analyses of Ca, P and total protein using relevant chemical kits and atomic absorption for silver were performed. ANOVA statistics was carried out. Phosphatic crystalline phases along with the presence of silk, chitosan and nano-silver were developed. The diameters of hydroxyapatite and octa calcium phosphate particles were ~ 8–17 nm and 15–22 nm respectively. Comparatively higher degradation of Octa composite possessing higher porosity proved in turn more osteoinduction with in situ apatitic development. - Highlights: • A bottom–up approach controlled the achieved in situ configurations. • The calculated (CI){sub XDR} and (CI){sub FTIR} for both phases are highly conformable. • Post-immersion trimodal pore system was achieved in OCP composite. • Silver nanoparticle inclusion expected to enhance composite bactericidal activity.

  2. Aqueous-Based Coaxial Electrospinning of Genetically Engineered Silk Elastin Core-Shell Nanofibers

    Jingxin Zhu

    2016-03-01

    Full Text Available A nanofabrication method for the production of flexible core-shell structured silk elastin nanofibers is presented, based on an all-aqueous coaxial electrospinning process. In this process, silk fibroin (SF and silk-elastin-like protein polymer (SELP, both in aqueous solution, with high and low viscosity, respectively, were used as the inner (core and outer (shell layers of the nanofibers. The electrospinnable SF core solution served as a spinning aid for the nonelectrospinnable SELP shell solution. Uniform nanofibers with average diameter from 301 ± 108 nm to 408 ± 150 nm were obtained through adjusting the processing parameters. The core-shell structures of the nanofibers were confirmed by fluorescence and electron microscopy. In order to modulate the mechanical properties and provide stability in water, the as-spun SF-SELP nanofiber mats were treated with methanol vapor to induce β-sheet physical crosslinks. FTIR confirmed the conversion of the secondary structure from a random coil to β-sheets after the methanol treatment. Tensile tests of SF-SELP core-shell structured nanofibers showed good flexibility with elongation at break of 5.20% ± 0.57%, compared with SF nanofibers with an elongation at break of 1.38% ± 0.22%. The SF-SELP core-shell structured nanofibers should provide useful options to explore in the field of biomaterials due to the improved flexibility of the fibrous mats and the presence of a dynamic SELP layer on the outer surface.

  3. Exploiting spiders’ silk

    Paula Gould

    2002-12-01

    As today’s film-goers and readers of the original Spiderman comic strip will know, spider silk makes an ideal tool for a modern day super-hero. The fine line can be produced at will, holds our hero’s weight as he swings between tall buildings, and then clumps together to trap dastardly villains in a tangled, sticky mess. Highly useful if your daily job description includes rescuing distressed damsels from improbable locations and preventing a thoroughly nasty goblin from wreaking havoc on your home city.

  4. Determination of the torsion angles of alanine and glycine residues of model compounds of spider silk (AGG){sub 10} using solid-state NMR methods

    Ashida, Jun; Ohgo, Kosuke; Komatsu, Kohei; Kubota, Ayumi; Asakura, Tetsuo [Tokyo University of Agriculture and Technology, Department of Biotechnology (Japan)], E-mail: asakura@cc.tuat.ac.jp

    2003-02-15

    Spiders synthesize several kinds of silk fibers. In the primary structure of spider silk, one of the major ampullate (dragline, frame) silks, spidroin 1, and flagelliform silk (core fibers of adhesive spiral), there are common repeated X-Gly-Gly (X = Ala, Leu, Pro, Tyr, Glu, and Arg) sequences, which are considered to be related to the elastic character of these fibers. In this paper, two dimensional spin diffusion solid-state NMR under off magic angle spinning (OMAS), {sup 13}C chemical shift contour plots, and Rotational Echo DOuble Resonance (REDOR) were applied to determine the torsion angles of one Ala and two kinds of Gly residues in the Ala-Gly-Gly sequence of {sup 13}C=O isotope-labeled (Ala-Gly-Gly){sub 10}. The torsion angles were determined to be ({phi}, {psi}) = (-90 deg., 150 deg.) within an experimental error of {+-}10 deg. for each residue. This conformation is characterized as 3{sub 1} helix which is in agreement with the structure proposed from the X-ray powder diffraction pattern of poly(Ala-Gly-Gly). The 3{sub 1} helix of (Ala-Gly-Gly){sub 10} does not change by formic acid treatment although (Ala-Gly){sub 15} easily changes from the silk I conformation (the structure of Bombyx mori silk fibroin before spinning in the solid state) to silk II conformation (the structure of the silk fiber after spinning) by such treatment. Thus, the 3{sub 1} helix conformation of (Ala-Gly-Gly){sub 10} is considered very stable. Furthermore, the torsion angles of the 16th Leu residue of (Leu-Gly-Gly){sub 10} were also determined as ({phi}, {psi}) = (-90 deg., 150 deg.) and this peptide is also considered to take 3{sub 1} helix conformation.

  5. Minor ampullate silks from Nephila and Argiope spiders: tensile properties and microstructural characterization.

    Guinea, G V; Elices, M; Plaza, G R; Perea, G B; Daza, R; Riekel, C; Agulló-Rueda, F; Hayashi, C; Zhao, Y; Pérez-Rigueiro, J

    2012-07-01

    The mechanical behavior and microstructure of minor ampullate gland silk (miS) of two orb-web spinning species, Argiope trifasciata and Nephila inaurata, were extensively characterized, enabling detailed comparison with other silks. The similarities and differences exhibited by miS when compared with the intensively studied major ampullate gland silk (MAS) and silkworm (Bombyx mori) silk offer a genuine opportunity for testing some of the hypotheses proposed to correlate microstructure and tensile properties in silk. In this work, we show that miSs of different species show similar properties, even when fibers spun by spiders that diverged over 100 million years are compared. The tensile properties of miS are comparable to those of MAS when tested in air, significantly in terms of work to fracture, but differ considerably when tested in water. In particular, miS does not show a supercontraction effect and an associated ground state. In this regard, the behavior of miS in water is similar to that of B. mori silk, and it is shown that the initial elastic modulus of both fibers can be explained using a common model. Intriguingly, the microstructural parameters measured in miS are comparable to those of MAS and considerably different from those found in B. mori. This fact suggests that some critical microstructural information is still missing in our description of silks, and our results suggest that the hydrophilicity of the lateral groups or the large scale organization of the sequences might be routes worth exploring. PMID:22668322

  6. Clinical Application of a Silk Fibroin Protein Biologic Scaffold for Abdominal Wall Fascial Reinforcement

    Mark W. Clemens, MD

    2014-11-01

    Conclusions: Postoperative complication rates after 18 months were low, and most surgical complications were managed nonoperatively on an outpatient basis without mesh removal. To our knowledge, this is the only series to report on a long-lasting, transitory SBS for abdominal wall repair and reinforcement. Procedure-specific outcome studies are warranted to delineate optimal patient selection and define potential device characteristic advantages.

  7. Incorporation of elastase inhibitor in silk fibroin nanoparticles for transdermal delivery

    Ferreira, Ana Vanessa Fernandes

    2013-01-01

    Elastase de Neutrfilos Humana, como resposta inflamatria, possui a capacidade de degradar os componentes da matriz extracelular, nomeadamente colagnio e elastina, estando envolvida na formao de rugas causadas pela radiao ultravioleta. De forma a desenvolver uma emulso cosmtica antirrugas, decidiu-se incorporar um potente inibidor sinttico de elastase, sivelestat (IC50 = 44 nM, Ki = 0,2 ?M), em nanopartculas biopolimricas formadas por mtodos de emulsificao de elev...

  8. A new route for silk

    Omenetto, Fiorenzo G.; Kaplan, David L.

    2008-11-01

    Famous for its use in clothing since early times, silk is now finding a new application as a useful biocompatible material in photonic devices. Thin films, diffraction gratings and organic photonic crystals are just a few of the exciting possibilities.

  9. Regeneration of Bombyx mori silk nanofibers and nanocomposite fibrils by the electrospinning process

    Ayutsede, Jonathan Eyitouyo

    In recent years, there has been significant interest in the utilization of natural materials for novel nanoproducts such as tissue engineered scaffolds. Silkworm silk fibers represent one of the strongest natural fibers known. Silkworm silk, a protein-based natural biopolymer, has received renewed interest in recent years due to its unique properties (strength, toughness) and potential applications such as smart textiles, protective clothing and tissue engineering. The traditional 10--20 mum diameter, triangular-shaped Bombyx mori fibers have remained unchanged over the years. However, in our study, we examine the scientific implication and potential applications of reducing the diameter to the nanoscale, changing the triangular shape of the fiber and adding nanofillers in the form of single wall carbon nanotubes (SWNT) by the electrospinning process. The electrospinning process preserves the natural conformation of the silk (random and beta-sheet). The feasibility of changing the properties of the electrospun nanofibers by post processing treatments (annealing and chemical treatment) was investigated. B. mori silk fibroin solution (formic acid) was successfully electrospun to produce uniform nanofibers (as small as 12 nm). Response Surface Methodology (RSM) was applied for the first time to experimental results of electrospinning, to develop a processing window that can reproduce regenerated silk nanofibers of a predictable size (d fibers were fabricated for the first time with anticipated properties (mechanical, thermal and electrically conductive) that may have scientific applications (nerve regeneration, stimulation of cell-scaffold interaction). In order to realize these applications, the following areas need to be addressed: a systematic investigation of the dispersion of the nanotubes in the silk matrix, a determination of new methodologies for characterizing the nanofiber properties and establishing the nature of the silk-SWNT interactions. A new visualization system was developed to characterize the transport properties of the nanofibrous assemblies. The morphological, chemical, structural and mechanical properties of the nanofibers were determined by field emission environmental scanning microscopy, Fourier transform infrared and Raman spectroscopy, wide angle x-ray diffraction and microtensile tester respectively.

  10. Silk coating as a novel delivery system and reversible adhesive for stiffening and shaping flexible probes

    Cinzia Metallo

    2015-02-01

    Full Text Available The performance of any implantable electrode depends not only on its recording or stimulation capabilities but also on its position in relation to the target site. Electrode displacement during or after implantation represents a major issue as it might result in tissue damage or incorrect recording or stimulation location, complicating the interpretation of experimental data. Although thin-film electrode arrays have overcome some of the main limitations of more traditional, stiffer probes, their intrinsic flexibility and unilateral contacts represent a new challenge: they tend to bend during insertion and are difficult to implant simultaneously while maintaining a specific relative position. Here, we present a method that addresses all these issues using a coating of silk fibroin, a versatile protein derived from silkworm cocoons. The method is demonstrated by acquiring electromyographic (EMG recordings in Manduca sexta, a soft-bodied animal that exemplifies the issues of electrode insertion and placement in delicate and deformable tissues.

  11. Optically probing torsional superelasticity in spider silks

    Kumar, Bhupesh; Thakur, Ashish; Panda, Biswajit; Singh, Kamal P. [Department of Physical Sciences, IISER Mohali, Sector 81, Manauli, Mohali 140306 (India)

    2013-11-11

    We investigate torsion mechanics of various spider silks using a sensitive optical technique. We find that spider silks are torsionally superelastic in that they can reversibly withstand great torsion strains of over 10{sup 2−3} rotations per cm before failure. Among various silks from a spider, we find the failure twist-strain is greatest in the sticky capture silk followed by dragline and egg-case silk. Our in situ laser-diffraction measurements reveal that torsional strains on the silks induce a nano-scale transverse compression in its diameter that is linear and reversible. These unique torsional properties of the silks could find applications in silk-based materials and devices.

  12. Endocrine Glands & Their Hormones

    ... Characteristics of Hormones Endocrine Glands & Their Hormones Pituitary & Pineal Glands Thyroid & Parathyroid Glands Adrenal Gland Pancreas Gonads Other ... hormone secretion. « Previous (Characteristics of Hormones) Next (Pituitary & Pineal Glands) » Contact Us | Privacy Policy | Accessibility | FOIA | File Formats ...

  13. A novel approach to fabricate silk nanofibers containing hydroxyapatite nanoparticles using a three-way stopcock connector

    Sheikh, Faheem A.; Ju, Hyung Woo; Moon, Bo Mi; Park, Hyun Jung; Kim, Jung Ho; Lee, Ok Joo; Park, Chan Hum

    2013-07-01

    Electrospinning technique is commonly used to produce micro- and/or nanofibers, which utilizes electrical forces to produce polymeric fibers with diameters ranging from several micrometers down to few nanometers. Desirably, electrospun materials provide highly porous structure and appropriate pore size for initial cell attachment and proliferation and thereby enable the exchange of nutrients. Composite nanofibers consisting of silk and hydroxyapatite nanoparticles (HAp) (NPs) had been considered as an excellent choice due to their efficient biocompatibility and bone-mimicking properties. To prepare these nanofiber composites, it requires the use of acidic solutions which have serious consequences on the nature of both silk and HAp NPs. It is ideal to create these nanofibers using aqueous solutions in which the physicochemical nature of both materials can be retained. However, to create those nanofibers is often difficult to obtain because of the fact that aqueous solutions of silk and HAp NPs can precipitate before they can be ejected into fibers during the electrospinning process. In this work, we had successfully used a three-way stopcock connector to mix the two different solutions, and very shortly, this solution is ejected out to form nanofibers due to electric fields. Different blend ratios consisting HAp NPs had been electrospun into nanofibers. The physicochemical aspects of fabricated nanofiber had been characterized by different state of techniques like that of FE-SEM, EDS, TEM, TEM-EDS, TGA, FT-IR, and XRD. These characterization techniques revealed that HAp NPs can be easily introduced in silk nanofibers using a stopcock connector, and this method favorably preserves the intact nature of silk fibroin and HAp NPs. Moreover, nanofibers obtained by this strategy were tested for cell toxicity and cell attachment studies using NIH 3 T3 fibroblasts which indicated non-toxic behavior and good attachment of cells upon incubation in the presence of nanofibers.

  14. Material properties of evolutionary diverse spider silks described by variation in a single structural parameter

    Madurga, Rodrigo; Plaza, Gustavo R.; Blackledge, Todd A.; Guinea, Gustavo.V.; Elices, Manuel; Pérez-Rigueiro, José

    2016-01-01

    Spider major ampullate gland silks (MAS) vary greatly in material properties among species but, this variation is shown here to be confined to evolutionary shifts along a single universal performance trajectory. This reveals an underlying design principle that is maintained across large changes in both spider ecology and silk chemistry. Persistence of this design principle becomes apparent after the material properties are defined relative to the true alignment parameter, which describes the orientation and stretching of the protein chains in the silk fiber. Our results show that the mechanical behavior of all Entelegynae major ampullate silk fibers, under any conditions, are described by this single parameter that connects the sequential action of three deformation micromechanisms during stretching: stressing of protein-protein hydrogen bonds, rotation of the β-nanocrystals and growth of the ordered fraction. Conservation of these traits for over 230 million years is an indication of the optimal design of the material and gives valuable clues for the production of biomimetic counterparts based on major ampullate spider silk. PMID:26755434

  15. Evidence from Multiple Species that Spider Silk Glue Component ASG2 is a Spidroin

    Matthew A. Collin; Clarke, Thomas H; Ayoub, Nadia A.; Hayashi, Cheryl Y.

    2016-01-01

    Spiders in the superfamily Araneoidea produce viscous glue from aggregate silk glands. Aggregate glue coats prey-capture threads and hampers the escape of prey from webs, thereby increasing the foraging success of spiders. cDNAs for Aggregate Spider Glue 1 (ASG1) and 2 (ASG2) have been previously described from the golden orb-weaver, Nephila clavipes, and Western black widow, Latrodectus hesperus. To further investigate aggregate glues, we assembled ASG1 and ASG2 from genomic target capture l...

  16. PEGylated Silk Nanoparticles for Anticancer Drug Delivery

    Wongpinyochit, Thidarat; Uhlmann, Petra; Urquhart, Andrew; Seib, F. Philipp

    2015-01-01

    especially necessary to support their evolution. The aim of this study was to develop and examine the potential of PEGylated silk nanoparticles as an anticancer drug delivery system. We first generated B. mori derived silk nanoparticles by driving β-sheet assembly (size 104 ± 1.7 nm, zeta potential −56 ± 5.......6 mV) using nanoprecipitation. We then surface grafted polyethylene glycol (PEG) to the fabricated silk nanoparticles and verified the aqueous stability and morphology of the resulting PEGylated silk nanoparticles. We assessed the drug loading and release behavior of these nanoparticles using...... to human breast cancer cells. In conclusion, these results, taken together with prior silk nanoparticle data, support a viable future for silk-based nanomedicines....

  17. Fabricating a reactive surface on the fibroin film by a room-temperature plasma jet array for biomolecule immobilization

    Chen, Guang-Liang; Zheng, Xu; L, Guo-Hua; Zhang, Zhao-Xia; Sylvain, Massey; Wilson, Smith; Michael, Tatoulian; Yang, Si-Ze

    2012-10-01

    A simple dielectric barrier discharge (DBD) jet array was designed with a liquid electrode and helium gas. The characteristics of the jet array discharge and the preliminary polymerization with acrylic acid (AA) monomer were presented. The plasma reactor can produce a cold jet array with a gas temperature lower than 315 K, using an applied discharge power between 6 W and 30 W (Vdis Idis). A silk fibroin film (SFF) was modified using the jet array and AA monomer, and the treated SFF samples were characterized by atomic force microscopy (AFM), scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and contact angle (CA). The deposition rate of the poly acrylic acid (PAA) was able to reach 300 nm/min, and the surface roughness and energy increased with the AA flow rate. The FTIR results indicate that the modified SFF had more carboxyl groups (-COOH) than the original SFF. This latter characteristic allowed the modified SFF to immobilize more quantities of antimicrobial peptide (AP, LL-37) which inhibited the Escherichia coli (E. Coli) effectively.

  18. Trends in World Silk Cocoons and Silk Production and Trade, 2007-2010

    AGATHA AGATHA POPESCU

    2013-10-01

    Full Text Available The paper aimed to analyze world production and trade for fresh silk cocoons and raw silk using FAO Stat data for  the  period 2007-2010. The use of index, share and comparison methods allowed to identify the major trends in the analyzed period. Silk decline was determined by the increased importance of cotton and artificial fibres in textile and clothing industry. Important changes are taking place on silk cocoons and silk market. While, the European market decreased, the Asian market has mainly developed fresh cocoons and raw silk, while the European market became more interested of clothes. Silk consumption declined because of consumer’s preference for synthetic fibres, except traditional consumers from Asia.  China is the main producer and exporter of fresh and dry cocoons, while raw silk is produced and exported by China, Brazil and Italy and imported by India, Japan and Italy. In Europe, Bulgaria is the top producer of fresh cocoons and raw silk and Italy is the main raw silk importer and the top producer and exporter of textile and fashion clothes. Silk will remain an important raw material for producing high quality and luxury clothes.

  19. Silkworms transformed with chimeric silkworm/spider silk genes spin composite silk fibers with improved mechanical properties

    The development of a spider silk manufacturing process is of great interest. piggyBac vectors were used to create transgenic silkworms encoding chimeric silkworm/spider silk proteins. The silk fibers produced by these animals were composite materials that included chimeric silkworm/spider silk prote...

  20. Unraveled mechanism in silk engineering: Fast reeling induced silk toughening

    Wu, Xiang; Liu, Xiang-Yang; Du, Ning; Xu, Gangqin; Li, Baowen

    2009-08-01

    We theoretically and experimentally study the mechanical response of silkworm and spider silks against stretching and the relationship with the underlying structural factors. It is found that the typical stress-strain profiles are predicted in good agreement with experimental measurements by implementing the "?-sheet splitting" mechanism we discovered and verified, primarily varying the secondary structure of protein macromolecules. The functions of experimentally observed structural factors responding to the external stress have been clearly addressed, and optimization of the microscopic structures to enhance the mechanical strength will be pointed out, beneficial to their biomedical and textile applications.

  1. Silkworms transformed with chimeric silkworm/spider silk genes spin composite silk fibers with improved mechanical properties.

    Teul, Florence; Miao, Yun-Gen; Sohn, Bong-Hee; Kim, Young-Soo; Hull, J Joe; Fraser, Malcolm J; Lewis, Randolph V; Jarvis, Donald L

    2012-01-17

    The development of a spider silk-manufacturing process is of great interest. However, there are serious problems with natural manufacturing through spider farming, and standard recombinant protein production platforms have provided limited progress due to their inability to assemble spider silk proteins into fibers. Thus, we used piggyBac vectors to create transgenic silkworms encoding chimeric silkworm/spider silk proteins. The silk fibers produced by these animals were composite materials that included chimeric silkworm/spider silk proteins integrated in an extremely stable manner. Furthermore, these composite fibers were, on average, tougher than the parental silkworm silk fibers and as tough as native dragline spider silk fibers. These results demonstrate that silkworms can be engineered to manufacture composite silk fibers containing stably integrated spider silk protein sequences, which significantly improve the overall mechanical properties of the parental silkworm silk fibers. PMID:22215590

  2. Thymus Gland Anatomy

    ... Search Quick Search Image Details Thymus Gland, Adult, Anatomy View/Download: Small: 720x576 View Download Add to My Pictures Title: Thymus Gland, Adult, Anatomy Description: Anatomy of the thymus gland; drawing shows ...

  3. Carotenoid silk coloration is controlled by a carotenoid-binding protein, a product of the Yellow blood gene

    Sakudoh, Takashi; Sezutsu, Hideki; Nakashima, Takeharu; Kobayashi, Isao; Fujimoto, Hirofumi; Uchino, Keiro; Banno, Yutaka; Iwano, Hidetoshi; Maekawa, Hideaki; Tamura, Toshiki; Kataoka, Hiroshi; Tsuchida, Kozo

    2007-01-01

    Mechanisms for the uptake and transport of carotenoids, essential nutrients for humans, are not well understood in any animal system. The Y (Yellow blood) gene, a critical cocoon color determinant in the silkworm Bombyx mori, controls the uptake of carotenoids into the intestinal mucosa and the silk gland. Here we provide evidence that the Y gene corresponds to the intracellular carotenoid-binding protein (CBP) gene. In the Y recessive strain, the absence of an exon, likely due to an incorrec...

  4. Molecular Orientation Enhancement of Silk by the Hot-Stretching-Induced Transition from α-Helix-HFIP Complex to β-Sheet.

    Yoshioka, Taiyo; Tashiro, Kohji; Ohta, Noboru

    2016-04-11

    Enhancing the molecular orientation of the regenerated silk fibroin (RF) up to a level comparable to the native silk is highly challenging. Our novel and promising strategy for the poststretching process is (1) creating at first an α-helix-HFIP complex with a hexagonal packing as an intermediate state and then (2) stretching it at a high temperature to induce the helix-to-sheet structural phase transition. Here we show for the first time the significantly high stretching efficiency of the proposed technique compared with the conventional wet-stretching techniques and the successful achievement of higher crystalline orientation and higher Young's modulus compared even with the native silk. The detailed structural analysis based on the time-resolved simultaneous measurement of stress-strain curve, synchrotron X-ray scatterings, and FTIR has revealed the structural transition mechanism from the hexagonally packed α-helix-HFIP complex to the highly oriented β-sheet crystalline state as well as the critical level of crystal orientation needed for the helix-to-sheet transition. PMID:26974170

  5. THE GREAT SILK ROAD AND TURKISH WORLD

    Mustafa ÖZDEMİR

    2011-04-01

    Full Text Available The expression of the great Silk Road first came from the famouse Chinese traveler Çjan Syan before the BC and is now known as Kervan Road. Later, this expression gained a new meaning between Asia, the Mediterranean and the European countries.An estimated of 20 000 km of total length of the Great Silk Road passed the Turkish peoples settled lands. The Silk Road that was considered as the Jugularvein major trade route that was connecting China and Byzantium was in Turks hands.

  6. Silk epoxy composite crash worthiness performance

    Over the past several decades, increasing focus has been paid to the impact of structures where energy, during the impact event, needs to be absorbed in a controlled manner. This has led to considerable research being carried out on energy absorbers. These devices are designed to dissipate energy during an impact event and hence protect the structure under consideration. This research focuses on the square tube composite energy absorber. The tested square tubes were featured by 24 layers of natural silk as woven fabric in thermosetting epoxy resin. The fiber orientation angle of woven silk in the over-wrap is 0 and 90 degree to the tube axis. In this research silk/ epoxy tubes were cut at various thickness and they have been subjected to quasi static compression test. Silk/ epoxy square tubes have been analysed according to the compression test and the behaviour of this composite have been explained.(author)

  7. Multiscale modeling of spider dragline silk

    Patil, Sandeep Parasharam

    2015-01-01

    Spider dragline silk features an unusual combination of high strength, extensibility and stiffness, which outperforms some of the best materials known in terms of its mechanical performance. It is as strong as high-carbon steel, has a higher extensibility than the best commercial nylon filaments, and is tougher than Kevlar. For these reasons, dragline silk serves as a benchmark of modern polymer fiber technology, and the mass-production of a biomimetic material is of high interest. Developing...

  8. Electrodeposited silk coatings for bone implants.

    Elia, Roberto; Michelson, Courtney D; Perera, Austin L; Brunner, Teresa F; Harsono, Masly; Leisk, Gray G; Kugel, Gerard; Kaplan, David L

    2015-11-01

    The aim of this study was to characterize the mechanical properties and drug elution features of silk protein-based electrodeposited dental implant coatings. Silk processing conditions were modified to obtain coatings with a range of mechanical properties on titanium studs. These coatings were assessed for adhesive strength and dissolution, with properties tuned using water vapor annealing or glycerol incorporation to modulate crystalline content. Coating reproducibility was demonstrated over a range of silk concentrations from 1% to 10%. Surface roughness of titanium substrates was altered using industry relevant acid etching and grit blasting, and the effect of surface topography on silk coating adhesion was assessed. Florescent compounds were incorporated into the silk coatings, which were modulated for crystalline content, to achieve four days of sustained release of the compounds. This silk electrogelation technique offers a safe and relatively simple approach to generate mechanically robust, biocompatible, and degradable implant coatings that can also be functionalized with bioactive compounds to modulate the local regenerative tissue environment. PMID:25545462

  9. Silk hydrogel for cartilage tissue engineering.

    Chao, Pen-Hsiu Grace; Yodmuang, Supansa; Wang, Xiaoqin; Sun, Lin; Kaplan, David L; Vunjak-Novakovic, Gordana

    2010-10-01

    Cartilage tissue engineering based on cultivation of immature chondrocytes in agarose hydrogel can yield tissue constructs with biomechanical properties comparable to native cartilage. However, agarose is immunogenic and nondegradable, and our capability to modify the structure, composition, and mechanical properties of this material is rather limited. In contrast, silk hydrogel is biocompatible and biodegradable, and it can be produced using a water-based method without organic solvents that enables precise control of structural and mechanical properties in a range of interest for cartilage tissue engineering. We observed that one particular preparation of silk hydrogel yielded cartilaginous constructs with biochemical content and mechanical properties matching constructs based on agarose. This finding and the possibility to vary the properties of silk hydrogel motivated this study of the factors underlying the suitability of hydrogels for cartilage tissue engineering. We present data resulting from a systematic variation of silk hydrogel properties, silk extraction method, gel concentration, and gel structure. Data suggest that silk hydrogel can be used as a tool for studies of the hydrogel-related factors and mechanisms involved in cartilage formation, as well as a tailorable and fully degradable scaffold for cartilage tissue engineering. PMID:20725950

  10. Silk Roads or Steppe Roads? The Silk Roads in World History.

    Christian, David

    2000-01-01

    Explores the prehistory of the Silk Roads, reexamines their structure and history in the classical era, and explores shifts in their geography in the last one thousand years. Explains that a revised understanding of the Silk Roads demonstrates how the Afro-Eurasian land mass has been linked by networks of exchange since the Bronze Age. (CMK)

  11. Characterization of polymer, DNA-based, and silk thin film resistivities and of DNA-based films prepared for enhanced electrical conductivity

    Yaney, Perry P.; Ouchen, Fahima; Grote, James G.

    2009-08-01

    DC resistivity studies were carried out on biopolymer films of DNA-CTMA and silk fibroin, and on selected traditional polymer films, including PMMA and APC. Films of DNA-CTMA versus molecular weight and with conductive dopants PCBM, BAYTRON P and ammonium tetrachloroplatinate are reported. The films were spin coated on glass slides configured for measurements of volume dc resistance. The measurements used the alternating polarity method to record the applied voltage-dependent current independent of charging and background currents. The Arrhenius equation plus a constant was fitted to the conductivity versus temperature data of the polymers and the non-doped DNA-based biopolymers with activation energies ranging from 0.8 to 1.4 eV.

  12. Doxorubicin loaded nanodiamond-silk spheres for fluorescence tracking and controlled drug release

    Khalid, Asma; Mitropoulos, Alexander N.; Marelli, Benedetto; Tomljenovic-Hanic, Snjezana; Omenetto, Fiorenzo G.

    2015-01-01

    Nanoparticle (NP) based technologies have proved to be considerably beneficial for advances in biomedicine especially in the areas of disease detection, drug delivery and bioimaging. Over the last few decades, NPs have garnered interest for their exemplary impacts on the detection, treatment, and prevention of cancer. The full potential of these technologies are yet to be employed for clinical use. The ongoing research and development in this field demands single multifunctional composite materials that can be employed simultaneously for drug delivery and biomedical imaging. In this manuscript, a unique combination of silk fibroin (SF) and nanodiamonds (NDs) in the form of nanospheres are fabricated and investigated. The spheres were loaded with the anthracyline Doxorubicin (DoX) and the drug release kinetics for these ND-SF-DoX (NDSX) spheres were studied. NDs provided the fluorescence modality for imaging while the degradable SF spheres stabilized and released the drug in a controlled manner. The emission and structural properties of the spheres were characterized during drug release. The degradability of SF and the subsequent release of DoX from the spheres were monitored through fluorescence of NDs inside the spheres. This research demonstrates the enormous potential of the ND-SF nanocomposite platforms for diagnostic and therapeutic purposes, which are both important for pharmaceutical research and clinical settings.

  13. High oxygen-reduction activity of silk-derived activated carbon

    Iwazaki, Tomoya [Department of Fine Materials Engineering, Faculty of Textile Science and Technology, Shinshu University, 3-15-1 Tokida, Ueda, Nagano 386-8567 (Japan)]|[Shinano Kenshi Co. Ltd., Engineered Materials Division, 1078, Kamimaruko, Ueda, Nagano 386-0498 (Japan); Obinata, Ryoujin; Sugimoto, Wataru; Takasu, Yoshio [Department of Fine Materials Engineering, Faculty of Textile Science and Technology, Shinshu University, 3-15-1 Tokida, Ueda, Nagano 386-8567 (Japan)

    2009-02-15

    Activated carbon prepared from silk fibroin, which is free of metal elements, showed a high catalytic activity for the oxygen-reduction reaction (ORR). The activated carbon had a very high onset potential of E{sub onset} 0.83 V (vs. RHE) in oxygen-saturated 0.5 M H{sub 2}SO{sub 4} at 60 C. The ORR on the activated carbon proceeded by a four-electron process in the high-electrode-potential region; this gradually decreased to a 3.5-electron reaction below about 0.6 V (vs. RHE). Only about 1% of nitrogen atoms (mostly quaternary) remained in the activated carbon by heat-treatment at up to 1200 C are responsible for the high catalytic activity. The open circuit voltage of a polymer electrolyte fuel cell using the activated carbon as the cathode and a platinum/carbon black anode under pure oxygen and hydrogen gases, respectively, both at one atmosphere, was 0.96 V at 27 C. (author)

  14. Enhancement of osseointegration of polyethylene terephthalate artificial ligament by coating of silk fibroin and depositing of hydroxyapatite

    Jiang J; Wan F; Yang JJ; Hao W; Wang YX; Yao JR; Shao ZZ; Zhang P; Chen J.; Zhou L; SY Chen

    2014-01-01

    Jia Jiang,1,2 Fang Wan,1 Jianjun Yang,1 Wei Hao,3 Yaxian Wang,3 Jinrong Yao,3 Zhengzhong Shao,3 Peng Zhang,1 Jun Chen,1 Liang Zhou,4 Shiyi Chen11Fudan University Sports Medicine Center and Department of Sports Medicine and Arthroscopy Surgery, Huashan Hospital, 2State Key Laboratory of Molecular Engineering of Polymers, 3Laboratory of Advanced Materials, National Key Laboratory of Molecular Engineering of Polymers, Department of Macromolecular Science, Fudan University, Shanghai, People&...

  15. N-Acetyl cysteine (NAC)-mediated reinforcement of alpha-tricalcium phosphate/silk fibroin (?-TCP/SF) cement.

    Feng, Tao; Pi, Bin; Li, Bin; Jiang, Lei; Wang, Yi-Meng; Zhu, Xue-Song; Yang, Hui-Lin

    2015-12-01

    Calcium phosphate cements (CPCs) are popular bone filling materials and drug carriers. However poor mechanical properties and lack of osteoinduction restrict their clinical applications. Recent studies suggested the osteogenic properties of NAC. In our study, we incorporated NAC with ?-TCP/SF. We found that the compressive strength of ?-TCP/SF-NAC composites increased with increase in NAC concentration, possibly due to complex three-dimensional networks of SF induced by NAC, which was large and chemically heterogeneous and induced compact oriented growth of HA crystals. However the setting time increased slightly with the addition of NAC, due to the ruptured disulfide bonds in SF. The ?-TCP/SF-NAC composites also showed decent biocompatibility in vitro. As a result, these composites hold great potential as bone filling materials for clinical applications, including minimally invasive surgeries. PMID:26547317

  16. Effect of nanofiber content on bone regeneration of silk fibroin/poly(?-caprolactone) nano/microfibrous composite scaffolds

    Kim, Beom Su; Park, Ko Eun; Kim, Min Hee; You, Hyung Keun; Lee, Jun; Park, Won Ho

    2015-01-01

    The broad application of electrospun nanofibrous scaffolds in tissue engineering is limited by their small pore size, which has a negative influence on cell migration. This disadvantage could be significantly improved through the combination of nano- and microfibrous structure. To accomplish this, different nano/microfibrous scaffolds were produced by hybrid electrospinning, combining solution electrospinning with melt electrospinning, while varying the content of the nanofiber. The morpholog...

  17. Silk from Crickets: A New Twist on Spinning

    Walker, Andrew A; Weisman, Sarah; Church, Jeffrey S.; Merritt, David J; Mudie, Stephen T.; Sutherland, Tara D

    2012-01-01

    Raspy crickets (Orthoptera: Gryllacrididae) are unique among the orthopterans in producing silk, which is used to build shelters. This work studied the material composition and the fabrication of cricket silk for the first time. We examined silk-webs produced in captivity, which comprised cylindrical fibers and flat films. Spectra obtained from micro-Raman experiments indicated that the silk is composed of protein, primarily in a beta-sheet conformation, and that fibers and films are almost i...

  18. Silkworms transformed with chimeric silkworm/spider silk genes spin composite silk fibers with improved mechanical properties

    Teul, Florence; Miao, Yun-Gen; Sohn, Bong-Hee; Kim, Young-Soo; Hull, J. Joe; Fraser, Malcolm J; Lewis, Randolph V; Jarvis, Donald L.

    2012-01-01

    The development of a spider silk-manufacturing process is of great interest. However, there are serious problems with natural manufacturing through spider farming, and standard recombinant protein production platforms have provided limited progress due to their inability to assemble spider silk proteins into fibers. Thus, we used piggyBac vectors to create transgenic silkworms encoding chimeric silkworm/spider silk proteins. The silk fibers produced by these animals were composite materials t...

  19. Defensive behavior associated with secretions from the prosternal paired glands of the larvae of Heliconius erato phyllis Fabricius (Lepidoptera, Nymphalidae

    Eliane de Oliveira Borges

    2014-06-01

    Full Text Available Defensive behavior associated with secretions from the prosternal paired glands of the larvae of Heliconius erato phyllis Fabricius (Lepidoptera, Nymphalidae. Our work presents for the first time, the defensive behavior associated with the release of the product of the prosternal paired glands of the larva of Heliconius erato phyllis Fabricius, 1775 (Lepidoptera, Nymphalidae, Heliconiinae. The prosternal glands were first described for larvae of H. erato phyllis. They are formed by two types of glandular structures: the impair gland and the paired glands. The prosternal glands are located within the conical integumentary sac, which in turn is situated on the individual's prosternum. The main goal of this study is to analyze the existence of any secretion from the prosternal paired glands, and check the action mode of this secretion. The methodology used for chemical analysis of the glands included the aeration and, analysis in gas chromatography and gas chromatography-mass spectrometry. The results show that the prosternal glands do not produce volatiles. Bioassays were conducted with simulated and natural attacks and revealed that the prosternal paired glands produce secretions of defense together with silk produced by labials glands as a defense strategy, described for the first time, against ants. The strategy consists in wrapping the ant with silk threads, the entire wrapped object moved to the end of the body, with the aid of the legs and prolegs, and possibly fixed in a nearby place. Evidence for the existence of a conical integumentary sac in larvae of other species and families of Lepidoptera allows us to propose the possibility of occurrence of prosternal paired glands with defensive function in these other groups as well.

  20. Bio-functionalized silk hydrogel microfluidic systems.

    Zhao, Siwei; Chen, Ying; Partlow, Benjamin P; Golding, Anne S; Tseng, Peter; Coburn, Jeannine; Applegate, Matthew B; Moreau, Jodie E; Omenetto, Fiorenzo G; Kaplan, David L

    2016-07-01

    Bio-functionalized microfluidic systems were developed based on a silk protein hydrogel elastomeric materials. A facile multilayer fabrication method using gelatin sacrificial molding and layer-by-layer assembly was implemented to construct interconnected, three dimensional (3D) microchannel networks in silk hydrogels at 100 μm minimum feature resolution. Mechanically activated valves were implemented to demonstrate pneumatic control of microflow. The silk hydrogel microfluidics exhibit controllable mechanical properties, long-term stability in various environmental conditions, tunable in vitro and in vivo degradability in addition to optical transparency, providing unique features for cell/tissue-related applications than conventional polydimethylsiloxane (PDMS) and existing hydrogel-based microfluidic options. As demonstrated in the work here, the all aqueous-based fabrication process at ambient conditions enabled the incorporation of active biological substances in the bulk phase of these new silk microfluidic systems during device fabrication, including enzymes and living cells, which are able to interact with the fluid flow in the microchannels. These silk hydrogel-based microfluidic systems offer new opportunities in engineering active diagnostic devices, tissues and organs that could be integrated in vivo, and for on-chip cell sensing systems. PMID:27077566

  1. Adrenal Gland Cancer

    ... either benign or malignant. Benign tumors aren't cancer. Malignant ones are. Most adrenal gland tumors are ... and may not require treatment. Malignant adrenal gland cancers are uncommon. Types of tumors include Adrenocortical carcinoma - ...

  2. Contrastive research on the waterproof and dustproof mechanism of wild silkworm silk and domestic silkworm silk

    CHEN, Rou-Xi.

    2014-12-01

    Full Text Available Wild silk has properties of waterproof and dustproof, but its domestic partner has neither. Scanning electron microscopy (SEM) was used to observe and compare their morphology difference so that the possible mechanism can be elucidated. By the contrastive research, this paper concludes that the mech [...] anism of waterproof and dustproof of wild silk is due to selective repulsion, each hierarchical cascade of nanoparticles can repel either water molecules or fine particles in air. The SEM study reveals that the different nanoparticles with hierarchical structure on the silk surface are main factors of the highly selective repulsion. This theory can also explain the waterproof property of lotus leaf. A better understanding of the repulsion mechanism of wild silk could help the further design of bio-mimetic waterproof/dustproof artificial materials.

  3. Weaving Lao Silk Into Indigo Nights

    Melody Kemp

    2010-12-01

    Full Text Available The air turned chilly as the sun sighed into the nearby hills. It picked up the smells of dust mixed with metallic and dung flavours. Miss Phaeng watched, holding her breath as the last sliver of red fell out of sight. Casting a quick mantra to the spirits of nature, she swallowed a glass of lao lao to start the evening.Leaning mindfully over her loom, Miss Phaeng raked her nails across the piano strings of silk warp, plucking each to test its tension. A black sheet of pin-straight hair fell over her face, hiding the claret birthmark shaped just like a spider, that crept over her right cheek, one leg disappearing into the fine hairs of her temple.The coarse ivory silk recently spooled from the cocoons gathered in her garden pushed back against her hand.She felt the fizz of anticipation low in her belly as she gathered all the many shuttles holding the weft silk and dumped them into an old blackened basket. Inhaling its heady stink of ash, grass and smoke, she placed the basket next to where she would sit.Melody Kemp offers a close look at the Lao silk industry.

  4. The Ancient Art of Silk Painting

    Yonker, Kim

    2010-01-01

    In this article, the author describes a silk-painting project with a sea-creature theme for eighth-grade students. Other themes can be used such as geometric quilt designs, tropical rain forest, large flowers, Art Nouveau motifs, portraits and more. (Contains 2 resources.)

  5. Constructing Knowledge with Silk Road Visuals

    Bisland, Beverly Milner

    2008-01-01

    In this study a group of elementary teachers use illustrations, rather than written text, to introduce their students to the peoples and places of the ancient silk routes. The illustrations are from two picture books; "Marco Polo," written by Gian Paolo Cesaerani and illustrated by Piero Ventura (1977), and "We're Riding on a Caravan: An Adventure…

  6. Ultrastructure of the Lyonet's glands in larvae of Diatraea saccharalis Fabricius (Lepidoptera: Pyralidae

    Eliane Victoriano

    2004-08-01

    Full Text Available The Lyonet's gland is found in Lepidoptera larvae, close to the excretory duct of the silk gland. The role played by this gland is still uncertain. This work aims to describe the ultrastructure of the Lyonet's gland in Diatraea saccharalis larvae, offering suggestions regarding its possible function. The insects were reared under laboratory-controlled conditions. The glands were conventionally prepared for transmission (TEM and scanning (SEM electron microscopy. SEM showed that Lyonet's glands are paired small structures located in the ventral side of the head. They are composed by clustered long cells resembling leaves. Under TEM observations, each cell is surrounded by a thin basal lamina and contains large stellate nucleus. The cytoplasm presents large and empty canaliculi with small microvilli. The basal plasma membrane forms numerous infoldings where numerous and well-developed mitochondria are concentrated. The cytoplasmic membrane system is poorly developed. Our ultrastructural results suggest that the Lyonet's gland in D. saccharalis larvae may be involved in the uptake of small molecules from the hemolymph; no morphological evidences of macromolecules synthesis and secretion were noticed. The detection of nerve fibers in the gland suggest a neural control for the glandular cell function.

  7. Bombyx mori silk protein films microprocessing with a nanosecond ultraviolet laser and a femtosecond laser workstation: theory and experiments

    Lazare, S.; Sionkowska, A.; Zaborowicz, M.; Planecka, A.; Lopez, J.; Dijoux, M.; Louména, C.; Hernandez, M.-C.

    2012-01-01

    Laser microprocessing of several biopolymers from renewable resources is studied. Three proteinic materials were either extracted from the extracellular matrix like Silk Fibroin/Sericin and collagen, or coming from a commercial source like gelatin. All can find future applications in biomedical experimentation, in particular for cell scaffolding. Films of ˜hundred of microns thick were made by aqueous solution drying and laser irradiation. Attention is paid to the properties making them processable with two laser sources: the ultraviolet and nanosecond (ns) KrF (248 nm) excimer and the infrared and femtosecond (fs) Yb:KGW laser. The UV radiation is absorbed in a one-photon resonant process to yield ablation and the surface foaming characteristics of a laser-induced pressure wave. To the contrary, resonant absorption of the IR photons of the fs laser is not possible and does not take place. However, the high field of the intense I>˜1012 W/cm2 femtosecond laser pulse ionizes the film by the multiphoton absorption followed by the electron impact mechanism, yielding a dense plasma capable to further absorb the incident radiation of the end of the pulse. The theoretical model of this absorption is described in detail, and used to discuss the presented experimental effects (cutting, ablation and foaming) of the fs laser. The ultraviolet laser was used to perform simultaneous multiple spots experiments in which energetic foaming yields melt ejection and filament spinning. Airborne nanosize filaments "horizontally suspended by both ends" (0.25 μm diameter and 10 μm length) of silk biopolymer were observed upon irradiation with large fluences.

  8. Salt-leached silk scaffolds with tunable mechanical properties

    Yao, Danyu; Dong, Sen; Lu, Qiang; Hu, Xiao; Kaplan, David L.; Zhang, Bingbo; Zhu, Hesun

    2012-01-01

    Substrate mechanical properties have remarkable influences on cell behavior and tissue regeneration. Although salt-leached silk scaffolds have been used in tissue engineering, applications in softer tissue regeneration can be encumbered with excessive stiffness. In the present study, silk-bound water interactions were regulated by controlling processing to allow the preparation of salt-leached porous scaffolds with tunable mechanical properties. Increasing silk-bound water interactions result...

  9. THE INHERENT ACIDIC CHARACTERISTICS OF SILK, PART II – WEIGHTED SILKS

    Paul Garside

    2010-01-01

    Full Text Available Silk is found in many textiles of cultural and historic significance,but these fibres may deteriorate rapidly underadverse conditions, so are of particular concern to conservatorsand curators. Measurement of pH is common practicewhen assessing condition, as enhanced acidity is oftenascribed to previous processing and treatment, adsorbedpollutants and oxidative deterioration, but recent researchhas demonstrated that the increased acidity of silk is anexpected consequence of ageing, irrespective of the causeof deterioration. The study presented here builds on thisearlier work to assess the role that common silk processingmethods (bleaching and metal salt weighting have onthis phenomenon. Tensile strength was used as a marker ofdeterioration, independent of mechanism, and was comparedwith pH values of saline extracts of various silk samples.These samples included surrogates, prepared usinghistorically accurate methods and subject to acceleratedageing, as well as a range of historic materials. It was seenthat most samples followed the same correlation betweenpH and strength as seen in the previous work. However,silks weighted with tin salts appeared to be buffered (in therange pH 6.75-7.05, irrespective of their mechanicalintegrity.

  10. Processing silk hydrogel and its applications in biomedical materials.

    Wang, Hai-Yan; Zhang, Yu-Qing

    2015-01-01

    This review mainly introduces the types of silk hydrogels, their processing methods, and applications. There are various methods for hydrogel preparation, and many new processes are being developed for various applications. Silk hydrogels can be used in cartilage tissue engineering, drug release materials, 3D scaffolds for cells, and artificial skin, among other applications because of their porous structure and high porosity and the large surface area for growth, migration, adhesion and proliferation of cells that the hydrogels provide. All of these advantages have made silk hydrogels increasingly attractive. In addition, silk hydrogels have wide prospects for application in the field of biomedical materials. PMID:25740113

  11. White-Light-Induced Collective Heating of Gold Nanocomposite/Bombyx mori Silk Thin Films with Ultrahigh Broadband Absorbance.

    Tsao, Shao Hsuan; Wan, Dehui; Lai, Yu-Sheng; Chang, Ho-Ming; Yu, Chen-Chieh; Lin, Keng-Te; Chen, Hsuen-Li

    2015-12-22

    This paper describes a systematic investigation of the phenomenon of white-light-induced heating in silk fibroin films embedded with gold nanoparticles (Au NPs). The Au NPs functioned to develop an ultrahigh broadband absorber, allowing white light to be used as a source for photothermal generation. With an increase of the Au content in the composite films, the absorbance was enhanced significantly around the localized surface plasmon resonance (LSPR) wavelength, while non-LSPR wavelengths were also increased dramatically. The greater amount of absorbed light increased the rate of photoheating. The optimized composite film exhibited ultrahigh absorbances of approximately 95% over the spectral range from 350 to 750 nm, with moderate absorbances (>60%) at longer wavelengths (750-1000 nm). As a result, the composite film absorbed almost all of the incident light and, accordingly, converted this optical energy to local heat. Therefore, significant temperature increases (ca. 100 C) were readily obtained when we irradiated the composite film under a light-emitting diode or halogen lamp. Moreover, such composite films displayed linear light-to-heat responses with respect to the light intensity, as well as great photothermal stability. A broadband absorptive film coated on a simple Al/Si Schottky diode displayed a linear, significant, stable photo-thermo-electronic effect in response to varying the light intensity. PMID:26552041

  12. Mammary Glands: Developmental Changes

    The mammary gland progresses from the accumulation of a few cells in the embryonic ectoderm to a highly arborescent tubulo-alveolar gland capable of secreting a highly nutritious product for consumption. Throughout this progression, various changes occur during each developmental stage: prenatal, pr...

  13. Salivary Gland Cancer Treatment

    ... Fast neutron radiation therapy to lymph nodes with cancer. Chemotherapy . A clinical trial of fast neutron radiation therapy to the tumor . A clinical ... Gland Cancer Treatment of stage IV salivary gland cancer may include the ... trial of chemotherapy with or without radiation therapy . Check the list ...

  14. Salivary Gland Cancer

    ... contains antibodies that can kill germs. Salivary gland cancer is a type of head and neck cancer. It is rare. It may not cause any ... pain in your face Doctors diagnose salivary gland cancer using a physical exam, imaging tests, and a ...

  15. Estradiol-17beta in Bombyx mori: possible significance and its effect on silk production.

    Keshan; Ray

    2000-06-01

    Although estrogen is well known as a vertebrate sex steroid, its presence in insects, including Bombyx mori, raises questions about its precise role in the physiology of insects. It was reported earlier that estradiol-17beta (E(2)) exerts a specific effect on silk-gland function in B. mori and that it may act in a nuclear-mediated way. To evaluate further the effect of E(2) on cocoon characters, larval growth and development, 1µg/g of E(2) was applied topically to the first and second day of fifth instar larvae. This resulted in a significant enhancement of cocoon characters, such as cocoon shell weight, silk filament length per cocoon, denier per filament and reelability of the cocoons, without any adverse effect on fecundity and hatchability. In the present study, E(2) levels in the haemolymph were quantified on different days of the fifth instar larvae and age-dependent changes in the endogenous E(2) titre have been demonstrated. These age-dependent variations in E(2) content coincide with physiological events occurring during the fifth instar. Such observations exclude the possibility of a dietary origin for E(2), as a sudden and sharp rise of the E(2) level in the haemolymph was observed on the 10th day of the fifth instar, preceded by a small increase on the ninth day after an eight-day feeding period. The increased level of estradiol in the haemolymph of larvae treated topically with E(2) indicates effective penetration of this hormone through the larval cuticle. Moreover, similar patterns of alteration of E(2) levels on different days of the fifth instar in both control and treated groups suggests the existence of some internal metabolic pathway in the silkworm body to regulate the hormone titre. Thus, the present investigation offers a system for investigating the unique function of E(2) in B. mori and offers potential for improvement of silk production. PMID:10802119

  16. Silk hydrogel for cartilage tissue engineering

    Chao, Pen-Hsiu Grace; Yodmuang, Supansa; Wang, Xiaoqin; Sun, Lin; Kaplan, David L.; Vunjak-Novakovic, Gordana

    2010-01-01

    Cartilage tissue engineering based on cultivation of immature chondrocytes in agarose hydrogel can yield tissue constructs with biomechanical properties comparable to native cartilage. However, agarose is immunogenic and non-degradable, and our capability to modify the structure, composition, and mechanical properties of this material is rather limited. In contrast, silk hydrogel is biocompatible and biodegradable, and it can be produced using a water-based method without organic solvents tha...

  17. Injectable Silk Foams for Soft Tissue Regeneration

    Bellas, E.; Lo, T.J.; Fournier, E.P.; Brown, J.E.; Abbott, R.D.; E. S. Gil; Marra, K.G.; Rubin, J.P.; Leisk, G.G.; Kaplan, D. L.

    2014-01-01

    Soft tissue fillers are needed for restoration of a defect or augmentation of existing tissues. Autografts and lipotransfer have been under study for soft tissue reconstruction but yield inconsistent results, often with considerable resorption of the grafted tissue. A minimally invasive procedure would reduce scarring and recovery time as well as allow for the implant and/or grafted tissue to be placed closer to existing vasculature. Here, we demonstrate the feasibility of an injectable silk ...

  18. Electricity from the Silk Cocoon Membrane

    Tulachan, Brindan; Meena, Sunil Kumar; Rai, Ratan Kumar; Mallick, Chandrakant; Kusurkar, Tejas Sanjeev; Teotia, Arun Kumar; Sethy, Niroj Kumar; Bhargava, Kalpana; Bhattacharya, Shantanu; Kumar, Ashok; Sharma, Raj Kishore; Sinha, Neeraj; Singh, Sushil Kumar; Das, Mainak

    2014-01-01

    Silk cocoon membrane (SCM) is an insect engineered structure. We studied the electrical properties of mulberry (Bombyx mori) and non-mulberry (Tussar, Antheraea mylitta) SCM. When dry, SCM behaves like an insulator. On absorbing moisture, it generates electrical current, which is modulated by temperature. The current flowing across the SCM is possibly ionic and protonic in nature. We exploited the electrical properties of SCM to develop simple energy harvesting devices, which could operate lo...

  19. Osteogenic signaling on silk-based matrices.

    Midha, Swati; Murab, Sumit; Ghosh, Sourabh

    2016-08-01

    Bone tissue engineering has mainly focused on generating 3D grafts to repair bone defects. However, the underlying signaling mechanisms responsible for development of such 3D bone equivalents have largely been ignored. Here we describe the crucial aspects of embryonic osteogenesis and bone development including cell sources and general signaling cascades that guide mesenchymal progenitors towards osteogenic lineage. Drawing from the knowledge of developmental biology, we then review how silk biomaterial can regulate osteogenic signaling by focusing on the expression of cell surface markers, functional genomic information (mRNA) of stem cells cultured on silk matrices. In an attempt to recapitulate exact in vivo microenvironment of osteogenesis, role of scaffold architecture and material chemistry in regulating cellular differentiation is elaborated. The generated knowledge will not only improve our understanding of cell-material interactions but reveal newer strategies beyond a conventional tissue engineering paradigm and open new prospects for developing silk-based therapies against clinically relevant bone disorders. PMID:27163625

  20. EFFECT OF METHANOLIC EXTRACT OF SWEET CORN SILK ON EXPERIMENTALLY INDUCED HYPERTHYROIDISM IN SWISS ALBINO RATS

    Thoudam Bhaigyabati

    2012-03-01

    Full Text Available Plant materials have been used as medicine for a wide variety of human ailments due to increase in cost of treatment, side effects of several allopathic drugs and development of resistance to currently used drugs for infectious diseases. In the present study, hyperthyroidism was induced to male Swiss albino rats orally using thyroxine for 14 days. The impact of methanolic extract of sweet corn silk in different concentrations (200, 300 & 400mg/kg for 21 days was assessed in these animals. The serum was analyzed for thyroid hormone level and lipid profile before and during the experiment. Results showed that 400mg/kg concentration of methanolic extract has the highest effectiveness which is equal to that of standard drug to revert the hyperthyroid condition in Swiss albino rats and were confirmed with the histopathological study of the thyroid gland.

  1. Visual Literacy with Picture Books: The Silk Road

    Bisland, Beverly Milner Lee

    2007-01-01

    The ancient Silk Routes connecting China to Europe across the rugged mountains and deserts of central Asia are one of the primary examples of transculturation in world history. Traders on these routes dealt not only in goods such as silk and horses but also made possible the spread of art forms as well as two major religions, Buddhism and Islam. …

  2. Geographic Perspectives with Elementary Students: The Silk Road

    Bisland, Beverly Milner

    2006-01-01

    The purpose of this study is to investigate elementary students' explanations of how physical features of the land influence the location of humanly defined structures including trade routes, such as the silk routes. The silk routes were a series of caravan trade routes that extended from Turkey to China and were located as far south as India and…

  3. Transgenic phenolic production in corn silks moderately enhances insect resistance

    Some phenolic compounds produced in corn silks, such as maysin, can promote resistance to caterpillar pests. We evaluated transgenic maize engineered to express a maize cDNA controlled by a putative silk specific promoter for secondary metabolite production and corn earworm resistance. Transgene e...

  4. Optical surface profiling of orb-web spider capture silks

    Much spider silk research to date has focused on its mechanical properties. However, the webs of many orb-web spiders have evolved for over 136 million years to evade visual detection by insect prey. It is therefore a photonic device in addition to being a mechanical device. Herein we use optical surface profiling of capture silks from the webs of adult female St Andrews cross spiders (Argiope keyserlingi) to successfully measure the geometry of adhesive silk droplets and to show a bowing in the aqueous layer on the spider capture silk between adhesive droplets. Optical surface profiling shows geometric features of the capture silk that have not been previously measured and contributes to understanding the links between the physical form and biological function. The research also demonstrates non-standard use of an optical surface profiler to measure the maximum width of a transparent micro-sized droplet (microlens).

  5. Optical surface profiling of orb-web spider capture silks

    Kane, D M; Joyce, A M; Staib, G R [Department of Physics, Macquarie University, Sydney, NSW 2109 (Australia); Herberstein, M E, E-mail: deb.kane@mq.edu.a [Department of Biological Sciences, Macquarie University, Sydney, NSW 2109 (Australia)

    2010-09-15

    Much spider silk research to date has focused on its mechanical properties. However, the webs of many orb-web spiders have evolved for over 136 million years to evade visual detection by insect prey. It is therefore a photonic device in addition to being a mechanical device. Herein we use optical surface profiling of capture silks from the webs of adult female St Andrews cross spiders (Argiope keyserlingi) to successfully measure the geometry of adhesive silk droplets and to show a bowing in the aqueous layer on the spider capture silk between adhesive droplets. Optical surface profiling shows geometric features of the capture silk that have not been previously measured and contributes to understanding the links between the physical form and biological function. The research also demonstrates non-standard use of an optical surface profiler to measure the maximum width of a transparent micro-sized droplet (microlens).

  6. Increased molecular mobility in humid silk fibers under tensile stress

    Seydel, Tilo; Knoll, Wiebke; Greving, Imke; Dicko, Cedric; Koza, Michael M.; Krasnov, Igor; Mller, Martin

    2011-01-01

    Silk fibers are semicrystalline nanocomposite protein fibers with an extraordinary mechanical toughness that changes with humidity. Diffusive or overdamped motion on a molecular level is absent in dry silkworm silk, but present in humid silk at ambient temperature. This microscopic diffusion distinctly depends on the externally applied macroscopic tensile force. Quasielastic and inelastic neutron-scattering data as a function of humidity and of tensile strain on humid silk fibers support the model that both the adsorbed water and parts of the amorphous polymers participate in diffusive motion and are affected by the tensile force. It is notable that the quasielastic linewidth of humid silk at 100% relative humidity increases significantly with the applied force. The effect of the tensile force is discussed in terms of an increasing alignment of the polymer chains in the amorphous fraction with increasing tensile stress which changes the geometrical restrictions of the diffusive motions.

  7. Study on silk yellowing induced by gamma-irradiation

    The changes in the yellow color of silk threads with total dose of irradiation applied were described and studied by a colorimetric method and by monochrome photography. The change into a yellow color of the specimen in the course of irradiation was clearly detected in photographs using filters, 2B and SC 56 under light conditions at the wavelength of 366 nm. The b/L value measured by colorimetry in undegummed and degummed silk fibers sharply increased in the early stage of irradiation. Yellow color indices (b/L) of the specimen subjected to gamma-irradiation continued to increase and the yellow color of the silk threads became more pronounced above a total dose of irradiation of 21 Mrad. The b/L value of the undegummed silk fiber which had deen irradiated was about 2 times that of the degummed silk fiber. (author)

  8. Chimeric spider silk proteins mediated by intein result in artificial hybrid silks.

    Lin, Senzhu; Chen, Gefei; Liu, Xiangqin; Meng, Qing

    2016-07-01

    Hybrid silks hold a great potential as specific biomaterials due to its controlled mechanical properties. To produce fibers with tunable properties, here we firstly made chimeric proteins in vitro, called W2C4CT and W2C8CT, with ligation of MaSp repetitive modules (C) with AcSp modules (W) by intein trans splicing technology from smaller precursors without final yield reduction. Intein mediated chimeric proteins form fibers at a low concentration of 0.4 mg/mL in 50 mM K3 PO4 pH 7.5 just drawn by hand. Hybrid fibers show smoother surface, and also have stronger chemical resistance as compared with fibers from W2CT (W fibers) and mixture of W2CT/C8CT (MHF8 fibers). Fibers from chimeric protein W2C4CT (HFH4) have improved mechanical properties than W fibers; however, with more C modules W2C8CT fibers (HFH8) properties decreased, indicates the length proportion of various modules is very important and should be optimized for fibers with specific properties. Generally, hybrid silks generated via chimeric proteins, which can be simplified by intein trans splicing, has greater potential to produce fibers with tunable properties. Our research shows that intein mediated directional protein ligation is a novel way to make large chimeric spider silk proteins and hybrid silks. © 2016 Wiley Periodicals, Inc. Biopolymers 105: 385-392, 2016. PMID:26948769

  9. The development project of electron irradiated silk material used for restoration of historical silk-mounted painting

    Many pictures have been painted on the silk materials in Japan. To repair the historical painting, the repairing materials were developed by new silk fibers with electron beam irradiation. Total exposure was about 2 MGy (170 to 230 Mrad). In order to be clear the degradation mechanism of the irradiated silk materials, the samples were irradiated by 254 nm and 365nm UV light and electron beam. The tensile strength of samples decreased with increasing exposure energy. The electron irradiated silk materials showed random clacking and small molecular weight. However, aromatic amino acid of the long wavelength UV light irradiated silk and peptide linkage of the short wavelength UV light irradiated silk were cracked. The historical silk-mounted painting in the Edo era showed decrease of aromatic amino acid peak and that in the Kamakura era not observable peak. Accordingly, the degradation process of historical silk-mounted painting seemed to be the same as the process depend on the long wavelength UV light. (S.Y.)

  10. The development project of electron irradiated silk material used for restoration of historical silk-mounted painting

    Sano, Chie [Independent Administrative Inst. National Research Inst. for cultural properties, Tokyo (Japan); Masuda, Katsuhiko

    2001-04-01

    Many pictures have been painted on the silk materials in Japan. To repair the historical painting, the repairing materials were developed by new silk fibers with electron beam irradiation. Total exposure was about 2 MGy (170 to 230 Mrad). In order to be clear the degradation mechanism of the irradiated silk materials, the samples were irradiated by 254 nm and 365nm UV light and electron beam. The tensile strength of samples decreased with increasing exposure energy. The electron irradiated silk materials showed random clacking and small molecular weight. However, aromatic amino acid of the long wavelength UV light irradiated silk and peptide linkage of the short wavelength UV light irradiated silk were cracked. The historical silk-mounted painting in the Edo era showed decrease of aromatic amino acid peak and that in the Kamakura era not observable peak. Accordingly, the degradation process of historical silk-mounted painting seemed to be the same as the process depend on the long wavelength UV light. (S.Y.)

  11. Thyroid gland removal - discharge

    ... surgery to remove part or all of your thyroid gland. This operation is called thyroidectomy . You probably ... in just a few weeks. If you had thyroid cancer, you may need to have radioactive iodine ...

  12. Undifferentiated salivary gland carcinomas

    Herbst, H.; Hamilton-Dutoit, S.; Jakel, K.T.; Niedobitek, G.

    2004-01-01

    , comprising malignant lymphomas, amelanotic melanomas, Merkel cell carcinomas, and adenoid cystic carcinomas, in particular in small biopsy materials. Because of the rarity of undifferentiated salivary gland carcinomas, the differential diagnosis should always include metastases of undifferentiated carcinomas...

  13. Pineal and Pituitary Glands

    ... Anatomy Neurons & Glial Cells Brain Meninges Spinal Cord Cranial Nerves Pineal and Pituitary Glands Abstracting, Coding, & Staging Topographic ... Anatomy & Physiology module on this Web site. « Previous (Cranial Nerves) Next (Abstracting, Coding, & Staging) » Contact Us | Privacy Policy | ...

  14. Pediatric salivary gland imaging

    Boyd, Zachary T. [University of Missouri-Kansas City School of Medicine, Kansas City, MO (United States); Goud, Asha R. [University of California Irvine Medical Center, Department of Radiology, Orange, CA (United States); Lowe, Lisa H. [Children' s Mercy Hospitals and Clinics and the University of Missouri-Kansas City, Department of Radiology, Kansas City, MO (United States); Shao, Lei [Children' s Mercy Hospitals and Clinics and the University of Missouri-Kansas City, Department of Pathology, Kansas City, MO (United States)

    2009-07-15

    A variety of lesions occur in the pediatric salivary glands. With modern imaging techniques such as Doppler sonography, helical CT, and MRI, identification of a specific etiology is often possible. Knowledge of clinical information, normal anatomy, and imaging characteristics of salivary gland pathology are essential for appropriate radiologic evaluation. This review illustrates the various congenital, neoplastic, and inflammatory entities that can occur within the parotid, submandibular, and sublingual spaces. (orig.)

  15. Pediatric salivary gland imaging

    A variety of lesions occur in the pediatric salivary glands. With modern imaging techniques such as Doppler sonography, helical CT, and MRI, identification of a specific etiology is often possible. Knowledge of clinical information, normal anatomy, and imaging characteristics of salivary gland pathology are essential for appropriate radiologic evaluation. This review illustrates the various congenital, neoplastic, and inflammatory entities that can occur within the parotid, submandibular, and sublingual spaces. (orig.)

  16. High-pressure gland

    A high-pressure gland is proposed for nuclear installations, operating at high temperatures. It consists of a stationary case with a stationary nonmetal sealing ring and of a rotary case with a rotary nonmetal sealing ring. The inner functional surfaces of the cases are provided with annular seating surfaces with semicircular grooves. Thus, the force and thermal deformations of the functional parts of the gland are eliminated. The sealing rings are manufactured from materials of identical hardness. (M.S.)

  17. Bioprospecting Finds the Toughest Biological Material: Extraordinary Silk from a Giant Riverine Orb Spider

    Agnarsson, Ingi; Kuntner, Matjaž; Todd A. Blackledge

    2010-01-01

    Background Combining high strength and elasticity, spider silks are exceptionally tough, i.e., able to absorb massive kinetic energy before breaking. Spider silk is therefore a model polymer for development of high performance biomimetic fibers. There are over 41.000 described species of spiders, most spinning multiple types of silk. Thus we have available some 200.000+ unique silks that may cover an amazing breadth of material properties. To date, however, silks from only a few tens of sp...

  18. In-vitro Characterization of Silk Sericin as an Anti-aging Agent

    Thitinan Kitisin; Pannamas Maneekan; Natthanej Luplertlop

    2013-01-01

    Silk sericin is a natural macromolecular protein derived from the silkworm, Bombyx mori. It is also a by-product of silk-making. From previous reports, many cosmeceuticals and other cosmetic products have been developed with silk sericin. This study aimed to investigate an anti-aging property of silk sericin by in-vitro characterization using fibroblast cell culture model. The results showed that silk sericin can stimulate collagen type I synthesis, suppress the regulation of nitrite, which ...

  19. The elaborate structure of spider silk: Structure and function of a natural high performance fiber

    Rmer, Lin; Scheibel, Thomas

    2008-01-01

    Biomaterials, having evolved over millions of years, often exceed man-made materials in their properties. Spider silk is one outstanding fibrous biomaterial which consists almost entirely of large proteins. Silk fibers have tensile strengths comparable to steel and some silks are nearly as elastic as rubber on a weight to weight basis. In combining these two properties, silks reveal a toughness that is two to three times that of synthetic fibers like Nylon or Kevlar. Spider silk is also antim...

  20. Radionuclide salivary gland imaging

    Salivary gland imaging with 99mTc as pertechnetate provides functional information concerning trapping and excretion of the parotid and submandibular glands. Anatomic information gained often adds little to clinical evaluation. On the other hand, functional information may detect subclinical involvement, which correlates well with biopsy of the minor labial salivary glands. Salivary gland abnormalities in systemic disease such as sarcoidosis, rheumatoid arthritis, lupus erythematosus, and other collagenvascular disorders may be detected before they result in the clinical manifestaions of Sjoegren's syndrome. Such glands, after initially demonstrating increased trapping in the acute phase, tend to have decreased trapping and failure to discharge pertechnetate in response to an appropriate physiologic stimulus. Increased uptake of gallium-67 citrate often accompanies these findings. Inflammatory parotitis can be suspected when increased perfusion is evident on radionuclide angiography with any agent. The ability of the salivary gland image to detect and categorize mass lesions, which result in focal areas of diminished activity such as tumors, cysts, and most other masses, is disappointing, while its ability to detect and categorize Warthin's tumor, which concentrates pertechnetate, is much more valuable, although not specific

  1. Radionuclide salivary gland imaging

    Mishkin, F.S.

    1981-10-01

    Salivary gland imaging with 99mTc as pertechnetate provides functional information concerning trapping and excretion of the parotid and submandibular glands. Anatomic information gained often adds little to clinical evaluation. On the other hand, functional information may detect subclinical involvement, which correlates well with biopsy of the minor labial salivary glands. Salivary gland abnormalities in systemic disease such as sarcoidosis, rheumatoid arthritis, lupus erythematosus, and other collagenvascular disorders may be detected before they result in the clinical manifestaions of Sjoegren's syndrome. Such glands, after initially demonstrating increased trapping in the acute phase, tend to have decreased trapping and failure to discharge pertechnetate in response to an appropriate physiologic stimulus. Increased uptake of gallium-67 citrate often accompanies these findings. Inflammatory parotitis can be suspected when increased perfusion is evident on radionuclide angiography with any agent. The ability of the salivary gland image to detect and categorize mass lesions, which result in focal areas of diminished activity such as tumors, cysts, and most other masses, is disappointing, while its ability to detect and categorize Warthin's tumor, which concentrates pertechnetate, is much more valuable, although not specific.

  2. Electricity from the Silk Cocoon Membrane

    Tulachan, Brindan; Meena, Sunil Kumar; Rai, Ratan Kumar; Mallick, Chandrakant; Kusurkar, Tejas Sanjeev; Teotia, Arun Kumar; Sethy, Niroj Kumar; Bhargava, Kalpana; Bhattacharya, Shantanu; Kumar, Ashok; Sharma, Raj Kishore; Sinha, Neeraj; Singh, Sushil Kumar; Das, Mainak

    2014-06-01

    Silk cocoon membrane (SCM) is an insect engineered structure. We studied the electrical properties of mulberry (Bombyx mori) and non-mulberry (Tussar, Antheraea mylitta) SCM. When dry, SCM behaves like an insulator. On absorbing moisture, it generates electrical current, which is modulated by temperature. The current flowing across the SCM is possibly ionic and protonic in nature. We exploited the electrical properties of SCM to develop simple energy harvesting devices, which could operate low power electronic systems. Based on our findings, we propose that the temperature and humidity dependent electrical properties of the SCM could find applications in battery technology, bio-sensor, humidity sensor, steam engines and waste heat management.

  3. Spider Silk: The Mother Nature's Biological Superlens

    Monks, James N; Wang, Zengbo

    2016-01-01

    This paper demonstrates a possible new microfiber bio near field lens that uses minor ampullate spider silk,spun from the Nephila edulis spider, to create a real time image of a surface using near field optical techniques. The microfiber bio lens is the world's first natural superlens created by exploring biological materials. The resolution of the surface image overcomes the diffraction limit, with the ability to resolve patterns at 100 nm under a standard white light source in reflection mode. This resolution offers further developments in superlens technology and paves the way for new bio optics.

  4. Inhibitory Effect of Corn Silk on Skin Pigmentation

    Sang Yoon Choi

    2014-03-01

    Full Text Available In this study, the inhibitory effect of corn silk on melanin production was evaluated. This study was performed to investigate the inhibitory effect of corn silk on melanin production in Melan-A cells by measuring melanin production and protein expression. The corn silk extract applied on Melan-A cells at a concentration of 100 ppm decreased melanin production by 37.2% without cytotoxicity. This was a better result than arbutin, a positive whitening agent, which exhibited a 26.8% melanin production inhibitory effect at the same concentration. The corn silk extract did not suppress tyrosinase activity but greatly reduced the expression of tyrosinase in Melan-A cells. In addition, corn silk extract was applied to the human face with hyperpigmentation, and skin color was measured to examine the degree of skin pigment reduction. The application of corn silk extract on faces with hyperpigmentation significantly reduced skin pigmentation without abnormal reactions. Based on the results above, corn silk has good prospects for use as a material for suppressing skin pigmentation.

  5. Biomimetic calcium phosphate coatings on recombinant spider silk fibres

    Yang Liang; Habibovic, Pamela; Van Blitterswijk, Clemens A [Department of Tissue Regeneration, University of Twente, PO Box 217, 7500 AE Enschede (Netherlands); Hedhammar, My; Johansson, Jan [Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, the Biomedical Centre, Box 575, 751 23 Uppsala (Sweden); Blom, Tobias; Leifer, Klaus [Department of Engineering Sciences, Uppsala University, Box 534, S-751 21 Uppsala (Sweden)

    2010-08-01

    Calcium phosphate ceramic coatings, applied on surfaces of metallic and polymeric biomaterials, can improve their performance in bone repair and regeneration. Spider silk is biocompatible, strong and elastic, and hence an attractive biomaterial for applications in connective tissue repair. Recently, artificial spider silk, with mechanical and structural characteristics similar to those of native spider silk, has been produced from recombinant minispidroins. In the present study, supersaturated simulated body fluid was used to deposit calcium phosphate coatings on recombinant spider silk fibres. The mineralization process was followed in time using scanning electron microscopy equipped with an energy dispersive x-ray (EDX) detector and Raman spectroscope. Focused ion beam technology was used to produce a cross section of a coated fibre, which was further analysed by EDX. Preliminary in vitro experiments using a culture of bone marrow-derived human mesenchymal stem cells (hMSCs) on coated fibres were also performed. This study showed that recombinant spider silk fibres were successfully coated with a homogeneous and thick crystalline calcium phosphate layer. In the course of the mineralization process from modified simulated body fluid, sodium chloride crystals were first deposited on the silk surface, followed by the deposition of a calcium phosphate layer. The coated silk fibres supported the attachment and growth of hMSCs.

  6. The effects of corn silk on glycaemic metabolism

    Han Linna

    2009-11-01

    Full Text Available Abstract Background Corn silk contains proteins, vitamins, carbohydrates, Ca, K, Mg and Na salts, fixed and volatile oils, steroids such as sitosterol and stigmasterol, alkaloids, saponins, tannins, and flavonoids. Base on folk remedies, corn silk has been used as an oral antidiabetic agent in China for decades. However, the hypoglycemic activity of it has not yet been understood in terms of modern pharmacological concepts. The purpose of this study is to investigate the effects of corn silk on glycaemic metabolism. Methods Alloxan and adrenalin induced hyperglycemic mice were used in the study. The effects of corn silk on blood glucose, glycohemoglobin (HbA1c, insulin secretion, damaged pancreatic β-cells, hepatic glycogen and gluconeogenesis in hyperglycemic mice were studied respectively. Results After the mice were orally administered with corn silk extract, the blood glucose and the HbA1c were significantly decreased in alloxan-induced hyperglycemic mice (p 0.05. Although corn silk extract increased the level of hepatic glycogen in the alloxan-induced hyperglycemic mice, there was no significant difference between them and that of the control group(p > 0.05. Conclusion Corn silk extract markedly reduced hyperglycemia in alloxan-induced diabetic mice. The action of corn silk extract on glycaemic metabolism is not via increasing glycogen and inhibiting gluconeogenesis but through increasing insulin level as well as recovering the injured β-cells. The results suggest that corn silk extract may be used as a hypoglycemic food or medicine for hyperglycemic people in terms of this modern pharmacological study.

  7. Submandibular gland excision

    Alexandre Sanfurgo de Carvalho

    2015-02-01

    Full Text Available OBJECTIVE: to analyze the value of fine needle aspiration and the rates of postoperative complications in patients undergoing resection of the submandibular gland. METHODS: we analyzed the records of patients treated with resection of the gland from January 1995 to December 2008. The data collected included age, gender, findings on clinical history, surgical procedure, results of fine needle aspiration (FNA, pathological diagnosis and complications. RESULTS: 117 patients were studied, aged 12-89 years (mean 48, 70 women and 47 men. Thirty-nine patients (33.3% were affected by inflammatory diseases (28 patients with lithiasis, 70 had benign tumors, and malignant tumors, eight. Regarding FNA, the sensitivity and specificity were 85.7% and 100%, respectively. Nine patients (7.7% had temporary paralysis of the marginal mandibular nerve and one had permanent paralysis. CONCLUSION: resection of the submandibular gland is a safe procedure, with low complication rates.

  8. Mammary gland stem cells

    Fridriksdottir, Agla J R; Petersen, Ole W; Rønnov-Jessen, Lone

    Distinct subsets of cells, including cells with stem cell-like properties, have been proposed to exist in normal human breast epithelium and breast carcinomas. The cellular origins of epithelial cells contributing to gland development, tissue homeostasis and cancer are, however, still poorly...... understood. The mouse is a widely used model of mammary gland development, both directly by studying the mouse mammary epithelial cells themselves and indirectly, by studying development, morphogenesis, differentiation and carcinogenesis of xenotransplanted human breast epithelium in vivo. While in early...... studies, human or mouse epithelium was implanted as fragments into the mouse gland, more recent technical progress has allowed the self-renewal capacity and differentiation potential of distinct cell populations or even individual cells to be interrogated. Here, we review and discuss similarities and...

  9. Malignant salivary gland tumours

    Thompson, S.H. (University of the Witwatersrand, Johannesburg (South Africa). Dept. of Oral Pathology)

    1982-08-01

    The most frequent malignant salivary gland tumours are the mucoepidermoid tumour, adenoid cystic carcinoma and adenocarcinoma. The major salivary glands and the minor glands of the mouth and upper respiratory tract may potentially develop any of these malignant lesions. Malignant lesions most frequently present as a palpable mass and tend to enlarge more rapidly than benign neoplasms. Pain, paresthesia, muscle paralysis and fixation to surrounding tissue are all ominous signs and symptoms. The only reliable means of differential diagnosis of these lesions is biopsy and histologic analysis. Therapy involves surgery or a combination of surgery and radiation therapy. The ultimate prognosis is governed by the intrinsic biologic behaviour of the neoplasms, the extent of disease and adequate clinical therapy.

  10. Malignant salivary gland tumours

    The most frequent malignant salivary gland tumours are the mucoepidermoid tumour, adenoid cystic carcinoma and adenocarcinoma. The major salivary glands and the minor glands of the mouth and upper respiratory tract may potentially develop any of these malignant lesions. Malignant lesions most frequently present as a palpable mass and tend to enlarge more rapidly than benign neoplasms. Pain, paresthesia, muscle paralysis and fixation to surrounding tissue are all ominous signs and symptoms. The only reliable means of differential diagnosis of these lesions is biopsy and histologic analysis. Therapy involves surgery or a combination of surgery and radiation therapy. The ultimate prognosis is governed by the intrinsic biologic behaviour of the neoplasms, the extent of disease and adequate clinical therapy

  11. Silk microfiber-reinforced silk hydrogel composites for functional cartilage tissue repair

    Yodmuang, Supansa; McNamara, Stephanie L.; Nover, Adam B.; Mandal, Biman B.; Agarwal, Monica; Kelly, Terri-Ann N.; Chao, Pen-Hsiu Grace; Hung, Clark; Kaplan, David L.; Vunjak-Novakovic, Gordana

    2014-01-01

    Cartilage tissue lacks an intrinsic capacity for self-regeneration due to slow matrix turnover, a limited supply of mature chondrocytes and insufficient vasculature. Although cartilage tissue engineering has achieved some success using agarose as a scaffolding material, major challenges of agarose-based cartilage repair, including non-degradability, poor tissue–scaffold integration and limited processing capability, have prompted the search for an alternative biomaterial. In this study, silk ...

  12. Thermodynamics Study of Lac Dyeing of Silk Yarn Coated with Chitosan

    Montra CHAIRAT

    2009-01-01

    Full Text Available A thermodynamic study of lac dyeing of silk pretreated with chitosan at pH 3.0 was investigated in a batch system. It was found that the adsorption of lac dye on silk pretreated with chitosan could be described by the Langmuir isotherm. The results showed that the pretreatment of silk with chitosan provided an enhancement of dye uptake on silk and also decreased the dye desorption from silk yarn compared with the results in the absence of chitosan. The negative value of the enthalpy change (DHº for the adsorption of lac dye on silk pretreated with chitosan indicated that the adsorption process was an exothermic one.

  13. The Silk Industry Of Assam Its Origin, Growth And Importance In The Economy Of Assam

    Rekha Kalita

    2013-01-01

    Silk has been recogniged as a textile fibre. Sericulture, on which the silk industry survives, is an agroindustry, the end product of which is silk. It provides jobs and generates income in the rural areas. Assam is the only place where muga silk is produced and is well known for silk textiles both mulberry,muga and endi textiles. The silk weavers possess the very highest skill in their craft and it is probable that under competent and energetic direction with the assistance of capital, the i...

  14. Review - Trade and Society along the Ancient Silk Road

    Bina Sengar

    2013-01-01

    Review of: Jacqueline H Fewkes. 2011. Trade and Contemporary Society Along the Silk Road: An Ethno-History of Ladakh. London: Routledge. xiii+196pp; list of plates, Ladakh in the twenty-first century is well known as a religious and touristic destination as well as an Indian border territory with Kargil and Siachin - significant military posts. In contrast, the commercial role of Ladakh in trade within the Himalayan region and along the Silk Route is less explored In this w...

  15. Bioengineered Chimeric Spider Silk-Uranium Binding Proteins

    Krishnaji, Sreevidhya Tarakkad; Kaplan, David L.

    2012-01-01

    Heavy metals constitute a source of environmental pollution. Here, novel functional hybrid biomaterials for specific interactions with heavy metals are designed by bioengineering consensus sequence repeats from spider silk of Nephila clavipes with repeats of a uranium peptide recognition motif from a mutated 33-residue of calmodulin protein from Paramecium tetraurelia. The self-assembly features of the silk to control nanoscale organic/inorganic material interfaces provides new biomaterials f...

  16. SilkDB: a knowledgebase for silkworm biology and genomics

    Wang, Jing; Xia, Qingyou; He, Ximiao; Dai, Mingtao; Ruan, Jue; Chen, Jie; Yu, Guo; Yuan, Haifeng; Hu, Yafeng; LI, RUIQIANG; Feng, Tao; Ye, Chen; Lu, Cheng; WANG, JUN; Li, Songgang

    2005-01-01

    The Silkworm Knowledgebase (SilkDB) is a web-based repository for the curation, integration and study of silkworm genetic and genomic data. With the recent accomplishment of a ?6X draft genome sequence of the domestic silkworm (Bombyx mori), SilkDB provides an integrated representation of the large-scale, genome-wide sequence assembly, cDNAs, clusters of expressed sequence tags (ESTs), transposable elements (TEs), mutants, single nucleotide polymorphisms (SNPs) and functional annotations of g...

  17. Ptychographic X-ray Tomography of Silk Fiber Hydration

    Esmaeili, Morteza; Fløystad, Jostein B.; Diaz, Ana; Høydalsvik, Kristin; Guizar-Sicairos, Manuel; Andreasen, Jens Wenzel; Breiby, Dag W.

    2013-01-01

    conditions, yield quantitative information about the spatial density variations in the form of detailed maps of the size, shape, and orientation distributions of the nanopores inside the silk fiber, revealing that the fiber swells anisotropically in humid conditions, with the expansion taking place solely...... normal to the fiber axis. Exploiting quantitative information on the fiber’s electron density, hydration was found to proceed through interaction with the silk protein rather than filling of pores....

  18. Canine mammary gland tumors.

    Sorenmo, Karin

    2003-05-01

    The National Consensus Group recommends that all women with tumors larger than 1 cm be offered chemotherapy regardless of tumor histology of lymph node status. This recommendation is to ensure that everyone at risk for failing, even though the risk may be low in women with relatively small tumors and favorable histology, has a choice and receives the benefit of adjuvant chemotherapy. This type of treatment recommendation may also be made in dogs based on recognized, well-accepted prognostic factors such as tumor size, stage, type, and histologic differentiation. Based on the limited clinical information available in veterinary medicine, the drugs that are effective in human breast cancer, such as cyclophosphamide, 5-fluorouracil, and doxorubicin, may also have a role in the treatment of malignant mammary gland tumors in dogs. Randomized prospective studies are needed, however, to evaluate the efficacy of chemotherapy in dogs with high-risk mammary gland tumors and to determine which drugs and protocols are the most efficacious. Until such studies are performed, the treatment of canine mammary gland tumors will be based on the individual oncologist's understanding of tumor biology, experience, interpretation of the available studies, and a little bit of gut-feeling. Table 2 is a proposal for treatment guidelines for malignant canine mammary gland tumors according to established prognostic factors, results from published veterinary studies, and current recommendations for breast cancer treatment in women. PMID:12852237

  19. Submandibular gland excision

    Springborg, Line Kanstrup; Møller, Martin Nue

    2013-01-01

    cosmetic result and all scars were ≤6 on the Vancouver Scar Scale. Problems with scarring were more common if there had beenpostoperative infection. We continue to use the lateral transcervical approach as standard in our institution for patients who cannot be managed by gland-sparing procedures....

  20. Formation of different gold nanostructures by silk nanofibrils.

    Fang, Guangqiang; Yang, Yuhong; Yao, Jinrong; Shao, Zhengzhong; Chen, Xin

    2016-07-01

    Metal nanostructures that have unique size- and shape-dependent electronic, optical and chemical properties gain more and more attention in modern science and technology. In this article, we show the possibility that we are able to obtain different gold nanostructures simply with the help of silk nanofibrils. We demonstrate that only by varying the pH of the reaction solution, we get gold nanoparticles, nano-icosahedrons, nanocubes, and even microplates. Particularly, we develop a practical method for the preparation of gold microplates in acid condition in the presence of silk nanofibrils, which is impossible by using other forms of silk protein. We attribute the role of silk nanofibrils in the formation of gold nanostructure to their reduction ability from several specific amino acid residues, and the suitable structural anisotropic features to sustain the crystal growth after the reduction process. Although the main purpose of this article is to demonstrate that silk nanofibrils are able to mediate the formation of different gold nanostructure, we show the potential applications of these resulting gold nanostructures, such as surface-enhanced Raman scattering (SERS) and photothermal transformation effect, as same as those produced by other methods. In conclusion, we present in this communication a facile and green synthesis route to prepare various gold nanostructures with silk nanofibrils by simply varying pH in the reaction system, which has remarkable advantages in future biomedical applications. PMID:27127067

  1. Preparation, structure and properties of multi-functional silk via ATRP method

    In order to develop multi-functional silk materials, silk was grafted using the first flame retardant monomer, dimethyl methacryloyloxyethyl phosphate (DMMEP), and the second antibacterial monomer, dimethylaminoethyl methacrylate (DMAEMA), via atom transfer radical polymerization (ATRP) method. The result of attenuated total reflection-Fourier transformed infrared spectroscopy (ATR-FTIR) indicated that two monomers were successfully grafted onto silk surface. X-ray diffraction (XRD) curves showed that grafting mainly occurred at the amorphous region of silk fibers. Thermal gravimetric analysis (TGA) and flame retardance measurements indicated that the Silk-grafted-poly(DMMEP) (Silk-g-PDMMEP) and Silk-grafted-poly(DMMEP)-blocked-poly(DMAEMA) (Silk-g-PDMMEP-b-PDMAEMA) both had good flame retardance. The LOI value could reach 31.2% and the char length was less than 6 cm for the Silk-g-PDMMEP sample with 22.21% of grafting percentage, and the LOI value was still over 28% after 50 times of washing. And the second grafts of DMAEMA monomer had no distinct effect on the flame retardance of Silk-g-PDMMEP. The inhibition rates of quaternized grafted silk to Staphylococcus aureu and Escherichia coli were both over 88%, and had excellent washing fastness. Multi-functional silk fabric with good flame retardance and anti-bacterial property could be obtained by properly controlling the grafting percentage, which had little effect on the intrinsic properties of silk fabric.

  2. Electricity from the silk cocoon membrane.

    Tulachan, Brindan; Meena, Sunil Kumar; Rai, Ratan Kumar; Mallick, Chandrakant; Kusurkar, Tejas Sanjeev; Teotia, Arun Kumar; Sethy, Niroj Kumar; Bhargava, Kalpana; Bhattacharya, Shantanu; Kumar, Ashok; Sharma, Raj Kishore; Sinha, Neeraj; Singh, Sushil Kumar; Das, Mainak

    2014-01-01

    Silk cocoon membrane (SCM) is an insect engineered structure. We studied the electrical properties of mulberry (Bombyx mori) and non-mulberry (Tussar, Antheraea mylitta) SCM. When dry, SCM behaves like an insulator. On absorbing moisture, it generates electrical current, which is modulated by temperature. The current flowing across the SCM is possibly ionic and protonic in nature. We exploited the electrical properties of SCM to develop simple energy harvesting devices, which could operate low power electronic systems. Based on our findings, we propose that the temperature and humidity dependent electrical properties of the SCM could find applications in battery technology, bio-sensor, humidity sensor, steam engines and waste heat management. PMID:24961354

  3. Silk Patterns: Conservation and Development of Traditional Thai silk Production for Added Commercial Value in Khon Kaen Province

    Kasinee Sawasdee; Boonsom Yodmalee; Kosit Phaengsoi

    2014-01-01

    Silk patterns are examples of fine art that show the valuable culture and identity of Thai communities. This is a qualitative research and the researchers used a purposive sampling technique to identify four districts in Khon Kaen province for assessment by means of survey, observation, interview, focus group discussion and workshop. The history and development of silk patterns in Khon Kaen province occurred from a process of pattern making called mudmee. The techniques were passed from gener...

  4. In vitro generation of a multilayered osteochondral construct with an osteochondral interface using rabbit bone marrow stromal cells and a silk peptide-based scaffold.

    Chen, Kelei; Shi, Pujiang; Teh, Thomas Kok Hiong; Toh, Siew Lok; Goh, James Ch

    2016-04-01

    Tissue engineering of a biological osteochondral multilayered construct with a cartilage-interface subchondral bone layer is a key challenge. This study presented a rabbit bone marrow stromal cell (BMSC)/silk fibroin scaffold-based co-culture approach to generate tissue-engineered osteochondral grafts with an interface. BMSC-seeded scaffolds were first cultured separately in osteogenic and chondrogenic stimulation media. The two differentiated pieces were then combined using an RADA self-assembling peptide and subsequently co-cultured. Gene expression, histological and biochemical analyses were used to evaluate the multilayered structure of the osteochondral graft. A complete osteochondral construct with a cartilage-subchondral bone interface was regenerated and BMSCs were used as the only cell source for the osteochondral construct and interface regeneration. Furthermore, in the intermediate region of co-cultured samples, hypertrophic chondrogenic gene markers type X collagen and MMP-13 were found on both chondrogenic and osteogenic section edges after co-culture. However, significant differences gene expression profile were found in distinct zones of the construct during co-culture and the section in the intermediate region had significantly higher hypertrophic chondrocyte gene expression. Biochemical analyses and histology results further supported this observation. This study showed that specific stimulation from osteogenic and chondrogenic BMSCs affected each other in this co-culture system and induced the formation of an osteochondral interface. Moreover, this system provided a possible approach for generating multilayered osteochondral constructs. Copyright © 2013 John Wiley & Sons, Ltd. PMID:23413023

  5. Genetic Mystery of Behcet's Disease Unfolds Along the Ancient Silk Road

    ... mystery of Behcet’s disease unfolds along the ancient Silk Road NIH researchers identify new gene variants associated with ... found predominantly in people with ancestors along the Silk Road. For nearly 2,000 years, traders used this ...

  6. Coiling and maturation of a high-performance fibre in hagfish slime gland thread cells.

    Winegard, Timothy; Herr, Julia; Mena, Carlos; Lee, Betty; Dinov, Ivo; Bird, Deborah; Bernards, Mark; Hobel, Sam; Van Valkenburgh, Blaire; Toga, Arthur; Fudge, Douglas

    2014-01-01

    The defensive slime of hagfishes contains thousands of intermediate filament protein threads that are manufactured within specialized gland thread cells. The material properties of these threads rival those of spider dragline silks, which makes them an ideal model for biomimetic efforts to produce sustainable protein materials, yet how the thread is produced and organized within the cell is not well understood. Here we show how changes in nuclear morphology, size and position can explain the three-dimensional pattern of thread coiling in gland thread cells, and how the ultrastructure of the thread changes as very young thread cells develop into large cells with fully mature coiled threads. Our model provides an explanation for the complex process of thread assembly and organization that has fascinated and perplexed biologists for over a century, and provides valuable insights for the quest to manufacture high-performance biomimetic protein materials. PMID:24698953

  7. Synthetic spider silk production on a laboratory scale.

    Hsia, Yang; Gnesa, Eric; Pacheco, Ryan; Kohler, Kristin; Jeffery, Felicia; Vierra, Craig

    2012-01-01

    As society progresses and resources become scarcer, it is becoming increasingly important to cultivate new technologies that engineer next generation biomaterials with high performance properties. The development of these new structural materials must be rapid, cost-efficient and involve processing methodologies and products that are environmentally friendly and sustainable. Spiders spin a multitude of different fiber types with diverse mechanical properties, offering a rich source of next generation engineering materials for biomimicry that rival the best manmade and natural materials. Since the collection of large quantities of natural spider silk is impractical, synthetic silk production has the ability to provide scientists with access to an unlimited supply of threads. Therefore, if the spinning process can be streamlined and perfected, artificial spider fibers have the potential use for a broad range of applications ranging from body armor, surgical sutures, ropes and cables, tires, strings for musical instruments, and composites for aviation and aerospace technology. In order to advance the synthetic silk production process and to yield fibers that display low variance in their material properties from spin to spin, we developed a wet-spinning protocol that integrates expression of recombinant spider silk proteins in bacteria, purification and concentration of the proteins, followed by fiber extrusion and a mechanical post-spin treatment. This is the first visual representation that reveals a step-by-step process to spin and analyze artificial silk fibers on a laboratory scale. It also provides details to minimize the introduction of variability among fibers spun from the same spinning dope. Collectively, these methods will propel the process of artificial silk production, leading to higher quality fibers that surpass natural spider silks. PMID:22847722

  8. Pituitary gland tumors

    This article gives an overview of the most common tumors of the pituitary gland and the differential diagnostics with special emphasis on radiological diagnostic criteria. A selective search of the literature in PubMed was carried out. Pituitary adenomas constitute 10-15 % of all intracranial tumors and are the most common tumors of the sellar region. Tumors smaller than 1 cm in diameter are called microadenomas while those larger than 1 cm in diameter are called macroadenomas. Approximately 65 % of pituitary gland adenomas secrete hormones whereby approximately 50 % secrete prolactin, 10 % secrete growth hormone (somatotropin) and 6 % secrete corticotropin. Other tumors located in the sella turcica can also cause endocrinological symptoms, such as an oversecretion of pituitary hormone or pituitary insufficiency by impinging on the pituitary gland or its stalk. When tumors spread into the space cranial to the sella turcica, they can impinge on the optic chiasm and cause visual disorders. A common differential diagnosis of a sellar tumor is a craniopharyngeoma. In children up to 10 % of all intracranial tumors are craniopharyngeomas. Other differential diagnoses for sellar tumors are metastases, meningiomas, epidermoids and in rare cases astrocytomas, germinomas or Rathke cleft cysts As these tumors are located in an anatomically complex region of the skull base and are often very small, a highly focused imaging protocol is required. The currently favored modality is magnetic resonance imaging (MRI) with the administration of a contrast agent. The sellar region should be mapped in thin slices. In cases of suspected microadenoma the imaging protocol should also contain a sequence with dynamic contrast administration in order to assess the specific enhancement characteristics of the tumor and the pituitary gland. (orig.)

  9. Sebaceous gland lipids

    Picardo, Mauro; Ottaviani, Monica; Camera, Emanuela; Mastrofrancesco, Arianna

    2009-01-01

    The principal activity of mature sebaceous glands is producing and secreting sebum, which is a complex mixture of lipids. Sebum composition is different among species and this difference is probably due to the function that sebum has to absolve. In human sebum there are unique lipids, such as squalene and wax esters not found anywhere else in the body nor among the epidermal surface lipids. Moreover, they correspond to major components supplying the skin with protection. However, the ultimate...

  10. A response to Dolliver's "Evaluating drug trafficking on the Tor Network: Silk Road 2, the sequel"

    Judith Aldridge, David Decary-Hetu

    2015-01-01

    The leading aim of Dolliver's (2015) paper Evaluating drug trafficking on the Tor Network: Silk Road 2, the sequel is to document changes in the size and structure of cryptomarkets following the demise of SILK ROAD 1 (SR1) using data she collected from SILK ROAD 2 (SR2), which she casts as successor to its namesake.

  11. [Engineered spider silk: the intelligent biomaterial of the future. Part I].

    Florczak, Anna; Piekoś, Konrad; Kaźmierska, Katarzyna; Mackiewicz, Andrzej; Dams-Kozłowska, Hanna

    2011-01-01

    The unique properties of spider silk such as strength, extensibility, toughness, biocompatibility and biodegradability are the reasons for the recent development in silk biomaterial technology. For a long time scientific progress was impeded by limited access to spider silk. However, the development of the molecular biology strategy was a breaking point in synthetic spider silk protein design. The sequences of engineered spider silk are based on the consensus motives of the corresponding natural equivalents. Moreover, the engineered silk proteins may be modified in order to gain a new function. The strategy of the hybrid proteins constructed on the DNA level combines the sequence of engineered silk, which is responsible for the biomaterial structure, with the sequence of polypeptide which allows functionalization of the silk biomaterial. The functional domains may comprise receptor binding sites, enzymes, metal or sugar binding sites and others. Currently, advanced research is being conducted, which on the one hand focuses on establishing the particular silk structure and understanding the process of silk thread formation in nature. On the other hand, there are attempts to improve methods of engineered spider silk protein production. Due to acquired knowledge and recent progress in synthetic protein technology, the engineered silk will turn into intelligent biomaterial of the future, while its industrial production scale will trigger a biotechnological revolution. PMID:21734322

  12. The Speed of Sound in Silk: Linking Material Performance to Biological Function

    Mortimer, Beth; Shira D Gordon; Holland, Chris; Siviour, Clive R; Vollrath, Fritz; Windmill, James F. C.

    2014-01-01

    Sonic properties of spider silks are measured independent of the web using laser vibrometry and ballistic impact providing insights into Nature's design of functionalized high-performance materials. Through comparison to cocoon silk and other industrial fibers, we find that major ampullate silk has the largest wavespeed range of any known material.

  13. UTILIZATION OF THE TOPICAL APPLICATION OF LIMONENE TO THE FIFTH INSTAR LARVAE OF THE SILKWORM, BOMBYX MORI (L (RACE: PM X CSR2 FOR THE PARAMETERS OF LARVAE, COCOON AND SILK FILAMENT

    Vitthalrao B Khyade*, Vivekanand V Khyade and Randy Wayne Schekman

    2015-01-01

    Full Text Available The attempt is concerned with topical application of the limonene on the fifth instar larvae of multivoltine silkworm, Bombyx mori (L. The ten microliters of one milligram per liter strength acetone solution of limonene was topically applied to the individual fifth instar larvae of multi voltine cross breed race of silkworm, Bombyx mori (L. (PM x CSR2 variously at 48, 54, 60 and 66 hours after the fourth moult. Topical application of acetone solution of limonene was found variously reflected into prolongation of larval age (23.129-38.775%; improvement in the tissue somatic index (TSI of silk glands (3.041-7.761; cocoon shell ratio (2.828-4.497; Tex of silk filament (0.038-0.107 and denier scale of silk filament (0.343- 0.962.  Topical application of acetone solution of limonene, thus chiefly lengthen the larval age in silkworm, Bombyx mori (L. Time required for eating and amount of mulberry leaves eaten had both been increased and they were practically reflected into the improvement in the quality of cocoon and the silk filament. The terpenoid nature of limonene and delay in the spinning among the larvae, recipient of this limonene in the attempt indicate the possibility of it’s insect juvenoid activity in silkworm, Bombyx mori (L.. Efficient utilization of limonene, the terpene compound through acetone solvent for topical application to the fifth instared larvae of silkworm, Bombyx mori (L may open a biotechnological avenue in the sericulture industries.

  14. Extracted sericin from silk waste for film formation

    Rungsinee Sothornvit

    2010-03-01

    Full Text Available Sericin is the second main component in cocoons, which are removed in the silk reeling process of the raw silk industry and in the silk waste degumming of the spun silk industry. The main amino acid of sericin, serine, exhibits a skin moisturing and antiwrinkle action, which is interesting to use for film formation in this study. The extraction conditions of sericin from two silk wastes, pieced cocoon and inferior knubbs were studied to find the optimum extraction conditions. Boiling water extraction was considered based on the response surface methodology (RSM in order to identify the important factors for the sericin extraction. The two factors considered were time and temperature. Both factors were needed to be independent parameters in the predicted equation in order to improve the model fit with R2 = 0.84. The components ofextracted sericin were 18.24% serine, 9.83% aspatate, and 5.51% glycine with a molecular weight of 132 kDa. Film formationfrom extracted sericin was carried out to find the optimum conditions. Extracted sericin could not form a stand-alonefilm. Therefore, polysaccharide polymers, such as glucomannan, were incorporated with glycerol to form a flexible film.Sericin-based films were characterized for its properties in terms of solubility and permeability before application. It wasfound that sericin-based films showed a film flexibility and solubility without an increasing film water vapor permeability.

  15. Airway Gland Structure and Function.

    Widdicombe, Jonathan H; Wine, Jeffrey J

    2015-10-01

    Submucosal glands contribute to airway surface liquid (ASL), a film that protects all airway surfaces. Glandular mucus comprises electrolytes, water, the gel-forming mucin MUC5B, and hundreds of different proteins with diverse protective functions. Gland volume per unit area of mucosal surface correlates positively with impaction rate of inhaled particles. In human main bronchi, the volume of the glands is ∼ 50 times that of surface goblet cells, but the glands diminish in size and frequency distally. ASL and its trapped particles are removed from the airways by mucociliary transport. Airway glands have a tubuloacinar structure, with a single terminal duct, a nonciliated collecting duct, then branching secretory tubules lined with mucous cells and ending in serous acini. They allow for a massive increase in numbers of mucus-producing cells without replacing surface ciliated cells. Active secretion of Cl(-) and HCO3 (-) by serous cells produces most of the fluid of gland secretions. Glands are densely innervated by tonically active, mutually excitatory airway intrinsic neurons. Most gland mucus is secreted constitutively in vivo, with large, transient increases produced by emergency reflex drive from the vagus. Elevations of [cAMP]i and [Ca(2+)]i coordinate electrolyte and macromolecular secretion and probably occur together for baseline activity in vivo, with cholinergic elevation of [Ca(2+)]i being mainly responsive for transient increases in secretion. Altered submucosal gland function contributes to the pathology of all obstructive diseases, but is an early stage of pathogenesis only in cystic fibrosis. PMID:26336032

  16. Silk moths in Madagascar: Biology, uses and challenges related to Borocera cajani (Boisduval, 1833) (Lepidoptera – Lasiocampidae)

    Razafimanantsoa, Tsiresy; Rajoelison, Gabrielle; Ramamonjisoa, Bruno; Raminosoa, Noromalala; Poncelet, Marc; Bogaert, Jan; Haubruge, Eric; Verheggen, François

    2012-01-01

    Borocera cajani (Lepidoptera, Lasiocampidae), also named Landibe, is main wild silk moth currently used to produce silk textiles in Madagascar. Silk production involve many member of the local population, from the wild silk harvesters, to the spinners, traders, dyers, weavers and the artisans who transform the silk into clothing, accessories and decorative items. Uapaca bojeri (Tapia) forests are the last remnants of highland primary forest, which are threatened by human destruction through b...

  17. Environmentally friendly surface modification of silk fiber: Chitosan grafting and dyeing

    In this paper, the surface modification of silk fiber using anhydrides to graft the polysaccharide chitosan and dyeing ability of the grafted silk were studied. Silk fiber was degummed and acylated with two anhydrides, succinic anhydride (SA) and phthalic anhydride (PA), in different solvents (dimethyl sulfoxide (DMSO) and N,N-dimethyl formamide (DMF)). The effects of anhydrides, solvents, anhydride concentration, liquor ratio (L:R) and reaction time on acylation of silk were studied. The polysaccharide chitosan was grafted to the acylated silk fiber and dyed by acid dye (Acid Black NB.B). The effects of pH, chitosan concentration, and reaction time on chitosan grafting of acylated silk were investigated. The physical properties show sensible changes regardless of weight gain. Scanning electron microscopy (SEM) analysis showed the presence of foreign materials firmly attached to the surface of silk. FTIR spectroscopy provided evidence that chitosan was grafted onto the acylated silk through the formation of new covalent bonds. The dyeing of the chitosan grafted-acylated silk fiber indicated the higher dye ability in comparison to the acylated and degummed silk samples. The mechanism of chitosan grafting over degummed silk through anhydride linkage was proposed. The findings of this research support the potential production of new environmentally friendly textile fibers. It is worthwhile to mention that the grafted samples have antibacterial potential due to the antibacterial property of chitosan molecules.

  18. Environmentally friendly surface modification of silk fiber: Chitosan grafting and dyeing

    Davarpanah, Saideh [Textile Engineering Department, Amirkabir University of Technology, Hafez St., Tehran (Iran, Islamic Republic of); Mahmoodi, Niyaz Mohammad [Textile Engineering Department, Amirkabir University of Technology, Hafez St., Tehran (Iran, Islamic Republic of); Department of Environmental Research, Institute for Colorants, Paint and Coatings, Tehran (Iran, Islamic Republic of)], E-mail: mahmoodi@icrc.ac.ir; Arami, Mokhtar [Textile Engineering Department, Amirkabir University of Technology, Hafez St., Tehran (Iran, Islamic Republic of)], E-mail: arami@aut.ac.ir; Bahrami, Hajir; Mazaheri, Firoozmehr [Textile Engineering Department, Amirkabir University of Technology, Hafez St., Tehran (Iran, Islamic Republic of)

    2009-01-15

    In this paper, the surface modification of silk fiber using anhydrides to graft the polysaccharide chitosan and dyeing ability of the grafted silk were studied. Silk fiber was degummed and acylated with two anhydrides, succinic anhydride (SA) and phthalic anhydride (PA), in different solvents (dimethyl sulfoxide (DMSO) and N,N-dimethyl formamide (DMF)). The effects of anhydrides, solvents, anhydride concentration, liquor ratio (L:R) and reaction time on acylation of silk were studied. The polysaccharide chitosan was grafted to the acylated silk fiber and dyed by acid dye (Acid Black NB.B). The effects of pH, chitosan concentration, and reaction time on chitosan grafting of acylated silk were investigated. The physical properties show sensible changes regardless of weight gain. Scanning electron microscopy (SEM) analysis showed the presence of foreign materials firmly attached to the surface of silk. FTIR spectroscopy provided evidence that chitosan was grafted onto the acylated silk through the formation of new covalent bonds. The dyeing of the chitosan grafted-acylated silk fiber indicated the higher dye ability in comparison to the acylated and degummed silk samples. The mechanism of chitosan grafting over degummed silk through anhydride linkage was proposed. The findings of this research support the potential production of new environmentally friendly textile fibers. It is worthwhile to mention that the grafted samples have antibacterial potential due to the antibacterial property of chitosan molecules.

  19. Environmentally friendly surface modification of silk fiber: Chitosan grafting and dyeing

    Davarpanah, Saideh; Mahmoodi, Niyaz Mohammad; Arami, Mokhtar; Bahrami, Hajir; Mazaheri, Firoozmehr

    2009-01-01

    In this paper, the surface modification of silk fiber using anhydrides to graft the polysaccharide chitosan and dyeing ability of the grafted silk were studied. Silk fiber was degummed and acylated with two anhydrides, succinic anhydride (SA) and phthalic anhydride (PA), in different solvents (dimethyl sulfoxide (DMSO) and N, N-dimethyl formamide (DMF)). The effects of anhydrides, solvents, anhydride concentration, liquor ratio (L:R) and reaction time on acylation of silk were studied. The polysaccharide chitosan was grafted to the acylated silk fiber and dyed by acid dye (Acid Black NB.B). The effects of pH, chitosan concentration, and reaction time on chitosan grafting of acylated silk were investigated. The physical properties show sensible changes regardless of weight gain. Scanning electron microscopy (SEM) analysis showed the presence of foreign materials firmly attached to the surface of silk. FTIR spectroscopy provided evidence that chitosan was grafted onto the acylated silk through the formation of new covalent bonds. The dyeing of the chitosan grafted-acylated silk fiber indicated the higher dye ability in comparison to the acylated and degummed silk samples. The mechanism of chitosan grafting over degummed silk through anhydride linkage was proposed. The findings of this research support the potential production of new environmentally friendly textile fibers. It is worthwhile to mention that the grafted samples have antibacterial potential due to the antibacterial property of chitosan molecules.

  20. Design and Optimization of Resorbable Silk Internal Fixation Devices

    Haas, Dylan S.

    Limitations of current material options for internal fracture fixation devices have resulted in a large gap between user needs and hardware function. Metal systems offer robust mechanical strength and ease of implantation but require secondary surgery for removal and/or result in long-term complications (infection, palpability, sensitivity, etc.). Current resorbable devices eliminate the need for second surgery and long-term complications but are still associated with negative host response as well as limited functionality and more difficult implantation. There is a definitive need for orthopedic hardware that is mechanically capable of immediate fracture stabilization and fracture fixation during healing, can safely biodegrade while allowing complete bone remodeling, can be resterilized for reuse, and is easily implantable (self-tapping). Previous work investigated the use of silk protein to produce resorbable orthopedic hardware for non- load bearing fracture fixation. In this study, silk orthopedic hardware was further investigated and optimized in order to better understand the ability of silk as a fracture fixation system and more closely meet the unfulfilled market needs. Solvent-based and aqueous-based silk processing formulations were cross-linked with methanol to induce beta sheet structure, dried, autoclaved and then machined to the desired device/geometry. Silk hardware was evaluated for dry, hydrated and fatigued (cyclic) mechanical properties, in vitro degradation, resterilization, functionalization with osteoinductive molecules and implantation technique for fracture fixation. Mechanical strength showed minor improvements from previous results, but remains comparable to current resorbable fixation systems with the advantages of self-tapping ability for ease of implantation, full degradation in 10 months, ability to be resterilized and reused, and ability to release molecules for osteoinudction. In vivo assessment confirmed biocompatibility, showed improved bone deposition and remodeling with functionalization and showed promising feasibility of fracture fixations with minor adjustments to geometry. The proposed silk orthopedic hardware exhibits high potential as a resorbable fixation system that can bridge the gap between the current materials for internal fixation devices.

  1. Sequential origin in the high performance properties of orb spider dragline silk

    Blackledge, Todd A.; Prez-Rigueiro, Jos; Plaza, Gustavo R.; Perea, Beln; Navarro, Andrs; Guinea, Gustavo V.; Elices, Manuel

    2012-01-01

    Major ampullate (MA) dragline silk supports spider orb webs, combining strength and extensibility in the toughest biomaterial. MA silk evolved ~376?MYA and identifying how evolutionary changes in proteins influenced silk mechanics is crucial for biomimetics, but is hindered by high spinning plasticity. We use supercontraction to remove that variation and characterize MA silk across the spider phylogeny. We show that mechanical performance is conserved within, but divergent among, major lineages, evolving in correlation with discrete changes in proteins. Early MA silk tensile strength improved rapidly with the origin of GGX amino acid motifs and increased repetitiveness. Tensile strength then maximized in basal entelegyne spiders, ~230?MYA. Toughness subsequently improved through increased extensibility within orb spiders, coupled with the origin of a novel protein (MaSp2). Key changes in MA silk proteins therefore correlate with the sequential evolution high performance orb spider silk and could aid design of biomimetic fibers. PMID:23110251

  2. Sequential origin in the high performance properties of orb spider dragline silk

    Blackledge, Todd A.; Pérez-Rigueiro, José; Plaza, Gustavo R.; Perea, Belén; Navarro, Andrés; Guinea, Gustavo V.; Elices, Manuel

    2012-10-01

    Major ampullate (MA) dragline silk supports spider orb webs, combining strength and extensibility in the toughest biomaterial. MA silk evolved ~376 MYA and identifying how evolutionary changes in proteins influenced silk mechanics is crucial for biomimetics, but is hindered by high spinning plasticity. We use supercontraction to remove that variation and characterize MA silk across the spider phylogeny. We show that mechanical performance is conserved within, but divergent among, major lineages, evolving in correlation with discrete changes in proteins. Early MA silk tensile strength improved rapidly with the origin of GGX amino acid motifs and increased repetitiveness. Tensile strength then maximized in basal entelegyne spiders, ~230 MYA. Toughness subsequently improved through increased extensibility within orb spiders, coupled with the origin of a novel protein (MaSp2). Key changes in MA silk proteins therefore correlate with the sequential evolution high performance orb spider silk and could aid design of biomimetic fibers.

  3. Imaging of salivary gland tumours

    Salivary gland neoplasms account for <3% of all tumors. Most of them are benign and parotid gland is the commonest site. As a general rule, the smaller the involved salivary gland, the higher is the possibility of the tumor being malignant. The role of imaging in assessment of salivary gland tumour is to define intra-glandular vs. extra-glandular location, detect malignant features, assess local extension and invasion, detect nodal metastases and systemic involvement. Image guided fine needle aspiration cytology provides a safe means to obtain cytological confirmation. For lesions in the superficial parotid and submandibular gland, ultrasound is an ideal tool for initial assessment. These are superficial structures accessible by high resolution ultrasound and FNAC which provides excellent resolution and tissue characterization without a radiation hazard. Nodal involvement can also be assessed. If deep tissue extension is suspected or malignancy confirmed on cytology, an MRI or CT is mandatory to evaluate tumour extent, local invasion and perineural spread. For all tumours in the sublingual gland, MRI should be performed as the risk of malignancy is high. For lesions of the deep lobe of parotid gland and the minor salivary glands, MRI and CT are the modalities of choice. Ultrasound has limited visualization of the deep lobe of parotid gland which is obscured by the mandible. Minor salivary gland lesions in the mucosa of oral cavity, pharynx and tracheo-bronchial tree, are also not accessible by conventional ultrasound. Recent study suggests that MR spectroscopy may differentiate malignant and benign salivary gland tumours as well as distinguishing Warthin's tumor from pleomorphic adenoma. However, its role in clinical practice is not well established. Similarly, the role of nuclear medicine and PET scan, in imaging of parotid masses is limited. Sialography is used to delineate the salivary ductal system and has limited role in assessment of tumour extent

  4. Structure and Properties of Nephila Clavipes Dragline Silk Polymer

    Mahoney, David Vincent

    Silk, spun from an aqueous state at room temperature by a variety of organisms, is the most commonly spun extracellular fibrous protein. It comprises polypeptide chains with regions which can crystallize and regions which are predominantly amorphous. The polymer chains in the crystalline regions form anti-parallel pleated sheet structures with an orthorhombic unit cell. Dragline silk is a structural material produced by a variety of spiders. It has been genetically tailored to meet a specific purpose. Dragline silk exhibits high extensibility and tensile strength approaching that of high-strength synthetic fibers. The specific energy to break it can exceed some steels and synthetic fibers. Samples of Nephila clavipes (golden orb-weaver) dragline silk were extracted from live specimens and examined with a series of experimental techniques including optical, scanning electron, and atomic force microscopy, wide and small angle X-ray diffraction and birefringence compensation. Computer modeling of the mechanical properties of the crystallite was also performed. An assortment of features at a variety of length scales was observed by microscopy. These occur on both the as-spun and abraded silk surfaces. The silk was observed to undergo large deformations without evidence of failure, suggesting the absence of a microfibrillar structure. There was no conclusive evidence for either a microfibrillar or a skin core structure. Meridional and equatorial SAXD peaks were observed at Bragg spacings of 79 AA and 250 AA, respectively. Analysis of the WAXD patterns indicated that the silk belongs in Warwicker's category 3b and that the minimum dimensions of the crystals are approximately 38 AA in the molecular direction and 16 x 23 AA in the transverse directions. The crystal modulus was determined with WAXD to be 16.7 GPa, applying the assumption of uniform stress. This is lower than the 200 GPa modulus calculated with molecular modeling. These results and other factors indicate the inapplicability of the widely used assumption of uniform stress. The X-ray data show increased alignment of the crystals along the fiber axis with stress. The birefringence of the silk fibers increases monotonically to failure which indicates increasing molecular alignment. The change in birefringence appears to be governed by the amorphous contribution.

  5. Presence of phosphorus in Nephila clavipes dragline silk.

    Michal, C A; Simmons, A H; Chew, B G; Zax, D B; Jelinski, L W

    1996-01-01

    Solid-state 31P-NMR of Nephila clavipes dragline silk indicates the presence of phosphorus in at least two chemically distinct environments. Amino acid analyses of acid-hydrolyzed silk confirm the presence of phosphotyrosine as one of the phosphorus-containing components. The unusual chemical shift (18.9 ppm downfield from 85% H3PO4), proton chemical shift, and acid lability of a second component suggest that it is part of a strained five-membered cyclic phosphate that might be found on a bet...

  6. The Algorithm to Detect Color Gradation on Silk

    Suyoto

    2012-03-01

    Full Text Available The process of silk dyeing with natural dye extracts will produce a certain color. Using extracts of wood, leaf and their combinations will give some color gradations. This paper aims to create a new algorithm which can help one, whose intention is to formulate the combination of coloring process to achieve the desired color through combining coloring materials on silk fabric. This algorithm will be expected to be able to formulate the combination of colors with more than 75 percent of accuracy. The natural dyes used were Ceriops candolleana arn wood for the red, Cudraina javanensis wood for the yellow, and indigofera leaf for the blue base color.

  7. Biosynthesis and characterization of typical fibroin crystalline polypeptides of silkworm Bombyx mori

    We aimed to investigate the self-organization/self-assembly mechanisms of silkworm fibroin-based material. In the present study, for the first time, we designed and multimerized four DNA 'monomer' sequences from structurally simple fibroin crystalline peptides or analog, [GAGAGX] (X = A, S, Y and V) to encode polypeptides [GAGAGX]16 (eGA, eGS, eGY and eGV) using a 'head-to-tail' construction strategy. Multimers were cloned into pGEX-KG and fusion proteins GST-[GAGAGX]16 (KGA, KGS, KGY and KGV) were efficiently expressed in Escherichia coli. These fusion proteins were isolated and purified by GST affinity chromatography and confirmed by SDS-PAGE and Western blot analysis using antibody reactive to GST. The polypeptides were cleavaged from GST fusion proteins by digesting with thrombin enzyme. The composition of the four polypeptides was confirmed by composition analysis of amino acids, and their abilities to form ?-sheet structure were determined by ThT fluorescence spectral analysis. The content of ?-sheet among the four polypeptides followed the order: eGS > eGV > eGY > eGA.

  8. Biosynthesis and characterization of typical fibroin crystalline polypeptides of silkworm Bombyx mori

    Wang Jiannan, E-mail: wangjn@suda.edu.cn [College of Material Engineering, Soochow University, Suzhou 215021 (China); Yan Shuqin [College of Material Engineering, Soochow University, Suzhou 215021 (China); Lu Changde [Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031 (China); Bai Lun [College of Material Engineering, Soochow University, Suzhou 215021 (China)

    2009-05-05

    We aimed to investigate the self-organization/self-assembly mechanisms of silkworm fibroin-based material. In the present study, for the first time, we designed and multimerized four DNA 'monomer' sequences from structurally simple fibroin crystalline peptides or analog, [GAGAGX] (X = A, S, Y and V) to encode polypeptides [GAGAGX]{sub 16} (eGA, eGS, eGY and eGV) using a 'head-to-tail' construction strategy. Multimers were cloned into pGEX-KG and fusion proteins GST-[GAGAGX]{sub 16} (KGA, KGS, KGY and KGV) were efficiently expressed in Escherichia coli. These fusion proteins were isolated and purified by GST affinity chromatography and confirmed by SDS-PAGE and Western blot analysis using antibody reactive to GST. The polypeptides were cleavaged from GST fusion proteins by digesting with thrombin enzyme. The composition of the four polypeptides was confirmed by composition analysis of amino acids, and their abilities to form {beta}-sheet structure were determined by ThT fluorescence spectral analysis. The content of {beta}-sheet among the four polypeptides followed the order: eGS > eGV > eGY > eGA.

  9. Treatment Option Overview (Salivary Gland Cancer)

    ... and Oropharyngeal Cancer Screening Research Salivary Gland Cancer Treatment (PDQ®)–Patient Version General Information About Salivary Gland ... in diagnosing salivary gland cancer. Certain factors affect treatment options and prognosis (chance of recovery). The treatment ...

  10. Treatment Options by Stage (Salivary Gland Cancer)

    ... and Oropharyngeal Cancer Screening Research Salivary Gland Cancer Treatment (PDQ®)–Patient Version General Information About Salivary Gland ... in diagnosing salivary gland cancer. Certain factors affect treatment options and prognosis (chance of recovery). The treatment ...

  11. Folding behavior of four silks of giant honey bee reflects the evolutionary conservation of aculeate silk proteins.

    Maitip, Jakkrawut; Trueman, Holly E; Kaehler, Benjamin D; Huttley, Gavin A; Chantawannakul, Panuwan; Sutherland, Tara D

    2015-04-01

    Multiple gene duplication events in the precursor of the Aculeata (bees, ants, hornets) gave rise to four silk genes. Whilst these homologs encode proteins with similar amino acid composition and coiled coil structure, the retention of all four homologs implies they each are important. In this study we identified, produced and characterized the four silk proteins from Apis dorsata, the giant Asian honeybee. The proteins were readily purified, allowing us to investigate the folding behavior of solutions of individual proteins in comparison to mixtures of all four proteins at concentrations where they assemble into their native coiled coil structure. In contrast to solutions of any one protein type, solutions of a mixture of the four proteins formed coiled coils that were stable against dilution and detergent denaturation. The results are consistent with the formation of a heteromeric coiled coil protein complex. The mechanism of silk protein coiled coil formation and evolution is discussed in light of these results. PMID:25712559

  12. Effects of the blended fibroin/aloe gel film on wound healing in streptozotocin-induced diabetic rats

    Delayed healing remains a major clinical problem and here we have sought to develop an improved dressing film comprising 1.95% w/v fibroin and 0.05% w/v aloe gel extract. The tensile strength of dry film was 21.1 ± 0.5 MPa and broke at 1.1 ± 0.2% elongation; corresponding values for wet film were 18.3 ± 1.3 MPa and 1.9 ± 0.1%. The film maintained its shape upon water immersion and the swelling ratio of the dry film was 0.8 ± 0.1 while the water uptake was 43.7 ± 2.6%. After 28 days of incubation in phosphate buffered saline (1 M, pH 7.4, 37 °C), the weight of film was reduced by 6.7 ± 1.1% and the tensile strength and elongation at breaking point (dry state) were 15.4 ± 0.6 MPa and 1.5 ± 0.2%, respectively. Compared to aloe-free fibroin film (2.0% fibroin extract only), the blended film enhanced the attachment and proliferation of skin fibroblasts. The bFGF immunofluorescence of fibroblasts cultured on the blended film appeared greater than those cultured on tissue culture plate or on aloe-free fibroin film while α-smooth muscle actin was maintained. In streptozotocin-induced diabetic rats, the wounds dressed with the blended film were smaller (p <0.05) by day 7 after wounding, compared to untreated diabetic wounds. Histology of repaired diabetic wounds showed the fibroblast distribution and collagen fiber organization to be similar to wounds in normal rats, and this was matched by enhanced hydroxyproline content. Thus, such accelerated wound healing by the blended fibroin/aloe gel films may find application in treatment of diabetic non-healing skin ulcers. (paper)

  13. Stages of Salivary Gland Cancer

    ... Fast neutron radiation therapy to lymph nodes with cancer. Chemotherapy . A clinical trial of fast neutron radiation therapy to the tumor . A clinical ... Gland Cancer Treatment of stage IV salivary gland cancer may include the ... trial of chemotherapy with or without radiation therapy . Check the list ...

  14. Structure to function: Spider silk and human collagen

    Rabotyagova, Olena S.

    Nature has the ability to assemble a variety of simple molecules into complex functional structures with diverse properties. Collagens, silks and muscles fibers are some examples of fibrous proteins with self-assembling properties. One of the great challenges facing Science is to mimic these designs in Nature to find a way to construct molecules that are capable of organizing into functional supra-structures by self-assembly. In order to do so, a construction kit consisting of molecular building blocks along with a complete understanding on how to form functional materials is required. In this current research, the focus is on spider silk and collagen as fibrous protein-based biopolymers that can shed light on how to generate nanostructures through the complex process of self-assembly. Spider silk in fiber form offers a unique combination of high elasticity, toughness, and mechanical strength, along with biological compatibility and biodegrability. Spider silk is an example of a natural block copolymer, in which hydrophobic and hydrophilic blocks are linked together generating polymers that organize into functional materials with extraordinary properties. Since silks resemble synthetic block copolymer systems, we adopted the principles of block copolymer design from the synthetic polymer literature to build block copolymers based on spider silk sequences. Moreover, we consider spider silk to be an important model with which to study the relationships between structure and properties in our system. Thus, the first part of this work was dedicated to a novel family of spider silk block copolymers, where we generated a new family of functional spider silk-like block copolymers through recombinant DNA technology. To provide fundamental insight into relationships between peptide primary sequence, block composition, and block length and observed morphological and structural features, we used these bioengineered spider silk block copolymers to study secondary structure, morphological features and assembly. Aside from fundamental perspectives, we anticipate that these results will provide a blueprint for the design of precise materials for a range of potential applications such as controlled release devices, functional coatings, components of tissue regeneration materials and environmentally friendly polymers in future studies. In the second part of this work, human collagen type I was studied as another representative of the family of fibrous proteins. Collagen type I is the most abundant extracellular matrix protein in the human body, providing the basis for tissue structure and directing cellular functions. Collagen has a complex structural hierarchy, organized at different length scales, including the characteristic triple helical feature. In the present study we assessed the relationship between collagen structure (native vs. denatured) and sensitivity to UV radiation with a focus on changes in the primary structure, conformation, microstructure and material properties. Free radical reactions are involved in collagen degradation and a mechanism for UV-induced collagen degradation related to structure was proposed. The results from this study demonstrated the role of collagen supramolecular organization (triple helix) in the context of the effects of electromagnetic radiation on extracellular matrices. Owing to the fact that both silks and collagens are proteins that have found widespread interest for biomaterial related needs, we anticipate that the current studies will serve as a foundation for future biomaterial designs with controlled properties. Furthermore, fundamental insight into self-assembly and environmentally-2mediated degradation, will build a foundation for fundamental understanding of the remodeling and functions of these types of fibrous proteins in vivo and in vitro. This type of insight is essential for many areas of scientific inquiry, from drug delivery, to scaffolds for tissue engineering, and to the stability of materials in space.

  15. Influence of Hybridization on the Traits of Silk Production and Filament Denier in Indian Tropical Tasar Silk Insect, Antheraea mylitta Drury

    A.K. Srivastava

    2010-01-01

    Full Text Available The aim of the present study is to explore hybridization influence to achieve the potential of silk yield and quality in commercial tasarculture, as against the current status of applying only a few wild and semi-domesticated ecoraces of Antheraea mylitta D to produce raw silk during commercial crop rearing season (September-December. The four F1 hybrid combinations generated using Daba, Jata and Raily tasar ecoraces of seed crop rearing season (July-August were evaluated during commercial crop season of 2007 and 2008 for the influence of hybridization as positive heterosis in the traits of economically important silk productivity and filament denier. The F1 hybrid, Daba x Jata has recorded positive heterosis in single shell weight (+25.6%, total silk yield (+79.0% and silk filament length (+68.1%, but for silk filament of high denier (11.98 d followed by its reciprocal hybrid, JataxDaba in shell weight (+18.8%, silk yield (+68.1%, filament length (+63.8% and filament of highest denier (12.32 d. Though, the RailyxDaba hybrid could show highest heterosis in shell weight (2.47 g with filament of least denier (10.86 d, the improvement in filament length was marginal (+44.7% and heterosis in silk yield was negative (-47.0%, while the performance of DabaxRaily hybrid was least among the hybrids. The study infers hybridization influence on silk related traits at F1 level as positive heterosis and the application feasibility of DabaxJata and JataxDaba hybrids during commercial crop rearing season for higher silk productivity with silk filament denier at least better than one of the parents of F1 hybrid combination.

  16. Pituitary gland tumors; Hypophysentumoren

    Jesser, J.; Schlamp, K.; Bendszus, M. [Radiologische Klinik, Universitaetsklinikum Heidelberg, Abteilung fuer Neuroradiologie, Heidelberg (Germany)

    2014-10-15

    This article gives an overview of the most common tumors of the pituitary gland and the differential diagnostics with special emphasis on radiological diagnostic criteria. A selective search of the literature in PubMed was carried out. Pituitary adenomas constitute 10-15 % of all intracranial tumors and are the most common tumors of the sellar region. Tumors smaller than 1 cm in diameter are called microadenomas while those larger than 1 cm in diameter are called macroadenomas. Approximately 65 % of pituitary gland adenomas secrete hormones whereby approximately 50 % secrete prolactin, 10 % secrete growth hormone (somatotropin) and 6 % secrete corticotropin. Other tumors located in the sella turcica can also cause endocrinological symptoms, such as an oversecretion of pituitary hormone or pituitary insufficiency by impinging on the pituitary gland or its stalk. When tumors spread into the space cranial to the sella turcica, they can impinge on the optic chiasm and cause visual disorders. A common differential diagnosis of a sellar tumor is a craniopharyngeoma. In children up to 10 % of all intracranial tumors are craniopharyngeomas. Other differential diagnoses for sellar tumors are metastases, meningiomas, epidermoids and in rare cases astrocytomas, germinomas or Rathke cleft cysts As these tumors are located in an anatomically complex region of the skull base and are often very small, a highly focused imaging protocol is required. The currently favored modality is magnetic resonance imaging (MRI) with the administration of a contrast agent. The sellar region should be mapped in thin slices. In cases of suspected microadenoma the imaging protocol should also contain a sequence with dynamic contrast administration in order to assess the specific enhancement characteristics of the tumor and the pituitary gland. (orig.) [German] Diese Arbeit ist eine Uebersicht ueber die haeufigsten Hypophysentumoren und deren Differenzialdiagnosen mit Augenmerk auf die radiologischen Diagnosekriterien. Selektive Literaturrecherche in PubMed. Hypophysenadenome umfassen 10-15 % aller intrakranieller Tumoren und die haeufigsten Tumoren der Hypophysenregion. Sind sie kleiner als 1 cm im Durchmesser, werden sie als Mikroadenome bezeichnet, bei ueber 1 cm im Durchmesser nennt man sie Makroadenome. Hypophysenadenome sind in der Mehrzahl der Faelle, in etwa 65 %, hormonaktiv. Die meisten, das sind fast 50 % aller Hypophysenadenome, sezernieren Prolaktin, seltener mit 10 % Somatotropin und mit 6 % Kortikotropin. Auch andere Tumoren mit Lage in der Sella turcica koennen durch die Verdraengung des Hypophysengewebes oder des Hypophysenstiels eine endokrinologische Symptomatik bedingen. Dies kann eine hypophysaere Hormonueberproduktion oder eine Hypophyseninsuffizienz sein. Breiten sich Tumoren ueber das sellaere Niveau nach kranial aus, kann es durch den Druck auf das Chiasma opticum zu Sehstoerungen kommen. Eine wichtige Differenzialdiagnose eines sellaeren Tumors ist das Kraniopharyngeom. Bei Kindern macht es bis zu 10 % aller intrakranieller Tumoren aus. Weitere Differenzialdiagnosen fuer sellaere Tumoren sind Metastasen, Meningeome, Epidermoide und seltener Astrozytome, Germinome und Rathke-Taschen-Zysten. Die Lage der Tumoren in einer anatomisch komplexen Region an der Schaedelbasis und die haeufig nur sehr kleinen Tumoren erfordern eine gezielte Bildgebung. Die Methode der Wahl ist die MRT mit Kontrastmittel, welche die Hypophysenregion in feiner Schichtung abdeckt. Bei Verdacht auf ein Mikroadenom sollte auch eine dynamische Kontrastmittelanflutung durchgefuehrt werden, um die tumor- und hypophysenspezifischen Anreicherungseigenschaften beurteilen zu koennen. (orig.)

  17. SILK: Higher Level Rules with Defaults and Semantic Scalability

    Grosof, Benjamin N.

    We overview the technical approach and motivations of the SILK system for semantic rules and ontologies, that radically extends the knowledge representation (KR) power of currently commercially important business rule systems, including not only Prologs but also production rules and event-condition-action rules, database systems, and semantic web.

  18. SilkDB: a knowledgebase for silkworm biology and genomics

    Wang, Jing; Xia, Qingyou; He, Ximiao; Dai, Mingtao; Ruan, Jue; Chen, Jie; Yu, Guo; Yuan, Haifeng; Hu, Yafeng; Li, Ruiqiang; Feng, Tao; Ye, Chen; Lu, Cheng; Wang, Jun; Li, Songgang; Wong, Gane Ka-Shu; Yang, Huanming; Wang, Jian; Xiang, Zhonghuai; Zhou, Zeyang; Yu, Jun

    displayed by using B.mori genome sequence as a reference framework. Designed as a basic platform, SilkDB strives to provide a comprehensive knowledgebase about the silkworm and present the silkworm genome and related information in systematic and graphical ways for the convenience of in-depth comparative...

  19. Nutrient deprivation induces property variations in spider gluey silk.

    Blamires, Sean J; Sahni, Vasav; Dhinojwala, Ali; Blackledge, Todd A; Tso, I-Min

    2014-01-01

    Understanding the mechanisms facilitating property variability in biological adhesives may promote biomimetic innovations. Spider gluey silks such as the spiral threads in orb webs and the gumfoot threads in cobwebs, both of which comprise of an axial thread coated by glue, are biological adhesives that have variable physical and chemical properties. Studies show that the physical and chemical properties of orb web gluey threads change when spiders are deprived of food. It is, however, unknown whether gumfoot threads undergo similar property variations when under nutritional stress. Here we tested whether protein deprivation induces similar variations in spiral and gumfoot thread morphology and stickiness. We manipulated protein intake for the orb web spider Nephila clavipes and the cobweb spider Latrodectus hesperus and measured the diameter, glue droplet volume, number of droplets per mm, axial thread width, thread stickiness and adhesive energy of their gluey silks. We found that the gluey silks of both species were stickier when the spiders were deprived of protein than when the spiders were fed protein. In N. clavipes a concomitant increase in glue droplet volume was found. Load-extension curves showed that protein deprivation induced glue property variations independent of the axial thread extensions in both species. We predicted that changes in salt composition of the glues were primarily responsible for the changes in stickiness of the silks, although changes in axial thread properties might also contribute. We, additionally, showed that N. clavipes' glue changes color under protein deprivation, probably as a consequence of changes to its biochemical composition. PMID:24523902

  20. Visual responses of corn silk flies (Diptera: Ulidiidae)

    Corn silk flies are major pests impacting fresh market sweet corn production in Florida and Georgia. Control depends solely on well-times applications of insecticides to protect corn ear development. Surveillance depends on visual inspection of ears with no effective trapping methods currently ava...