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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. LIM-homeodomain transcription factor Awh is a key component activating all three fibroin genes, fibH, fibL and fhx, in the silk gland of the silkworm, Bombyx mori

    Kimoto, Mai; Tsubota, Takuya; Uchino, Keiro; Sezutsu, Hideki; Takiya, Shigeharu

    2015-01-01

    In the silkworm Bombyx mori, three fibroin genes, fibroin-heavy-chain (fibH), fibroin-light-chain (fibL) and fibrohexamerin (fhx), are coexpressed only in the posterior silk gland (PSG) cells, while the sericin genes encoding silk glue proteins are expressed in the middle silk gland (MSG) cells. Silk gland factor-2 (SGF-2) is a PSG-specific activator complex of fibH, composed of a LIM-homeodomain protein, Awh, and its cofactors, Ldb and Lcaf. We investigated whether SGF-2 can activate other f...

  3. LIM-homeodomain transcription factor Awh is a key component activating all three fibroin genes, fibH, fibL and fhx, in the silk gland of the silkworm, Bombyx mori.

    Kimoto, Mai; Tsubota, Takuya; Uchino, Keiro; Sezutsu, Hideki; Takiya, Shigeharu

    2015-01-01

    In the silkworm Bombyx mori, three fibroin genes, fibroin-heavy-chain (fibH), fibroin-light-chain (fibL) and fibrohexamerin (fhx), are coexpressed only in the posterior silk gland (PSG) cells, while the sericin genes encoding silk glue proteins are expressed in the middle silk gland (MSG) cells. Silk gland factor-2 (SGF-2) is a PSG-specific activator complex of fibH, composed of a LIM-homeodomain protein, Awh, and its cofactors, Ldb and Lcaf. We investigated whether SGF-2 can activate other fibroin genes using transgenic silkworms. The genes for Ldb and Lcaf were expressed ubiquitously in various tissues, while the gene for Awh was expressed strictly specific in PSG of the wild type silkworms. Misexpression of Awh in transgenic silkworms induced ectopic expression of fibL and fhx as well as fibH in MSG. Coincidently with the induction of fibL and fhx by Awh, binding of SGF-2 to the promoter of fibL and fhx was detected in vitro, and SGF-2 binds directly to the fhx core promoter. Ectopic expression of the fibroin genes was observed at high levels in the middle part of MSG. Moreover, fibL and fhx were induced in the anterior silk gland (ASG) of the transgenic silkworms, but fibH was not. These results indicate that Awh is a key activator of all three fibroin genes, and the activity is probably regulated in conjunction with additional factors. PMID:25449130

  4. 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

  5. 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.

  6. Lithium-free processing of silk fibroin.

    Zheng, Zhaozhu; Guo, Shaozhe; Liu, Yawen; Wu, Jianbing; Li, Gang; Liu, Meng; Wang, Xiaoqin; Kaplan, David

    2016-09-01

    Silk fibroin protein was purified from Bombyx mori silkworm cocoons using a novel dialysis strategy to avoid fibroin aggregation and pre-mature formation of β-sheets. The degummed silk fibers were dissolved in Ajisawa's reagent, a mixture of CaCl2-EtOH-H2O, that is less expensive than lithium bromide. The dissolved solutions were dialyzed against either water or urea solution with a stepwise decrease in concentration. When the steps of 4 M-2 M-1 M-0 M urea (referred to as silk-TS-4210) were adopted, the purified silk fibroin had smaller aggregates (lithium bromide (silk-Li-0) method. Polyvinyl alcohol-emulsified silk microspheres generated using the purified solution had a similar size distribution and morphology when compared to lithium bromide dissolved solutions, while glycerol-blended silk films showed different mechanical properties. The silk-Li-0 generated films with the highest breaking strength (5.7 MPa ± 0.3) while the silk-TS-4210 had the highest extension at break (215.1% ± 12.5). The films prepared from silk-TS-4210 were cytocompatible to support the adhesion and proliferation of human mesenchymal stem cells, with improvements compared to the other samples likely due to the porous morphology of these films. PMID:27298185

  7. Silver nanoparticle containing silk fibroin bionanotextiles

    Calamak, Semih; Aksoy, Eda Ayse [Hacettepe University, Department of Basic Pharmaceutical Sciences, Faculty of Pharmacy (Turkey); Erdogdu, Ceren; Sagıroglu, Meral [Hacettepe University, Department of Pharmaceutical Microbiology, Faculty of Pharmacy (Turkey); Ulubayram, Kezban, E-mail: ukezban@hacettepe.edu.tr [Hacettepe University, Department of Basic Pharmaceutical Sciences, Faculty of Pharmacy (Turkey)

    2015-02-15

    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.

  8. 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

  9. The effect of sterilization on silk fibroin biomaterial properties.

    Rnjak-Kovacina, Jelena; DesRochers, Teresa M; Burke, Kelly A; Kaplan, David L

    2015-06-01

    The effects of common sterilization techniques on the physical and biological properties of lyophilized silk fibroin sponges are described. Sterile silk fibroin sponges were cast using a pre-sterilized silk fibroin solution under aseptic conditions or post-sterilized via autoclaving, γ radiation, dry heat, exposure to ethylene oxide, or hydrogen peroxide gas plasma. Low average molecular weight and low concentration silk fibroin solutions could be sterilized via autoclaving or filtration without significant loses of protein. However, autoclaving reduced the molecular weight distribution of the silk fibroin protein solution, and silk fibroin sponges cast from autoclaved silk fibroin were significantly stiffer compared to sponges cast from unsterilized or filtered silk fibroin. When silk fibroin sponges were sterilized post-casting, autoclaving increased scaffold stiffness, while decreasing scaffold degradation rate in vitro. In contrast, γ irradiation accelerated scaffold degradation rate. Exposure to ethylene oxide significantly decreased cell proliferation rate on silk fibroin sponges, which was rescued by leaching ethylene oxide into PBS prior to cell seeding. PMID:25761231

  10. Silk fibroin microtubes for blood vessel engineering.

    Lovett, Michael; Cannizzaro, Christopher; Daheron, Laurence; Messmer, Brady; Vunjak-Novakovic, Gordana; Kaplan, David L

    2007-12-01

    Currently available synthetic grafts demonstrate moderate success at the macrovascular level, but fail at the microvascular scale (inner diameter). We report on the development of silk fibroin microtubes for blood vessel repair with several advantages over existing scaffold materials/designs. These microtubes were prepared by dipping straight lengths of stainless steel wire into aqueous silk fibroin, where the addition of poly(ethylene oxide) (PEO) enabled control of microtube porosity. The microtube properties were characterized in terms of pore size, burst strength, protein permeability, enzymatic degradation, and cell migration. Low porosity microtubes demonstrated superior mechanical properties in terms of higher burst pressures, but displayed poor protein permeability; whereas higher porosity tubes had lower burst strengths but increased permeability and enhanced protein transport. The microtubes also exhibited cellular barrier functions as low porosity tubes prevented outward migration of GFP-transduced HUVECs, while the high porosity microtubes allowed a few cells per tube to migrate outward during perfusion. When combined with the biocompatible and suturability features of silk fibroin, these results suggest that silk microtubes, either implanted directly or preseeded with cells, are an attractive biomaterial for microvascular grafts. PMID:17727944

  11. Ras1CA overexpression in the posterior silk gland improves silk yield

    Li Ma; Hanfu Xu; Jinqi Zhu; Sanyuan Ma; Yan Liu; Rong-Jing Jiang; Qingyou Xia; Sheng Li

    2011-01-01

    Sericulture has been greatly advanced by applying hybrid breeding techniques to the domesticated silkworm,Bombyx mori,but has reached a plateau during the last decades. For the first time,we report improved silk yield in a GAL4/UAS transgenic silkworm. Overexpression of the Ras1CA oncogene specifically in the posterior silk gland improved fibroin production and silk yield by 60%,while increasing food consumption by only 20%. Ras activation by Ras1CA overexpression in the posterior silk gland enhanced phosphorylation levels of Ras downstream effector proteins,up-regulated fibroin mRNA levels,increased total DNA content,and stimulated endoreplication. Moreover,Rasl activation increased cell and nuclei sizes,enriched subcellular organelles related to protein synthesis,and stimulated ribosome biogenesis for mRNA translation. We conclude that Rasl activation increases cell size and protein synthesis in the posterior silk gland,leading to silk yield improvement.

  12. 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.

  13. Gelation behavior of Antheraea pernyi silk fibroin

    2010-01-01

    The sol-gel transition behavior of Antherae pernyi silk fibroin(Ap-SF) has not been systematically investigated.In this work,the influence of environmental temperature,pH,the concentration of Ap-SF,K+ and Ca2+ on the gelation time,and the structural changes of Ap-SF in sol-gel transformation were studied.The results indicated that the gelation time of the Ap-SF aqueous solution decreased with the increase of the Ap-SF concentration and environmental temperature.The sol-gel transformation of Ap-SF was much more rapid than that of Bombyx mori silk fibroin under the same conditions.The Ap-SF was sensitive to changes in the concentration of Ca2+ and K+.Upon gelation,the random coil structure of the Ap-SF was significantly transformed into the β-sheet structure.

  14. Orange IV stabilizes silk fibroin microemulsions

    Ferreira, A. V.; Volkov, Vadim; Abreu, Ana S; Azóia, Nuno G.; Botelho, C. M.; Paulo, Artur Cavaco

    2015-01-01

    Silk fibroin (SF) is a natural biopolymer that has been extensively studied in various applications due to its impressive mechanical properties and biocompatibility. Recently, SF-based particles have been proposed as controlled drug delivery systems. A new and efficient method to prepare SF microemulsions (SF-MEs) was developed by oil-in-water emulsions using high-pressure homogenization to promote emulsification. During SF-ME production, the secondary structure of SF changed to a more stable...

  15. Production of silk sericin/silk fibroin blend nanofibers

    Zhang Xianhua; Tsukada Masuhiro; Morikawa Hideaki; Aojima Kazuki; Zhang Guangyu; Miura Mikihiko

    2011-01-01

    Abstract Silk sericin (SS)/silk fibroin (SF) blend nanofibers have been produced by electrospinning in a binary SS/SF trifluoroacetic acid (TFA) solution system, which was prepared by mixing 20 wt.% SS TFA solution and 10 wt.% SF TFA solution to give different compositions. The diameters of the SS/SF nanofibers ranged from 33 to 837 nm, and they showed a round cross section. The surface of the SS/SF nanofibers was smooth, and the fibers possessed a bead-free structure. The average diameters o...

  16. Silk fibroin nanostructured materials for biomedical applications

    Mitropoulos, Alexander N.

    Nanostructured biopolymers have proven to be promising to develop novel biomedical applications where forming structures at the nanoscale normally occurs by self-assembly. However, synthesizing these structures can also occur by inducing materials to transition into other forms by adding chemical cross-linkers, changing pH, or changing ionic composition. Understanding the generation of nanostructures in fluid environments, such as liquid organic solvents or supercritical fluids, has not been thoroughly examined, particularly those that are based on protein-based block-copolymers. Here, we examine the transformation of reconstituted silk fibroin, which has emerged as a promising biopolymer due to its biocompatibility, biodegradability, and ease of functionalization, into submicron spheres and gel networks which offer applications in tissue engineering and advanced sensors. Two types of gel networks, hydrogels and aerogels, have small pores and large surface areas that are defined by their structure. We design and analyze silk nanoparticle formation using a microfluidic device while offering an application for drug delivery. Additionally, we provide a model and characterize hydrogel formation from micelles to nanoparticles, while investigating cellular response to the hydrogel in an in vitro cell culture model. Lastly, we provide a second model of nanofiber formation during near-critical and supercritical drying and characterize the silk fibroin properties at different drying pressures which, when acting as a stabilizing matrix, shows to improve the activity of entrapped enzymes dried at different pressures. This work has created new nanostructured silk fibroin forms to benefit biomedical applications that could be applied to other fibrous proteins.

  17. Controlling silk fibroin particle features for drug delivery

    Lammel, Andreas; Hu, Xiao; Park, Sang-Hyug; Kaplan, David L.; Scheibel, Thomas

    2010-01-01

    Silk proteins are a promising material for drug delivery due to their aqueous processability, biocompatibility, and biodegradability. A simple aqueous preparation method for silk fibroin particles with controllable size, secondary structure and zeta potential is reported. The particles were produced by salting out a silk fibroin solution with potassium phosphate. The effect of ionic strength and pH of potassium phosphate solution on the yield and morphology of the particles was determined. Se...

  18. Silk Fibroin Encapsulated Powder Reservoirs for Sustained Release of Adenosine

    Pritchard, Eleanor M.; Szybala, Cory; Boison, Detlev; Kaplan, David L.

    2010-01-01

    Due to its unique properties, silk fibroin was studied as a biodegradable polymer vehicle for sustained, local delivery of the anticonvulsant adenosine from encapsulated reservoirs. Silk is a biologically derived protein polymer that is biocompatible, mechanically strong and degrades to non-toxic products in vivo. To achieve local, sustained, controlled adenosine release from fully degradable implants, solid adenosine powder reservoirs were coated with silk fibroin. Material properties of the...

  19. 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

  20. 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

  1. Silk Fibroin for Flexible Electronic Devices.

    Zhu, Bowen; Wang, Hong; Leow, Wan Ru; Cai, Yurong; Loh, Xian Jun; Han, Ming-Yong; Chen, Xiaodong

    2016-06-01

    Flexible electronic devices are necessary for applications involving unconventional interfaces, such as soft and curved biological systems, in which traditional silicon-based electronics would confront a mechanical mismatch. Biological polymers offer new opportunities for flexible electronic devices by virtue of their biocompatibility, environmental benignity, and sustainability, as well as low cost. As an intriguing and abundant biomaterial, silk offers exquisite mechanical, optical, and electrical properties that are advantageous toward the development of next-generation biocompatible electronic devices. The utilization of silk fibroin is emphasized as both passive and active components in flexible electronic devices. The employment of biocompatible and biosustainable silk materials revolutionizes state-of-the-art electronic devices and systems that currently rely on conventional semiconductor technologies. Advances in silk-based electronic devices would open new avenues for employing biomaterials in the design and integration of high-performance biointegrated electronics for future applications in consumer electronics, computing technologies, and biomedical diagnosis, as well as human-machine interfaces. PMID:26684370

  2. 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...

  3. Silk Fibroin as Edible Coating for Perishable Food Preservation

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

    2016-05-01

    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.

  4. 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

  5. Study on Electrospinning Silk Fibroin Solution

    LI Ni; QIN Xiao-hong; WANG Shan-yuan

    2007-01-01

    A new method of preparing silk fibroin (SF) solution used in the electerospinning was introduced in this paper. According to the method, SF was dissolved in the LiBr/CH2O2 solution directly at room temperature. The method was compared with the traditional method---SF was dissolved in CaCl2 ternary solution. The structure of SF films and the morphology of SF nanofibers were examined by attenuated total reflectance fourier transform intrared (ATR- FrlR) spectroscopy, Scanning electron microscope (SEM) and optical polarizing microscope. The result of this study shows that the new method is a faster, more convenient and high efficient way to get the SF solution and the characteristics of SF fibet made by the new method is much betty.

  6. 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.

  7. 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.

  8. 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

  9. Production of silk sericin/silk fibroin blend nanofibers

    Zhang Xianhua

    2011-01-01

    Full Text Available Abstract Silk sericin (SS/silk fibroin (SF blend nanofibers have been produced by electrospinning in a binary SS/SF trifluoroacetic acid (TFA solution system, which was prepared by mixing 20 wt.% SS TFA solution and 10 wt.% SF TFA solution to give different compositions. The diameters of the SS/SF nanofibers ranged from 33 to 837 nm, and they showed a round cross section. The surface of the SS/SF nanofibers was smooth, and the fibers possessed a bead-free structure. The average diameters of the SS/SF (75/25, 50/50, and 25/75 blend nanofibers were much thicker than that of SS and SF nanofibers. The SS/SF (100/0, 75/25, and 50/50 blend nanofibers were easily dissolved in water, while the SS/SF (25/75 and 0/100 blend nanofibers could not be completely dissolved in water. The SS/SF blend nanofibers could not be completely dissolved in methanol. The SS/SF blend nanofibers were characterized by Fourier transform infrared (FTIR spectroscopy, differential scanning calorimetry, and differential thermal analysis. FTIR showed that the SS/SF blend nanofibers possessed a random coil conformation and ß-sheet structure.

  10. Production of silk sericin/silk fibroin blend nanofibers

    Zhang, Xianhua; Tsukada, Masuhiro; Morikawa, Hideaki; Aojima, Kazuki; Zhang, Guangyu; Miura, Mikihiko

    2011-08-01

    Silk sericin (SS)/silk fibroin (SF) blend nanofibers have been produced by electrospinning in a binary SS/SF trifluoroacetic acid (TFA) solution system, which was prepared by mixing 20 wt.% SS TFA solution and 10 wt.% SF TFA solution to give different compositions. The diameters of the SS/SF nanofibers ranged from 33 to 837 nm, and they showed a round cross section. The surface of the SS/SF nanofibers was smooth, and the fibers possessed a bead-free structure. The average diameters of the SS/SF (75/25, 50/50, and 25/75) blend nanofibers were much thicker than that of SS and SF nanofibers. The SS/SF (100/0, 75/25, and 50/50) blend nanofibers were easily dissolved in water, while the SS/SF (25/75 and 0/100) blend nanofibers could not be completely dissolved in water. The SS/SF blend nanofibers could not be completely dissolved in methanol. The SS/SF blend nanofibers were characterized by Fourier transform infrared (FTIR) spectroscopy, differential scanning calorimetry, and differential thermal analysis. FTIR showed that the SS/SF blend nanofibers possessed a random coil conformation and ß-sheet structure.

  11. 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)

  12. 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)

  13. Surface Modification and Characterisation of Silk Fibroin Fabric Produced by the Layer-by-Layer Self-Assembly of Multilayer Alginate/Regenerated Silk Fibroin.

    Gaotian Shen

    Full Text Available Silk-based medical products have a long history of use as a material for surgical sutures because of their desirable mechanical properties. However, silk fibroin fabric has been reported to be haemolytic when in direct contact with blood. The layer-by-layer self-assembly technique provides a method for surface modification to improve the biocompatibility of silk fibroin fabrics. Regenerated silk fibroin and alginate, which have excellent biocompatibility and low immunogenicity, are outstanding candidates for polyelectrolyte deposition. In this study, silk fabric was degummed and positively charged to create a silk fibroin fabric that could undergo self-assembly. The multilayer self-assembly of the silk fibroin fabric was achieved by alternating the polyelectrolyte deposition of a negatively charged alginate solution (pH = 8 and a positively charged regenerated silk fibroin solution (pH = 2. Finally, the negatively charged regenerated silk fibroin solution (pH = 8 was used to assemble the outermost layer of the fabric so that the surface would be negatively charged. A stable structural transition was induced using 75% ethanol. The thickness and morphology were characterised using atomic force microscopy. The properties of the self-assembled silk fibroin fabric, such as the bursting strength, thermal stability and flushing stability, indicated that the fabric was stable. In addition, the cytocompatibility and haemocompatibility of the self-assembled silk fibroin fabrics were evaluated. The results indicated that the biocompatibility of the self-assembled multilayers was acceptable and that it improved markedly. In particular, after the self-assembly, the fabric was able to prevent platelet adhesion. Furthermore, other non-haemolytic biomaterials can be created through self-assembly of more than 1.5 bilayers, and we propose that self-assembled silk fibroin fabric may be an attractive candidate for anticoagulation applications and for promoting

  14. Correlation between fibroin amino acid sequence and physical silk properties.

    Fedic, Robert; Zurovec, Michal; Sehnal, Frantisek

    2003-09-12

    The fiber properties of lepidopteran silk depend on the amino acid repeats that interact during H-fibroin polymerization. The aim of our research was to relate repeat composition to insect biology and fiber strength. Representative regions of the H-fibroin genes were sequenced and analyzed in three pyralid species: wax moth (Galleria mellonella), European flour moth (Ephestia kuehniella), and Indian meal moth (Plodia interpunctella). The amino acid repeats are species-specific, evidently a diversification of an ancestral region of 43 residues, and include three types of regularly dispersed motifs: modifications of GSSAASAA sequence, stretches of tripeptides GXZ where X and Z represent bulky residues, and sequences similar to PVIVIEE. No concatenations of GX dipeptide or alanine, which are typical for Bombyx silkworms and Antheraea silk moths, respectively, were found. Despite different repeat structure, the silks of G. mellonella and E. kuehniella exhibit similar tensile strength as the Bombyx and Antheraea silks. We suggest that in these latter two species, variations in the repeat length obstruct repeat alignment, but sufficiently long stretches of iterated residues get superposed to interact. In the pyralid H-fibroins, interactions of the widely separated and diverse motifs depend on the precision of repeat matching; silk is strong in G. mellonella and E. kuehniella, with 2-3 types of long homogeneous repeats, and nearly 10 times weaker in P. interpunctella, with seven types of shorter erratic repeats. The high proportion of large amino acids in the H-fibroin of pyralids has probably evolved in connection with the spinning habit of caterpillars that live in protective silk tubes and spin continuously, enlarging the tubes on one end and partly devouring the other one. The silk serves as a depot of energetically rich and essential amino acids that may be scarce in the diet. PMID:12816957

  15. 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)

  16. 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

  17. 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.

  18. 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

  19. Nanolayer Biomaterial Coatings of Silk Fibroin for Controlled Release

    Wang, Xianyan; Hu, Xiao; Daley, Andrea; Rabotyagova, Olena; Cebe, Peggy; Kaplan, David L.

    2007-01-01

    An all-aqueous, stepwise deposition process with silk fibroin protein for the assembly of nanoscale layered controlled release coatings was exploited. Model compounds, Rhodamine B, Even Blue and Azoalbumin, representing small molecule drugs and therapeutically relevant proteins were incorporated in the nanocoating process and their loading and release behavior was quantified. In addition, the structure and morphology of the coatings were characterized. Release studies in vitro showed that con...

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

    Shetty, G. Rajesha [Department of Physics, Govt. First Grade College Hiriadka, Udupi - 576113 (India); Kumar, R. Madhu; Rao, B. Lakshmeesha; Asha, S.; Sangappa, E-mail: syhalabhavi@yahoo.co.in [Department of Studies in Physics, Mangalore University, Mangalagangotri, Mangalore - 574199 (India)

    2015-06-24

    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.

  1. 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

  2. Silk Fibroin as Edible Coating for Perishable Food Preservation

    B. Marelli; M. A. Brenckle; 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-proc...

  3. 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

  4. 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.

  5. 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.

  6. Interactions between fibroin and sericin proteins from Antheraea pernyi and Bombyx mori silk fibers.

    Du, Shan; Zhang, Jin; Zhou, Wei T; Li, Quan X; Greene, George W; Zhu, Hai J; Li, Jing L; Wang, Xun G

    2016-09-15

    Silkworm silk fibers are core-shell composites of fibroin and sericin proteins. Studying the interactions between fibroin and sericin is essential for understanding the properties of these composites. It is observed that compared to the domestic silk cocoon Bombyx mori (B. mori), the adhesion between fibroin and sericin from the wild silk cocoon, Antheraea pernyi (A. pernyi), is significantly stronger with a higher degree of heterogeneity. The adsorption of A. pernyi sericin on its fibroin is almost twice the value for B. mori sericin on fibroin, both showing a monolayer Langmuir adsorption. (1)H NMR and FTIR studies demonstrate on a molecular level the stronger interactions and the more intensive complex formation between A. pernyi fibroin and sericin, facilitated by the hydrogen bonding between glycine and serine. The findings of this study may help the design of composites with superior interfacial adhesion between different components. PMID:27314644

  7. 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.

  8. Silk fibroin/pullulan blend films: Preparation and characterization

    Shivananda, C. S.; Rao, B. Lakshmeesha; Madhukumar, R.; Sarojini, B. K.; Somashekhar, R.; Asha, S.; Sangappa, Y.

    2016-05-01

    In this work silk fibroin/pullulan blend films have been prepared by solution casting method. The blend films were examined for structural, and thermal properties using X-ray diffraction (XRD), thermogravimatric (TGA) and differential scanning calorimetry (DSC) analysis. The XRD results indicate that with the introduction of pullulan, the interaction between SF and pullulan in the blend films induced the conformation transition of SF films and amorphous phase increases with increasing pullulan ratio. The thermal properties of the blend films were improved significantly in the blend films.

  9. Fibroin silk proteins from the nonmulberry silkworm Philosamia ricini are biochemically and immunochemically distinct from those of the mulberry silkworm Bombyx mori.

    Ahmad, Raies; Kamra, Anita; Hasnain, Seyed Ehtesham

    2004-03-01

    Silk proteins were isolated from the cocoons of the nonmulberry silkworm, Philosamia ricini. Three polypeptides of 97, 66, and 45 kDa were identified. The 66-kDa molecule represented sericin, whereas the 97-kDa and the 45-kDa polypeptides linked together through a disulfide bond constituted the fibroin protein. Antibodies raised against the 97-kDa P. ricini fibroin heavy chain reacted specifically with this molecule and did not recognize fibroin heavy chain from another nonmulberry silkworm, Antheraea assama or from the mulberry silkworm, Bombyx mori, suggesting the presence of P. ricini species-specific determinants in this heavy chain. Antibodies generated against fibroin light chain of P. ricini also showed similar reactivity pattern. Immunoblot analysis with proteins isolated from the silk glands of P. ricini at different stages of larval development showed that the expression of fibroin heavy chain was developmentally and spatially regulated. The protein was most abundant in the 5th instar larva, and could be detected in the middle and the posterior but not the anterior silk glands. The amino acid composition of the 97-kDa fibroin protein showed abundance of glutamic acid and did not contain (Gly-Ala)(n) motifs, a characteristic feature of B. mori fibroin heavy chain. Our study reveals significant differences between the nonmulberry silkworm P. ricini and the mulberry silkworm B. mori in the biochemical composition and immunochemical characteristics of fibroin heavy chain. These differences might be responsible for the differences seen in the quality of silk produced by these two silkworms. PMID:15068584

  10. 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.

  11. 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 μm 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 β-sheet structure.

  12. Silk fibroin diaphragm-based fiber-tip Fabry-Perot pressure sensor.

    Cheng, Linghao; Wang, Cengzhong; Huang, Yunyun; Liang, Hao; Guan, Bai-Ou

    2016-08-22

    A miniature fiber-optic Fabry-Perot is built on the tip of a single mode fiber with a thin silk fibroin film as the diaphragm for pressure measurement. The silk fibroin film is regenerated from aqueous silk fibroin solution obtained by an environmentally benign fabrication process, which exhibits excellent optical and physicochemical properties, such as transparency in visible and near infrared region, membrane-forming ability, good adhesion, and high mechanical strength. The resulted Fabry-Perot pressure sensor is therefore highly biocompatible and shows good airtightness with a response of 12.3 nm/kPa in terms of cavity length change. PMID:27557238

  13. 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 (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

  14. Preparation and characterization of noble metal nanocolloids by silk fibroin in situ reduction

    CHEN; Wenxing(陈文兴); WU; Wen(吴雯); CHEN; Haixiang(陈海相); SHEN; Zhiquan(沈之荃)

    2003-01-01

    Noble metal nanocolloids are prepared from their precursors by in situ reduction of a silk fibroin solution at room temperature without any reducing agent. The mechanism, the effects of pH and the molar ratio of the reactants on the reduction reaction are studied by UV-Vis spectroscopy. The structure of the colloids is characterized by FT-IR, TEM and AFM. According to the TEM images, the gold-silk fibroin colloid is a nanostructured bioconjugate with novel core-shell, while the silver-silk fibroin colloid tends to be congregated as clusters having more than ten nanoparticles of silver-silk fibroin. The gold colloid is highly dispersed and stable while the silver colloid is less dispersed and stable than the gold colloid.

  15. 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

  16. 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

  17. 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)

  18. 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

  19. 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.

  20. 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)

  1. 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)

  2. 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.

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

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

    2016-03-01

    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

  4. 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.

  5. High-Q silk fibroin whispering gallery microresonator on a flexible chip

    Xu, Linhua; Zhao, Guangming; Liu, Zhiwen; Yang, Lan

    2016-01-01

    We have experimentally demonstrated on-chip all silk fibroin whispering gallery mode microresonator by using the molding and solution casting technique. The quality factors of the fabricated silk protein microresonators are up to 0.9*10^5. A high-sensitivity thermal sensor was realized in this silk fibroin microtoroid with the sensitivity of 1.17 nm/K, 8 times higher than previous WGM resonantor based thermal sensors. This opens the way to fabricate the biodegradable and biocompatible protein based microresonators on a flexible chip for biophotonics applications in vivo.

  6. Activation of the Ubiquitin Proteasome Pathway by Silk Fibroin Modified Chitosan Nanoparticles in Hepatic Cancer Cells

    Ming-Hui Yang; Tze-Wen Chung; Yi-Shan Lu; Yi-Ling Chen; Wan-Chi Tsai; Shiang-Bin Jong; Shyng-Shiou Yuan; Pao-Chi Liao; Po-Chiao Lin; Yu-Chang Tyan

    2015-01-01

    Silk fibroin (SF) is a protein with bulky hydrophobic domains and can be easily purified as sericin-free silk-based biomaterial. Silk fibroin modified chitosan nanoparticle (SF-CSNP), a biocompatible material, has been widely used as a potential drug delivery system. Our current investigation studied the bio-effects of the SF-CSNP uptake by liver cells. In this experiment, the characterizations of SF-CSNPs were measured by particle size analysis and protein assay. The average size of the SF-C...

  7. 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.

  8. 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 samples were greater than 80 % when hydrothermal degradation was performed in the sodium hydroxide solution at an appropriate concentration of 0.05 M. However, the solubility of irradiated fibroin was greater than that of unirradiated sample. The soluble silk protein content increased from 0.462 to 0.653 mg protein/mg silk fibroin 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)

  9. Effect of Na2CO3 degumming concentration on LiBr-formic acid-silk fibroin solution properties

    Liu Zhi

    2016-01-01

    Full Text Available Salt-acid system has been proved to be of high efficiency for silk fibroin dissolution. Using salt-acid system to dissolve silk, native silk fibrils can be preserved in the regenerated solution. Increasing experiments indicate that acquirement of silk fibrils in solution is strongly associated with the degumming process. In this study, the effect of sodium carbonate degumming concentration on solution properties based on lithium bromide-formic acid dissolution system was systematically investigated. Results showed that the morphology transformation of silk fibroin in solution from nanospheres to nanofibrils is determined by sodium carbonate concentration during the degumming process. Solutions containing different silk fibroin structure exhibited different rheological behaviors and different electrospinnability, leading to different electrospun nanofibre properties. The results have guiding significance for preparation and application of silk fibroin solutions.

  10. 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

  11. 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

  12. Improvements of anticoagulant activities of silk fibroin films with fucoidan

    2008-01-01

    Fucoidan (FC),an effective anticoagulant constituent extracted from brown algae,was introduced into silk fibroin (SF) for improving its blood compatibility.The SF and SF/FC blend films were characterized by attenuated total reflectance Fourier-transform infrared (ATR-FTIR),X-ray photoelectron spectroscopy (XPS),scanning electron microscopy (SEM) and dynamic contact angle determinator (CA).The in vitro anticoagulant activities of the films were evaluated by activated partial thromboplastin time (APTT),thrombin time (TT) and prothrombin time (PT) measurements.The endothelial cell attachment and proliferation viability on the film were assessed by micropipette aspiration technique and MTT assay,respectively.The testing results indicated that the introduction of FC increased the roughness,hydrophilicity and sulfate component of the film surface without impeding the formation of β-sheet conformation in SF.More important,FC brought excellent anticoagulant activity and better endothelial cell affinity to SF.The SF/FC blend film was hopeful to be used as blood-contacting biomaterials.

  13. 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.

  14. Sonication induced silk fibroin cryogels for tissue engineering applications

    Kadakia, P. U.; Jain, E.; Hixon, K. R.; Eberlin, C. T.; Sell, S. A.

    2016-05-01

    In this study, we report a method to form macroporous silk fibroin (SF) scaffolds through a combination of ultrasonication followed by cryogelation at subzero temperatures. The resultant sonication induced SF cryogels encompassed larger pore sizes (151 ± 56 μm) and higher mechanical stability (127.15 ± 24.71 kPa) than their hydrogel counterparts made at room temperature. Furthermore, the addition of dopants like Manuka honey and bone char in SF cryogels did not affect cryogel synthesis but decreased the pore size in a concentration dependent manner. With no crack propagation at 50% strain and promising stability under cyclic loads, mineralization and cellular infiltration potential were analyzed for bone tissue engineering purposes. Although the scaffolds showed limited mineralization, encouraging cellular infiltration results yield promise for other tissue engineering applications. The use of mild processing conditions, a simplistic procedure, and the lack of organic solvents or chemical cross-linkers renders the combination of sonication and cryogelation as an attractive fabrication technique for 3D SF macroporous scaffolds.

  15. 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.

  16. 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

  17. 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)

  18. Silks produced by insect labial glands

    Sehnal, František; Sutherland, T.

    Austin: Landes Bioscience, 2008 - (Scheibel, T.), s. 106-120 ISBN 978-1-58706-316-9 R&D Projects: GA AV ČR IAA5007402 Institutional research plan: CEZ:AV0Z50070508 Keywords : silks * Insect * labial glands Subject RIV: ED - Physiology

  19. 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.

  20. Rheological and Mechanical Behavior of Silk Fibroin Reinforced Waterborne Polyurethane

    Yongzhen Tao

    2016-03-01

    Full Text Available Waterborne polyurethane (WPU is a versatile and environment-friendly material with growing applications in both industry and academia. Silk fibroin (SF is an attractive material known for its structural, biological and hemocompatible properties. The SF reinforced waterborne polyurethane (WPU is a promising scaffold material for tissue engineering applications. In this work, we report synthesis and characterization of a novel nanocomposite using SF reinforced WPU. The rheological behaviors of WPU and WPU-SF dispersions with different solid contents were investigated with steady shear and dynamic oscillatory tests to evaluate the formation of the cross-linked gel structure. The average particle size and the zeta potential of WPU-SF dispersions with different SF content were examined at 25 °C to investigate the interaction between SF and WPU. FTIR, SEM, TEM and tensile testing were performed to study the effects of SF content on the structural morphology and mechanical properties of the resultant composite films. Experimental results revealed formation of gel network in the WPU dispersions at solid contents more than 17 wt %. The conjugate reaction between the WPU and SF as well as the hydrogen bond between them helped in dispersing the SF powder into the WPU matrix as small aggregates. Addition of SF to the WPU also improved the Young’s modulus from 0.30 to 3.91 MPa, tensile strength from 0.56 to 8.94 MPa, and elongation at break from 1067% to 2480%, as SF was increased up to 5 wt %. Thus, significant strengthening and toughening can be achieved by introducing SF powder into the WPU formulations.

  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. 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

  3. Effects of sterilization methods on the physical, chemical, and biological properties of silk fibroin membranes.

    de Moraes, Mariana Agostini; Weska, Raquel Farias; Beppu, Marisa Masumi

    2014-05-01

    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 jeopardize the functionality of medical devices. The aim of this study was to analyze the influence of different sterilization methods in the physical, chemical, and biological characteristics of dense and porous silk fibroin membranes. Silk fibroin membranes were treated by several procedures: immersion in 70% ethanol solution, ultraviolet radiation, autoclave, ethylene oxide, and gamma radiation, and were analyzed by scanning electron microscopy, Fourier-transformed infrared spectroscopy (FTIR), X-ray diffraction, tensile strength and in vitro cytotoxicity to Chinese hamster ovary cells. The results indicated that the sterilization methods did not cause perceivable morphological changes in the membranes and the membranes were not toxic to cells. The sterilization methods that used organic solvent or an increased humidity and/or temperature (70% ethanol, autoclave, and ethylene oxide) increased the silk II content in the membranes: the dense membranes became more brittle, while the porous membranes showed increased strength at break. Membranes that underwent sterilization by UV and gamma radiation presented properties similar to the nonsterilized membranes, mainly for tensile strength and FTIR results. PMID:24259492

  4. Modulated Degradation of Transient Electronic Devices through Multilayer Silk Fibroin Pockets.

    Brenckle, Mark A; Cheng, Huanyu; Hwang, Sukwon; Tao, Hu; Paquette, Mark; Kaplan, David L; Rogers, John A; Huang, Yonggang; Omenetto, Fiorenzo G

    2015-09-16

    The recent introduction of transient, bioresorbable electronics into the field of electronic device design offers promise for the areas of medical implants and environmental monitors, where programmed loss of function and environmental resorption are advantageous characteristics. Materials challenges remain, however, in protecting the labile device components from degradation at faster than desirable rates. Here we introduce an indirect passivation strategy for transient electronic devices that consists of encapsulation in multiple air pockets fabricated from silk fibroin. This approach is investigated through the properties of silk as a diffusional barrier to water penetration, coupled with the degradation of magnesium-based devices in humid air. Finally, silk pockets are demonstrated to be useful for controlled modulation of device lifetime. This approach may provide additional future opportunities for silk utility due to the low immunogenicity of the material and its ability to stabilize labile biotherapeutic dopants. PMID:26305434

  5. 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.

  6. 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.

  7. 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.

  8. 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

  9. 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...

  10. Effect of freezing methods on the properties of lyophilized porous silk fibroin membranes

    Raquel Farias Weska; Wellington Carlos Vieira Jr.; Grínia Michelle Nogueira; Marisa Masumi Beppu

    2009-01-01

    Silk fibroin is a fibrous protein that has been extensively studied for application in the biomedical field, and has been used as a scaffold for bone tissue engineering. Biomaterials made of proteins are prone to physical and chemical degradation during storage; lyophilization, a drying method that consists of freezing and drying steps, is known to promote minimal changes in structure and biological activity of biomaterials. This study evaluates the effect of freezing methods on the propertie...

  11. 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...

  12. 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)

  13. Silk fibroin nanoparticles for drug delivery purposes : stabilization, incorporation and release design

    Ferreira, A. V.; Volkov, Vadim; Vasconcelos, Andreia; Abreu, Aana S.; Botelho, C. M.; Paulo, Artur Cavaco

    2014-01-01

    Silk Fibroin (SF) has been extensively studied for various applications due to its impressive mechanical properties and biocompatibility. Recently, SF based-particles have been proposed as controlled drug delivery systems. A new and efficient method was developed to prepare SF nanoparticles (SF-NPs) by high pressure homogenization (HPH) emulsification, in oil-in-water emulsions (o/w). During the NPs production by HPH emulsification process, the secondary SF structure changed from ran...

  14. Chemically cross-linked silk fibroin hydrogel with enhanced elastic properties, biodegradability, and biocompatibility

    Park, Won Ho

    2016-01-01

    Min Hee Kim, Won Ho Park Department of Advanced Organic Materials and Textile Engineering System, Chungnam National University, Daejeon, Korea Abstract: In this study, the synthesis of silk fibroin (SF) hydrogel via chemical cross-linking reactions of SF due to gamma-ray (γ-ray) irradiation was investigated, as were the resultant hydrogel’s properties. Two different hydrogels were investigated: physically cross-linked SF hydrogel and chemically cross-linked SF hydrogel i...

  15. 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.

  16. 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

  17. Coimmobilization of Naringinases on Silk Fibroin Nanoparticles and Its Application in Food Packaging

    Min-Hui Wu

    2013-01-01

    Full Text Available Bombyx mori silk fibroin is a macromolecular biopolymer with remarkable biocompatibility. It was degummed and subjected to a series of treatments, including dissolution and dialysis, to yield an aqueous solution of silk fibroin, which was introduced rapidly into excess acetone to produce crystalline silk fibroin nanoparticles (SFNs, which were conjugated covalently with naringinase using glutaraldehyde as the cross-linking reagent. The SFN naringinases are easily recovered by centrifugation and can be used repeatedly. Naringinase is a bienzyme consisting of α-L-rhamnosidase and flavonoid-β-glucosidase. The enzyme activity and its kinetics were similar to those of the native form, and the optimum reactive temperature for both is 55°C. In our study, centrifugation allowed the separation of enzyme and substrate; after eight cycles the SFN naringinases retained >70% residual activity. The highly efficient processing technology and the use of SFN as a novel vector for a bienzyme have great potential for research and the development of food processing such as the debittering of naringin-containing juices.

  18. Antihypertensive effects of silk fibroin hydrolysate by alcalase and purification of an ACE inhibitory dipeptide.

    Zhou, Fengjuan; Xue, Zhaohui; Wang, Jiehua

    2010-06-01

    Silk fibroin, which is normally discarded as an industrial byproduct in clothing plants, was hydrolyzed with alcalase. Angiotensin I converting enzyme (ACE) inhibitory activities of the silk fibroin hydrolysates (SFH) were investigated, and the hydrolysate with hydrolysis degree of 17% exhibited the highest ACE inhibitory activity. At the tested 600 mg/kg.d and 1200 mg/kg x d doses, SFH significantly lowered blood pressure of spontaneously hypertensive rats (SHR) after chronic oral administration. SFH was further purified using consecutive chromatographic methods on Sephadex G-15 column and reverse phase high-performance liquid chromatography (RP-HPLC) on an octadecylsilane column. After its purity was confirmed by analytical RP-HPLC and capillary electrophoresis, one ACE inhibitory dipeptide was isolated, and its molecular mass and amino acid sequence were determined as 238.2 Da and Gly-Tyr, respectively, by LC-ESI/MS. The results of this study suggest that silk fibroin byproducts have the possibility to become an effective source for ACE inhibitory peptides. PMID:20481470

  19. 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.

  20. In vitro and in vivo research on using Antheraea pernyi silk fibroin as tissue engineering tendon scaffolds

    In this paper, the feasibility of using Antheraea pernyi silk fibroin as tissue engineering tendon scaffold was investigated in vitro and in vivo, respectively, utilizing tenocytes and animal model. The animal model used here was an adult New Zealand White rabbit with a 15-mm gap defect in both sides of the Achilles tendon. The Achilles tendon defects in one side of hind legs were repaired using the braided A. pernyi silk fibroin scaffold in experimental group (n = 24), while the other side left untreated as negative group (n = 24). The recovery of the defect tendons were evaluated postoperatively at the 2nd, 6th, 12th, and 16th week using macroscopic, histological, immunohistochemical, scanning electron micrograph and biomechanical test techniques. In vitro results examined by scanning electron micrograph showed that A. pernyi silk fibroin promote the adhesion and propagation of the tenocytes. In vivo, at 16 weeks after implantation, morphological results showed that neo-tendons were formed, and bundles of collagen fibers in the neo-tendons were uniform and well oriented. Immunohistochemical results showed that collagen type in the regenerated tendons was predominantly type I. The maximum load of regenerated tendon at 16 weeks reached 55.46% of the normal tendon values. Preliminary, we concluded that A. pernyi silk fibroin promoted the recovery of Achilles tendon defect of rabbit and the application of A. pernyi silk fibroin as tissue engineering tendon scaffold is feasible.

  1. Bioconjugation of neutral protease on silk fibroin nanoparticles and application in the controllable hydrolysis of sericin.

    Zhu, Lin; Hu, Ren-Ping; Wang, Hai-Yan; Wang, Yuan-Jing; Zhang, Yu-Qing

    2011-09-28

    Bombyx mori silk fibroin is a protein-based macromolecular biopolymer with remarkable biocompatibility. Silk fiber was degummed and subjected to a series of treatments, including dissolution and dialysis, to yield an aqueous solution of silk fibroin, which was introduced rapidly into excess acetone to produce crystalline silk fibroin nanoparticles (SFNs). The SFNs were conjugated covalently with a neutral protease (NP) using glutaraldehyde as the cross-linking reagent. The objective of this study was to determine the optimal conditions for biosynthesis of the SFN-NP bioconjugates. First, SFN-NP was obtained by covalent cross-linking of SFN and NP at an SFN/NP ratio of 5-8 mg:1 IU with 0.75% glutaraldehyde for 6 h at 25 °C. When adding 50 IU of the enzyme, the residual activity of biological conjugates was increased to 31.45%. Studies on the enzyme activity of SFN-NP and its kinetics showed that the stability of SFN-NP bioconjugates was greater than that of the free enzyme, the optimum reactive temperature range was increased by 5-10 °C, and the optimum pH value range was increased to 6.5-8.0. Furthermore, the thermal stability was improved to some extent. A controlled hydrolysis test using the poorly water-soluble protein sericin as a substrate and SFN-NP as the enzyme showed that the longer the reaction time (within 1 h), the smaller the molecular mass (<30 kDa) is of the sericin peptide produced. The SFN-NP bioconjugate is easily recovered by centrifugation and can be used repeatedly. The highly efficient processing technology and the use of SFN as a novel vector for a protease has great potential for research and the development of food processing. PMID:21846144

  2. 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.

  3. 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...

  4. Activation of the Ubiquitin Proteasome Pathway by Silk Fibroin Modified Chitosan Nanoparticles in Hepatic Cancer Cells

    Ming-Hui Yang

    2015-01-01

    Full Text Available Silk fibroin (SF is a protein with bulky hydrophobic domains and can be easily purified as sericin-free silk-based biomaterial. Silk fibroin modified chitosan nanoparticle (SF-CSNP, a biocompatible material, has been widely used as a potential drug delivery system. Our current investigation studied the bio-effects of the SF-CSNP uptake by liver cells. In this experiment, the characterizations of SF-CSNPs were measured by particle size analysis and protein assay. The average size of the SF-CSNP was 311.9 ± 10.7 nm, and the average zeta potential was +13.33 ± 0.3 mV. The SF coating on the SF-CSNP was 6.27 ± 0.17 μg/mL. Moreover, using proteomic approaches, several proteins involved in the ubiquitin proteasome pathway were identified by analysis of differential protein expressions of HepG2 cell uptake the SF-CSNP. Our experimental results have demonstrated that the SF-CSNP may be involved in liver cancer cell survival and proliferation.

  5. Activation of the ubiquitin proteasome pathway by silk fibroin modified chitosan nanoparticles in hepatic cancer cells.

    Yang, Ming-Hui; Chung, Tze-Wen; Lu, Yi-Shan; Chen, Yi-Ling; Tsai, Wan-Chi; Jong, Shiang-Bin; Yuan, Shyng-Shiou; Liao, Pao-Chi; Lin, Po-Chiao; Tyan, Yu-Chang

    2015-01-01

    Silk fibroin (SF) is a protein with bulky hydrophobic domains and can be easily purified as sericin-free silk-based biomaterial. Silk fibroin modified chitosan nanoparticle (SF-CSNP), a biocompatible material, has been widely used as a potential drug delivery system. Our current investigation studied the bio-effects of the SF-CSNP uptake by liver cells. In this experiment, the characterizations of SF-CSNPs were measured by particle size analysis and protein assay. The average size of the SF-CSNP was 311.9 ± 10.7 nm, and the average zeta potential was +13.33 ± 0.3 mV. The SF coating on the SF-CSNP was 6.27 ± 0.17 μg/mL. Moreover, using proteomic approaches, several proteins involved in the ubiquitin proteasome pathway were identified by analysis of differential protein expressions of HepG2 cell uptake the SF-CSNP. Our experimental results have demonstrated that the SF-CSNP may be involved in liver cancer cell survival and proliferation. PMID:25588218

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

    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

  7. Effect of Sodium Carbonate Concentrations on the Formation and Mechanism of Regenerated Silk Fibroin Nanofibers by Electrospinning

    Hao Dou

    2014-01-01

    Full Text Available Degumming is the first process for the preparation of all silk-based products. In this paper, effect of sodium carbonate concentrations for silk degumming on the formation of electrospun silk fibroin nanofibers was investigated and the reason for the silk electrospinning process was explained for the first time by differences from the microstructure of regenerated silk fibroin. With increasing the sodium carbonate concentration, microstructure both in the aqueous solutions and in the electrospinning solutions transformed from nanofibrils to nanoparticles, leading to obvious changes on rheological property; electrospinning solutions with nanofibrils behaved like the native silk dope and owned remarkably higher viscosity than the solutions with nanoparticles showing very low viscosity. More interestingly, nanofibrils favored the formation of silk nanofibers with ease, and even nanofibers could be electrospun at concentration 2%. However, nanoparticles were completely unable to generate nanofibers at high spinning concentration 8%. Importance of sodium carbonate concentrations is heavily emphasized for impacting the microstructure types and further influencing the electrospinning performance of regenerated silk. Hence, sodium carbonate concentrations provide a controllable choice for the preparation of silk-based electrospun biomaterials with desired properties.

  8. Thermal Properties of Silk Fibroin Using Fast Scanning Calorimetry

    Cebe, Peggy; Partlow, Benjamin; Kaplan, David; Wurm, Andreas; Zhuravlev, Evgeny; Schick, Christoph

    We performed fast scanning chip-based calorimetry of silk protein using the Mettler Flash DSC1. We suggest the methodology by which to obtain quantitative information on the very first scan to high temperature, including the melting endotherm of the beta pleated sheets. For proteins, this first scan is the most important one, because the crystalline secondary structural features, the beta pleated sheets, melt after the first heating and cannot be thermally reintroduced. To obtain high quality data, the samples must be treated to drying and enthalpy relaxation sequences. The heat flow rates in heating and cooling must be corrected for asymmetric heat loses. We evaluate methods to obtain an estimate of the sample mass, finally choosing internal calibration using the known heat capacity increment at the glass transition. We report that even heating at rates of 2000 K/s, thermal degradation of silk cannot be totally avoided, though it can be minimized. Using a set of nineteen samples, we successfully determine the liquid state heat capacity of silk as: Cpliquid (T) = (1.98 +0.06) J/gK + T (6.82 +1.4) x10-4 J/gK2. Methods for estimation of the sample mass will be presented and compared. National Science Foundation, Polymers Program DMR-1206010; DAAD; Tufts Faculty Supported Leave.

  9. 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.

  10. 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

  11. Chemically cross-linked silk fibroin hydrogel with enhanced elastic properties, biodegradability, and biocompatibility

    Kim MH

    2016-06-01

    Full Text Available Min Hee Kim, Won Ho Park Department of Advanced Organic Materials and Textile Engineering System, Chungnam National University, Daejeon, Korea Abstract: In this study, the synthesis of silk fibroin (SF hydrogel via chemical cross-linking reactions of SF due to gamma-ray (γ-ray irradiation was investigated, as were the resultant hydrogel’s properties. Two different hydrogels were investigated: physically cross-linked SF hydrogel and chemically cross-linked SF hydrogel irradiated at different doses of γ-rays. The effects of the irradiation dose and SF concentration on the hydrogelation of SF were examined. The chemically cross-linked SF hydrogel was compared with the physically cross-linked one with regard to secondary structure and gel strength. Furthermore, the swelling behavior, crystallinity, and biodegradation of the SF hydrogels were characterized. To assay cell proliferation, the cell viability of human mesenchymal stem cells on the lyophilized SF hydrogel scaffolds was evaluated, and no significant cytotoxicity against human mesenchymal stem cells was observed. Keywords: silk fibroin, hydrogels, biodegradation rate, gamma irradiation, cross-linking

  12. 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

  13. Fibroin and sericin from Bombyx mori silk stimulate cell migration through upregulation and phosphorylation of c-Jun.

    Celia Martínez-Mora

    Full Text Available Wound healing is a biological process directed to the restoration of tissue that has suffered an injury. An important phase of wound healing is the generation of a basal epithelium able to wholly replace the epidermis of the wound. A broad range of products derived from fibroin and sericin from Bombyx mori silk are used to stimulate wound healing. However, so far the molecular mechanism underlying this phenomenon has not been elucidated. The aim of this work was to determine the molecular basis underlying wound healing properties of silk proteins using a cell model. For this purpose, we assayed fibroin and sericin in a wound healing scratch assay using MDA-MB-231 and Mv1Lu cells. Both proteins stimulated cell migration. Furthermore, treatment with sericin and fibroin involved key factors of the wound healing process such as upregulation of c-Jun and c-Jun protein phosphorylation. Moreover, fibroin and sericin stimulated the phosphorylation of ERK 1/2 and JNK 1/2 kinases. All these experiments were done in the presence of specific inhibitors for some of the cell signalling pathways referred above. The obtained results revealed that MEK, JNK and PI3K pathways are involved in fibroin and sericin stimulated cells migration. Inhibition of these three kinases prevented c-Jun upregulation and phosphorylation by fibroin or sericin. Fibroin and sericin were tested in the human keratinocyte cell line, HaCaT, with similar results. Altogether, our results showed that fibroin and sericin initiate cell migration by activating the MEK, JNK and PI3K signalling pathways ending in c-Jun activation.

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

    Bai, Liqiang; Zhu, Liangjun; Min, Sijia; Liu, Lin; Cai, Yurong; Yao, Juming

    2008-03-01

    The Bombyx mori silk fibroin films (SFFs) were modified by a Cecropin B ( CB) antimicrobial peptide, (NH 2)-NGIVKAGPAIAVLGEAAL-CONH 2, 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).

  15. 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)

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

    Bai Liqiang [Key Laboratory of Advanced Textile Materials and Manufacturing Technology of Ministry of Education, College of Materials and Textile, Zhejiang Sci-Tech University, Xiasha Higher Education Park, Hangzhou 310018 (China); Zhu Liangjun; Min Sijia [College of Animal Sciences, Zhejiang University, Hangzhou 310029 (China); Liu Lin; Cai Yurong [Key Laboratory of Advanced Textile Materials and Manufacturing Technology of Ministry of Education, College of Materials and Textile, Zhejiang Sci-Tech University, Xiasha Higher Education Park, Hangzhou 310018 (China); Yao Juming [Key Laboratory of Advanced Textile Materials and Manufacturing Technology of Ministry of Education, College of Materials and Textile, Zhejiang Sci-Tech University, Xiasha Higher Education Park, Hangzhou 310018 (China)], E-mail: yaoj@zstu.edu.cn

    2008-03-15

    The Bombyx mori silk fibroin films (SFFs) were modified by a Cecropin B (CB) antimicrobial peptide, (NH{sub 2})-NGIVKAGPAIAVLGEAAL-CONH{sub 2}, 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)

  17. Potential of inherent RGD containing silk fibroin-poly (Є-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 (Є-caprolactone) to act as an ideal scaffold for bone regeneration. The use of poly (Є-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 (Є-caprolactone) solution (10 wt%) to fabricate nanofibers. The nanofibers possess an average diameter of 152 ± 18 nm (2 % fibroin/PCL) and 175 ± 15 nm (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 (Є-caprolactone) blended nanofibrous scaffolds have an excellent prospect as scaffolding material in bone tissue engineering. PMID:26174955

  18. 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

  19. 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

  20. Strong composite films with layered structures prepared by casting silk fibroin-graphene oxide hydrogels.

    Huang, Liang; Li, Chun; Yuan, Wenjing; Shi, Gaoquan

    2013-05-01

    Composite films of graphene oxide (GO) sheets and silk fibroin (SF) with layered structures have been prepared by facile solution casting of SF-GO hydrogels. The as-prepared composite film containing 15% (by weight, wt%) of SF shows a high tensile strength of 221 ± 16 MPa and a failure strain of 1.8 ± 0.4%, which partially surpass those of natural nacre. Particularly, this composite film also has a high modulus of 17.2 ± 1.9 GPa. The high mechanical properties of this composite film can be attributed to its high content of GO (85 wt%), compact layered structure and the strong hydrogen bonding interaction between SF chains and GO sheets. PMID:23538717

  1. On the growth morphous of capillaries and tissue in porous silk fibroin films

    Lun BAI; Bao-qi ZUO; Guo-ping GUAN; Li-xing DAI; Yong-zhen CHEN; Zheng-yu ZHOU; Jian-mei XU; Zhen-yu WU

    2008-01-01

    In this study, the porous silk fibroin film (PSFF) is implanted into the body of a rat. Ten days later, the growing state of the capillaries in the material was observed, and the growing law of the capillaries in porous material is discussed to better understand the growing state of the peripheral tissue and cells around the material. The fact that the PSFF creates a beneficial environment for the growth of the capillaries in the tissue is confirmed, and the good growing states of the new skeleton muscle and the hypodermal tissue are also observed. This pro-vides basic experimental results for the design of the PSFFs, which is a new medical biomaterial.

  2. Mineralization of HA crystals regulated by terephthaloyl chloride-modified silk fibroin films

    Rong Li; Guang Mei Chen; Xin Lan Ma; Qiao Yan Chen; Ge Wen Xu; Yi Ping Huang

    2011-01-01

    Terephthaloyl chloride (DB)-modified silk fibroin (SF) films were immersed into 1.5 times simulated body fluid (1.5 SBF) to regulate the mineralization of hydroxyapatite (HA) crystals at about 36.5 ℃ for 24 h. UV was used to prove that the new bonds form between the DB and SF. The structure and morphology of the SF/HA were investigated by FTIR, ICP, XRD and SEM. The results showed that the apatite deposited on the matrix of SF mainly was HA. HA was self-assembled on the matrix of SF and formed three-dimensional framework when the weight ratio of DB/SF was 0.30. The content of DB affected the structure and morphology of the apatite composites deposited on the SF films.

  3. Tuning the Refractive Index and Optical Band Gap of Silk Fibroin Films by Electron Irradiation

    S. Asha

    2015-01-01

    Full Text Available The Bombyx mori silk fibroin (SF films were prepared by solution casting method and effects of electron beam on the optical properties and optical constants of the films have been studied by using UV-Visible spectrophotometer. Optical properties like optical band gap Eg, refractive index n, extinction coefficient k, optical conductivity σopt, and dielectric constants ε∗ of virgin and electron irradiated films were determined by using UV-Visible absorption and transmission spectra. It was found that the reduction in optical band gap and increase in refractive index with increasing radiation dosage was observed. It is also observed from results that there is increase in dielectric constants with increasing photon energy. The observed optical changes have been tried to be correlated with the structural changes, revealed through FT-IR spectroscopy. The present study is quite important for tailoring the optical responses of SF films as per specific requirements.

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

    Yaowalak Srisuwan

    2013-01-01

    Full Text Available 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 depended on the SF/alginate blend ratio. The in vitro drug release profile of the blend films was determined by plotting the cumulative drug release versus time. It was found that the drug release significantly decreased as the SF/alginate blend ratio increased. The results demonstrated that the SF/alginate blend films should be a useful controlled-release delivery system for water-soluble drugs.

  5. 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

  6. 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

  7. Regulation of Silk Genes by Hox and Homeodomain Proteins in the Terminal Differentiated Silk Gland of the Silkworm Bombyx mori

    Shigeharu Takiya

    2016-05-01

    Full Text Available The silk gland of the silkworm Bombyx mori is a long tubular organ that is divided into several subparts along its anteroposterior (AP axis. As a trait of terminal differentiation of the silk gland, several silk protein genes are expressed with unique regional specificities. Most of the Hox and some of the homeobox genes are also expressed in the differentiated silk gland with regional specificities. The expression patterns of Hox genes in the silk gland roughly correspond to those in embryogenesis showing “colinearity”. The central Hox class protein Antennapedia (Antp directly regulates the expression of several middle silk gland–specific silk genes, whereas the Lin-1/Isl-1/Mec3 (LIM-homeodomain transcriptional factor Arrowhead (Awh regulates the expression of posterior silk gland–specific genes for silk fiber proteins. We summarize our results and discuss the usefulness of the silk gland of Bombyx mori for analyzing the function of Hox genes. Further analyses of the regulatory mechanisms underlying the region-specific expression of silk genes will provide novel insights into the molecular bases for target-gene selection and regulation by Hox and homeodomain proteins.

  8. In situ synthesis and photoluminescence of QD-CdS on silk fibroin fibers at room temperature

    A convenient room-temperature bioinspired technique has been developed to synthesize hybrid nanocomposites consisting of well-dispersed CdS quantum dots (QD) and the substrate silk fibroin fibers (SFF). The biomaterial SFF provides both a supporting substrate and functional sites for the in situ generation of QD-CdS, which is supported by FTIR and PL measurements. The solid QD-CdS/SFF nanocomposites could be useful in photocatalyst, novel luminescence and photoelectron transfer devices. The QD-CdS/silk fibroin (SF) colloid, in which SF acts as both an inherent biocompatibilizer and an efficient passivator of trap sites on the QD-CdS surface, is also available for some potential applications in the biological fields. The bioinspired method and relevant ideas could extend to fabricating other functional hybrid materials

  9. 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)

  10. Fibroin and Sericin from Bombyx mori Silk Stimulate Cell Migration through Upregulation and Phosphorylation of c-Jun

    Celia Martínez-Mora; Anna Mrowiec; Eva María García-Vizcaíno; Antonia Alcaraz; José Luis Cenis; Francisco José Nicolás

    2012-01-01

    Wound healing is a biological process directed to the restoration of tissue that has suffered an injury. An important phase of wound healing is the generation of a basal epithelium able to wholly replace the epidermis of the wound. A broad range of products derived from fibroin and sericin from Bombyx mori silk are used to stimulate wound healing. However, so far the molecular mechanism underlying this phenomenon has not been elucidated. The aim of this work was to determine the molecular bas...

  11. Square Wave Voltammetric Label-free Determination of the Natural Protein Material Silk Fibroin

    MA Ming-Ming; SONG Jun-Feng

    2008-01-01

    The electrochemical behavior of silk fibroin(SF)was investigated by cyclic voltammetry and square wave voltammetry in 0.01 mol/L HCI for the first time.Within the potential scan range of 0.0 to1.2 V(vs.SCE),two oxi-dative peaks at 0.91 V(Pa,1)and 0.43 V(Pa,2)as well as one reductive peak at 0.24 V(Pc)were observed on cyclic voltammogram at scan rate of 0.2 V/s.The peak current of the peak Pa,1 was linear with SF concentration in the range of 5.8×10-8 to 1.1×10-6 mol/L,with the limit of detection 3.0×10-8 mol/L(SIN=3).The proposed method was of high selectivity without the interferences from the coexisting substances such as another natural protein material sericin and other small molecular substances.It was applied to the determination of SF in raw silk liquid samples without any pre-separation and pre-purification.

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

    Luan, Xi-Ying; Wang, Yong; Duan, Xiang; Duan, Qiao-Yan; Li, Ming-Zhong; Lu, Shen-Zhou; Zhang, Huan-Xiang; Zhang, Xue-Guang

    2006-12-01

    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. PMID:18458403

  13. 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.

  14. 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

  15. 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

  16. Silks produced by insect labial glands

    Sehnal, František; Sutherland, T.

    2008-01-01

    Roč. 2, č. 4 (2008), s. 145-153. ISSN 1933-6896 R&D Projects: GA MŠk ME 907 Institutional research plan: CEZ:AV0Z50070508 Keywords : silk * proteinaceous polymers * alfa-helices Subject RIV: ED - Physiology Impact factor: 0.875, year: 2008 http://www.landesbioscience.com/journals/prion/article/7489

  17. Carbonic anhydrase generates a pH gradient in Bombyx mori silk glands.

    Domigan, L J; Andersson, M; Alberti, K A; Chesler, M; Xu, Q; Johansson, J; Rising, A; Kaplan, D L

    2015-10-01

    Silk is a protein of interest to both biological and industrial sciences. The silkworm, Bombyx mori, forms this protein into strong threads starting from soluble silk proteins using a number of biochemical and physical cues to allow the transition from liquid to fibrous silk. A pH gradient has been measured along the gland, but the methodology employed was not able to precisely determine the pH at specific regions of interest in the silk gland. Furthermore, the physiological mechanisms responsible for the generation of this pH gradient are unknown. In this study, concentric ion selective microelectrodes were used to determine the luminal pH of B. mori silk glands. A gradient from pH 8.2 to 7.2 was measured in the posterior silk gland, with a pH 7 throughout the middle silk gland, and a gradient from pH 6.8 to 6.2 in the beginning of the anterior silk gland where silk processing into fibers occurs. The small diameter of the most anterior region of the anterior silk gland prevented microelectrode access in this region. Using a histochemical method, the presence of active carbonic anhydrase was identified in the funnel and anterior silk gland of fifth instar larvae. The observed pH gradient collapsed upon addition of the carbonic anhydrase inhibitor methazolamide, confirming an essential role for this enzyme in pH regulation in the B. mori silk gland. Plastic embedding of whole silk glands allowed clear visualization of the morphology, including the identification of four distinct epithelial cell types in the gland and allowed correlations between silk gland morphology and silk stages of assembly related to the pH gradient. B. mori silk glands have four different epithelial cell types, one of which produces carbonic anhydrase. Carbonic anhydrase is necessary for the mechanism that generates an intraluminal pH gradient, which likely regulates the assembly of silk proteins and then the formation of fibers from soluble silk proteins. These new insights into native silk

  18. 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)

  19. 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

  20. 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

  1. 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)

  2. Direct electrochemistry and electrocatalysis of heme-proteins in regenerated silk fibroin film

    A biocompatible silk fibroin (SF) film provided a feasible microenvironment for heme-proteins to direct electron transfer on graphite electrodes (GE). Myoglobin (Mb), hemoglobin (Hb), horseradish peroxidase (HRP), and catalase (Cat) incorporated in SF films exhibited a pair of well-defined, nearly reversible cyclic voltammetric peaks, corresponding to the reaction of hemeFe (III) + e → hemeFe (II). The formal potential (E 0), the apparent coverage (Γ) and the electron transfer rate constant (k s) of four proteins in SF films were evaluated by analyzing the cyclic voltammograms (CVs) of heme-proteins. The formal potential was pH dependent, suggesting that proton ion was involved in the reaction. Ultraviolet visible (UV-vis) spectra and reflectance absorbance infrared (RAIR) spectra indicated that heme-proteins in SF films were not grossly denatured. The structure of heme-proteins-SF films was investigated using scanning electron microscopy (SEM) and RAIR. It indicated that there existed intermolecular interaction between heme-proteins and SF and this governed their different morphology in SF films. Hydrogen peroxide and nitric oxide were catalytically reduced by the heme-proteins in SF films, showing the potential applicability of the heme-proteins-SF films as the new type of biosensors based on the protein film voltammetry

  3. Fabrication and characterization of bioactive silk fibroin/wollastonite composite scaffolds

    Composite scaffolds of silk fibroin (SF) with bioactive wollastonite were prepared by freeze-drying. X-ray diffraction (XRD) and Fourier transform infrared (FT-IR) spectroscopy analysis showed that random coil and β-sheet structure co-existed in the SF scaffold. The mechanical performance, surface hydrophilicity and water-uptake capacity of the composite scaffolds were improved compared with those of pure SF scaffold. The bioactivity of the composite scaffold was evaluated by soaking in a simulated body fluid (SBF), and formation of a hydroxycarbonate apatite (HCA) layer was determined by FT-IR and XRD. The results showed that the SF/wollastonite composite scaffold was bioactive as it induced the formation of HCA on the surface of the composite scaffold after soaking in SBF for 5 days. In vitro cell attachment and proliferation tests showed that the composite scaffold was a good matrix for the growth of L929 mouse fibroblast cells. Consequently, the incorporation of wollastonite into the SF scaffold can enhance both the mechanical strength and bioactivity of the scaffold, which suggests that the SF/wollastonite composite scaffold may be a potential biomaterial for tissue engineering.

  4. 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.

  5. Electrospun silk fibroin-hydroxybutyl chitosan nanofibrous scaffolds to biomimic extracellular matrix.

    Zhang, Kuihua; Qian, Yongfang; Wang, Hongsheng; Fan, Linpeng; Huang, Chen; Mo, Xiumei

    2011-01-01

    Silk fibroin (SF)-hydroxybutyl chitosan (HBC) blend nanofibrous scaffolds were fabricated using 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP) and trifluoroacetic acid (TFA) as solvents to biomimic the native ECM by electrospinning. SEM results showed that the average nanofibrous diameter increased when the content of HBC was raised from 20% to 100%. Whereas water contact angle measurements confirmed that SF/HBC nanofibrous scaffolds with different weight ratios were of good hydrophilicity. Both the tensile strength and the elongation at break were improved obviously when the weight ratio of SF to HBC was 20:80. (13)C-NMR clarified that SF and HBC molecules existed in H-bond interactions, but HBC did not induce SF conformation to transform from random coil form to β-sheet structure. Moreover, the use of genipin vapour not only induced conformation of SF to convert from random coil to β-sheet structure but also acted as a cross-linking agent for SF and HBC. Cell viability studies demonstrated that SF/HBC nanofibrous scaffolds presented good cellular compatibility. Thus, electrospun SF/HBC blended nanofibres may provide an ideal biomimic tissue-engineering scaffold. PMID:20615313

  6. 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.

  7. Mechanism of Conformational Transition of Silk Fibroin in Alcohol-water Mixtures

    Ma Lin; He Weiren; Huang Aimin; Li Lishuo; Wei Qiaona; Huang Zilun

    2011-01-01

    Circular dichroism, intrinsic fluorescence of protein and exogenous fluorescence probe of 8-anilino-l-naphtha-lenesulfonic acid hemimagnesium salt (ANS) was used to investigate the mechanism of conformational change of silk fibroin (SF) in aqueous alcohol including methanol and ethanol. The conformational transition of SF from ran-dom coil to β-sheet was found to be of a close relationship with the microstructure of the solvent. The alcohol-water mixture at low concentration had little effect on the solvation of the peptide unit, as the inherent water structure was conserved. At high alcohol concentration, the transition from the tetrahedral-like water structure to the chain-like alcohol structure in the mixtures induced a β-sheet conformation of SF, as a result of the formation of intramolecu-lar hydrogen bond between the peptide units in order to eliminate the thermodynamic unfavorite from the contact to the solvent molecules. Meanwhile, the aggregating of hydrophobic side chains was decreased by the alcohol via the destruction of hydrogen bond network of water by alcohol and the binding of alcohol to hydrophobic group.

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

    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. (paper)

  9. 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)

  10. Three dimensional poly(ε-caprolactone) and silk fibroin nanocomposite fibrous matrix for artificial dermis.

    Lee, Jung Min; Chae, Taesik; Sheikh, Faheem A; Ju, Hyung Woo; Moon, Bo Mi; Park, Hyun Jung; Park, Ye Ri; Park, Chan Hum

    2016-11-01

    Ideal dermal substitutes should have comparable physicochemical and biological properties to the natural skin tissue. In this study, we report a novel strategy to "engineer" controlled 3D nanocomposite fibrous matrix of poly(ε-caprolactone) (PCL) and silk fibroin (SF) for an artificial dermis application. Using a custom-designed cold-plate electrospinning and automatic magnet agitation system, up to 6mm of the thickness was achieved resulting from the accumulation of ice crystal layers on the PCL nanofibers surface-modified with the SF particles. The sacrificed ice crystals induced interconnected macro-pores ranging from tens to hundreds μm. The agitation system introduced uniform distribution of the SF protein within/on the nanofibers, preventing the particles from precipitation and agglomeration. NIH 3T3 fibroblasts proliferated in vitro on the PCL and PCL/SF scaffolds for 7days, but there was no statistical difference between the groups. Conversely, In vivo rat model studies revealed that the wound healing rate and collagen deposition increased with the SF content within the nanocomposites. The unique 3D construct with the PCL/SF nanocomposite fibers provided desirable spatial cues, surface topography, and surface chemistry for the native cells to infiltrate into the scaffolds. The wound healing potential of the nanocomposites was comparable to the commercial Matriderm® artificial dermis. PMID:27524077

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

    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

  12. 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)

  13. Effect of sterilization on structural and material properties of 3-D silk fibroin scaffolds.

    Hofmann, Sandra; Stok, Kathryn S; Kohler, Thomas; Meinel, Anne J; Müller, Ralph

    2014-01-01

    The development of porous scaffolds for tissue engineering applications requires the careful choice of properties, as these influence cell adhesion, proliferation and differentiation. Sterilization of scaffolds is a prerequisite for in vitro culture as well as for subsequent in vivo implantation. The variety of methods used to provide sterility is as diverse as the possible effects they can have on the structural and material properties of the three-dimensional (3-D) porous structure, especially in polymeric or proteinous scaffold materials. Silk fibroin (SF) has previously been demonstrated to offer exceptional benefits over conventional synthetic and natural biomaterials in generating scaffolds for tissue replacements. This study sought to determine the effect of sterilization methods, such as autoclaving, heat-, ethylene oxide-, ethanol- or antibiotic-antimycotic treatment, on porous 3-D SF scaffolds. In terms of scaffold morphology, topography, crystallinity and short-term cell viability, the different sterilization methods showed only few effects. Nevertheless, mechanical properties were significantly decreased by a factor of two by all methods except for dry autoclaving, which seemed not to affect mechanical properties compared to the native control group. These data suggest that SF scaffolds are in general highly resistant to various sterilization treatments. Nevertheless, care should be taken if initial mechanical properties are of interest. PMID:24013025

  14. 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.

  15. Biocompatibility studies of silk fibroin-based artificial nerve grafts in vitro and in vivo

    2007-01-01

    Silk fibroin (SF) has been used extensively in the biomedical field including tissue engineering for the generation of artificial bones, skins or ligaments. We have previously reported on good in vitro biocompatibility of SF fibers with peripheral nerve tissues and cells. In the present study, we developed a novel design of the SF-based artificial nerve graft (SF graft) which was composed of a SF-nerve guidance conduit (NGC) inserted with SF fibers. MTT assay was performed to determine the cytotoxicity of the SF-NGC extract fluid on the cultured L929 cells derived from an immortalized mouse fibroblast cell line. In addition, this SF graft was implanted into adult rats for bridging a 10-mm long sciatic nerve defect. The following-up experiments at initial stage (1-4 week) of nerve regeneration including routine blood tests and histochemical investigation were conducted to evaluate the in vivo biocompatibility of the SF graft with peripheral nerves. The results demonstrated that the SF-NGC graft was biocompatible with the surrounding tissues and cells due to its low inflammatory potential with a grade O under the U. S. Pharmacopeia guidelines and it was generally suitable to a certain degree for bridging peripheral nerve defects in virtue of supporting Schwann cell adherence, expansion and migration. Therefore the SF graft is a promising alternative to classical autografts for peripheral nerve repair.

  16. 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.

  17. 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)

  18. 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.

  19. 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

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

    Xue, Rui; Kang, Tian-Fang; Lu, Li-Ping; Cheng, Shui-Yuan

    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 × 10-6 to 2.0 × 10-3 M with a detection limit of 5.0 × 10-7 M. Similarly, the linearly response range of carbaryl was from 1.0 × 10-7 to 3.0 × 10-5 M with a detection limit of 6.0 × 10-8 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.

  1. 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)

  2. Effect of Strongly Alkaline Electrolyzed Water on Silk Degumming and the Physical Properties of the Fibroin Fiber.

    Ting-Ting Cao

    Full Text Available Strongly alkaline electrolyzed water (SAEW was prepared by electrolysis of tap water in a laboratory-made water electrolyzer. The pH of stored SAEW was stable for more than one month. The hardness of the electrolyzed water was 30% lower and the Na(+ concentration was 18% higher than those of the tap water. Silkworm cocoon shells were boiled in pH 11.50 SAEW at a ratio of 1∶40∼80 (W/V for 20 min and the sericin layers around the silk fibroin fibers were removed completely. The tensile properties and thermal decomposition temperature of a single filament of silk fibroin obtained by the SAEW method were almost the same as those for the fiber obtained by the neutral soap, and much higher than those for the fiber obtained by Na2CO3 degumming. The results demonstrate that SAEW is an environmentally friendly and pollution-free silk degumming agent that allows highly efficient, low cost recovery of sericin.

  3. Thin films of silk fibroin and its blend with chitosan strongly promote biofilm growth of Synechococcus sp. BDU 140432.

    Kaushik, Sharbani; Sarma, Mrinal K; Thungon, Phurpa Dema; Santhosh, Mallesh; Goswami, Pranab

    2016-10-01

    The activating role of different polymer thin films coated over polystyrene support on the Synechococcus sp. biofilm growth was examined concurrently by measuring biofilm florescence using a dye and by measuring cell density in the isolated biofilm. Compared to blank (no coating), the increase in biofilm formation (%) on silk, chitosan, silk-chitosan (3:2) blend, polyaniline, osmium, and Nafion films were 27.73 (31.16), 21.55 (23.74), 37.21 (38.34), 5.35 (8.96), 6.70 (6.55) and (nil), respectively with corresponding cell density (%) shown in the parentheses. This trend of biofilm formation on the films did not significantly vary for Escherichia coli and Lactobacillus plantarum strains. The films of 20 residues long each of glycine-alanine repeat peptide, which mimics a silk fibroin motif, and a hydrophobic glycine-valine repeat peptide, increased the biofilm growth by 13.53 % and 26.08 %, respectively. Silk and blend films showed highest adhesion unit (0.48-0.49), adhesion rate ((4.2-4.8)×10(-6), m/s) and Gibbs energy of adhesion (-8.5 to -8.6kT) with Synechococcus sp. The results confirmed interplay of electrostatic and hydrophobic interaction between cell-surface and polymer films for promoting rapid biofilm growth. This study established that the thin films of silk and the blend (3:2) promote rapid biofilm growth for all the tested microorganisms. PMID:27393887

  4. Tunable Structures and Properties of Electrospun Regenerated Silk Fibroin Mats Annealed in Water Vapor at Different Times and Temperatures

    Xiangyu Huang

    2014-01-01

    Full Text Available Regenerated silk fibroin (SF mats were fabricated using electrospinning technique, followed by mild water vapor annealing to effectively tune the structures and improve the mechanical properties of the mats at different annealing times and temperatures. The breaking strength and the breaking energy of the mats treated with water vapor at 65°C for 12 h reached 6.0 MPa and 171.7 J/kg, respectively. The conformational transition of the SF mats was significantly influenced by the treating temperature, while the influence of time was comparatively limited. The influence is consistent with the time-temperature equivalent principle and would be helpful for the preparation of water-vapor-annealed silk-based biomaterials for various applications.

  5. 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)

  6. Fabrication and characterization of silk fibroin/bioactive glass composite films

    Composite films of silk fibroin (SF) with nano bioactive glass (NBG) were prepared by the solvent casting method, and the structures and properties of the composite films were characterized. Fourier transform infrared (FT-IR) spectroscopy analysis shows that the random coil and β-sheet structure co-exist in the SF films. Results of field emission scanning electron microscope (FESEM) indicate that the NBG particles are uniformly dispersed in the SF films. The measurements of the water contact angles suggest that the incorporation of NBG into SF can improve the hydrophilicity of the composites. The bioactivity of the composite films was evaluated by soaking in 1.5 times simulated body fluid (1.5 × SBF), and formation of a hydroxycarbonate apatite (HCA) layer was determined by XRD and FESEM. The results show that the SF/NBG composite film is bioactive as it induces the formation of HCA on the surface of the composite film after soaking in 1.5 × SBF for 7 days. In vitro osteoblasts attachment and proliferation tests show that the composite film is a good matrix for the growth of osteoblasts. Consequently, the incorporation of NBG into the SF film can enhance both the bioactivity and biocompatibility of the film, which suggests that the SF/NBG composite film may be a potential biomaterial for bone tissue engineering. - Highlights: ► The incorporation of NBG into SF can improve the hydrophilicity of the SF/NBG composite films. ► The SF/NBG composite films show the better bioactivity than the pure SF film. ► The SF/NBG composite films facilitate cell growth and promote cell proliferation and differentiation.

  7. 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.

  8. 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

  9. 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.

  10. 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)

  11. 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.

  12. 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

  13. Expression of the Japanese oak silkworm Antheraea yamamai fibroin gene in the domesticated silkworm Bombyx mori

    Isao Kobayashi; Katsura Kojima; Hideki Sezutsu; Keiro Uchino; Toshiki Tamura

    2009-01-01

    To understand the evolutionary conservation of the gene expression mechanism and secretion machinery between Antheraea and Bombyx fibroins, we introduced the genomic A. yamamai fibroin gene into the domesticated silkworm, B. mori. The spliced A. yamamai fibroin mRNA appeared only in the posterior region of the silk gland of the transgenic silkworm, suggesting that the functions of the fibroin promoter region and the splicing machinery are conserved between these two species. The A. yamamai fibroin protein was detected in the lumen of the silk gland of the transgenic silkworm, albeit at lower levels compared with the B. mori-type fibroin. We found a strong degeneration of the posterior region of the silk gland of the transgenic silkworm. As a result, the cocoon shell weight was much lower in the transgenic silkworm than in the non-transgenic line. These results indicate that the promoter function and splicing machinery are well conserved between A. yamamai and B. mori but that the secretion mechanism of fibroin is diversified between the two.

  14. Tissue-specific transcription enhancement of the fibroin gene characterized by cell-free systems.

    Suzuki, Y.; Tsuda, M.; Takiya, S; Hirose, S; Suzuki, E; Kameda, M; Ninaki, O

    1986-01-01

    Six cell-free extracts have been used to characterize the nature of DNA signals and trans-acting factors responsible for the transcription enhancement of the Bombyx mori fibroin gene. The upstream element of the fibroin gene involved in the enhancement can be divided into two regions. The proximal region, -72 to -32, is recognized as a common enhancing signal by all B. mori extracts from the posterior silk gland, the middle silk gland, the ovarian tissue, and an embryonic cell line. It is wea...

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

    Paşcu, Elena I; Stokes, Joseph; McGuinness, Garrett B

    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 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

  16. Treatment of Silk Fibroin with Poly(ethylene glycol for the Enhancement of Corneal Epithelial Cell Growth

    Shuko Suzuki

    2015-05-01

    Full Text Available A silk protein, fibroin, was isolated from the cocoons of the domesticated silkworm (Bombyx mori and cast into membranes to serve as freestanding templates for tissue-engineered corneal cell constructs to be used in ocular surface reconstruction. In this study, we sought to enhance the attachment and proliferation of corneal epithelial cells by increasing the permeability of the fibroin membranes and the topographic roughness of their surface. By mixing the fibroin solution with poly(ethylene glycol (PEG of molecular weight 300 Da, membranes were produced with increased permeability and with topographic patterns generated on their surface. In order to enhance their mechanical stability, some PEG-treated membranes were also crosslinked with genipin. The resulting membranes were thoroughly characterized and compared to the non-treated membranes. The PEG-treated membranes were similar in tensile strength to the non-treated ones, but their elastic modulus was higher and elongation lower, indicating enhanced rigidity. The crosslinking with genipin did not induce a significant improvement in mechanical properties. In cultures of a human-derived corneal epithelial cell line (HCE-T, the PEG treatment of the substratum did not improve the attachment of cells and it enhanced only slightly the cell proliferation in the longer term. Likewise, primary cultures of human limbal epithelial cells grew equally well on both non-treated and PEG-treated membranes, and the stratification of cultures was consistently improved in the presence of an underlying culture of irradiated 3T3 feeder cells, irrespectively of PEG-treatment. Nevertheless, the cultures grown on the PEG-treated membranes in the presence of feeder cells did display a higher nuclear-to-cytoplasmic ratio suggesting a more proliferative phenotype. We concluded that while the treatment with PEG had a significant effect on some structural properties of the B. mori silk fibroin (BMSF membranes, there were

  17. 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.

  18. 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.

  19. 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

  20. Comparative proteomic analysis of the silkworm middle silk gland reveals the importance of ribosome biogenesis in silk protein production.

    Li, Jian-ying; Ye, Lu-peng; Che, Jia-qian; Song, Jia; You, Zheng-ying; Yun, Ki-chan; Wang, Shao-hua; Zhong, Bo-xiong

    2015-08-01

    The silkworm middle silk gland (MSG) is the sericin synthesis and secretion unique sub-organ. The molecular mechanisms of regulating MSG protein synthesis are largely unknown. Here, we performed shotgun proteomic analysis on the three MSG subsections: the anterior (MSG-A), middle (MSG-M), and posterior (MSG-P) regions. The results showed that more strongly expressed proteins in the MSG-A were involved in multiple processes, such as silk gland development and silk protein protection. The proteins that were highly expressed in the MSG-M were enriched in the ribosome pathway. MSG-P proteins with stronger expression were mainly involved in the oxidative phosphorylation and citrate cycle pathways. These results suggest that the MSG-M is the most active region in the sericin synthesis. Furthermore, comparing the proteome of the MSG with the posterior silk gland (PSG) revealed that the specific and highly expressed proteins in the MSG were primarily involved in the ribosome and aminoacyl-tRNA biosynthesis pathways. These results indicate that silk protein synthesis is much more active as a result of the enhancement of translation-related pathways in the MSG. These results also suggest that enhancing ribosome biogenesis is important to the efficient synthesis of silk proteins. PMID:26051239

  1. 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

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

    Tatyana Dyakonov

    2012-01-01

    Full Text Available 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 compositions. SF-based films, spray-dried microparticles, and matrixes loaded with naproxen were prepared. Both FTIR spectra and in vitro dissolution data demonstrated that SF β-sheet conformation regulates the release profile of naproxen. The controlled release characteristics of the SF-containing compositions were evaluated as a function of SF concentration, temperature, and exposure to dehydrating solvents. The results suggest that SF may be an attractive polymer for use in controlled drug delivery systems.

  3. 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

  4. SURFACE MODIFICATION OF BLEND FILMS COMPOSED OF SILK FIBROIN AND POLY(ETHYLENE GLYCOL) MACROMER AND THEIR IN VITRO ANTITHROMBOGENICITY

    Hua-xin Sun; Song Wang; He-sun Zhu

    2004-01-01

    In order to improve the blood compatibility of silk fibroin (SF), poly(ethylene glycol) macromer (PEGM) in different amounts was added to the SF film to incorporate C=C group into the surface of blend films which were then modified by SO2 gas plasma treatment. ATR-FITR and XPS were used to analyze the chemical change which had occurred on the film's surface. When the content of sulfur on the surface of blend films surpasses 1.59%, the antithrombogenicity of plasma treated films increases remarkably due to surface sulfonation. This result implies that SF with blend of PEGM after SO2 plasma treatment have potential use for making blood-contacting biomaterials.

  5. 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.

  6. 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

  7. An implantable and controlled drug-release silk fibroin nanofibrous matrix to advance the treatment of solid tumour cancers.

    Xie, Maobin; Fan, Dejun; Chen, Yufeng; Zhao, Zheng; He, Xiaowen; Li, Gang; Chen, Aizheng; Wu, Xiaojian; Li, Jiashen; Li, Zhi; Hunt, John A; Li, Yi; Lan, Ping

    2016-10-01

    The development of more effective cancer therapeutic strategies are still critically required. The maximization of the therapeutic effect in combination with avoiding the severe side effects on normal tissues when using chemotherapy drugs is still an urgent problem that requires improvements urgently. Here we provide implantable and controllable drug-release that utilises silk fibroin (SF) as a nanofibrous drug delivery system (DDS) for cancer treatment. A nanofibrous structure with controllable fibre diameter (curcumin (CM)-SF nanofibrous matrix had a superior anti-cancer potential when the concentration was >5 μg/mL. The mechanism could be explained by the cell cycle being held in the S phase. The toxic effect on normal cells (NCM460) was minimized by using a treatment concentration range (5-20 μg/mL). Implantation of this DDS into the tumour site inhibited the growth of solid tumour; this offers an alternative approach for novel cancer therapy. PMID:27376557

  8. 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.

  9. 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.

  10. Bioprintable, cell-laden silk fibroin-gelatin hydrogel supporting multilineage differentiation of stem cells for fabrication of three-dimensional tissue constructs.

    Das, Sanskrita; Pati, Falguni; Choi, Yeong-Jin; Rijal, Girdhari; Shim, Jin-Hyung; Kim, Sung Won; Ray, Alok R; Cho, Dong-Woo; Ghosh, Sourabh

    2015-01-01

    Bioprinting has exciting prospects for printing three-dimensional (3-D) tissue constructs by delivering living cells with appropriate matrix materials. However, progress in this field is currently extremely slow due to limited choices of bioink for cell encapsulation and cytocompatible gelation mechanisms. Here we report the development of clinically relevant sized tissue analogs by 3-D bioprinting, delivering human nasal inferior turbinate tissue-derived mesenchymal progenitor cells encapsulated in silk fibroin-gelatin (SF-G) bioink. Gelation in this bioink was induced via in situ cytocompatible gelation mechanisms, namely enzymatic crosslinking by mushroom tyrosinase and physical crosslinking via sonication. Mechanistically, tyrosinases oxidize the accessible tyrosine residues of silk and/or gelatin into reactive o-quinone moieties that can either condense with each other or undergo nonenzymatic reactions with available amines of both silk and gelatin. Sonication alters the hydrophobic interaction and accelerates self-assembly of silk fibroin macromolecules to form β-sheet crystals, which physically crosslink the hydrogel. However, sonication has no effect on the conformation of gelatin. The effect of optimized rheology, secondary conformations of silk-gelatin bioink, temporally controllable gelation strategies and printing parameters were assessed to achieve maximum cell viability and multilineage differentiation of the encapsulated human nasal inferior turbinate tissue-derived mesenchymal progenitor cells. This strategy offers a unique path forward in the direction of direct printing of spatially customized anatomical architecture in a patient-specific manner. PMID:25242654

  11. 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.

  12. 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

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

    Buga, Mihaela-Ramona [National Research and Development Institute for Cryogenics and Isotopic Technologies, ICIT Rm. Valcea, 240050 Rm. Valcea, Uzinei 4, CP7, Raureni, Valcea (Romania); Zaharia, Cătălin, E-mail: zaharia.catalin@gmail.com [Advanced Polymer Materials Group, University Politehnica of Bucharest, 1-7, Gh. Polizu Street, Sector 1, 011061 Bucharest (Romania); Bălan, Mihai [National Research and Development Institute for Cryogenics and Isotopic Technologies, ICIT Rm. Valcea, 240050 Rm. Valcea, Uzinei 4, CP7, Raureni, Valcea (Romania); Bressy, Christine [Université de Toulon, MAPIEM, EA 4323, 83957 La Garde (France); Ziarelli, Fabio [Fédération des Sciences Chimiques de Marseille, CNRS-FR1739, Spectropole, 13397 Marseille (France); Margaillan, André [Université de Toulon, MAPIEM, EA 4323, 83957 La Garde (France)

    2015-06-01

    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, {sup 13}C, {sup 29}Si 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.

  14. 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

  15. 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.

  16. Cloning and expression of Bombyx mori silk gland elongation factor 1gamma in Escherichia coli.

    Kamiie, Katsuyoshi; Nomura, Yoshitaka; Kobayashi, Satoru; Taira, Hideharu; Kobayashi, Kohmei; Yamashita, Tetsuro; Kidou, Shin-ichiro; Ejiri, Shin-ichiro

    2002-03-01

    Elongation factor 1 (EF-1) from the silk gland of Bombyx mori consists of alpha-, beta-, gamma-, and delta-subunits. EF-1alpha GTP catalyzes the binding of aminoacyl-tRNA to ribosomes concomitant with the hydrolysis of GTP. EF-1betagammadelta catalyzes the exchange of EF-1alpha-bound GDP for exogenous GTP and stimulates the EF-1alpha-dependent binding of aminoacyl-tRNA to ribosomes. EF-1gamma cDNA, which contains an open reading frame (ORF) encoding a polypeptide of 423 amino acid residues, was amplified and cloned by PCR from a silk gland cDNA library. The calculated molecular mass and predicted pI of the product were 48,388 Da and 5.84, respectively. The silk gland EF-1gamma shares 67.3% amino acid identity with Artemia salina EF-lgamma. The N-terminal domain (amino acid residues 1-211) of silk gland EF-lgamma is 29.3% identical to maize glutathione S-transferase. We demonstrated that silk gland EF-lgamma bound to glutathione Sepharose, suggesting that the N-terminal domain of EF-1gamma may have the capacity to bind to glutathione. PMID:12005049

  17. 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

  18. Silk fibroin nanoparticles constitute a vector for controlled release of resveratrol in an experimental model of inflammatory bowel disease in rats

    Lozano-Pérez, Antonio Abel; Rodriguez-Nogales, Alba; Ortiz-Cullera, Víctor; Algieri, Francesca; Garrido-Mesa, José; Zorrilla, Pedro; Rodriguez-Cabezas, M Elena; Garrido-Mesa, Natividad; Utrilla, M Pilar; De Matteis, Laura; de la Fuente, Jesús Martínez; Cenis, José Luis; Gálvez, Julio

    2014-01-01

    Purpose We aimed to evaluate the intestinal anti-inflammatory properties of silk fibroin nanoparticles, around 100 nm 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 264.7 cells (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 3 hours, which was maintained over a period of 80 hours. 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. PMID:25285004

  19. 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

  20. 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.

  1. Hox transcription factor Antp regulates sericin-1 gene expression in the terminal differentiated silk gland of Bombyx mori

    Kimoto, Mai; Tsubota, Takuya; Uchino, Keiro; Sezutsu, Hideki; Takiya, Shigeharu

    2014-01-01

    Hox genes are well-known master regulators in developmental morphogenesis along the anteroposterior axis of animals. However, the molecular mechanisms by which Hox proteins regulate their target genes and determine cell fates are not fully understood. The silk gland of Bombyx mori is a tubular tissue divided into several subparts along the anteroposterior axis, and the silk genes are expressed with specific patterns. The sericin-1 gene (ser1) is expressed in the middle silk gland (MSG) with s...

  2. 丝素蛋白-氧化石墨烯共混薄膜热处理红外光谱研究%Study on Structure Variation of Silk Fibroin-Graphene Oxide Composite Film During Heat Treatment by FTIR

    2015-01-01

    Recently, silk fibroin-graphene oxide composites were studied, which showed that graphene oxide can improve mechanical properties of silk fibroin greatly. In this research, Fourier transform infrared microspectroscopy was used for in-suit examination of the heat treatment process of the silk fibroin-graphene oxide composite film. Secondary structures of the pure silk fibroin film and the silk fibroin-graphene oxide composite film containing 10%graphene oxide before and after the heat treatment were analyzed. Effect of graphene oxide and heat treatment on secondary structures of silk fibroin was discussed.%近些年,人们开始对丝素蛋白-氧化石墨烯这一体系进行了研究,发现氧化石墨烯可以大大提高丝素蛋白的力学性能。在本实验中,利用显微红外对丝素蛋白-氧化石墨烯共混薄膜的热处理过程进行了在线研究。通过对丝素蛋白酰胺 I 区的分峰拟合,分析了热处理前后纯丝素蛋白薄膜和含有10%氧化石墨烯的丝素蛋白共混薄膜中丝素蛋白二级结构的转变,初步解释了共混薄膜中氧化石墨烯和热处理对丝素蛋白二级结构的影响。

  3. 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

  4. 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

  5. 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; Cantürk, M; Aksakal, B; Alekperov, V; Bülbül, 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

  6. 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

  7. 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.

  8. 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.

  9. 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

  10. 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

  11. 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

  12. Electrophoretic-deposited novel ternary silk fibroin/graphene oxide/hydroxyapatite nanocomposite coatings on titanium substrate for orthopedic applications

    Li, Ming; Xiong, Pan; Mo, Maosong; Cheng, Yan; Zheng, Yufeng

    2016-09-01

    The combination of graphene oxide (GO) with robust mechanical property, silk fibroin (SF) with fascinating biological effects and hydroxyapatite (HA) with superior osteogenic activity is a competitive approach to make novel coatings for orthopedic applications. Herein, the feasibility of depositing ternary SF/GO/HA nanocomposite coatings on Ti substrate was firstly verified by exploiting electrophoretic nanotechnology, with SF being used as both a charging additive and a dispersion agent. The surface morphology, microstructure and composition, in vitro hemocompatibility and in vitro cytocompatibility of the resulting coatings were investigated by SEM, Raman, FTIR spectra and biocompatibility tests. Results demonstrated that GO, HA and SF could be co-deposited with a uniform, smooth thin-film morphology. The hemolysis rate analysis and the platelet adhesion test indicated good blood compatibility of the coatings. The human osteosarcoma MG63 cells displayed well adhesion and proliferation behaviors on the prepared coatings, with enhanced ALP activities. The present study suggested that SF/GO/HA nanocomposite coatings could be a promising candidate for the surface functionalization of biomaterials, especially as orthopedic implant coating.

  13. Anti-EGFR-iRGD recombinant protein conjugated silk fibroin nanoparticles for enhanced tumor targeting and antitumor efficiency

    Bian, Xinyu; Wu, Puyuan; Sha, Huizi; Qian, Hanqing; Wang, Qing; Cheng, Lei; Yang, Yang; Yang, Mi; Liu, Baorui

    2016-01-01

    In this study, we report a novel kind of targeting with paclitaxel (PTX)-loaded silk fibroin nanoparticles conjugated with iRGD–EGFR nanobody recombinant protein (anti-EGFR-iRGD). The new nanoparticles (called A-PTX-SF-NPs) were prepared using the carbodiimide-mediated coupling procedure and their characteristics were evaluated. The cellular cytotoxicity and cellular uptake of A-PTX-SF-NPs were also investigated. The results in vivo suggested that NPs conjugated with the recombinant protein exhibited more targeting and anti-neoplastic property in cells with high EGFR expression. In the in vivo antitumor efficacy assay, the A-PTX-SF-NPs group showed slower tumor growth and smaller tumor volumes than PTX-SF-NPs in a HeLa xenograft mouse model. A real-time near-infrared fluorescence imaging study showed that A-PTX-SF-NPs could target the tumor more effectively. These results suggest that the anticancer activity and tumor targeting of A-PTX-SF-NPs were superior to those of PTX-SF-NPs and may have the potential to be used for targeted delivery for tumor therapies.

  14. Electrophoretic-deposited novel ternary silk fibroin/graphene oxide/hydroxyapatite nanocomposite coatings on titanium substrate for orthopedic applications

    Li, Ming; Xiong, Pan; Mo, Maosong; Cheng, Yan; Zheng, Yufeng

    2016-06-01

    The combination of graphene oxide (GO) with robust mechanical property, silk fibroin (SF) with fascinating biological effects and hydroxyapatite (HA) with superior osteogenic activity is a competitive approach to make novel coatings for orthopedic applications. Herein, the feasibility of depositing ternary SF/GO/HA nanocomposite coatings on Ti substrate was firstly verified by exploiting electrophoretic nanotechnology, with SF being used as both a charging additive and a dispersion agent. The surface morphology, microstructure and composition, in vitro hemocompatibility and in vitro cytocompatibility of the resulting coatings were investigated by SEM, Raman, FTIR spectra and biocompatibility tests. Results demonstrated that GO, HA and SF could be co-deposited with a uniform, smooth thin-film morphology. The hemolysis rate analysis and the platelet adhesion test indicated good blood compatibility of the coatings. The human osteosarcoma MG63 cells displayed well adhesion and proliferation behaviors on the prepared coatings, with enhanced ALP activities. The present study suggested that SF/GO/HA nanocomposite coatings could be a promising candidate for the surface functionalization of biomaterials, especially as orthopedic implant coating.

  15. Nature-Derived Aloe Vera Gel Blended Silk Fibroin Film Scaffolds for Cornea Endothelial Cell Regeneration and Transplantation.

    Kim, Do Kyung; Sim, Bo Ra; Khang, Gilson

    2016-06-22

    The goal of this study was to fabricate an appropriate replacement for cadaveric corneas to overcome a shortage of cadaveric corneas for transplantation. In this study, we fabricated transparent ultrathin film scaffolds with nature-derived aloe vera (AV) gel and silk fibroin (SF) for corneal endothelial cells (CECs). The scaffolds were subjected to analysis of transparency and contact angle using field emission scanning electron microscopy (FESEM) and Fourier transform infrared spectroscopy to determine their physical and chemical properties. FESEM images revealed that the critical morphology of CECs was formed on the AV gel in the blend with SF rather than in the scaffold with SF alone. The cell proliferation, phenotype, and specific gene marker expressions for CECs were determined by MTT assays, immunofluorescence, and reverse transcription polymerase chain reactions. Incorporation of a small amount of AV gel increased the cell viability and maintained its functions well. The scaffolds were easily handled for transplantation into rabbit eyes with small incisions and examined by their transparency after transplantation and histological staining. The scaffolds attached to the surface of the corneal stroma and integrated with surrounding corneal tissue without a significant inflammatory reaction. These results indicate that AV blended SF film scaffolds might be a suitable substitute for alternative corneal grafts for transplantation. PMID:27243449

  16. Electrospun and woven silk fibroin/poly(lactic-co- glycolic acid nerve guidance conduits for repairing peripheral nerve injury

    Ya-ling Wang

    2015-01-01

    Full Text Available We have designed a novel nerve guidance conduit (NGC made from silk fibroin and poly(lactic-co-glycolic acid through electrospinning and weaving (ESP-NGCs. Several physical and biological properties of the ESP-NGCs were assessed in order to evaluate their biocompatibility. The physical properties, including thickness, tensile stiffness, infrared spectroscopy, porosity, and water absorption were determined in vitro. To assess the biological properties, Schwann cells were cultured in ESP-NGC extracts and were assessed by morphological observation, the MTT assay, and immunohistochemistry. In addition, ESP-NGCs were subcutaneously implanted in the backs of rabbits to evaluate their biocompatibility in vivo. The results showed that ESP-NGCs have high porosity, strong hydrophilicity, and strong tensile stiffness. Schwann cells cultured in the ESP-NGC extract fluids showed no significant differences compared to control cells in their morphology or viability. Histological evaluation of the ESP-NGCs implanted in vivo indicated a mild inflammatory reaction and high biocompatibility. Together, these data suggest that these novel ESP-NGCs are biocompatible, and may thus provide a reliable scaffold for peripheral nerve repair in clinical application.

  17. Electrospun and woven silk fibroin/poly(lactic-co-glycolic acid) nerve guidance conduits for repairing peripheral nerve injury.

    Wang, Ya-Ling; Gu, Xiao-Mei; Kong, Yan; Feng, Qi-Lin; Yang, Yu-Min

    2015-10-01

    We have designed a novel nerve guidance conduit (NGC) made from silk fibroin and poly(lactic-co-glycolic acid) through electrospinning and weaving (ESP-NGCs). Several physical and biological properties of the ESP-NGCs were assessed in order to evaluate their biocompatibility. The physical properties, including thickness, tensile stiffness, infrared spectroscopy, porosity, and water absorption were determined in vitro. To assess the biological properties, Schwann cells were cultured in ESP-NGC extracts and were assessed by morphological observation, the MTT assay, and immunohistochemistry. In addition, ESP-NGCs were subcutaneously implanted in the backs of rabbits to evaluate their biocompatibility in vivo. The results showed that ESP-NGCs have high porosity, strong hydrophilicity, and strong tensile stiffness. Schwann cells cultured in the ESP-NGC extract fluids showed no significant differences compared to control cells in their morphology or viability. Histological evaluation of the ESP-NGCs implanted in vivo indicated a mild inflammatory reaction and high biocompatibility. Together, these data suggest that these novel ESP-NGCs are biocompatible, and may thus provide a reliable scaffold for peripheral nerve repair in clinical application. PMID:26692862

  18. 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.

  19. 多孔丝素材料组织相容性的初步研究%A preliminary study on the histocompatibility of silk fibroin

    陆艳; 赵霞; 邵正中; 曹正兵; 蔡丽慧

    2011-01-01

    背景:丝素蛋白支架材料被植入生物体内后会发生降解且无法完全与宿主组织分离,这类材料生物相容性的研究大多为体外实验,其体内的组织相容性和降解过程的研究结果仍不充分.目的:初步观察多孔丝素材料的体内组织相容性.方法:将多孔丝素支架埋藏于SD大鼠背部皮下,术后2,4,6,8周分别取材,对伤口局部及材料情况大体观察,然后材料切片苏木精-伊红染色行组织学观察.结果与结论:动物伤口愈合良好,多孔丝素表面形成极薄的纤维包裹,周围组织反应轻微.组织切片见炎细胞浸润,以巨噬细胞为主,支架材料边缘孔隙内有成纤维细胞和毛细血管长入.8周时材料边缘部分可见支架结构崩解现象,而材料内部变化不大.结果显示组织细胞可以沿多孔丝素支架表面贴附生长,提示支架材料具有较好的组织相容性.%BACKGROUND: As one kind of naturally occurring polymers, silk fi broin has been widely used in tissue engeering research,which can be made into different forms for application. Scientists find many difficulties in histocompatibility study of silk fibroin,because it is made of protein and biodegradable. And there are few reports on the study on histocompatibility of silk fibroin.OBJECTIVE: To perform a preliminary in vivo study on the histocompatibility of silk fibroin.METHODS: Porous silk fibroin scaffolds were buried into the subcutaneous part of the back of SD rats. At 4, 6, 8 weeks afteroperation, general observation and histological examination were performed.RESULTS AND CONCLUSION: The wounds healed up well after operation. Only thin connective tissue wrap was found on silkfibroin scaffold. Macrophages were observed in tissue section, and fibroblasts and capillary vessels could grow into the porespaces. We found the border of the scaffolds break down at 8 weeks postoperatively, while the inside part remained as it was.Porous silk fibroin scaffold shows

  20. 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.

  1. 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)

  2. Entrapment of both glucose oxidase and peroxidase in regenerated silk fibroin membraneCharacterization of the membrane structure and its application to an amperometric glucose sensor employing methylene green as an electron transfer mediato.

    Liu, Y; Liu, H; Qian, J; Deng, J; Yu, T

    1996-04-01

    Two enzmyes, glucose oxidase and peroxidase, were for the first time simultaneously immobilized in regenerated silk fibroin membrane. The structure and morphology of the regenerated silk fibroin membrane containing both glucose oxidase and peroxidase were investigated with IR spectra and SEM. The bienzymes do not change the structures of the regenerated silk fibroin in the membrane, which has an islands-sea structure. For the first time, an amperometric methylene green mediating sensor for glucose based on co-immobilization of both glucose oxidase and peroxidase in regenerated silk fibroin was constructed. Cyclic voltammetry and amperometry were used to test the suitability of methylene green shuttling electrons between peroxidase and the glassy carbon electrode. The bienzyme-based system offers fast response and high sensitivity of the sensor to glucose. The effects of pH, temperature, and the concentration of the mediator on the response current were evaluated, and the dependence of the Michaelis-Menten constant K(m)(app) on the concentration of the mediator was investigated. PMID:15045461

  3. A biomimetic multilayer nanofiber fabric fabricated by electrospinning and textile technology from polylactic acid and Tussah silk fibroin as a scaffold for bone tissue engineering.

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

    2016-10-01

    To engineer bone tissue, a scaffold with good biological properties should be provided to approximate the hierarchical structure of collagen fibrils in natural bone. In this study, we fabricated a novel scaffold consisting of multilayer nanofiber fabrics (MLNFFs) by weaving nanofiber yarns of polylactic acid (PLA) and Tussah silk fibroin (TSF). The yarns were fabricated by electrospinning, and we found that spinnability, as well as the mechanical properties of the resulting scaffold, was determined by the ratio between polylactic acid and Tussah silk fibroin. In particular, a 9:1 mixture can be spun continuously into nanofiber yarns with narrow diameter distribution and good mechanical properties. Accordingly, woven scaffolds based on this mixture had excellent mechanical properties, with Young's modulus 417.65MPa and tensile strength 180.36MPa. For nonwoven scaffolds fabricated from the same materials, the Young's modulus and tensile strength were 2- and 4-fold lower, respectively. Woven scaffolds also supported adhesion and proliferation of mouse mesenchymal stem cells, and promoted biomineralization via alkaline phosphatase and mineral deposition. Finally, the scaffolds significantly enhanced the formation of new bone in damaged femoral condyle in rabbits. Thus, the scaffolds are potentially suitable for bone tissue engineering because of biomimetic architecture, excellent mechanical properties, and good biocompatibility. PMID:27287159

  4. 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.

  5. 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

  6. 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)

  7. 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)

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

    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.

  9. 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 numerou...

  10. When inordinate tissue growth is beneficial: Improving silk production by increasing silk gland size

    Xavier Bellés

    2011-01-01

    @@ Some 5000 years ago, the legendary Chinese emperor Huang-Ti ordered his wife, Hsi-Ling-Shi, to find out what was damaging his mulberry trees.HsiLing-Shi found white worms eating the mulberry leaves and spinning shiny cocoons.She accidentally dropped a cocoon into her tea cup and saw a delicate, slender thread unwind itself from the cocoon.The young princess had discovered silk [1, 2].

  11. Comparative transcriptome analyses on silk glands of six silkmoths imply the genetic basis of silk structure and coloration

    Dong, Yang; Dai, Fangyin; Ren, Yandong; Liu, Hui; Chen, Lei; Yang, Pengcheng; Liu, Yanqun; LI, XIN; Wang, Wen; Xiang, Hui

    2015-01-01

    Background Silk has numerous unique properties that make it a staple of textile manufacturing for several thousand years. However, wider applications of silk in modern have been stalled due to limitations of traditional silk produced by Bombyx mori. While silk is commonly produced by B. mori, several wild non-mulberry silkmoths--especially members of family Saturniidae--produce silk with superior properties that may be useful for wider applications. Further utilization of such silks is hamper...

  12. Fabrication of freestanding silk fibroin films containing Ag nanowires/NaYF4:Yb,Er nanocomposites with metal-enhanced fluorescence behavior.

    Zhao, Bing; Qi, Ning; Zhang, Ke-Qin; Gong, Xiao

    2016-06-01

    Solar cells containing upconversion nanoparticles (UCNPs) used as a power source in biomedical nanosystems have attracted great interest. However, such solar cells further need to be developed because their substrate materials should be biocompatible, flexible and highly luminescent. Here, we report that freestanding silk fibroin (SF) films containing a mesh of silver nanowires (AgNWs) and β-NaYF4:Yb,Er nanocrystals with metal-enhanced fluorescence behavior can be fabricated. The freestanding composite films exhibit properties such as good optical transparency, conductivity and flexibility. Furthermore, they show significantly enhanced upconversion fluorescence due to surface plasmon polaritons (SPPs) of AgNWs compared to the SF-UCNP films without AgNWs. The freestanding composite films with metal-enhanced fluorescence behavior show great promise for future applications in self-powered nanodevices such as cardiac pacemakers, biosensors and nanorobots. PMID:27210511

  13. Biocompatibility of plasma-treated poly(3-hydroxybutyrate-co-3-hydroxyvalerate) nanofiber mats modified by silk fibroin for bone tissue regeneration.

    Unalan, Irem; Colpankan, Oylum; Albayrak, Aylin Ziylan; Gorgun, Cansu; Urkmez, Aylin Sendemir

    2016-11-01

    The objective of this study was to produce biocompatible plasma-treated and silk-fibroin (SF) modified poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) nanofiber mats. The mats were plasma-treated using O2 or N2 gas to increase their hydrophilicity followed by SF immobilization for the improvement of biocompatibility. Contact angle measurements and SEM showed increased hydrophilicity and no disturbed morphology, respectively. Cell proliferation assay revealed that SF modification together with N2 plasma (PS/N2) promoted higher osteoblastic (SaOs-2) cell viability. Although, O2 plasma triggered more mineral formation on the mats, it showed poor cell viability. Consequently, the PS/N2 nanofiber mats would be a potential candidate for bone tissue engineering applications. PMID:27524087

  14. 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

  15. 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.

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

    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 Cu2+ 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

  17. Silk fibroin nanoparticles constitute a vector for controlled release of resveratrol in an experimental model of inflammatory bowel disease in rats

    Lozano-Pérez AA

    2014-09-01

    Full Text Available Antonio Abel Lozano-Pérez,1 Alba Rodriguez-Nogales,2 Víctor Ortiz-Cullera,1 Francesca Algieri,2 José Garrido-Mesa,2 Pedro Zorrilla,2 M Elena Rodriguez-Cabezas,2 Natividad Garrido-Mesa,2 M Pilar Utrilla,2 Laura De Matteis,3 Jesús Martínez de la Fuente,3 José Luis Cenis,1 Julio Gálvez2 1Instituto Murciano de Investigación y Desarrollo Agrario y Alimentario, Murcia, Spain; 2Centro de Investigaciones Biomédicas en Red – Enfermedades Hepáticas y Digestivas, Department of Pharmacology, ibs Granada, Center for Biomedical Research, University of Granada, Granada, Spain; 3Instituto de Nanociencia de Aragón, Universidad de Zaragoza, Zaragoza, Spain Purpose: We aimed to evaluate the intestinal anti-inflammatory properties of silk fibroin nanoparticles, around 100 nm 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 264.7 cells (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 3 hours, which was maintained over a period of 80 hours. 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

  18. 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

  19. Progress of electrospun silk fibroin based scaffolds for tissue engineering%静电纺丝素蛋白及其应用于组织工程的研究进展

    黄继伟; 张锋; 左保齐

    2011-01-01

    The researches related to electrospinning of silk, including electrospim solvents, blends, and electrospin device are reviewed, and the application of electrospun silk fibroin based scaffolds in tissue engineering is introduced.%从丝素蛋白静电纺丝的溶剂开发、共混纺丝及纺丝装置3个方面回顾了丝素蛋白静电纺丝的研究进展,重点介绍了静电纺丝素蛋白微纳米纤维支架材料在组织工程领域的应用研究.

  20. 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)

  1. 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 ...

  2. Recognition of signal peptide by protein translocation machinery in middle silk gland of silkworm Bombyx mori

    Xiuyang Guo; Yi Zhang; Xue Zhang; Shengpeng Wang; Changde Lu

    2008-01-01

    To investigate the functions of signal peptide in protein secretion in the middle silk gland of silkworm Bombyx mori,a series of recombinant Autographa californica multiple nucleopolyhedroviruses containing enhanced green fluorescent protein (egfp) gene,led by sericin-1 promoter and mutated signal peptide coding sequences,were constructed by region-deletions or single amino acid residue deletions.The recombinant Autographa californica multiple nucleopolyhedroviruses were injected into the hemocoele of newly ecdysed fifth-instar silkworm larvae.The expression and secretion of EGFP in the middle silk gland were examined by fluorescence microscopy and Western blot analysis.Results showed that even with a large part (up to 14 amino acid residues) of the ser-1 signal peptide deleted,the expressed EGFP could still be secreted into the cavity of the silk gland.Western blot analysis showed that shortening of the signal peptide from the C-terminal suppressed the maturation of pro-EGFP to EGFP.When 8 amino acid residues were deleted from the C-terminal of the signal peptide (mutant 13 aa),the secretion of EGFP was incomplete,implicating the importance of proper coupling of the h-region and c-region.The deletion of amino acid residue(s) in the h-region did not affect the secretion of EGFP,indicating that the recognition of signal peptide by translocation machinery was mainly by a structural domain,but not by special amino acid residue(s).Furthermore,the deletion of Arg2 or replacement with Asp in the n-region of the signal peptide did not influence secretion of EGFP,suggesting that a positive charge is not crucial.

  3. Some autophagic and apoptotic features of programmed cell death in the anterior silk glands of the silkworm, Bombyx mori.

    Goncu, Ebru; Parlak, Osman

    2008-11-01

    Programmed cell death has been subdivided into two major groups: apoptosis and autophagic cell death. The anterior silk gland of Bombyx mori degenerates during larval-pupal metamorphosis. Our findings indicate that two types of programmed cell death features are observed during this physiological process. During the prepupal period, pyknosis of the nucleus, cell detachment,and membrane blebbing occur and they are the first signs of programmed cell death in the anterior silk glands. According to previous studies, all of these morphological appearances are common for both cell-death types. Autophagy features are also exhibited during the prepupal period. Levels of one of the lysosomal marker enzymes-acid phosphatase-are high during this period then decrease gradually. Vacuole formation begins to appear first at the basal surface of the cell, then expands to the apical surface just before the larval pupal ecdysis. After larval-pupal ecdysis, DNA fragmentation, which is the obvious biochemical marker of apoptosis, is detected in agarose gel electrophoresis, which also shows that caspase-like enzyme activities occur during the programmed cell death process of the anterior silk glands. Apoptosis and autophagic cell death interact with each other during the degeneration process of the anterior silk gland in Bombyx mori and this interaction occurs at a late phase of cell death. We suggest that apoptotic cell death only is not enough for whole gland degeneration and that more effective degeneration occurs with this cooperation. PMID:18838861

  4. 丝素共混膜在化妆品保湿效果评价中的应用初探%The Preliminary Study on Application of Silk Fibroin in Moisturizing Efficacy Evaluation of Cosmetics

    李晓芹; 郑利; 倪梦嘉

    2013-01-01

    MMV and TEWL on sile fibroin were applied for assessing the moisturizing efficacy of cosmetics with different glycerol content cosmetics, which were determined by skin moisture content tester and water loss tester, respectively. Instead of human skin, silk fibroin was utilized in moisturizing efficacy evaluation of cosmetics. The purpose of this experiment was to explore the value of of silk fibroin in moisturizing efficacy evaluation of cosmetics and give a guidance on the appropriate percentage of moisturizing ingredients. The results showed that the silk fibroin model system and human body studies have similar trends, which provides the possibility of surrogate experimental evaluation for cosmetic moisturizing effect.%利用皮肤水分含量测试仪和水分散失测定仪,对添加不同质量分数甘油的保湿化妆品作用前后丝素共混膜的水分含量值(MMV)和经表皮水分散失值(TEWL)进行测定,以人体试验为参比,创造性地探索丝素共混膜在化妆品保湿效果评价中的应用价值,并对保湿类化妆品中保湿剂的最佳添加量做出指导.结果发现,丝素共混膜模拟体系的试验结果和人体试验具有相似的趋势,以其作为化妆品保湿效果评价的人体替代试验具有一定的可能性.

  5. 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

  6. Effect of Processing on Silk-Based Biomaterials: Reproducibility and Biocompatibility

    Wray, Lindsay S.; Hu, Xiao; Gallego, Jabier; Georgakoudi, Irene; Omenetto, Fiorenzo G.; Schmidt, Daniel; Kaplan, David L.

    2011-01-01

    Silk fibroin has been successfully used as a biomaterial for tissue regeneration. In order to prepare silk fibroin biomaterials for human implantation a series of processing steps are required to purify the protein. Degumming to remove inflammatory sericin is a crucial step related to biocompatibility and variability in the material. Detailed characterization of silk fibroin degumming is reported. The degumming conditions significantly affected cell viability on the silk fibroin material and ...

  7. 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

  8. 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%.

  9. 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.

  10. 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%.

  11. Porous nitrogen-doped carbon derived from silk fibroin protein encapsulating sulfur as a superior cathode material for high-performance lithium-sulfur batteries

    Zhang, Jiawei; Cai, Yurong; Zhong, Qiwei; Lai, Dongzhi; Yao, Juming

    2015-10-01

    The features of a carbon substrate are crucial for the electrochemical performance of lithium-sulfur (Li-S) batteries. Nitrogen doping of carbon materials is assumed to play an important role in sulfur immobilisation. In this study, natural silk fibroin protein is used as a precursor of nitrogen-rich carbon to fabricate a novel, porous, nitrogen-doped carbon material through facile carbonisation and activation. Porous carbon, with a reversible capacity of 815 mA h g-1 at 0.2 C after 60 cycles, serves as the cathode material in Li-S batteries. Porous carbon retains a reversible capacity of 567 mA h g-1, which corresponds to a capacity retention of 98% at 1 C after 200 cycles. The promising electrochemical performance of porous carbon is attributed to its mesoporous structure, high specific surface area and nitrogen doping into the carbon skeleton. This study provides a general strategy to synthesise nitrogen-doped carbons with a high specific surface area, which is crucial to improve the energy density and electrochemical performance of Li-S batteries.

  12. 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.

  13. Influence of the mechanical environment on the engineering of mineralised tissues using human dental pulp stem cells and silk fibroin scaffolds.

    Anna Woloszyk

    Full Text Available Teeth constitute a promising source of stem cells that can be used for tissue engineering and regenerative medicine purposes. Bone loss in the craniofacial complex due to pathological conditions and severe injuries could be treated with new materials combined with human dental pulp stem cells (hDPSCs that have the same embryonic origin as craniofacial bones. Optimising combinations of scaffolds, cells, growth factors and culture conditions still remains a great challenge. In the present study, we evaluate the mineralisation potential of hDPSCs seeded on porous silk fibroin scaffolds in a mechanically dynamic environment provided by spinner flask bioreactors. Cell-seeded scaffolds were cultured in either standard or osteogenic media in both static and dynamic conditions for 47 days. Histological analysis and micro-computed tomography of the samples showed low levels of mineralisation when samples were cultured in static conditions (0.16±0.1 BV/TV%, while their culture in a dynamic environment with osteogenic medium and weekly µCT scans (4.9±1.6 BV/TV% significantly increased the formation of homogeneously mineralised structures, which was also confirmed by the elevated calcium levels (4.5±1.0 vs. 8.8±1.7 mg/mL. Molecular analysis of the samples showed that the expression of tooth correlated genes such as Dentin Sialophosphoprotein and Nestin were downregulated by a factor of 6.7 and 7.4, respectively, in hDPSCs when cultured in presence of osteogenic medium. This finding indicates that hDPSCs are able to adopt a non-dental identity by changing the culture conditions only. Also an increased expression of Osteocalcin (1.4x and Collagen type I (1.7x was found after culture under mechanically dynamic conditions in control medium. In conclusion, the combination of hDPSCs and silk scaffolds cultured under mechanical loading in spinner flask bioreactors could offer a novel and promising approach for bone tissue engineering where appropriate and

  14. 酶解丝素蛋白制备ACE抑制肽的研究%Enzymatic Hydrolysis of Silk Fibroin for Production of ACE Inhibitors

    吴晖; 罗美琪; 唐语谦; 肖性龙; 袁坤; 李晓凤

    2011-01-01

    Silk fibroin was hydrolyzed by Alcalase to produce the peptide with high ACE-inhibition activity. The effects of several influential factors on the enzymatic hydrolysis were investigated and the optimum reaction conditions were determined as follows: pH 8.5, temperature 60 "C, substrate concentration 5%, enzyme dosage 1000 U/g protein, and reaction time 240 min, under which the DH. ACE-inhibition rate, peptide yield and protein recovery of the product were 17.12%, 74.07%, 50.31% and 77.37%, respectively. MALDI-TOF mass spectrometry of the hydrolysate showed that the molecular weight of (he peptides was below 2.4kDa.%本研究首次探讨利用Alcalase酶解丝素蛋白制备高活性ACE抑制肽.以水解度为主要评价指标,研究了温度、浓度、pH、加酶量以及反应时间对酶解反应的影响.分析了不同水解度下的酶解产物与ACE体外抑制活性、肽得率和蛋白回收率的关系.确定了制备ACE抑制肽的最佳酶解反应DH值为17.12%,此时所得丝素蛋白肽的肽得率为50.31%,蛋白回收率为77.37%,ACE抑制率可达74.07%.基质辅助激光解吸电离-飞行时间质谱仪(MALDI-TOF)分析表明,所制备的丝素蛋白肽的分子量小于2.4 kDa.

  15. Antheraea pernyi Silk Fibroin-Coated PEI/DNA Complexes for Targeted Gene Delivery in HEK 293 and HCT 116 Cells

    Yu Liu

    2014-04-01

    Full Text Available Polyethylenimine (PEI has attracted much attention as a DNA condenser, but its toxicity and non-specific targeting limit its potential. To overcome these limitations, Antheraea pernyi silk fibroin (ASF, a natural protein rich in arginyl-glycyl-aspartic acid (RGD peptides that contains negative surface charges in a neutral aqueous solution, was used to coat PEI/DNA complexes to form ASF/PEI/DNA ternary complexes. Coating these complexes with ASF caused fewer surface charges and greater size compared with the PEI/DNA complexes alone. In vitro transfection studies revealed that incorporation of ASF led to greater transfection efficiencies in both HEK (human embryonic kidney 293 and HCT (human colorectal carcinoma 116 cells, albeit with less electrostatic binding affinity for the cells. Moreover, the transfection efficiency in the HCT 116 cells was higher than that in the HEK 293 cells under the same conditions, which may be due to the target bonding affinity of the RGD peptides in ASF for integrins on the HCT 116 cell surface. This result indicated that the RGD binding affinity in ASF for integrins can enhance the specific targeting affinity to compensate for the reduction in electrostatic binding between ASF-coated PEI carriers and cells. Cell viability measurements showed higher cell viability after transfection of ASF/PEI/DNA ternary complexes than after transfection of PEI/DNA binary complexes alone. Lactate dehydrogenase (LDH release studies further confirmed the improvement in the targeting effect of ASF/PEI/DNA ternary complexes to cells. These results suggest that ASF-coated PEI is a preferred transfection reagent and useful for improving both the transfection efficiency and cell viability of PEI-based nonviral vectors.

  16. Transcription modulation in vitro of the fibroin gene exerted by a 200-base-pair region upstream from the "TATA" box.

    Tsuda, M; Suzuki, Y

    1983-01-01

    We have previously reported that the 5'-flanking sequence upstream from the "TATA" box modulates the faithful transcription initiation of the fibroin gene in a homologous whole cell extract prepared from the silk glands, whereas such a modulating effect is not observed in a HeLa cell extract. Subsequently we have determined that major signals responsible for the modulating effect are located within a 200-base-pair region upstream from the TATA box, mainly in a distal region between nucleotide...

  17. 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

    2016-02-01

    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

  18. 气流辅助高压静电纺丝法制备蚕丝蛋白纳米纤维的研究%The Preparation of Silk-fibroin Nano-fibers with Air-assisted Electrospinning

    李罡; 李艳红; 刘娟; 杜江; 姚勇毅

    2011-01-01

    目的 研究蚕丝丝素蛋白亚微米和纳米纤维二者在超细纤维加工方面的特点.方法 通过高压静电纺丝和气流辅助高压静电纺丝工艺制备蚕丝蛋白亚微米、纳米纤维.结果 当气体流量达到4L/min时,气流辅助高压静电纺丝可以制的更细的纳米纤维,结论 气流辅助高压静电纺丝工艺有助于获得更细的纳米纤维.%Objective To compare the difference between electrospinning and air-assisted electrospinning in preparation of sub-micron and Nano silk-fibroin fibers. Method The silk-fibroin fibers were prepared with electrospinning and air-assisted electrospinning. Result The thinner fibers could be obtained by air-assisted electrnapinning, when air flow rate was increased to 4 L/min. Conclusion With the air-assisted electrospinning, the thinner fibers can be prepared, compared with electrospinning.

  19. Effect of Silk Protein Processing on Drug Delivery from Silk Films

    Pritchard, Eleanor M.; Hu, Xiao; Finley, Violet; Kuo, Catherine K.; Kaplan, David L.

    2013-01-01

    Sericin removal from the core fibroin protein of silkworm silk is a critical first step in the use of silk for biomaterial-related applications, but degumming can affect silk biomaterial properties, including molecular weight, viscosity, diffusivity and degradation behavior. Increasing the degumming time (10, 30, 60 and 90 min) decreases the average molecular weight of silk protein in solution, silk solution viscosity, and silk film glass transition temperature, and increases the rate of degr...

  20. Screen-Printing Enzyme Electrode-Immobilizing the Enzyme with Silk Fibroin%丝素膜固定化丝网印刷酶电极

    屠一锋; 付志强; 李明忠

    2001-01-01

    This paper reports the technique to develop a disposable screen-printing base carbon electrode for the utilization in the study of enzyme electrode. The silk fibroin was applied to immobilize the mushroom extracted protein, which contained abundant polyphenol oxidase, on the surface of base electrode when it was chemically moldified by ferrocene. The voltammetric current of this enzyme electrode responded to the concentration of substrate such as catechol or dopamine. The linear range is from 2.0× 10-8 mol/L to 2.0× 10-2 mol/L, it reaches to a 95%steady value of current within less than 30 seconds, and the RSD equals to 2.7%. The service life of this enzyme electrode is at least 10 days and it will be preserved for a longer period at a humid and cool condition. All of these performances of the enzyme electrode make clear that its commercial development would be very possible.%采用丝网印刷技术制作的一次性薄层碳电极具有良好的分析性能。经化学修饰二茂铁后,以丝素为固定化载体固定含有多酚氧化酶的蘑菇萃取蛋白,所得酶电极对其底物邻苯二酚和多巴胺有良好响应,线性范围为2.0×10-8~2.0 × 10-2mol/L,在30s内即可达到95%稳态电流。该电极的RSD为2.7%,且可保持10天以上的工作寿命。该酶电极在低温湿润环境中可保存较长时间,因此有较好的商品化前景。

  1. 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

  2. Variation and Characterization Analysis of Partial Fragment of Fibroin Gene From Silkworm, Antheraea pernyi

    Li Wenli(李文利); Jin Liji; An Lijia

    2003-01-01

    A 1.4Kb DNA fragment containing 3' flanking sequence of fibroin gene of silkworm, Antheraea pernyi, was obtained from the silk gland's mRNA of 5th larva. Analysis of this sequence with another A.pernyi fibroin protein (accession No. D83241) revealed that it consists of a completely open reading frame (ORF), which includes 14 polyalanine-containing units (motifs) and 100bp 3'-UTR. The sequence of the predicted amino acid reveals the highest level of overall identity (90%) with D83241. It was found that it loses a repeat region at the upstream of TAA codon and some mutations. A putative polyadenylation signal AATAAA tail was found in position 1300, which follows the termination codon.

  3. Preparation and Properties of the Blended Films of Polyurethane and Silk Fibroin Powder by the Method of Sol Mixing-Hot Pressure%丝素粉体与医用聚氨酯溶胶共混-热压共混膜的制备及性能

    鄢芸; 陶咏真; 徐卫林; 李利燕

    2011-01-01

    采用溶胶共混-热压法成功制备出一系列丝素粉体/聚氨酯共混膜。利用扫描电子显微镜、红外光谱、X射线衍射、动态力学分析和拉伸实验对共混膜的形貌、结构和性能进行了表征,详细研究了丝素含量对共混膜形貌、结构及性能的影响。结果表明,聚氨酯和丝素粉体间存在一定的氢键相互作用,丝素小颗粒均匀分散在聚氨酯中,形成结构较致密的共混膜。溶胶共混-热压成膜法能够较好地提高二者的共混相容性,其工作提供了一种制备丝素粉体/聚氨酯生物医用材料的新途径。%The blended films of polyurethane and Silk fibroin powder(PU/SF) were successfully prepared by sol mixture-hot pressure method.The effects of the Silk fibroin ratio on the morphology、structure and properties of the blends were studied by scanning electron microscopy,infrared spectroscopy,wide-angle X-ray diffraction,dynamic mechanical thermal analysis,and tensile testing.The results reveal that intermolecular hydrogen bonding exists between Silk fibroin and polyurethane,leading to a homogeneous dispersion of Silk powder into polyurethane and a denser structure of blends.The method of sol mixture-hot pressure can better increase the compatibility of Silk fibroin and polyurethane in the blended films.This work provided a new pathway to prepare biomaterials made of polyurethane and Silk fibroin powder.

  4. Structure, evolution, and expression of antimicrobial silk proteins, seroins in Lepidoptera.

    Dong, Zhaoming; Song, Qianru; Zhang, Yan; Chen, Shiyi; Zhang, Xiaolu; Zhao, Ping; Xia, Qingyou

    2016-08-01

    The silks of silkworm and waxworm contain abundant antimicrobial proteins, including protease inhibitors and seroins. Protease inhibitors have antifungal activities, whereas seroins have antiviral and antibacterial activities. In order to obtain insights into the structure, evolution, and expression of seroins, we performed an extensive survey based on the available genome, transcriptome, and expressed sequence tags datasets. Sixty-four seroins were identified in 32 lepidopteran species. The phylogenetic and structural analyses revealed that seroins can be classified into five subfamilies: seroin 1, seroin 2, seroin 3, seroin 2 + 1, and seroin 3 + 3. It is interesting that seroin 2 + 1 contains two tandem seroin domains, seroin 2 and seroin 1, whereas seroin 3 + 3 has two tandem seroin 3 domains. Each seroin domain contains a proline-rich N-terminal motif and a conserved C-terminal motif. The transcriptome and EST data indicated that seroin 1 and seroin 2 genes were expressed in the silk gland but seroin 3 genes were not. Semi-quantitative RT-PCR and western blot analyses suggested that seroin 1 and seroin 2 were constantly accumulated in the silk gland of silkworm during the fifth instar, and then secreted into cocoon silk during spinning. Immunofluorescence analyses indicated that seroin 1 was secreted into the fibroin and sericin layers, whereas seroin 2 protein was only secreted into the sericin layer. However, the antimicrobial activity of seroin 2 was more effective than that of seroin 1. The presence of seroin 1 in the fibroin layer suggested that this protein not only acts as an antimicrobial protein, but might also play a role in the assembly and secretion of fibroins. Seroin 3, which was first identified here, might be related to pheromone synthesis or recognition, as it was highly expressed in male antennae and in the pheromone gland. PMID:27180727

  5. Effects of Pectin, Xanthan Gum, Sodium Alginate and Acid-resistant CMC on Lactobacillus Contents of Silk Fibroin Yoghurt%果胶、黄原胶、海藻酸钠、耐酸性CMC对丝素酸奶乳酸菌数的影响

    刘冠卉; 屠洁; 燕薇

    2012-01-01

    采用两因素三水平析因试验设计,考察了4种常用的酸奶稳定剂(果胶、黄原胶、海藻酸钠、耐酸性CMC)对搅拌型丝素酸奶乳酸菌数的影响.结果显示:果胶、黄原胶、海藻酸钠、耐酸性CMC对丝素酸奶的乳酸菌数均有极显著的影响(P<0.01),其中黄原胶、海藻酸钠和丝素对乳酸菌数有极显著的交互作用(P<0.01),果胶和丝素对乳酸菌含量有显著的交互作用(P<0.05),耐酸性CMC和丝素对酸奶的乳酸菌数无显著的交互作用(P>0.05).4种稳定剂中,黄原胶和海藻酸钠可提高搅拌型丝素酸奶的乳酸菌数,其中添加1%丝素和0.03%海藻酸钠的酸奶的乳酸菌数最高,达到1.20×1011 mL-1;其次为添加1%丝素和0.05%黄原胶的酸奶,乳酸菌数为1.07×1011mL-1.%Two factors and three levels factorial experiment design was used to study the effects of pectin, xanthan gum, sodium alginate, acidresistant carboxymethyl-cellulose on the lactobacillus contents of silk fibroin stirred yoghurt. Results showed pectin, xanthate gum, sodium alginate and acid-resistant carboxymethyl-cellulose had significant effects on lactobacillus contents of silk fibroin yogurt (P0.05). About three four stabilizers, xanthan gum and sodium alginate raised lactobacillus contents of silk fibroin stirred yoghurt, and the lactobacillus contents of stirred yoghurt including l%silk fibroin and 0.03% sodium alginate were highest(1.20×l011 mL-1), then that of the samples(l% silk fibroin and 0.05% xanthate gum) were 1.07× 1011 mL-1.

  6. Silk structure and degradation.

    Liu, Bin; Song, Yu-wei; Jin, Li; Wang, Zhi-jian; Pu, De-yong; Lin, Shao-qiang; Zhou, Chan; You, Hua-jian; Ma, Yan; Li, Jin-min; Yang, Li; Sung, K L Paul; Zhang, Yao-guang

    2015-07-01

    To investigate the structure of silk and its degradation properties, we have monitored the structure of silk using scanning electron microscopy and frozen sections. Raw silk and degummed raw silk were immersed in four types of degradation solutions for 156 d to observe their degradation properties. The subcutaneous implants in rats were removed after 7, 14, 56, 84, 129, and 145 d for frozen sectioning and subsequent staining with hematoxylin and eosin (H.E.), DAPI, Beta-actin and Collagen I immunofluorescence staining. The in vitro weight loss ratio of raw silk and degummed raw silk in water, PBS, DMEM and DMEM containing 10% FBS (F-DMEM) were, respectively, 14%/11%, 12.5%/12.9%, 11.1%/14.3%, 8.8%/11.6%. Silk began to degrade after 7 d subcutaneous implantation and after 145 d non-degraded silk was still observed. These findings suggest the immunogenicity of fibroin and sericin had no essential difference. In the process of in vitro degradation of silk, the role of the enzyme is not significant. The in vivo degradation of silk is related to phagocytotic activity and fibroblasts may be involved in this process to secrete collagen. This study also shows the developing process of cocoons and raw silk. PMID:25982316

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

    2009-01-01

    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 μg/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 5×105 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.

  8. 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.

  9. Sericin Composition in the Silk of Antheraea yamamai.

    Zurovec, Michal; Yonemura, Naoyuki; Kludkiewicz, Barbara; Sehnal, František; Kodrik, Dalibor; Vieira, Ligia Cota; Kucerova, Lucie; Strnad, Hynek; Konik, Peter; Sehadova, Hana

    2016-05-01

    The silks produced by caterpillars consist of fibroin proteins that form two core filaments, and sericin proteins that seal filaments into a fiber and conglutinate fibers in the cocoon. Sericin genes are well-known in Bombyx mori (Bombycidae) but have received little attention in other insects. This paper shows that Antheraea yamamai (Saturniidae) contains five sericin genes very different from the three sericin genes of B. mori. In spite of differences, all known sericins are characterized by short exons 1 and 2 (out of 3-12 exons), expression in the middle silk gland section, presence of repeats with high contents of Ser and charged amino acid residues, and secretion as a sticky silk component soluble in hot water. The B. mori sericins represent tentative phylogenetic lineages (I) BmSer1 and orthologs in Saturniidae, (II) BmSer2, and (III) BmSer3 and related sericins of Saturniidae and of the pyralid Galleria mellonella. The lineage (IV) seems to be limited to Saturniidae. Concerted evolution of the sericin genes was apparently associated with gene amplifications as well as gene loses. Differences in the silk fiber morphology indicate that the cocktail of sericins linking the filaments and coating the fiber is modified during spinning. Silks are composite biomaterials of conserved function in spite of great diversity of their composition. PMID:27049111

  10. 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)

  11. 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)

  12. 4种常用稳定剂和丝素蛋白对搅拌型酸奶黏度和保水性的影响%Effects of Four Common Stabilizers Combined with Silk Fibroin on Viscosity and Water-Holding Capacity of Stirred Yogurt

    屠洁; 刘冠卉; 燕薇

    2012-01-01

    为改善丝素酸奶的品质,采用二因素三水平析因试验设计,考察4种常用稳定剂(果胶、黄原胶、海藻酸钠、耐酸性羧甲基纤维素钠(CMC))与丝素蛋白对搅拌型酸奶黏度、保水性的影响。结果显示:果胶、黄原胶、海藻酸钠和丝素蛋白对搅拌型酸奶的黏度均有极显著的影响(P〈0.01),上述3种稳定剂和丝素蛋白对酸奶的黏度均有极显著的交互作用(P〈0.01);果胶、黄原胶、耐酸性CMC和丝素蛋白对酸奶的保水性均有极显著的影响(P〈0.01),其中黄原胶和丝素蛋白对酸奶的保水性有极显著的交互作用(P〈0.01)。获得4个优化组合,分别为0.5%丝素蛋白和0.01%果胶,酸奶的黏度和保水性分别为903.8mPa.s、65.5%;1%丝素蛋白和0.03%黄原胶,酸奶的黏度和保水性分别为1196.2mPa.s、68.0%;1%丝素蛋白和0.03%海藻酸钠,酸奶的黏度和保水性分别为1320.6mPa.s、68.3%;0.5%丝素蛋白和0.05%酸性CMC,酸奶的黏度和保水性分别为962.1mPa.s、65.0%。%In order to improve the quality of silk fibroin yogurt, a two-variable, three-level factorial experimental design was used to explore the effects of silk fibroin in combination with one of four common stabilizers including pectin, xanthan gum, sodium alginate, acid-resistant carboxymethyl-cellulose (CMC) on the viscosity and water-holding capacity of stirred yogurt. The results showed that combined with pectin, xanthan gum or sodium alginate, silk fibroin had a highly significant on the viscosity of stirred yogurt (P〈0.01). Moreover, there was a highly significant interaction of silk fibroin with each of the three stabilizers (P〈0.01). All the combinations of silk fibroin with pectin, xanthan gum or acid-resistant CMC had a highly significant effect on water- holding capacity of stirred yogurt (P〈0.01), and a highly significant interaction between silk fibroin and xanthan gum was found (P〈 0.01). Four optimized

  13. Changes in growth and lipid profiles of silk gland, mid-gut biochemical composition of silkworm, Bombyx mori L. on exposure to prostaglandin F2alpha.

    Miao, Yun-gen; Jiang, Li-jun

    2003-01-01

    The growth of the silkworm is influenced by the outside and inside environment. Among them, the category of various endocrine hormone of inside is the main factors that adjust the characters such as growth and propagate. In this experiment, we applied different dosage of prostaglandin to the fourth and fifth instar silkworm to observe the effects of prostaglandin F2alpha (PGF2alpha) on silk gland growth, mid-gut biochemical constituents and the lipid profiles of silkworm larva, Bombyx mori L. The weight of the posterior silk gland increased significantly (P mori L. indicate that PGF2alpha favored stimulatory effect on physiology of digestion, absorption and transportation of nutrients which might influence on the growth and development of larva. PMID:12611497

  14. Structural Analysis of Fibroin Heavy Chain Signal Peptide of Silkworm Bombyx mori

    Sheng-Peng WANG; Ting-Qing GUO; Xiu-Yang GUO; Jun-Ting HUANG; Chang-De LU

    2006-01-01

    To study the minimal length required for the secretion of recombinant proteins and silk proteins in posterior silk gland, the signal peptide (SP) of the fibroin heavy chain (FibH) of silkworm Bombyx mori was systematically shortened from the C-terminal. Its effect on the secretion of protein was observed using enhanced green fluorescent protein (EGFP) as a reporter. Secretion of EGFP fusion proteins was examined under fluorescence microscope. FibH SPs with lengths of 20, 18, 16 and 12 a.a. can direct the secretion of the reporter, yet those with lengths of 11, 10, 9, 8 and 1 a.a. can not. When the FibH SP was shortened to 12 a.a., the secretion efficiency was decreased slightly and cleavage occurred within EGFP.When 16 a.a. of the FibH SP were used, the secretion of fusion protein was normal and the cleavage site was between the Gly-Ser linker and Met, the starting amino acid of EGFP. These findings are applicable for the expression of foreign proteins in silkworm silk gland. The cleavage site of the SP is discussed and compared with the predictive results of the SignalP 3.0 online prediction program.

  15. Effect of silk protein surfactant on silk degumming and its properties.

    Wang, Fei; Cao, Ting-Ting; Zhang, Yu-Qing

    2015-10-01

    The silk protein surfactant (SPS) first used as a silk degumming agent in this study is an amino acid-type anionic surfactant that was synthesized using silk fibroin amino acids and lauroyl chloride. We studied it systematically in comparison with the traditional degumming methods such as sodium carbonate (Na2CO3) and neutral soap (NS). The experimental results showed that the sericin can be completely removed from the silk fibroin fiber after boiling the fibers three times for 30 min and using a bath ratio of 1:80 (g/mL) and a concentration of 0.2% SPS in an aqueous solution. The results of the tensile properties, thermal analysis, and SEM all show that SPS is similar to the NS, far superior to Na2CO3. In short, SPS may be used as an environmentally friendly silk degumming/refining agent in the silk textile industry and in the manufacture of silk floss quilts. PMID:26117747

  16. Novel molecular and mechanical properties of egg case silk from wasp spider, Argiope bruennichi.

    Zhao, Ai-Chun; Zhao, Tian-Fu; Nakagaki, Koichi; Zhang, Yuan-Song; Sima, Yang-Hu; Miao, Yun-Gen; Shiomi, Kunihiro; Kajiura, Zenta; Nagata, Yoko; Takadera, Masayuki; Nakagaki, Masao

    2006-03-14

    Araneoid spiders use specialized abdominal glands to produce up to seven different protein-based silks/glues that have various mechanical properties. To date, the fibroin sequences encoding egg case fibers have not been fully determined. To gain further understanding of a recently reported spider silk protein gene family, several novel strategies were utilized in this study to isolate two full-length cDNAs of egg case silk proteins, cylindrical silk protein 1 (CySp1, 9.1 kb) and cylindrical silk protein 2 (CySp2, 9.8 kb), from the wasp spider, Argiope bruennichi. Northern blotting analysis demonstrated that CySp1 and CySp2 are selectively expressed in the cylindrical glands. The amino acid composition of raw egg case silk was closely consistent with the deduced amino acid composition based on the sequences of CySp1 and CySp2, which supports the assertion that CySp1 and CySp2 represent two major components of egg case silk. CySp1 and CySp2 are primarily composed of remarkable homogeneous assemble repeats that are 180 residues in length and consist of several complex subrepeats, and they contain highly homologous C-termini and markedly different N-termini. Our results suggest a possible link between CySp1 and CySp2. In addition, comparisons of stress/strain curves for dragline and egg case silk from Argiope bruennichi showed obvious differences in ultimate strength and extensibility, and similarities in toughness. PMID:16519529

  17. Nanocomposite gold-silk nanofibers

    Cohen-Karni, Tzahi; Jeong, Kyung Jae; Tsui, Jonathan H.; Reznor, Gally; Mustata, Mirela; Wanunu, Meni; Graham, Adam; Marks, Carolyn; Bell, David C.; Langer, Robert S; Kohane, Daniel S.

    2012-01-01

    Cell-biomaterial interactions can be controlled by modifying the surface chemistry or nanotopography of the material, to induce cell proliferation and differentiation if desired. Here we combine both approaches in forming silk nanofibers (SNFs) containing gold nanoparticles (AuNPs) and subsequently chemically modifying the fibers. Silk fibroin mixed with gold seed nanoparticles was electrospun to form SNFs doped with gold seed nanoparticles (SNFseed). Following gold reduction, there was a two...

  18. 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

  19. Formulation of Biologically-Inspired Silk-Based Drug Carriers for Pulmonary Delivery Targeted for Lung Cancer

    Sally Yunsun Kim; Deboki Naskar; Kundu, Subhas C.; Bishop, David P.; Doble, Philip A.; Boddy, Alan V.; Hak-Kim Chan; Ivan B. Wall; Wojciech Chrzanowski

    2015-01-01

    The benefits of using silk fibroin, a major protein in silk, are widely established in many biomedical applications including tissue regeneration, bioactive coating and in vitro tissue models. The properties of silk such as biocompatibility and controlled degradation are utilized in this study to formulate for the first time as carriers for pulmonary drug delivery. Silk fibroin particles are spray dried or spray-freeze-dried to enable the delivery to the airways via dry powder inhalers. The a...

  20. SILK FIBRE DEGRADATION AND ANALYSIS BY PROTEOMICS

    YUKSELOGLU S. Muge; CANOGLU Suat

    2016-01-01

    Silk is one of the promising natural fibres and has a long established history in textile production throughout the centuries. Silk is produced by cultured silk worms, spiders, scorpions, mites and flies. It is extracellular proteinaceous fibres which consist of highly crystalline and insoluble proteins, the fibroins glued with sericin and an amourphous protein. On the other hand, understanding and controlling the degradation of protein materials are important for determining quality and the...

  1. Study on electrospun Silk Fibroin-Hyaluronic acid blends nanofbers and its structure%静电纺丝素-透明质酸共混纳米纤维形态结构研究

    陈梅; 张锋; 明津法; 高艳菲; 左保齐

    2012-01-01

    Silk fibroin and different molecular hyaluronic acid blend nanofibers (SF-HA) were obtained by electro spinning and using formic acid as solvent. Morphological structure and molecular structure of the SF-HA blend nanofibers were examinedby SEM, FTIR, XRD, TG-DTA. It shows that the average diameters of SF-HA blend nanofibers was about 100 nm; the surface of blend fibers was smooth and the shape was cylindrical; with the addition of high molecular hyaluronic acid, the morphology of SF-HA became worse and the nanofibers had serious connection with each other. However, the result proved that the addition of little hyaluronic acid had no significant effect on the molecular conformation and crystal structure.%以甲酸为溶剂,采用静电纺丝方法制备丝素蛋白(SF)与不同相对分子质量的透明质酸(HA)共混纳米纤维,并通过扫描电镜(SEM)、红外光谱(FTIR)、X射线衍射(XRD)和热分析(TG-DTA)研究了SF-HA共混纳米纤维非织造网的形态、结构和性能.结果表明:通过静电纺丝,丝素与低分子量的透明质酸共混纳米纤维直径在100m左右,纤维表面光滑,呈圆柱状:高分子量透明质酸的加入使得纤维形态变差,出现严重的粘连现象:透明质酸的加入未对丝素分子构象和结晶结构产生明显影响.

  2. Sensitization to silk allergen among workers of silk filatures in India: a comparative study

    Gowda, Giriyanna; Shivalingaiah, Anwith Huluvadi; Vijayeendra, Anagha Manakari; Sarkar, Nivedita; Nagaraj, Chitra; Masthi, Nugehally Raju Ramesh

    2016-01-01

    Background Sericulture plays an eminent role in development of rural economy in India. Silk filature is a unit where silk is unwound from the cocoons and the strands are collected into skeins. During the process workers are exposed to the high molecular weight proteins like Sericin and Fibroin which are potent allergens leading to sensitization over a period of time and subsequently occupational related health disorders. Objective To identify and compare the magnitude of silk allergen sensiti...

  3. 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

  4. 静电纺丝素/聚丁二酸丁二醇酯血管材料的结构与性能%Structure and properties of electrospun silk fibroin-poly( butylene succinate) artificial blood vessel

    吴斌伟; 朱海霖; 张乐伟; 冯新星; 陈建勇

    2011-01-01

    为制备组织工程血管支架,以丝素蛋白(SF)和聚丁二酸丁二醇酯(PBS)为原料,通过静电纺丝法,以具有三维结构的收集模板取代传统的二维平板作为静电纺丝收集基板,构建丝素/PBS血管支架材料.研究纺丝条件和三维收集模板对管状支架形貌的影响,采用FT-IR对丝素蛋白二级结构进行表征,测试血管支架材料的孔隙率和力学性能.结果表明,通过改变电压、纺丝间距以及三维收集模板的宏观结构,可制备出具有不同直径和长度的血管支架材料.这种支架材料的孔隙率达84.6%,拉伸应力为4.31 MPa,断裂伸长率为46.21%,爆破压力为358kPa.%To develop a small-diameter tissue engineering blood vessel, the silk fibroin (SF)/poly( butylene succinate) (PBS) tubular scaffolds were fabricated via electrospinning. During the electrospinning process, the electrospun SF/PBS composite tubular scaffolds were collected on a 3-D collector instead of traditional 2-D collector. The effects of 3-D collector and process parameters of electrospinning on the morphologies of the tubular scaffolds were investigated. FT-IR was used to study the secondary structure of SF. The mechanical properties and porosities of the tubular fiber materials were characterized. The results showed that SF/PBS composite tubular materials with different sizes and shapes could be prepared by changing the voltage, TCD and the structure of 3-D collectors. The porosity, tensile stress, elongation at break and burst pressure of the composite tubular scaffold were 84. 6%, 4. 31 MPa,46. 21% and 358 kPa, respectively.

  5. Characterization of Partial Coding Region Fibroin Gene on Wild Silkmoth Cricula trifenestrata Helfer (Lepidoptera: Saturniidae

    Suriana

    2011-04-01

    Full Text Available The study was conducted to characterize coding region of wild silkmoth C. trifenestrata partial fibroin gene, and detect these gene potential as molecular marker. A total of six larvae C. trifenestrata were collected from Bogor, Purwakarta and Bantul Regency. Genomic DNA was extracted from silk gland individual larvae, then amplified by PCR method and sequenced. DNA sequenced result was 986 nucleotide partial fibroin gene of C. trifenestrata, which are comprising complete coding region of first exon (42 nucleotide, an intron (113 nucleotide, and partial of second was exon (831 nucleotide. Only coding region was characterized. Results showed that first exon very conserved in C. trifenestrata. These gene consisted of 31%, thymine, 28% guanine, 21% cytosine, and 19% adenine. Cytosine and thymine (sites of 25th and 35th respectively were marker for C. trifenestrata species. The first exon encoding 14 amino acids. Valine amino acid (12th site was marker to the species C. trifenestrata. The partial second exon consisted of guanine (32.7%, alanine (26.5%, thymine (21% and cytosine (19.7%. These region encoded 277 amino acids, which were dominated by the alanine (27.8% and glycine (21.66%. Alanine formed polyalanine sequence with different motifs namely: AAAAAAASS, AAAAAAAAAAAGSSG, AAAAAAAAAAAAGSGTGFGGYDS, AAAAAAAAAAGSSGRGGYDGVDGGYGSGSS, and AAAAAAAAAAAAGSSGRGLGGYDGWVDDGYGSGSGS.

  6. 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.

  7. Silk Microgels Formed by Proteolytic Enzyme Activity

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

    2013-01-01

    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 (SMG) at physiological temperature. These microgels consist of lamellar crystals in the micrometer scale, in contrast to the nanometer scaled crystals in native silkworm fibers. SDS-PAGE and zeta potential results demonstrated that α-chymotrypsin utilized only the nonamorphous domains or segments of the heavy chai...

  8. 高解舒率家蚕品种中部丝腺腔内容物的蛋白质组分析%A Proteomic Analysis to the Substance in Middle Silk Gland Lumen of Silkworm Varieties with High Reelability

    杜鑫; 陈玉银; 刘培刚; 李军; 王永强

    2013-01-01

    Cocoon filament reelability is one of the important economic traits of silkworm (Bombyx mori).To find specific proteins related to high cocoon filament reelability,the silkworm strain B84 (yellow cocoon color) with high cocoon filament reelability and Fengyi (white cocoon color) with normal cocoon filament reelability were used as materials to conduct proteomic analysis and identification on the substance in middle silk gland lumen from the mature larvae of these two silkworm strains by utilizing two-dimensional electrophoresis (2-DE) and matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS).341 and 363 protein spots were detected from the substance in middle silk gland lumen of Fengyi and B84 respectively.These proteins included a large number of functional proteins which have no relationship with sericin.For instance,there were serine protease inhibitors that have protective effect to silk fibroin.In the substance of middle silk gland lumen of B84,apart from carotenoid binding proteins that are involved in cocoon color formation,specific Sericin 2 protein and carbohydrate metabolism related enzymes were also identified.It was inferred that these proteins might be the underlying reason for high cocoon filament reelability of B84 due to the existence of Sericin 2 in cocoon shell as well and the influence of glycan chain on protein adhesion.%茧丝解舒率是家蚕的重要经济性状之一.为了寻找与高解舒性状相关的特异性蛋白质,以高解舒率家蚕品系B84(黄茧)和普通解舒率家蚕品种丰一(白茧)为材料,通过双向电泳(2-DE)和基质辅助激光解析电离飞行时间质谱(MALDITOF-MS)技术对2个家蚕品种(品系)熟蚕中部丝腺腔内容物进行蛋白质组学分析鉴定.分别在丰一与B84的中部丝腺腔内容物中检测到341和363个蛋白点,其中包括大量非丝胶蛋白质成分的功能蛋白质,如对丝素蛋白具有保护作用的丝氨

  9. Biomedical Applications of Mulberry Silk and its Proteins: A Review

    Nivedita, S.; Sivaprasad, V.

    2014-04-01

    Silk is a natural fibre used mainly for aesthetic purposes. It has also been used for making surgical sutures for centuries. The recent rediscovery of silk's biological properties have led to new areas of research and utilization in cosmetic, health and medical fields. The silk proteins, fibroin and sericin are processed into biomaterials because of bio-compatibility, bio-degradability, excellent mechanical properties, thermo tolerance and UV protective properties. Silk proteins could be obtained as pure liquids and regenerated in different forms suitable for tissue engineering applications. This paper presents some of the biomedical products and biomaterials made from native, degraded and regenerated silk and their fabrication techniques.

  10. Using FTIR spectroscopy to detect sericin on historic silk

    WYETH; Paul

    2010-01-01

    Silks represent some of the most precious ancient and historic textile artefacts in collections worldwide.Their optimum preservation demands an appreciation of their characteristics.One important concern,especially with regard to ancient Chinese silks,is whether the fabrics have been degummed.Silks with remnant sericin gum coating the fibroin fibres would require different conservation protocol.In previous research on aged silks,the presence of sericin has been inferred from amino acid analysis of hydrolysates.In the study reported here,the potential of FTIR spectroscopy to provide a simpler and rapid method of detecting sericin on silk has been investigated.Both fibroin and sericin exhibit singular IR absorptions.Attenuated total reflectance spectroscopy was found to highlight the sericin coating more effectively than transmission and reflectance spectroscopy.Three particular peak intensity ratios were identified which might provide a quantitative estimate of the sericin content of new silk,to a sensitivity of 1%-2%.These were also shown to be valid indicators for the presence of sericin on artificially aged and archaeological silks,although quantitation was now not possible.Besides the peak intensity ratios,two signature peaks were also seen to be useful markers for silk fibroin,and their presence in a spectrum could be used to infer a degummed silk.

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

    Baran, E. T.; Tuzlakoglu, K.; Mano, J. F.; Reis, R. L.

    2012-01-01

    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 α-amylase was more ...

  12. Electrospun silk-based nanofibrous scaffolds: fiber diameter and oxygen transfer

    Chomachayi, Masoud Dadras; Solouk, Atefeh; Mirzadeh, Hamid

    2016-01-01

    In this study, silk fibroin was extracted from cocoons of silkworms and fabricated into nonwoven mats by electrospinning method. A new model based on the group method of data handling (GMDH) and artificial neural network (ANN) was developed for estimation of the average diameter of electrospun silk fibroin nanofibers. In this regard, concentration, flow rate, voltage, distance, and speed of collector were used as input parameters and average diameter of the fibers was considered as output par...

  13. Production of fine powder from silk by radiation

    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)

  14. 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)

  15. Vibrational spectroscopic study of sulphated silk proteins

    Monti, P.; Freddi, G.; Arosio, C.; Tsukada, M.; Arai, T.; Taddei, P.

    2007-05-01

    Degummed Bombyx mori ( B. m.) silk fibroin fabric and mutant naked pupa cocoons (Nd-s) consisting of almost pure silk sericin were treated with chlorosulphonic acid in pyridine and investigated by FT-IR and FT-Raman spectroscopies. Untreated silk fibroin and sericin displayed typical spectral features due to characteristic amino acid composition and molecular conformation (prevailing β-sheet with a less ordered structure in sericin). Upon sulphation, the degree of molecular disorder increased in both proteins and new bands appeared. The IR bands at 1049 and 1014 cm -1 were attributed to vibrations of sulphate salts and that at 1385 cm -1 to the νasSO 2 mode of organic covalent sulphates. In the 1300-1180 cm -1 range various contributions of alkyl and aryl sulphate salts, sulphonamides, sulphoamines and organic covalent sulphates, fell. Fibroin covalently bound sulphate groups through the hydroxyl groups of tyrosine and serine, while sericin through the hydroxyl groups of serine, since the δOH vibrations at 1399 cm -1 in IR and at 1408 cm -1 in Raman disappeared almost completely. Finally, the increase of the I850/ I830 intensity ratio of Raman tyrosine doublet in fibroin suggested a change towards a more exposed state of tyrosine residues, in good agreement with the more disordered conformation taken upon sulphation.

  16. 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.

  17. 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)

  18. Silk-mediated synthesis and modification of photoluminescent ZnO nanoparticles

    Han Jie; Su Huilan, E-mail: hlsu@sjtu.edu.cn; Xu Jia; Song Weiqiang; Gu Yu; Chen Ying [Shanghai Jiao Tong University, State Key Lab of Metal Matrix Composites (China); Moon, Won-Jin [Gwangju Center, Korea Basic Science Institute (Korea, Republic of); Zhang Di, E-mail: zhangdi@sjtu.edu.cn [Shanghai Jiao Tong University, State Key Lab of Metal Matrix Composites (China)

    2012-02-15

    In this article, a bio-inspired silk-mediated method was established to produce natural material-modified photoluminescent zinc oxide nanoparticles (nano-ZnO). Silk fibroin fibers were employed as the reactive substrates to synthesize nano-ZnO, and silk fibroins (SF) were taken as the biocompatible stabilizers to modify dispersed nano-ZnO. As-prepared nano-ZnO were mainly hexagonal phase particles with diameter around 13 nm. The resulting nano-ZnO/SF hybrids displayed orange emission and good biocompatibility in aqueous system.

  19. Preparation and Characterization of a Novel Hybrid Hydrogel Composed of Bombyx mori Fibroin and Poly(N-isopropylacrylamide)

    A novel hybrid hydrogel was prepared and investigated based on silkworm silk fibroin and poly(N-isopropylacrylamide) (PNIPAAm). PNIPAAm was introduced to silk fibroin, the resultant composite hydrogel was examined, and freeze-dried SF/PNIPAAm scaffold was analyzed using LB-550 dynamic light scattering particle-size analyzer, circular dichroism (CD), and scanning electron microscopy (SEM). Our results suggested that the hybrid hydrogels owned the porous sponge-like structures, and the gelation time of SF/PNIPAAm hybrids decreased with an increase in temperature and concentration of each polymer. Results of rheological analysis suggested that the rheological property of resultant SF/PNIPAAm gel depended on the concentration combinations as well as the aging time, which elapsed after mixing the two polymers. Results of CD spectra demonstrated that ph showed little influence on the secondary structure of silk fibroin, and significant changes of G', Gand G* as surrounding increase temperature above the lower critical solution temperature (LCST)

  20. 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.

  1. Nanoscale Control of Silks for Nanofibrous Scaffold Formation with Improved Porous Structure

    Lin, Shasha; Lu, Guozhong; Liu, Shanshan; Bai, Shumeng; Liu, Xi; Lu, Qiang; Zuo, Baoqi; Kaplan, David L.; Zhu, Hesun

    2014-01-01

    Silk-based porous scaffolds have been used extensively in tissue engineering because of their excellent biocompatibility, tunable biodegradability and robust mechanical properties. Although many silk-based scaffolds have been prepared through freeze-drying, a challenge remains to effectively control porous structures during this process. In the present study silk fibroin with different nanostructures were self-assembled in aqueous solution by repeated drying-dissolving process and then used t...

  2. 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...

  3. Differential Scanning Fluorimetry provides high throughput data on silk protein transitions

    Vollrath, Fritz; Hawkins, Nick; Porter, David; Holland, Chris; Boulet-Audet, Maxime

    2014-01-01

    Here we present a set of measurements using Differential Scanning Fluorimetry (DSF) as an inexpensive, high throughput screening method to investigate the folding of silk protein molecules as they abandon their first native melt conformation, dehydrate and denature into their final solid filament conformation. Our first data and analyses comparing silks from spiders, mulberry and wild silkworms as well as reconstituted ‘silk' fibroin show that DSF can provide valuable insights into details of...

  4. SILK FIBRE DEGRADATION AND ANALYSIS BY PROTEOMICS

    YUKSELOGLU S.Muge

    2016-05-01

    Full Text Available Silk is one of the promising natural fibres and has a long established history in textile production throughout the centuries. Silk is produced by cultured silk worms, spiders, scorpions, mites and flies. It is extracellular proteinaceous fibres which consist of highly crystalline and insoluble proteins, the fibroins glued with sericin and an amourphous protein. On the other hand, understanding and controlling the degradation of protein materials are important for determining quality and the value of appearance retention in textiles. Hence, for silk textiles, appearance retention is critical value for the quality. And this is one of the key properties directly related to the degree and nature of protein degradation. It is therefore necessary to understand the silk composition and damage to obtain good conservation treatments and long-term preservation especially for the historical silk fabrics. In this study, silk fibre and its properties are briefly introduced along with images on their fibre damages. Additionally, proteomics method which helps to understand the degradation at the molecular level in textiles is introduced. Finally, proteomic evaluation of silk is summarized according to the researchers carried out in the literature.

  5. 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

  6. 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...

  7. 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

  8. Role of humidity on the structures and properties of regenerated silk fibers

    Qingfa Peng

    2015-10-01

    Full Text Available Silk fiber was processed from highly concentrated spinning dope to solid fibers along with water removal. To understand the mechanism of water removal during silk fiber spinning process, a microfluidic chip was designed and applied to investigate the structures and mechanical properties of two kinds of regenerated silk fibroin fibers dry-spun at different relative humidity. The experimental results showed that the diameters of the fibers spun at 40% RH are always larger than the fibers spun at 50% RH due to different removal rates of water. The fibers spun at low humidity contain more β-sheet structure and lower degree of chain orientation and crystalline orientation. These results indicate that the fast phase transition of silk fibroin from sol–gel to silk fiber undergoes with rapid water removal and higher fiber orientation relates to more residue water and drawing force.

  9. Silk elasticity as a potential constraint on spider body size

    Rodríguez-Gironés, Miguel A; Corcobado, Guadalupe; Moya-Laraño, Jordi

    2010-01-01

    Abstract Silk is known for its strength and extensibility and has played a key role in the radiation of spiders. Individual spiders use different glands to produce silk types with unique sets of proteins. Most research has studied the properties of major ampullate and capture spiral silks and their ecological implications, while little is known about minor ampullate silk, the type used by those spider species studied to date for bridging displacements. A biomechanical model paramet...

  10. 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

  11. 胶原/丝素导管介导雪旺细胞联合神经干细胞修复坐骨神经缺损%Collagen/silk fibroin conduit combined with the co-culture of Schwann cells and neural stem cells for bridging sciatic nerve defect

    徐云强; 张振辉; 余欣; 陈旭义; 李东; 李瑞欣; 冯世庆

    2015-01-01

    目的 观察胶原/丝素导管介导雪旺细胞联合神经干细胞所构建的组织工程化神经修复大鼠10 mm坐骨神经缺损的效果.方法 体外分离培养乳鼠坐骨神经雪旺细胞(SCs),并做S-100蛋白免疫荧光鉴定;从孕14 ~ 15 d的SD大鼠体内取出子鼠,分离纯化获得原代神经干细胞(NSCs)进行体外培养.实验分为4组,每组10只:自体神经移植组(A组)、胶原/丝素导管介导雪旺细胞联合神经干细胞移植组(B组)、胶原/丝素导管移植组(C组)、单纯损伤组(D组).SCs-NSCs与胶原/丝素导管联合培养14 d后,行扫描电镜观察.分别将3种不同移植物桥接于大鼠坐骨神经10 cm缺损处,并在12周后进行大体观察、电生理学检测、形态学观察及计量学分析.结果 术后12周,桥接组都不同程度地实现了坐骨神经缺损再通,且实验动物未出现明显排斥及炎性反应.神经电生理学检测坐骨神经复合肌动作电位(CMAPs)波幅分别为:未手术正常侧(21.00±1.83) mV,A组(15.00±1.12) mV,B组(13.00±1.06) mV,C组(6.00±0.58) mV,透射电子显微镜再生神经纤维髓鞘厚度分别为:A组(0.80±0.15) μm,B组(0.70±0.11) μm,C组(0.30±0.07) μm,D组(0.25±0.06) μm.统计结果表明:A组与B组之间差异无统计学意义(P>0.05),但两者相对C组及D组差异有统计学意义(P<0.05).结论 SCs和NSCs能够在胶原/丝素导管上共同生长分化,生物相容性良好.胶原/丝素导管介导雪旺细胞联合神经干细胞所构建的组织工程化神经对坐骨神经缺损的修复具有良好的桥接和促神经生长作用.%Objective To observe the repair effect of collagen/silk fibroin conduit transplanted with co-cultured Schwann cells (SCs) and neural stem cells (NSCs) for bridging 10 mm sciatic nerve defect by tissue engineering technique in rats.Methods SCs were isolated and purified from the sciatic nerve of neonatal SD rats,and were identified by immunocytochemical labelling for S100

  12. 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.

  13. 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.

  14. 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.

  15. 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...

  16. Tissue Regeneration: A Silk Road.

    Jao, Dave; Mou, Xiaoyang; Hu, Xiao

    2016-01-01

    Silk proteins are natural biopolymers that have extensive structural possibilities for chemical and mechanical modifications to facilitate novel properties, functions, and applications in the biomedical field. The versatile processability of silk fibroins (SF) into different forms such as gels, films, foams, membranes, scaffolds, and nanofibers makes it appealing in a variety of applications that require mechanically superior, biocompatible, biodegradable, and functionalizable biomaterials. There is no doubt that nature is the world's best biological engineer, with simple, exquisite but powerful designs that have inspired novel technologies. By understanding the surface interaction of silk materials with living cells, unique characteristics can be implemented through structural modifications, such as controllable wettability, high-strength adhesiveness, and reflectivity properties, suggesting its potential suitability for surgical, optical, and other biomedical applications. All of the interesting features of SF, such as tunable biodegradation, anti-bacterial properties, and mechanical properties combined with potential self-healing modifications, make it ideal for future tissue engineering applications. In this review, we first demonstrate the current understanding of the structures and mechanical properties of SF and the various functionalizations of SF matrices through chemical and physical manipulations. Then the diverse applications of SF architectures and scaffolds for different regenerative medicine will be discussed in detail, including their current applications in bone, eye, nerve, skin, tendon, ligament, and cartilage regeneration. PMID:27527229

  17. 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.

  18. 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.

  19. Inhibitory Effect of Curcumin on Metal Ions-induced Conformational Transition of Silk Fibroin and Its Mechanism%姜黄素抑制金属离子诱导的丝素蛋白构象转变及其作用机理

    章月虹; 江腾; 赵学舟; 周平

    2013-01-01

    以丝素蛋白(SF)为神经退行性疾病相关蛋白的模型蛋白,分析了姜黄素(Curcumin)对Zn(Ⅱ)和Cu(Ⅱ)离子诱导的丝素蛋白构象转变的干预作用及其作用机理,试图探讨姜黄素在神经退行性疾病中的预防与治疗作用.结果表明,姜黄素可通过与Cu(Ⅱ)和Zn(Ⅱ)离子络合,干预金属离子诱导的丝素蛋白构象转变,并且所形成的络合物Cu(Ⅱ)-Curcumin可抑制丝素蛋白的构象转变,但是Zn(Ⅱ)-Curcumin络合物不具有这种能力.因此,基于姜黄素对于金属离子诱导的丝素蛋白构象转变具有良好的干预作用,可以将姜黄素作为神经退行性疾病预防和治疗的潜在候选药物.%The inhibitory effect of curcumin on the Cu ( II ) and Zn ( Ⅱ) -induced conformation transition of silk fibroin(SF) as a model protein of neurodegenerative diseases, was investigated for aiming at investigating the potency of curcumin in the prevention and treatment of neurodegenerative diseases. The results indicate that curcumin can inhibit Cu ( II ) and Zn ( II ) -induced conformational transition of SF through chelating Cu ( II ) and Zn ( II ), meanwhile, Cu ( Ⅱ ) -curcumin complex also has ability to inhibit the conformation transition of SF, but Zn ( II ) -curcumin complex does not. Therefore, based on those results about the .effective interference of curcumin on Cu ( II ) and Zn ( Ⅱ ) -induced conformation transition of SF, curcumin could be suggested as a potential agent for the prevention and treatment of neurodegenerative diseases.

  20. Unravelling the biodiversity of nanoscale signatures of spider silk fibres.

    Silva, Luciano P; Rech, Elibio L

    2013-01-01

    Living organisms are masters at designing outstanding self-assembled nanostructures through a hierarchical organization of modular proteins. Protein-based biopolymers improved and selected by the driving forces of molecular evolution are among the most impressive archetypes of nanomaterials. One of these biomacromolecules is the myriad of compound fibroins of spider silks, which combine surprisingly high tensile strength with great elasticity. However, no consensus on the nano-organization of spider silk fibres has been reached. Here we explore the biodiversity of spider silk fibres, focusing on nanoscale characterization with high-resolution atomic force microscopy. Our results reveal an evolution of the nanoroughness, nanostiffness, nanoviscoelastic, nanotribological and nanoelectric organization of microfibres, even when they share similar sizes and shapes. These features are related to unique aspects of their molecular structures. The results show that combined nanoscale analyses of spider silks may enable the screening of appropriate motifs for bioengineering synthetic fibres from recombinant proteins. PMID:24345771

  1. Biopatterning of Silk Proteins for Soft Micro-optics.

    Pal, Ramendra K; Kurland, Nicholas E; Wang, Congzhou; Kundu, Subhas C; Yadavalli, Vamsi K

    2015-04-29

    Silk proteins from spiders and silkworms have been proposed as outstanding candidates for soft micro-optic and photonic applications because of their optical transparency, unique biological properties, and mechanical robustness. Here, we present a method to form microstructures of the two constituent silk proteins, fibroin and sericin for use as an optical biomaterial. Using photolithography, chemically modified silk protein photoresists are patterned in 2D arrays of periodic patterns and Fresnel zone plates. Angle-dependent iridescent colors are produced in these periodic micropatterns because of the Bragg diffraction. Silk protein photolithography can used to form patterns on different substrates including flexible sheets with features of any shape with high fidelity and resolution over large areas. Finally, we show that these mechanically stable and transparent iridescent architectures are also completely biodegradable. This versatile and scalable technique can therefore be used to develop biocompatible, soft micro-optic devices that can be degraded in a controlled manner. PMID:25853731

  2. Silk-microfluidics for advanced biotechnological applications: A progressive review.

    Konwarh, Rocktotpal; Gupta, Prerak; Mandal, Biman B

    2016-01-01

    Silk based biomaterials have not only carved a unique niche in the domain of regenerative medicine but new avenues are also being explored for lab-on-a-chip applications. It is pertinent to note that biospinning of silk represents nature's signature microfluidic-maneuver. Elucidation of non-Newtonian flow of silk in the glands of spiders and silkworms has inspired researchers to fabricate devices for continuous extrusion and concentration of silk. Microfluidic channel networks within porous silk scaffolds ensure optimal nutrient and oxygen supply apart from serving as precursors for vascularization in tissue engineering applications. On the other hand, unique topographical features and surface wettability of natural silk fibers have inspired development of a number of simple and cost-effective devices for applications like blood typing and chemical sensing. This review mirrors the recent progress and challenges in the domain of silk-microfluidics for prospective avant-garde applications in the realm of biotechnology. PMID:27165254

  3. 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

  4. 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

  5. Tuning Molecular Weights of Bombyx mori (B. mori) Silk Sericin to Modify Its Assembly Structures and Materials Formation

    Yang, Mingying; Shuai, Yajun; Zhou, Guanshan; Mandal, Namita; Zhu, Liangjun; Mao, Chuanbin

    2014-01-01

    Bombyx mori (B. mori) silk sericin is a protein with features desirable as a biomaterial, such as increased hydrophilicity and biodegradation, as well as resistance to oxidation, bacteria, and ultraviolet light. In contrast to other widely studied B. mori silk proteins such as fibroin, sericin is still unexplored as a building block for fabricating biomaterial, and thus a facile technique of processing it into a material is needed. Here, electrospinning technology was used to fabricate it int...

  6. 聚乙烯醇/柞蚕丝素蛋白材料作为肌腱组织工程支架材料的初步研究%PRELIMINARY STUDY ON POLYVINYL ALCOHOL/WILD ANTHERAEA PERNYI SILK FIBROIN AS NANOFIBER SCAFFOLDS FOR TISSUE ENGINEERED TENDON

    吴琳琳; 李敏; 赵晋; 陈登龙; 周志华

    2011-01-01

    Objective To investigate the cellular compatibility of polyvinyl alcohol (PVA)/wild antheraea pernyi silk fibroin (WSF), and to explore the feasibility for tendon tissue engineering scaffold in vitro.Methods The solutions of WSF (11%), PVA (11%), and PVA/WSF (11%) were prepared with 98% formic acid (mass fraction) at a mass ratio of 9: 1.The electrospinning membranes of WSF, PVA, and PVA/WSF were prepared by electrostatic spinning apparatus.The morphologies of scaffolds were evaluated using scanning electronic microscope (SEM).The tendon cells were isolated from tail tendon of 3-dayold Sprague Dawley rats in vitro.The experiment was performed using the 3rd generation cells.The tendon cells (1 × 106/mL)were cocultured with PVA and PVA/WSF electrospinning film, respectively, and MTT test was used to assess the cell adhesion rate 4, 12 hours after coculture.The tendon cells were cultured in PVA and PVA/WSF extraction medium of different concentration (1, 1/2, and 1/4), respectively; and the absorbance (A) values were detected at 1, 3, 5, and 7 days to evaluate the cytotoxicity.The composite of tendon cells and the PVA or PVA/WSF scaffold were observed by HE staining at 7 days and characterized by SEM at 1,3,5, and 7 days.Results The solution of WSF could not be used to electrospin; and the solution of PVA and PVA/WSF could be electrospun.After coculture of tendon and PVA or PVA/WSF electrospinning membranes, the cell adhesion rates were 26.9% ±0.4% and 87.0% ± 1.0%, respectively for 4 hours, showing significant difference (t=100.400, P=0.000); the cell adhesion rates were 35.2% ± 0.6% and 110.0% ± 1.7%, respectively for 12 hours, showing significant difference (t=42.500, P=0.000).The cytotoxicity of PVA/WSF was less significantly than that of PVA (P < 0.05) and significant difference was observed between 1/2 PVA and 1/4 PVA (P < 0.05).HE staining and SEM images showed that the tendon cells could adhere to PVA and PVA/WSF scaffolds, but

  7. Relationships between physical properties and sequence in silkworm silks

    Malay, Ali D.; Sato, Ryota; Yazawa, Kenjiro; Watanabe, Hiroe; Ifuku, Nao; Masunaga, Hiroyasu; Hikima, Takaaki; Guan, Juan; Mandal, Biman B.; Damrongsakkul, Siriporn; Numata, Keiji

    2016-06-01

    Silk has attracted widespread attention due to its superlative material properties and promising applications. However, the determinants behind the variations in material properties among different types of silk are not well understood. We analysed the physical properties of silk samples from a variety of silkmoth cocoons, including domesticated Bombyx mori varieties and several species from Saturniidae. Tensile deformation tests, thermal analyses, and investigations on crystalline structure and orientation of the fibres were performed. The results showed that saturniid silks produce more highly-defined structural transitions compared to B. mori, as seen in the yielding and strain hardening events during tensile deformation and in the changes observed during thermal analyses. These observations were analysed in terms of the constituent fibroin sequences, which in B. mori are predicted to produce heterogeneous structures, whereas the strictly modular repeats of the saturniid sequences are hypothesized to produce structures that respond in a concerted manner. Within saturniid fibroins, thermal stability was found to correlate with the abundance of poly-alanine residues, whereas differences in fibre extensibility can be related to varying ratios of GGX motifs versus bulky hydrophobic residues in the amorphous phase.

  8. 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.

  9. A simple model of multiphoton micromachining in silk hydrogels

    Applegate, Matthew B.; Alonzo, Carlo; Georgakoudi, Irene; Kaplan, David L.; Omenetto, Fiorenzo G.

    2016-06-01

    High resolution three-dimensional voids can be directly written into transparent silk fibroin hydrogels using ultrashort pulses of near-infrared (NIR) light. Here, we propose a simple finite-element model that can be used to predict the size and shape of individual features under various exposure conditions. We compare predicted and measured feature volumes for a wide range of parameters and use the model to determine optimum conditions for maximum material removal. The simplicity of the model implies that the mechanism of multiphoton induced void creation in silk is due to direct absorption of light energy rather than diffusion of heat or other photoproducts, and confirms that multiphoton absorption of NIR light in silk is purely a 3-photon process.

  10. 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

  11. Decoration of silk fibroin by click chemistry for biomedical application.

    Zhao, Hongshi; Heusler, Eva; Jones, Gabriel; Li, Linhao; Werner, Vera; Germershaus, Oliver; Ritzer, Jennifer; Luehmann, Tessa; Meinel, Lorenz

    2014-06-01

    Silkfibroin (SF) has an excellent biocompatibility and its remarkable structure translates into exciting mechanical properties rendering this biomaterial particularly fascinating for biomedical application. To further boost the material's biological/preclinical impact, SF is decorated with biologics, typically by carbodiimide/N-hydroxysuccinimide coupling (EDC/NHS). For biomedical application, this chemistry challenges the product risk profile due to the formation of covalent aggregates, particularly when decoration is with biologics occurring naturally in humans as these aggregates may prime for autoimmunity. Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC; click chemistry) provides the necessary specificity to avoid such intermolecular, covalent aggregates. We present a blueprint outlining the necessary chemistry rendering SF compatible with CuAAC and with a particular focus on structural consequences. For that, the number of SF carboxyl groups (carboxyl-SF; required for EDC/NHS chemistry) or azido groups (azido-SF; required for click chemistry) was tailored by means of diazonium coupling of the SF tyrosine residues. Structural impact on SF and decorated SF was characterized by Fourier transform infrared spectroscopy (FTIR). The click chemistry yielded a better controlled product as compared to the EDC/NHS chemistry with no formation of inter- and intramolecular crosslinks as demonstrated for SF decorated with fluorescent model compounds or a biologic, fibroblast growth factor 2 (FGF2), respectively. In conclusion, SF can readily be translated into a scaffold compatible with click chemistry yielding decorated products with a better risk profile for biomedical application. PMID:24576682

  12. Transmission Electron Microscopy of Bombyx Mori Silk Fibers

    Shen, Y.; Martin, D. C.

    1997-03-01

    The microstructure of B. Mori silk fibers before and after degumming was examined by TEM, selected area electron diffraction (SAED), WAXS and low voltage SEM. SEM micrographs of the neat cocoon revealed a network of pairs of twisting filaments. After degumming, there were only individual filaments showing a surface texture consistent with an oriented fibrillar structure in the fiber interior. WAXS patterns confirmed the oriented beta-sheet crystal structure common to silkworm and spider silks. Low dose SAED results were fully consistent with the WAXS data, and revealed that the crystallographic texture did not vary significantly across the fiber diameter. TEM observations of microtomed fiber cross sections indicated a somewhat irregular shape, and also revealed a 0.5-2 micron sericin coating which was removed by the degumming process. TEM observations of the degummed silk fiber showed banded features with a characteristic spacing of nominally 600 nm along the fiber axis. These bands were oriented in a roughly parabolic or V-shape pointing along one axis within a given fiber. We hypothesize that this orientation is induced by the extrusion during the spinning process. Equatorial DF images revealed that axial and lateral sizes of the β-sheet crystallites in silk fibroin ranged from 20 to 170 nm and from 1 to 24 nm, respectively. Crazes developed in the degummed silk fiber parallel to the fiber direction. The formation of these crazes suggests that there are significant lateral interactions between fibrils in silk fibers.

  13. 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)

  14. 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 (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

  15. Functional expression of a Bombyx mori cocoonase: potential application for silk degumming

    Prangprapai Rodbumrer; Dumrongkiet Arthan; Utai Uyen; Jirundon Yuvaniyama; Jisnuson Svasti; Pramvadee Y.Wongsaengchantra

    2012-01-01

    Cocoon,a shelter for larva development to silk moth,contains the fibrous protein fibroin,which is coated by the globular protein sericin.Emergence of the silk moth requires the action of cocoonase,a protease secreted by the pupa.The full-length prococoonase cDNA,with 780 bp open reading frame encoding 260 amino acids,was cloned by reverse transcription from total RNA of the head of 6-day-old Thai-silk Bombyx mori pupa.Only the gene fragment lacking the propeptide encoding sequence was successfully expressed in Pichia pastoris,yielding an extracellularly active cocoonase.The recombinant cocoonase was purified to homogeneity by 80% ammonium-suffate fractionation and CM-Sepharose chromatography,and its internal peptide sequences were analyzed by nano liquid chromatographymass spectrometry/mass spectrometry.This monomeric protein has native molecular weight of 26 kDa by gel exclusion analysis and 25 kDa subunit size by sodium dodecyl sulphate-polyacrylamide gel electrophoresis.The enzyme hydrolyses sericin but does not hydrolyse fibroin,as shown by radial diffusion on thin-layer enzyme assay (RD-TEA).Scanning electron microscopy showed that purified recombinant cocoonase could remove sericin from natural silk completely in 24 h,without damaging fibroin,using only 1immobilized sericin unit (ISU) of enzyme as determined by RD-TEA.Natural cocoonase isolated from B.mori pupa could also digest sericin effectively,but required more enzymes (2 ISU) and longer time (48 h).In comparison,a commercial enzyme,alcalase,with the same activity not only showed less complete digestion of sericin but also caused damage of fibroin.These results suggest that recombinant B.mori cocoonase is potentially useful for silk degumming.

  16. 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

  17. 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 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 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 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

  18. 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

  19. 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

  20. 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.

  1. 纤维素-丝素复合膜的制备与表征%Preparation and characterization of cellulose/silk composite membrane

    李娟; 何建新; 余燕平

    2011-01-01

    The cellulose-silk fibroin composite film is prepared by mixing cellulose solution dissolved primary wood pulp in solution containing of NaOH/urea/thiourea/water and silk fibroin solution. The composite film is characterized by scanning electron microscope (SEM) , Fourier transform infrared spectrometer (FT-IR) , X-ray diffraction (XRD). SEM shows the surface of the composite is uniform and rough. The as-prepared composite film may be a potential biomedical material. The results of IR and XRD show that composite film is composed of cellulose and silk fibroin molecules and exists an interaction including hydrogen bond between the molecular of cellulose and silk fibroin.%通过NaOH/尿素/硫脲/水新型溶剂溶解原生木浆纤维素得到纤维素溶液,并与丝素溶液混合制备纤维素-丝素复合膜.利用扫描电镜、红外光谱、X-射线衍射对复合材料的结构进行表征.SEM结果表明复合材料表面粗糙,比表面积较大,可以作为潜在的生物医用材料.IR和X-衍射结果表明再生纤维素与丝素分子之间存在着强烈的氢键作用,且二者相容性较好.

  2. Effects of pH and initial Ca2+-H2PO4-concentration on fibroin mineralization

    PEN Yongjuan; SUN Xiaodan; CUI Fuzhai; KONG Xiangdong

    2007-01-01

    In the present study,the effects of pH and initial Ca2+-H2PO4-(Ca-P) concentration on fibroin mineralization were studied.The crystal growth of calcium phosphates was regulated by regenerated silk fibroin for 8 h(at pH 4.0.7.0and 1 0.0,respectively).Meanwhile,different concentrations of Ca2+ were employed at a certain pH value,keeping the initial Ca-P molar ratio constant at 1.67,i.e.,the stoichiometry of hydroxyapatite[Ca10(PO4)6(OH)2,HAP].The products were characterized by X-ray diffractometry(XRD),scanning electron microscopy(SEM) and thermogravimetric analysis (TGA).The results demonstrated that,compared to PH 4.0 and 10.0.pH 7.0 promoted the transformation of brushite alized fibroin,DCPD is the main inorganic phase at both relatively lowW and high pH,while HAP is the main inorganic phase at PH 7.0.Additionally,the initial Ca-P concentration does not afrect the kind of inorganic phase in the synthesized mineralized fibroin,but induce to different contents of inorganic mineral and different morphology of DCPD at PH 4.0 and pH 10.0.

  3. Advanced silk material spun by a transgenic silkworm promotes cell proliferation for biomedical application.

    Wang, Feng; Xu, Hanfu; Wang, Yuancheng; Wang, Riyuan; Yuan, Lin; Ding, Huan; Song, Chunnuan; Ma, Sanyuan; Peng, Zhixin; Peng, Zhangchuan; Zhao, Ping; Xia, Qingyou

    2014-12-01

    Natural silk fiber spun by the silkworm Bombyx mori is widely used not only for textile materials, but also for biofunctional materials. In the present study, we genetically engineered an advanced silk material, named hSFSV, using a transgenic silkworm, in which the recombinant human acidic fibroblast growth factor (hFGF1) protein was specifically synthesized in the middle silk gland and secreted into the sericin layer to surround the silk fiber using our previously optimized sericin1 expression system. The content of the recombinant hFGF1 in the hSFSV silk was estimated to be approximate 0.07% of the cocoon shell weight. The mechanical properties of hSFSV raw silk fiber were enhanced slightly compared to those of the wild-type raw silk fiber, probably due to the presence of the recombinant of hFGF1 in the sericin layer. Remarkably, the hSFSV raw silk significantly stimulated the cell growth and proliferation of NIH/3T3 mouse embryonic fibroblast cells, suggesting that the mitogenic activity of recombinant hFGF1 was well maintained and functioned in the sericin layer of hSFSV raw silk. These results show that the genetically engineered raw silk hSFSV could be used directly as a fine biomedical material for mass application. In addition, the strategy whereby functional recombinant proteins are expressed in the sericin layer of silk might be used to create more genetically engineered silks with various biofunctions and applications. PMID:24980060

  4. Biomimetic spinning of silk fibers and in situ cell encapsulation.

    Cheng, Jie; Park, DoYeun; Jun, Yesl; Lee, JaeSeo; Hyun, Jinho; Lee, Sang-Hoon

    2016-07-01

    In situ embedding of sensitive materials (e.g., cells and proteins) in silk fibers without damage presents a significant challenge due to the lack of mild and efficient methods. Here, we report the development of a microfluidic chip-based method for preparation of meter-long silk fibroin (SF) hydrogel fibers by mimicking the silkworm-spinning process. For the spinning of SF fibers, alginate was used as a sericin-like material to induce SF phase separation and entrap liquid SFs, making it possible to shape the outline of SF-based fibers under mild physicochemical conditions. L929 fibroblasts were encapsulated in the fibric hydrogel and displayed excellent viability. Cell-laden SF fibric hydrogels prepared using our method offer a new type of SF-based biomedical device with potential utility in biomedicine. PMID:27296229

  5. A Hox Gene, Antennapedia, Regulates Expression of Multiple Major Silk Protein Genes in the Silkworm Bombyx mori.

    Tsubota, Takuya; Tomita, Shuichiro; Uchino, Keiro; Kimoto, Mai; Takiya, Shigeharu; Kajiwara, Hideyuki; Yamazaki, Toshimasa; Sezutsu, Hideki

    2016-03-25

    Hoxgenes play a pivotal role in the determination of anteroposterior axis specificity during bilaterian animal development. They do so by acting as a master control and regulating the expression of genes important for development. Recently, however, we showed that Hoxgenes can also function in terminally differentiated tissue of the lepidopteranBombyx mori In this species,Antennapedia(Antp) regulates expression of sericin-1, a major silk protein gene, in the silk gland. Here, we investigated whether Antpcan regulate expression of multiple genes in this tissue. By means of proteomic, RT-PCR, and in situ hybridization analyses, we demonstrate that misexpression of Antpin the posterior silk gland induced ectopic expression of major silk protein genes such assericin-3,fhxh4, and fhxh5 These genes are normally expressed specifically in the middle silk gland as is Antp Therefore, the evidence strongly suggests that Antpactivates these silk protein genes in the middle silk gland. The putativesericin-1 activator complex (middle silk gland-intermolt-specific complex) can bind to the upstream regions of these genes, suggesting that Antpdirectly activates their expression. We also found that the pattern of gene expression was well conserved between B. moriand the wild species Bombyx mandarina, indicating that the gene regulation mechanism identified here is an evolutionarily conserved mechanism and not an artifact of the domestication of B. mori We suggest that Hoxgenes have a role as a master control in terminally differentiated tissues, possibly acting as a primary regulator for a range of physiological processes. PMID:26814126

  6. 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.

  7. 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.

  8. Role of chondroitin sulphate tethered silk scaffold in cartilaginous disc tissue regeneration.

    Bhattacharjee, Maumita; Chawla, Shikha; Chameettachal, Shibu; Murab, Sumit; Bhavesh, Neel Sarovar; Ghosh, Sourabh

    2016-04-01

    Strategies for tissue engineering focus on scaffolds with tunable structure and morphology as well as optimum surface chemistry to simulate the anatomy and functionality of the target tissue. Silk fibroin has demonstrated its potential in supporting cartilaginous tissue formation both in vitro and in vivo. In this study, we investigate the role of controlled lamellar organization and chemical composition of biofunctionalized silk scaffolds in replicating the structural properties of the annulus region of an intervertebral disc using articular chondrocytes. Covalent attachment of chondroitin sulfate (CS) to silk is characterized. CS-conjugated silk constructs demonstrate enhanced cellular metabolic activity and chondrogenic redifferentiation potential with significantly improved mechanical properties over silk-only constructs. A matrix-assisted laser desorption ionization-time of flight analysis and protein-protein interaction studies help to generate insights into how CS conjugation can facilitate the production of disc associated matrix proteins, compared to a silk-only based construct. An in-depth understanding of the interplay between such extra cellular matrix associated proteins should help in designing more rational scaffolds for cartilaginous disc regeneration needs. PMID:27068621

  9. 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)

  10. 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

  11. 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,...

  12. 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.

  13. 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.

  14. 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

  15. 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

  16. Alleged silk spigots on tarantula feet: electron microscopy reveals sensory innervation, no silk.

    Foelix, Rainer; Erb, Bruno; Rast, Bastian

    2013-05-01

    Several studies on tarantulas have claimed that their tarsi could secrete fine silk threads which would provide additional safety lines for maintaining a secure foot-hold on smooth vertical surfaces. This interpretation was seriously questioned by behavioral experiments, and more recently morphological evidence indicated that the alleged spigots ("ribbed hairs") were not secretory but most likely sensory hairs (chemoreceptors). However, since fine structural studies were lacking, the sensory nature was not proven convincingly. By using transmission electron microscopy we here present clear evidence that these "ribbed hairs" contain many dendrites inside the hair lumen - as is the case in the well-known contact chemoreceptors of spiders and insects. For comparison, we also studied the fine structure of regular silk spigots on the spinnerets and found them distinctly different from sensory hairs. Finally, histological studies of a tarantula tarsus did not reveal any silk glands, which, by contrast, are easily found within the spinnerets. In conclusion, the alleged presence of silk spigots on tarantula feet is refuted. PMID:23474440

  17. Modifying the mechanical properties of silk nanofiber scaffold by knitted orientation for regenerative medicine applications.

    Dodel, M; Hemmati Nejad, N; Bahrami, S H; Soleimani, M; Hanaee-Ahvaz, H

    2016-01-01

    Tissue reconstruction is among the increasing applications of polymer nanofibers. Fibrous scaffolds (mats) can be easily produced using the electrospinning method with structure and biomechanical properties similar to those of a cellular matrix. Electrospinning is widely used in the production of nanofibers and the GAP-method electrospinning is one of the means of producing fully aligned nanofibers. In this research, using the GAP-method, knitted fibrous scaffolds were made of silk fibroin, which is a biocompatible and biodegradable polymer. To extract fibroin from cocoons, the sodium chloride solution as well as dialysis and freeze-drying techniques were employed. The molecular weight of the extracted fibroin was measured with the SDS-Page electrophoresis technique. Moreover, the pure fibroin structure was examined using the ATR-FTIR method, and the viscosity of the solution used for electrospinning was measured with the Brookfield rotational viscometer. The scaffolds were prepared through electrospinning of the silk fibroin in pure formic acid solution. The following three structures were electrospun: 1) a random structure; 2) a knitted structure with an interstitial angle of 60 degrees; 3) a knitted structure with an interstitial angle of 90 degrees. Morphology of the resulting fibers was studied with a SEM (scanning electron microscope). Fibroin scaffolds are degradable in water. Therefore, they were fixated through immersion in methanol to be prepared for assays. The mechanical properties of the scaffolds were also studied using a tensile strength test device. The effect of methanol on the strength properties of the samples was also assessed. The hydrophilic potential of the samples was measured via a contact angle test. To increase the hydrophilicity of the scaffold surfaces, the cold oxygen plasma technique was employed. Finally, the biocompatibility and cell adhesion of the resulting scaffolds were examined through a HEK 293 cell culture, and the results

  18. The Tao of Silk

    VALERIE; SARTOR

    2007-01-01

    China’s most ancient gift to the world, silk, arrived long before gunpowder, paper and printmaking. This coveted fabric predated Christ and Buddha. Silk startled the world: Lenient ancient Romans banned it as sexy and immoral. Durable, useful and elegant, it is one of the oldest fibers known to man. Production remained a Chinese secret until 550 when two Nestorian monks, risking their lives,

  19. 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.

  20. 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

    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. (paper)

  1. 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

  2. The molecular structures of major ampullate silk proteins of the wasp spider, Argiope bruennichi: a second blueprint for synthesizing de novo silk.

    Zhang, Yang; Zhao, Ai-Chun; Sima, Yang-Hu; Lu, Cheng; Xiang, Zhong-Huai; Nakagaki, Masao

    2013-03-01

    The dragline silk of orb-weaving spiders possesses extremely high tensile strength and elasticity. To date, full-length sequences of only two genes encoding major ampullate silk protein (MaSp) in Latrodectus hesperus have been determined. In order to further understand this gene family, we utilized in this study a variety of strategies to isolate full-length MaSp1 and MaSp2 cDNAs in the wasp spider Argiope bruennichi. A. bruennichi MaSp1 and MaSp2 are primarily composed of remarkably homogeneous ensemble repeats containing several complex motifs, and both have highly conserved C-termini and N-termini. Two novel amino acid motifs, GGF and SGR, were found in MaSp1 and MaSp2, respectively. Amino acid composition analysis of silk, luminal contents and predicted sequences indicates that MaSp1 and MaSp2 are two major components of major ampullate glands and that the ratio of MaSp1 to MaSp2 is approximately 3:2 in dragline silk. Furthermore, both the MaSp1:MaSp2 ratio and the conserved termini are closely linked with the production of high quality synthetic fibers. Our results make an important contribution to our understanding of major ampullate silk protein structure and provide a second blueprint for creating new composite silk which mimics natural spider dragline silk. PMID:23262065

  3. 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 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.

  4. 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

  5. Optically switchable natural silk

    An optically active bio-material is created by blending natural silk fibers with photoisomerizable chromophore molecules—azobenzenebromide (AzBr). The material converts the energy of unpolarized light directly into mechanical work with a well-defined direction of action. The feasibility of the idea to produce optically driven microsized actuators on the basis of bio-material (silk) is proven. The switching behavior of the embedded AzBr molecules was studied in terms of UV/Vis spectroscopy. To test the opto-mechanical properties of the modified fibers and the structural changes they undergo upon optically induced switching, single fiber X-ray diffraction with a micron-sized synchrotron radiation beam was combined in situ with optical switching as well as with mechanical testing and monitoring. The crystalline regions of silk are not modified by the presence of the guest molecules, hence occupy only the amorphous part of the fibers. It is shown that chromophore molecules embedded into fibers can be reversibly switched between the trans and cis conformation by illumination with light of defined wavelengths. The host fibers respond to this switching with a variation of the internal stress. The amplitude of the mechanical response is independent of the applied external stress and its characteristic time is shorter than the relaxation time of the usual mechanical response of silk

  6. 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

  7. 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.

  8. 丝素蛋白支架的制备及其在组织工程中的应用%Preparation and Application of Silk Fibrion Scaffolds for Tissue Engineering

    孔令玲; 李延报

    2013-01-01

    作为一种天然的纤维蛋白,蚕丝具有良好的力学性能、生物相容性和可降解性,这使其在生物医学方面的应用不断拓展.多孔支架能够为细胞的粘附和增殖提供有利的微环境,在重塑和修复组织的过程中起着至关重要的作用.本文综述了丝素蛋白支架的多种制备方法,如静电纺丝法、冷冻干燥法、沥除法和发泡法等,阐述了丝素蛋白支架的表面改性方法,总结了丝素蛋白支架在组织工程中的应用.%As a natural fibrous protein,silk fibroin has remarkable mechanical properties,biocompatibility and biodegradability,which makes it widely used in biomedical area.Porous scaffolds provide a favorable microenvironment for cell adhesion and proliferation and play a key role in the process of remodeling and repairing tissue.In this paper,a variety of preparation methods of silk fibroin scaffolds including electrospinning,freeze-drying,leaching and foaming.The surface modification of the silk fibroin scaffolds is described and the applications of silk fibroin scaffolds in tissue engineering are summarized.

  9. The silk protein, sericin, protects against cell death caused by acute serum deprivation in insect cell culture.

    Takahashi, Masakazu; Tsujimoto, Kazuhisa; Yamada, Hideyuki; Takagi, Hiroshi; Nakamori, Shigeru

    2003-11-01

    Sericin is the silk protein that covers fibroin fibers and functions as a 'glue' in the cocoons of silkworms, and its most abundant component, Ser1, contains repeats of Ser- and Thr-rich 38 amino acid residues. The viability of Sf9 insect cells was 20, 57 and 49% on the fifth day and 41, 91 and 70% on the ninth day after serum deprivation in the presence of no additives, 3000 microg sericin hydrolysate and 350 microg SerD (the peptide containing the two repetitive units) ml(-1), respectively. Thus, the sericin samples were useful in preventing cell death and promoting cellular growth after acute serum deprivation. PMID:14677702

  10. 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

  11. 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.

  12. Complementary effects of two growth factors in multifunctionalized silk nanofibers for nerve reconstruction.

    Tony M Dinis

    Full Text Available With the aim of forming bioactive guides for peripheral nerve regeneration, silk fibroin was electrospun to obtain aligned nanofibers. These fibers were functionalized by incorporating Nerve Growth Factor (NGF and Ciliary NeuroTrophic Factor (CNTF during electrospinning. PC12 cells grown on the fibers confirmed the bioavailability and bioactivity of the NGF, which was not significantly released from the fibers. Primary neurons from rat dorsal root ganglia (DRGs were grown on the nanofibers and anchored to the fibers and grew in a directional fashion based on the fiber orientation, and as confirmed by growth cone morphology. These biofunctionalized nanofibers led to a 3-fold increase in neurite length at their contact, which was likely due to the NGF. Glial cell growth, alignment and migration were stimulated by the CNTF in the functionalized nanofibers. Organotypic culture of rat fetal DRGs confirmed the complementary effect of both growth factors in multifunctionalized nanofibers, which allowed glial cell migration, alignment and parallel axonal growth in structures resembling the 'bands of Bungner' found in situ. Graftable multi-channel conduits based on biofunctionalized aligned silk nanofibers were developed as an organized 3D scaffold. Our bioactive silk tubes thus represent new options for a biological and biocompatible nerve guidance conduit.

  13. 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

  14. 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.

  15. 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 ...

  16. Anterior cruciate ligament reconstruction in a rabbit model using silk-collagen scaffold and comparison with autograft.

    Fanggang Bi

    Full Text Available The objective of the present study was to perform an in vivo assessment of a novel silk-collagen scaffold for anterior cruciate ligament (ACL reconstruction. First, a silk-collagen scaffold was fabricated by combining sericin-extracted knitted silk fibroin mesh and type I collagen to mimic the components of the ligament. Scaffolds were electron-beam sterilized and rolled up to replace the ACL in 20 rabbits in the scaffold group, and autologous semitendinosus tendons were used to reconstruct the ACL in the autograft control group. At 4 and 16 weeks after surgery, grafts were retrieved and analyzed for neoligament regeneration and tendon-bone healing. To evaluate neoligament regeneration, H&E and immunohistochemical staining was performed, and to assess tendon-bone healing, micro-CT, biomechanical test, H&E and Russell-Movat pentachrome staining were performed. Cell infiltration increased over time in the scaffold group, and abundant fibroblast-like cells were found in the core of the scaffold graft at 16 weeks postoperatively. Tenascin-C was strongly positive in newly regenerated tissue at 4 and 16 weeks postoperatively in the scaffold group, similar to observations in the autograft group. Compared with the autograft group, tendon-bone healing was better in the scaffold group with trabecular bone growth into the scaffold. The results indicate that the silk-collagen scaffold has considerable potential for clinical application.

  17. Anterior cruciate ligament reconstruction in a rabbit model using silk-collagen scaffold and comparison with autograft.

    Bi, Fanggang; Shi, Zhongli; Liu, An; Guo, Peng; Yan, Shigui

    2015-01-01

    The objective of the present study was to perform an in vivo assessment of a novel silk-collagen scaffold for anterior cruciate ligament (ACL) reconstruction. First, a silk-collagen scaffold was fabricated by combining sericin-extracted knitted silk fibroin mesh and type I collagen to mimic the components of the ligament. Scaffolds were electron-beam sterilized and rolled up to replace the ACL in 20 rabbits in the scaffold group, and autologous semitendinosus tendons were used to reconstruct the ACL in the autograft control group. At 4 and 16 weeks after surgery, grafts were retrieved and analyzed for neoligament regeneration and tendon-bone healing. To evaluate neoligament regeneration, H&E and immunohistochemical staining was performed, and to assess tendon-bone healing, micro-CT, biomechanical test, H&E and Russell-Movat pentachrome staining were performed. Cell infiltration increased over time in the scaffold group, and abundant fibroblast-like cells were found in the core of the scaffold graft at 16 weeks postoperatively. Tenascin-C was strongly positive in newly regenerated tissue at 4 and 16 weeks postoperatively in the scaffold group, similar to observations in the autograft group. Compared with the autograft group, tendon-bone healing was better in the scaffold group with trabecular bone growth into the scaffold. The results indicate that the silk-collagen scaffold has considerable potential for clinical application. PMID:25938408

  18. Incorporation of elastase inhibitor in silk fibroin nanoparticles for transdermal delivery

    Ferreira, Ana Vanessa Fernandes

    2013-01-01

    Dissertação de mestrado em Bioengenharia Human neutrophil elastase, as inflammatory response, has the capacity to degrade collagen and elastin component of extracellular matrix, being extensively involved in wrinkles formation induced by UV radiation damage. In order to develop a cosmetic antiwrinkling emulsion, we incorporated a potent synthetic HNE inhibitor, sivelestat (IC50 = 44 nM, Ki = 0.2 μM), into biopolymeric nanoparticles prepared by high-energy emulsification meth...

  19. Alcalase水解丝素蛋白的特性%Properties of Alcalase hydrolyzing silk fibroin

    周凤娟; 许时婴; 杨瑞金; 王璋

    2007-01-01

    研究了Alcalase水解丝素蛋白的特性.结果表明,Alcalase水解丝素蛋白的最佳条件是:丝素蛋白质量浓度50 mg/mL,pH值9.0,温度60℃;丝素肽中主要氨基酸(甘氨酸、丙氨酸、丝氨酸和酪氨酸)的含量之和占其氨基酸总量的85%左右;水解度为17%的丝素蛋白水解产物中主要组分的相对分子质量分别为249 Da和762 Da,含量分别约占72.9%和15.5%,水解度为21%的丝素蛋白水解产物中主要组分的相对分子质量分别为209 Da和668 Da,含量分别约占84.4%和7.2%.

  20. Ultra-Lightweight Resistive Switching Memory Devices Based on Silk Fibroin.

    Wang, Hong; Zhu, Bowen; Wang, Hua; Ma, Xiaohua; Hao, Yue; Chen, Xiaodong

    2016-07-01

    Ultra-lightweight resistive switching memory based on protein has been demonstrated. The memory foil is 0.4 mg cm(-2) , which is 320-fold lighter than silicon substrate, 20-fold lighter than office paper and can be sustained by a human hair. Additionally, high resistance OFF/ON ratio of 10(5) , retention time of 10(4) s, and excellent flexibility (bending radius of 800 μm) have been achieved. PMID:27315137

  1. Analytical markers for silk degradation: comparing historic silk and silk artificially aged in different environments

    Vilaplana, Francisco; Nilsson, Johanna; Sommer, Dorte V. P.; Karlsson, Sigbritt

    2014-01-01

    Suitable analytical markers to assess the degree of degradation of historic silk textiles at molecular and macroscopic levels have been identified and compared with silk textiles aged artificially in different environments, namely (i) ultraviolet (UV) exposure, (ii) thermo-oxidation, (iii) controlled humidity and (iv) pH. The changes at the molecular level in the amino acid composition, the formation of oxidative moieties, crystallinity and molecular weight correlate well with the changes in ...

  2. 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.

  3. Production of Silk & Wool Blended Worsted Cloth

    段亚峰; 许琳

    2004-01-01

    Wool & silk blended fancy suiting is desinged. Through trial-production with silk sliver and Australian wool top, the spinning technology is investigated, and the relationship of spinning technology, blending ratio and yam construction is discussed.

  4. Recombinant DNA production of spider silk proteins

    Tokareva, Olena; Michalczechen-Lacerda, Valquíria A; Rech, Elíbio L; Kaplan, David L.

    2013-01-01

    Spider dragline silk is considered to be the toughest biopolymer on Earth due to an extraordinary combination of strength and elasticity. Moreover, silks are biocompatible and biodegradable protein-based materials. Recent advances in genetic engineering make it possible to produce recombinant silks in heterologous hosts, opening up opportunities for large-scale production of recombinant silks for various biomedical and material science applications. We review the current strategies to produce...

  5. PEGylated Silk Nanoparticles for Anticancer Drug Delivery

    Wongpinyochit, Thidarat; Uhlmann, Petra; Urquhart, Andrew; Seib, F. Philipp

    2015-01-01

    Silk has a robust clinical track record and is emerging as a promising biopolymer for drug delivery, including its use as nanomedicine. However, silk-based nanomedicines still require further refinements for full exploitation of their potential; the application of “stealth” design principals is e...... to human breast cancer cells. In conclusion, these results, taken together with prior silk nanoparticle data, support a viable future for silk-based nanomedicines....

  6. Identification of conserved microRNAs in Bombyx mori (silkworm) and regulation of fibroin L chain production by microRNAs in heterologous system.

    Cao, Jun; Tong, Chuanzhou; Wu, Xiaojie; Lv, Jianning; Yang, Zhaolin; Jin, Yongfeng

    2008-12-01

    MicroRNAs (miRNAs) are an extensive class of small non-coding RNAs with important roles in the regulation of gene expression. Little study has been performed on identifying miRNAs in Bombyx mori (silkworm). Here, 41 conserved miRNAs were identified by a computational homology search approach and some of them were selected and confirmed experimentally. Some characteristics such as position-specific nucleotide distribution, evolutionary conservation and clustered organization have been described. Through general method of miRNA target identification, we also find that at least 4 miRNAs (miR-33/-190/-276/-7) target fibroin L chain transcripts, suggesting that miRNAs might play an important role in the regulation of silk protein production. In order to validate this, 3'-UTR of fibroin L chain mRNA was cloned and several expression vectors were constructed. Assays of semi-RT-PCR, histochemical and biochemical assays after agroinfiltration indicated that miRNAs can repress reporter gene (GUS) expression with sequence complement. Clearly, agroinfiltration proved a highly efficient method to identify the function of miRNAs inhibiting their targets mRNA in heterologous system. PMID:18977439

  7. 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.

  8. Analytical markers for silk degradation: comparing historic silk and silk artificially aged in different environments.

    Vilaplana, Francisco; Nilsson, Johanna; Sommer, Dorte V P; Karlsson, Sigbritt

    2015-02-01

    Suitable analytical markers to assess the degree of degradation of historic silk textiles at molecular and macroscopic levels have been identified and compared with silk textiles aged artificially in different environments, namely (i) ultraviolet (UV) exposure, (ii) thermo-oxidation, (iii) controlled humidity and (iv) pH. The changes at the molecular level in the amino acid composition, the formation of oxidative moieties, crystallinity and molecular weight correlate well with the changes in the macroscopic properties such as brightness, pH and mechanical properties. These analytical markers are useful to understand the degradation mechanisms that silk textiles undergo under different degradation environments, involving oxidation processes, hydrolysis, chain scission and physical arrangements. Thermo-oxidation at high temperatures proves to be the accelerated ageing procedure producing silk samples that most resembled the degree of degradation of early seventeenth-century silk. These analytical markers will be valuable to support the textile conservation tasks currently being performed in museums to preserve our heritage. PMID:25492090

  9. Fabricating a reactive surface on the fibroin film by a room-temperature plasma jet array for biomolecule immobilization

    Chen Guang-Liang; Zheng Xu; Lü Guo-Hua; Zhang Zhao-Xia; Sylvain Massey; Wilson Smith; Michael Tatoulian; Yang Si-Ze

    2012-01-01

    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.

  10. Sunlight-Induced Coloration of Silk.

    Yao, Ya; Tang, Bin; Chen, Wu; Sun, Lu; Wang, Xungai

    2016-12-01

    Silk fabrics were colored by gold nanoparticles (NPs) that were in situ synthesized through the induction of sunlight. Owing to the localized surface plasmon resonance (LSPR) of gold NPs, the treated silk fabrics presented vivid colors. The photo-induced synthesis of gold NPs was also realized on wet silk through adsorbing gold ions out of solution, which provides a water-saving coloration method for textiles. Besides, the patterning of silk was feasible using this simple sunlight-induced coloration approach. The key factors of coloration including gold ion concentration, pH value, and irradiation time were investigated. Moreover, it was demonstrated that either ultraviolet (UV) light or visible light could induce the generation of gold NPs on silk fabrics. The silk fabrics with gold NPs exhibited high light resistance including great UV-blocking property and excellent fastness to sunlight. PMID:27297220

  11. Thymus Gland Anatomy

    ... historical Searches are case-insensitive Thymus Gland, Adult, Anatomy Add to My Pictures View /Download : Small: 720x576 ... Large: 3000x2400 View Download Title: Thymus Gland, Adult, Anatomy Description: Anatomy of the thymus gland; drawing shows ...

  12. Revival of the Silk Road

    2006-01-01

    China's Xinjiang Uygur Autonomous Region, bordering eight countries and providing a corridor to the Central Asian region, was once famous for the Silk Road. But now this region is often reported by Western media as having the problems of "political separatism, economic exclusion and religious fundamentalism." Professor David Gosset with the Shanghai-based China Europe International Business School analyzed Xinjiang's development, which he framed as "Xinjiang's experience." According to him, Xinjiang has ...

  13. 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.

  14. Extracted sericin from silk waste for film formation

    Rungsinee Sothornvit; Rungsima Chollakup; Potjanart Suwanruji

    2010-01-01

    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 ex...

  15. Drömmen om svenskt silke

    Johansson Åbonde, Anders

    2010-01-01

    The grandiose, but failed, attempts to produce silk in Sweden in the past have been largely ignored by historians. This thesis describes the history of sericulture in Sweden, including three periods of practical trials in 1735-1765, 1830-1898 and 1913-1918. For a long time, the secrets of sericulture were closely guarded in China but by the beginning of the 16th century, knowledge of silk cultivation techniques had reached a number of European countries. The pursuit of domestic silk productio...

  16. 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.

  17. Silk as an innovative biomaterial for cancer therapy.

    Jastrzebska, Katarzyna; Kucharczyk, Kamil; Florczak, Anna; Dondajewska, Ewelina; Mackiewicz, Andrzej; Dams-Kozlowska, Hanna

    2015-01-01

    Silk has been used for centuries in the textile industry and as surgical sutures. In addition to its unique mechanical properties, silk possesses other properties, such as biocompatibility, biodegradability and ability to self-assemble, which make it an interesting material for biomedical applications. Although silk forms only fibers in nature, synthetic techniques can be used to control the processing of silk into different morphologies, such as scaffolds, films, hydrogels, microcapsules, and micro- and nanospheres. Moreover, the biotechnological production of silk proteins broadens the potential applications of silk. Synthetic silk genes have been designed. Genetic engineering enables modification of silk properties or the construction of a hybrid silk. Bioengineered hybrid silks consist of a silk sequence that self-assembles into the desired morphological structure and the sequence of a polypeptide that confers a function to the silk biomaterial. The functional domains can comprise binding sites for receptors, enzymes, drugs, metals or sugars, among others. Here, we review the current status of potential applications of silk biomaterials in the field of oncology with a focus on the generation of implantable, injectable and targeted drug delivery systems and the three-dimensional cancer models based on silk scaffolds for cancer research. However, the systems described could be applied in many biomedical fields. PMID:25859397

  18. Structure and post-translational modifications of the web silk protein spidroin-1 from Nephila spiders.

    dos Santos-Pinto, José Roberto Aparecido; Lamprecht, Günther; Chen, Wei-Qiang; Heo, Seok; Hardy, John George; Priewalder, Helga; Scheibel, Thomas Rainer; Palma, Mario Sergio; Lubec, Gert

    2014-06-13

    Spidroin-1 is one of the major ampullate silk proteins produced by spiders for use in the construction of the frame and radii of orb webs, and as a dragline to escape from predators. Only partial sequences of spidroin-1 produced by Nephila clavipes have been reported up to now, and there is no information on post-translational modifications (PTMs). A gel-based mass spectrometry strategy with ETD and CID fragmentation methods were used to sequence and determine the presence/location of any PTMs on the spidroin-1. Sequence coverage of 98.06%, 95.05%, and 98.37% were obtained for N. clavipes, Nephila edulis and for Nephila madagascariensis, respectively. Phosphorylation was the major PTM observed with 8 phosphorylation sites considered reliable on spidroin-1 produced by N. clavipes, 4 in N. madagascariensis and 2 for N. edulis. Dityrosine and 3,4-dihydroxyphenylalanine (formed by oxidation of the spidroin-1) were observed, although the mechanism by which they are formed (i.e. exposure to UV radiation or to peroxidases in the major ampullate silk gland) is uncertain. Herein we present structural information on the spidroin-1 produced by three different Nephila species; these findings may be valuable for understanding the physicochemical properties of the silk proteins and moreover, future designs of recombinantly produced spider silk proteins. Biotechnological significance The present investigation shows for the first time spidroin structure and post-translational modifications observed on the major ampullate silk spidroin-1. The many site specific phosphorylations (localized within the structural motifs) along with the probably photoinduction of hydroxylations may be relevant for scientists in material science, biology, biochemistry and environmental scientists. Up to now all the mechanical properties of the spidroin have been characterized without any consideration about the existence of PTMs in the sequence of spidroins. Thus, these findings for major ampullate silk

  19. 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 - ...

  20. Effect of solute concentration on fibroin regulated biomineralization of calcium phosphate

    In this paper we used UV/Visible method to study the effect of solute concentration on fibroin regulated biomineralization of calcium phosphate. During the reaction process, ultraviolet-visible spectrophotometer was used to track the extinction in the reaction solution. It is found that kinetics of the biomineralization can be strongly affected by the presence of fibroin. Fibroin with higher concentration has more positive effect on the biomineralization process. Under the appropriate reaction conditions, wave crest and wave trough appear in the kinetic curves of fibroin biomineralization. The wave crest and wave trough phenomenon is mainly related with the process of phase separation. X-ray Diffraction (XRD) result shows the calcium phosphate before the wave trough is mainly amorphous calcium phosphate, while after the wave trough crystal of hydroxyapatite (HA) and brushite (DCPD) are the mainly ingredients

  1. Capacity of Nylon Industrial Silk Is Surplus

    2007-01-01

    @@ The total nylon (polyamide) industrial silk output worldwide was 1.025 million tons in 2005, mostly provided by the producers in Asia, Europe and America, in which the ratio of nylon 6 to nylon 66 was 6:4.

  2. 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.

  3. Modified fiber qualities of the transgenic cotton expressing a silkworm fibroin gene

    LI FeiFei; WU ShenJie; L(U) FenNi; CHEN TianZi; JU Ming; WANG HaiHai; JIANG YanJie; ZHANG Jie; GUO WangZhen; ZHANG TianZhen

    2009-01-01

    A silkworm gene for fibroin was introduced into the upland cotton WC line by Agrobacterium-mediated transformation. PCR detection for fibroin, nptll and gus genes, Kanamycin (Km)-resistance analysis and GUS-histochemical assay were conducted on 30 regenerated plants from 9 callus lines, and 17 positive plants were obtained by these 5 screening methods. By Km-resistance analysis and PCR for fibroin, 6 homozygous lines in T3 were obtained. Southern blot and Northern bolt demonstrated that the fibroin gene was inserted into the genome of these 6 lines, stably inherited and expressed. Compared to the control, the surface structure of mature fiber in the 6 lines was significantly distorted and an increased number of convolution was observed by scanning electron microscopy (SEM). Fiber quality traits analysis indicated that fiber elongation of the 6 homozygous lines was all increased and fiber strength of 3 lines was enhanced. These results indicated that fibroin expression influenced cotton fiber structure and quality, suggesting that fibroin has great potential for improving cotton fiber quality by genetic engineering.

  4. 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

  5. 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)

  6. New Evidence for Early Silk in the Indus Civilization

    Good, I. L.; Kenoyer, J. M.; Meadow, Richard Henry

    2009-01-01

    Silk is an important economic fibre, and is generally considered to have been the exclusive cultural heritage of China. Silk weaving is evident from the Shang period c. 1600–1045 bc, though the earliest evidence for silk textiles in ancient China may date to as much as a millennium earlier. Recent microscopic analysis of archaeological thread fragments found inside copper-alloy ornaments from Harappa and steatite beads from Chanhu-daro, two important Indus sites, have yielded silk fibres, dat...

  7. Silk as an innovative biomaterial for cancer therapy

    Jastrzebska, Katarzyna; Kucharczyk, Kamil; Florczak, Anna; Dondajewska, Ewelina; Mackiewicz, Andrzej; Dams-Kozlowska, Hanna

    2014-01-01

    Silk has been used for centuries in the textile industry and as surgical sutures. In addition to its unique mechanical properties, silk possesses other properties, such as biocompatibility, biodegradability and ability to self-assemble, which make it an interesting material for biomedical applications. Although silk forms only fibers in nature, synthetic techniques can be used to control the processing of silk into different morphologies, such as scaffolds, films, hydrogels, microcapsules, an...

  8. Speeding up the Internationalization of Raw Silk Standard

    Hu Zuozhong; Du Zhouhe

    2005-01-01

    @@ China is the main silk producer in the world,whose RSS (Raw Silk Standard) is widely approved in international trade. After entering WTO, China needs to revise the RSS according to the changes of silk industry and develop advanced electronic testing system and improve testing means, so as to promote the internationlization of the RSS.

  9. Post-secretion processing influences spider silk performance.

    Blamires, Sean J; Wu, Chung-Lin; Blackledge, Todd A; Tso, I-Min

    2012-10-01

    Phenotypic variation facilitates adaptations to novel environments. Silk is an example of a highly variable biomaterial. The two-spidroin (MaSp) model suggests that spider major ampullate (MA) silk is composed of two proteins-MaSp1 predominately contains alanine and glycine and forms strength enhancing β-sheet crystals, while MaSp2 contains proline and forms elastic spirals. Nonetheless, mechanical properties can vary in spider silks without congruent amino acid compositional changes. We predicted that post-secretion processing causes variation in the mechanical performance of wild MA silk independent of protein composition or spinning speed across 10 species of spider. We used supercontraction to remove post-secretion effects and compared the mechanics of silk in this 'ground state' with wild native silks. Native silk mechanics varied less among species compared with 'ground state' silks. Variability in the mechanics of 'ground state' silks was associated with proline composition. However, variability in native silks did not. We attribute interspecific similarities in the mechanical properties of native silks, regardless of amino acid compositions, to glandular processes altering molecular alignment of the proteins prior to extrusion. Such post-secretion processing may enable MA silk to maintain functionality across environments, facilitating its function as a component of an insect-catching web. PMID:22628213

  10. Woven silk fabric-reinforced silk nanofibrous scaffolds for regenerating load-bearing soft tissues.

    Han, F; Liu, S; Liu, X; Pei, Y; Bai, S; Zhao, H; Lu, Q; Ma, F; Kaplan, D L; Zhu, H

    2014-02-01

    Although three-dimensional (3-D) porous regenerated silk scaffolds with outstanding biocompatibility, biodegradability and low inflammatory reactions have promising application in different tissue regeneration, the mechanical properties of regenerated scaffolds, especially suture retention strength, must be further improved to satisfy the requirements of clinical applications. This study presents woven silk fabric-reinforced silk nanofibrous scaffolds aimed at dermal tissue engineering. To improve the mechanical properties, silk scaffolds prepared by lyophilization were reinforced with degummed woven silk fabrics. The ultimate tensile strength, elongation at break and suture retention strength of the scaffolds were significantly improved, providing suitable mechanical properties strong enough for clinical applications. The stiffness and degradation behaviors were then further regulated by different after-treatment processes, making the scaffolds more suitable for dermal tissue regeneration. The in vitro cell culture results indicated that these scaffolds maintained their excellent biocompatibility after being reinforced with woven silk fabrics. Without sacrifice of porous structure and biocompatibility, the fabric-reinforced scaffolds with better mechanical properties could facilitate future clinical applications of silk as matrices in skin repair. PMID:24090985

  11. 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

  12. Spinning gland transcriptomics from two main clades of spiders (order: Araneae--insights on their molecular, anatomical and behavioral evolution.

    Francisco Prosdocimi

    Full Text Available Characterized by distinctive evolutionary adaptations, spiders provide a comprehensive system for evolutionary and developmental studies of anatomical organs, including silk and venom production. Here we performed cDNA sequencing using massively parallel sequencers (454 GS-FLX Titanium to generate ∼80,000 reads from the spinning gland of Actinopus spp. (infraorder: Mygalomorphae and Gasteracantha cancriformis (infraorder: Araneomorphae, Orbiculariae clade. Actinopus spp. retains primitive characteristics on web usage and presents a single undifferentiated spinning gland while the orbiculariae spiders have seven differentiated spinning glands and complex patterns of web usage. MIRA, Celera Assembler and CAP3 software were used to cluster NGS reads for each spider. CAP3 unigenes passed through a pipeline for automatic annotation, classification by biological function, and comparative transcriptomics. Genes related to spider silks were manually curated and analyzed. Although a single spidroin gene family was found in Actinopus spp., a vast repertoire of specialized spider silk proteins was encountered in orbiculariae. Astacin-like metalloproteases (meprin subfamily were shown to be some of the most sampled unigenes and duplicated gene families in G. cancriformis since its evolutionary split from mygalomorphs. Our results confirm that the evolution of the molecular repertoire of silk proteins was accompanied by the (i anatomical differentiation of spinning glands and (ii behavioral complexification in the web usage. Finally, a phylogenetic tree was constructed to cluster most of the known spidroins in gene clades. This is the first large-scale, multi-organism transcriptome for spider spinning glands and a first step into a broad understanding of spider web systems biology and evolution.

  13. 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 ...

  14. 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...

  15. Actinomycosis of Submandibular Gland

    Actinomycosis is defined as a chronic, specific, suppurative, granulomatous disease caused mainly by the anaerobic, gram positive organism, Actinomyces israelii. Actinomycosis in the salivary gland is a rare disease that is caused by an inhabitant of the normal flora. We report the case of the actinomycosis of submandibular gland. A 53-year old man presented with the swelling on left submandiblar area. The lesion was not painful but had been increasing for about 10 days. In the CT view, the internal portion of the mass showed homogeneous moderate signal. The mass had continuities with the inferior portion of the left enlarged submandibular gland. In the MRI, there was a mass that showed a buldging pattern inferiorly in the left submandibular gland without bony invasion sign. The biopsy shows the colony of special organism. Many filaments are discovered with clubbed ends diffused from center of colony. We diagnosed this disease as actinomycosis in the submandibular gland by the postoperational biopsy.

  16. 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.)

  17. 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…

  18. The Application of Tea Dyeing to Silk

    金成嬉

    2001-01-01

    Vegetable dyes are eco-friendly throughout the full production process. A study is conducted with the purpose of assessing the properties of dye extracted from green tea, black tea and the tea tree cultivated and used in Jiang Nan area of China. The extracted dyes are applied with and without mordants on silk fabric and the dyeing properties are evaluated.

  19. Silk-Screening a la Andy.

    Mathes, Len

    2000-01-01

    Describes a project that was used with advanced 11th and 12th grade art students in which they created silk-screen self-portraits in the style of Andy Warhol. Discusses the process of creating the portraits and the activities that concluded the project. Lists the needed materials. (CMK)

  20. China-Indian Silk Trade: Current Production and Future Prospects

    Liu Feng; Sun Shimin; Qiao Xianjuan

    2009-01-01

    As an important traditional labor-inteusive industry of both India and China, the cocoon silk industry has long made great contributions to the ecological environment protection, rural economic development and the increase in export income of both countries. India is not only a very important cocoon silk trading partner, but an important production competitor of China.In recent years, there has been a large increase in the production and trade of the cocoon silk between China and India; however,China relies heavily on Indian market, which leads to a tendency of further deterioration in the silk trade environment between both countries. The present article makes an empirical study of the cocoon silk resources of the two countries and the scale, product mix and market structure of China-Indian silk trade from 2001 to 2007. Overall silk trading volumes from China to India and market concentration rate are on the increase because of the superiority of Chinese cocoon silk production over that of India. Owing to scat-tered market share and export that mainly focused on raw materi-als product, there has been a phenomenon of price reduction and quantity increase. India carries out fierce competition with China in the international market and even imposes antidumping sanction on Chinese silk, which are key factors restricting further increase between China-India trade. Based on the abovementioned facts, the authors aim to put forward suggestions for steadily developing the production and trade of China's silk.

  1. 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.

  2. 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 ...

  3. Thyroid gland removal - discharge

    ... this page: //medlineplus.gov/ency/patientinstructions/000293.htm Thyroid gland removal - discharge To use the sharing features ... surgery. This will make your scar show less. Thyroid Hormone Replacement You may need to take thyroid ...

  4. Thyroid gland removal

    ... small thyroid growth ( nodule or cyst) A thyroid gland that is so overactive it is dangerous ( thyrotoxicosis ) Cancer of the thyroid Noncancerous (benign) tumors of the thyroid that are causing symptoms Thyroid ...

  5. Pituitary Gland Disorders Overview

    ... Thyroid-stimulating hormone (TSH) - TSH stimulates the thyroid gland to produce thyroid hormones, which regulate the body's metabolism, energy balance, growth, and nervous system activity. Read about TSH-secreting tumors . Luteinizing hormone (LH) - LH stimulates testosterone production in ...

  6. Laparoscopic Adrenal Gland Removal

    ... adrenal tumors that appear malignant. What are the Advantages of Laparoscopic Adrenal Gland Removal? In the past, ... of procedure and the patients overall condition. Common advantages are: Less postoperative pain Shorter hospital stay Quicker ...

  7. Pineal and Pituitary Glands

    ... Registries Review Brain & CNS Tumors Signs & Symptoms Anatomy Neurons & Glial Cells Brain Meninges Spinal Cord Cranial Nerves Pineal and Pituitary Glands Abstracting, Coding, & Staging Topographic Sites Histological Types Grading Staging Multiple Primaries Malignant Transformation Coding Sequence ...

  8. Mammary gland stem cells

    Fridriksdottir, Agla J R; Petersen, Ole W; Rønnov-Jessen, Lone

    2011-01-01

    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...... develops and is maintained, significant discrepancies exist between the mouse and human gland which should be taken into consideration in current and future models of mammary stem cell biology....

  9. 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.

  10. 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

  11. Identification and classification of silks using infrared spectroscopy

    M. Boulet-Audet; Vollrath, F.; Holland, C.

    2015-01-01

    ABSTRACT Lepidopteran silks number in the thousands and display a vast diversity of structures, properties and industrial potential. To map this remarkable biochemical diversity, we present an identification and screening method based on the infrared spectra of native silk feedstock and cocoons. Multivariate analysis of over 1214 infrared spectra obtained from 35 species allowed us to group silks into distinct hierarchies and a classification that agrees well with current phylogenetic data an...

  12. Elasto-capillary windlass : from spider silk to smart actuators

    Elettro, Hervé

    2015-01-01

    This PhD work aimed to understand and recreate artificially a self-assembling mechanism involving capillarity and elasticity present in spider silk. The primary function of the micronic glue droplets that exist on spider capture silk is to provide the spider web with adhesive properties. These droplets play yet another role: the dramatic enhancement of silk mechanical properties, as well as the preservation of the integrity of the web structure. The localization of the buckling instability wi...

  13. Blended Yarns of Modacrylic Fibers with Silk Protein

    江慧

    2001-01-01

    The fiber properties of modacrylic fibers with silk protein and spinning technology for blended knitting yarns are studied. By testing the fiber properties, fiber spinnability is analysed. Modacrylic fibers with silk protein are brittle and have poor cohesion, so the key to process modacrylic fibers with silk protein lies in forming lap and sliver. During opening and carding, low speed is used to decrease fiber damage and the proper static resistant oil is applied to eliminate lapping fibers. Besides, the temperature and relative humidity have to be strictly controlled. The applications of modacrylic fibers with silk protein are also put forward.

  14. The Consolidation Behavior of Silk Hydrogels

    Kluge, Jonathan A.; Rosiello, Nicholas C.; Leisk, Gary G.; Kaplan, David L.; Dorfmann, A. Luis

    2009-01-01

    Hydrogels have mechanical properties and structural features that are similar to load bearing soft tissues including intervertebral disc and articular cartilage, and can be implanted for tissue restoration or for local release of therapeutic factors. To help predict their performance, mechanical characterization and mathematical modeling are available methods for use in tissue engineering and drug delivery settings. In this study, confined compression creep tests were performed on silk hydrog...

  15. Antioxidant Activities of Iranian Corn Silk

    Ebrahimzadeh, Mohammad Ali; POURMORAD, Fereshteh; HAFEZI, Samira

    2008-01-01

    Traditionally corn silk (CS) has been used as diuretic, antilithiasic, uricosuric, and antiseptic. It is used for the treatment of edema as well as for cystitis, gout, kidney stones, nephritis, and prostatitis. In the present study, the antioxidant properties of ethanol-water extract from CS were estimated by different methods. Also phenol and flavonoid content of the extract were measured by Folin Ciocalteu and AlCl3 assays. CS extract contained a significant amount of phenol and flavonoids....

  16. Injectable Silk Foams for Soft Tissue Regeneration

    Bellas, E.; Lo, T.J.; Fournier, E. P.; Brown, J E; Abbott, R.D.; Gil, E.S.; 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 ...

  17. From Nano to Micro: Importance of Structure and Architecture in Spider Silk Adhesives

    Sahni, Vasav; Dhinojwala, Ali

    2012-02-01

    Spiders have developed outstanding adhesives over millions of years of evolution for prey capture and locomotion. We show that the structure and architecture of these adhesives play an important role in the adhesion. The adhesive produced by orb-weaving spiders to capture prey (viscid glue) is laid on a pair of silk fibers as micron-size glue drops composed of salts and glycoproteins. By stretching single drops, we show that viscid glue behaves like a viscoelastic solid and that elasticity is critical in enhancing adhesion caused by specific adhesive ligands by over 100 times. Comparing viscid glue with gumfoot glue, the glue produced by cob-weavers, the evolutionary descendants of orb-weavers, showed that, in spite of being produced in homologous aggregate glands, gumfoot glue behaves like a viscoelastic liquid. Moreover, gumfoot glue is humidity-resistant and viscid glue is humidity-sensitive. We use a synthetic strategy to spin beads-on-a-string (BOAS) architecture to mimic the adhesive properties of spider silk. Using these mimic threads, we show that the BOAS structure adheres more than a cylindrical structure during contact (collision of prey) and during separation (escape attempt of prey). These results inspire design of novel tunable adhesives.

  18. 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

  19. Recombinant protein blends: silk beyond natural design.

    Dinjaski, Nina; Kaplan, David L

    2016-06-01

    Recombinant DNA technology and new material concepts are shaping future directions in biomaterial science for the design and production of the next-generation biomaterial platforms. Aside from conventionally used synthetic polymers, numerous natural biopolymers (e.g., silk, elastin, collagen, gelatin, alginate, cellulose, keratin, chitin, polyhydroxyalkanoates) have been investigated for properties and manipulation via bioengineering. Genetic engineering provides a path to increase structural and functional complexity of these biopolymers, and thereby expand the catalog of available biomaterials beyond that which exists in nature. In addition, the integration of experimental approaches with computational modeling to analyze sequence-structure-function relationships is starting to have an impact in the field by establishing predictive frameworks for determining material properties. Herein, we review advances in recombinant DNA-mediated protein production and functionalization approaches, with a focus on hybrids or combinations of proteins; recombinant protein blends or 'recombinamers'. We highlight the potential biomedical applications of fibrous protein recombinamers, such as Silk-Elastin Like Polypeptides (SELPs) and Silk-Bacterial Collagens (SBCs). We also discuss the possibility for the rationale design of fibrous proteins to build smart, stimuli-responsive biomaterials for diverse applications. We underline current limitations with production systems for these proteins and discuss the main trends in systems/synthetic biology that may improve recombinant fibrous protein design and production. PMID:26686863

  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. 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.

  2. 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

  3. Undifferentiated salivary gland carcinomas

    Herbst, H.; Hamilton-Dutoit, S.; Jakel, K.T.;

    2004-01-01

    Undifferentiated salivary gland carcinomas may be divided into small cell and large cell types. Among large cell undifferentiated carcinomas, lymphoepithelial carcinomas have to be distinguished, the latter of which are endemic in the Arctic regions and southern China where virtually all cases of...... these tumors are associated with the Epstein-Barr virus (EBV). Association with EBV may also be observed in sporadic cases, and detection of EBV gene products may aid their diagnosis. Immunohistology may be employed to resolve the differential diagnosis of undifferentiated salivary gland carcinomas......, 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...

  4. Radioiodine and thyroid gland

    Takavar A

    1994-05-01

    Full Text Available Administration of a precise amount of activity in thyroid gland radioiodine (¹³¹I therapy is of prime importance. Several methods for this purpose have been suggested and used over many years. In this paper, a method for required ¹³¹I activity calculation for each case has been proposed. Major factors affecting ¹³¹I calculation such as gland volume, integral dose, and prescription index have been described. Using the method, ¹³¹I activity or any other radioisotope activity required for a certain case can be determined with a good estimate.

  5. Salivary gland carcinomas

    Therkildsen, M H; Andersen, L J; Christensen, M;

    1998-01-01

    The prognosis of salivary gland carcinomas is difficult to assess. Simple mucin-type carbohydrates (T and sialosyl-T antigens, Tn and sialosyl-Tn antigens) have been shown to be of value in predicting prognosis for carcinomas in other locations. We studied the prognostic significance of the...... expression of these structures in a retrospective study of 133 patients with salivary gland carcinomas, using immunohistochemistry and a panel of well-defined monoclonal antibodies (MAbs) on formalin-fixed paraffin-embedded tissues. Sialosyl-Tn, T and sialosyl-T antigens were not correlated with prognosis...

  6. 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. …

  7. 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…

  8. 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...

  9. Photoelectric Measurement of the Fineness of Raw Silk

    Wan-chun FEI

    2010-01-01

    In order to precisely measure the diameters for obtaining the fineness of rolling raw silk, the physical features of raw silk are analyzed. By means of Fresnel principle, diffractions caused by different transparent raw silk filaments are analyzed and simulated. Image data of raw silk filament measured by digital CMOS camera are analyzed and processed for obtaining the precise diameters of the filament with the relative error of less than 1%. On the assumption of appropriate elliptic cross-section of the filament, the cross-section area is calculated as the fineness of the filament. Measurement experiments are carried out. Finally, some suggestions are proposed for photoelectric measuring the fineness of raw silk.

  10. 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

  11. 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.)

  12. 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....

  13. 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.

  14. Fabrication of robust Antheraea assama fibroin nanofibrous mat using ionic liquid for skin tissue engineering.

    Srivastava, Chandra Mohan; Purwar, Roli

    2016-11-01

    Electrospinning is an emerging technique used for fabrication of nanofibrous mats for skin tissue engineering applications. The aim of this study centered on fabrication of muga fibroin electrospun mats by using ionic liquid and its characterizations. The muga fibroin extracted from cocoon of Antheraea assama is dissolved in 1-butyl, 3-methyl imidazolium acetate (BMIMAc), a green solvent, to prepare a dope solution for electrospinning. The molecular weight, rheology and structural properties of dope solution are characterized. The process parameters of electrospinning machine such as voltage and concentration of dope solution are varied to obtain nanofibrous mats. The nanofibrous mat having average fiber diameter of 160nm are obtained from 10% w/v concentration of muga fibroin in BMIMAc with an applied voltage of 20KV. The mechanical, structural, physical and thermal properties of muga nanofibrous mat (MNF) are analyzed and compare with muga cast film. The cytocompatibility test is performed using L929 fibroblast cells. It is observed that muga nanofibrous mat support higher growth of fibroblast cells (pefficiency of gentamycin sulphate loaded muga nanofibrous mat are found to be significantly higher (pmaterial for skin tissue engineering. PMID:27524022

  15. 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.

  16. Biomimetic calcium phosphate coatings on recombinant spider silk fibres

    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.

  17. 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.

  18. Binding Quantum Dots to Silk Biomaterials for Optical Sensing

    Disi Lu

    2015-01-01

    Full Text Available Quantum dots (QDs, have great potential for fabricating optical sensing devices and imaging biomaterial degradation in vivo. In the present study, 2-mercaptoethylamine- (MEA- and mercaptopropionic acid- (MPA- capped CdTe-QDs were physically incorporated in silk films that contained a high content (>30% of crystalline beta-sheet structure. The beta-sheets were induced by the addition of glycerol, water annealing, glycerol/annealing, or treatment with methanol. Incorporation of QDs did not influence the formation of beta-sheets. When the films were extracted with water, most QDs remained associated with the silk, based on the retention of photoluminescence in the silk films and negligible photoluminescence in the extracts. Compared to the solution state, photoluminescence intensity significantly decreased for MEA-QDs but not for MPA-QDs in the silk films, while the emission maximum blue shifted (≈4 nm slightly for both. Further film digestion using protease XIV, alpha-chymotrypsin, and the combination of the two proteases suggested that QDs may be bound to the silk beta-sheet regions but not the amorphous regions. QDs photoluminescence in silk films was quenched when the concentration of hydrogen peroxide (H2O2 was above 0.2-0.3 mM, indicating the QDs-incorporated silk films can be used to report oxidation potential in solution.

  19. 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.

  20. Nephila clavipes Flagelliform Silk-like GGX Motifs Contribute to Extensibility and Spacer Motifs Contribute to Strength in Synthetic Spider Silk Fibers

    Adrianos, Sherry L.; Teulé, Florence; Hinman, Michael B.; Jones, Justin A.; Weber, Warner S.; Yarger, Jeffery L.; Lewis, Randolph V.

    2013-01-01

    Flagelliform spider silk is the most extensible silk fiber produced by orb weaver spiders, though not as strong as the dragline silk of the spider. The motifs found in the core of the Nephila clavipes flagelliform Flag protein are: GGX, spacer, and GPGGX. Flag does not contain the polyalanine motif known to provide the strength of dragline silk. To investigate the source of flagelliform fiber strength, four recombinant proteins were produced containing variations of the three core motifs of t...

  1. 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.)

  2. Evaluation on Gambiered Guangdong Silk's Ecological Properties

    LI Wei-xian; ZHAO Yao-ming; SHI Yan-ming

    2008-01-01

    To evaluate gambiered Guangdong silk's ecology properties,the raw materials ecology,prttluctien ecology and use ecology are analyzed;and the safety to htmaan is tested according to GB/T18885-2002.Gambiered Gnangdong silk is a kind ofnatural product.All its raw materials are reproducible and degradable natural resources,and it can decompose completely when being discarded.It is comfortable to wear,and easy to dean and dry.It is mainly manufactured by dip-dyeing in dye yam solution,and insohting under the burning sun.The preduction energy is solar,a kind of dean and reproducible energy.The production ptocess is clean and eavironmental friendly.There is neither the utilization of synthetic dyestuff and chemical auxiliary,nor the discharge of environment pollutant,and even the dreg of dye yam is used as fuel.The safety test results show that pH value of water extract,heavy metal content(except for Pb content),color fastness and odor meet the requirouents of GB/T18885-2002 and Okeo-tex standard 100.

  3. 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

    2014-04-01

    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.

  4. Zibo Daranfang Silk Group:Big Dye House Shines over Silk Road

    Liu Yi

    2011-01-01

    Zibo Daranfang Silk Group is a large-scale joint-stock enterprise,which integrates scientific research,production and trade.It owns ten stock holding and participating companies.It is the only enterprise of weaving,printing and dyeing within China,whose capacity of weaving,scouring and bleaching,printing and dyeing is in the first place of the north area of the Yangze River.

  5. 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.

  6. Studies on Application of Aroma Finish on Silk Fabric

    Hipparagi, Sanganna Aminappa; Srinivasa, Thirumalappa; Das, Brojeswari; Naik, Subhas Venkatappa; Purushotham, Serampur Parappa

    2016-06-01

    Aromatic treatments on textiles have gained importance in the recent years. In the present article work has been done on fragrance finish application on silk material. Silk is an expensive natural fibre used for apparel purpose and known for its feel and appeal. Incorporation of fragrance material in silk product, will add more value to it. Present work focuses to impart durable aroma finish for silk products to be home washed or subjected to dry cleaning. Microencapsulated aroma chemical has been used for the treatment. Impregnation method, Exhaust method, Dip-Pad-Dry method and Spray method have been used to see the influence of application method on the uptake and performance. Evaluation of the aroma treated material has been done through subjective evaluation as per Odor Intensity Reference Scaling (OIRS). Effect of the aroma finishing on the physical properties of the fabric has also been studied. No adverse effect has been observed on the stiffness of the fabric after the aroma treatment.

  7. 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....

  8. Characterization of sericin protein recovered from silk wastewaters

    Gökşen Çapar; Seylan Saniye Aygün

    2015-01-01

    INTRODUCTION[|]This study aims to determine the characteristics of sericin protein recovered from silk wastewaters.[¤]METHODS[|]Sericin protein was recovered from silk wastewaters by membrane technology in Engineering Sciences Department of the Middle East Technical University between 2007 and 2008. The protein characterization study was completed in Ankara University Water Management Institute in 2012. The recovered protein was characterized in terms of molecular w...

  9. 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...

  10. 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

  11. 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.

  12. In vitro transcription of eukaryotic genes is affected differently by the degree of DNA supercoiling.

    Hirose, S; Suzuki, Y

    1988-01-01

    In a posterior silk gland extract, covalently closed circular (ccc) DNA is in a superhelical state that supports more transcription of fibroin gene than does linear DNA. A HeLa cell extract showed neither the supercoiling activity nor the preference for the transcription of ccc DNA over linear DNA. These activities could be added to the HeLa cell extract. Phosphocellulose fractionation of the posterior silk gland extract yielded a flow-through fraction and a 0.6 M KCl eluate fraction that wer...

  13. The potential of silk and silk-like proteins as natural mucoadhesive biopolymers for controlled drug delivery

    Brooks, Amanda

    2015-11-01

    Drug delivery across mucus membranes is a particularly effective route of administration due to the large surface area. However, the unique environment present at the mucosa necessitates altered drug formulations designed to (1) deliver sensitive biologic molecules, (2) promote intimate contact between the mucosa and the drug, and (3) prolong the drug’s local residence time. Thus, the pharmaceutical industry has an interest in drug delivery systems formulated around the use of mucoadhesive polymers. Mucoadhesive polymers, both synthetic and biological, have a history of use in local drug delivery. Prominently featured in the literature are chitosan, alginate, and cellulose derivatives. More recently, silk and silk-like derivatives have been explored for their potential as mucoadhesive polymers. Both silkworms and spiders produce sticky silk-like glue substances, sericin and aggregate silk respectively, that may prove an effective, natural matrix for drug delivery to the mucosa. This mini review will explore the potential of silk and silk-like derivatives as a biocompatible mucoadhesive polymer matrix for local controlled drug delivery.

  14. The potential of silk and silk-like proteins as natural mucoadhesive biopolymers for controlled drug delivery

    Amanda E Brooks

    2015-11-01

    Full Text Available Drug delivery across mucus membranes is a particularly effective route of administration due to the large surface area. However, the unique environment present at the mucosa necessitates altered drug formulations designed to (1 deliver sensitive biologic molecules, (2 promote intimate contact between the mucosa and the drug, and (3 prolong the drug’s local residence time. Thus, the pharmaceutical industry has an interest in drug delivery systems formulated around the use of mucoadhesive polymers. Mucoadhesive polymers, both synthetic and biological, have a history of use in local drug delivery. Prominently featured in the literature are chitosan, alginate, and cellulose derivatives. More recently, silk and silk-like derivatives have been explored for their potential as mucoadhesive polymers. Both silkworms and spiders produce sticky silk-like glue substances, sericin and aggregate silk respectively, that may prove an effective, natural matrix for drug delivery to the mucosa. This mini review will explore the potential of silk and silk-like derivatives as a biocompatible mucoadhesive polymer matrix for local controlled drug delivery.

  15. The Hagfish Gland Thread Cell: A Fiber-Producing Cell Involved in Predator Defense

    Douglas S. Fudge

    2016-05-01

    Full Text Available Fibers are ubiquitous in biology, and include tensile materials produced by specialized glands (such as silks, extracellular fibrils that reinforce exoskeletons and connective tissues (such as chitin and collagen, as well as intracellular filaments that make up the metazoan cytoskeleton (such as F-actin, microtubules, and intermediate filaments. Hagfish gland thread cells are unique in that they produce a high aspect ratio fiber from cytoskeletal building blocks within the confines of their cytoplasm. These threads are elaborately coiled into structures that readily unravel when they are ejected into seawater from the slime glands. In this review we summarize what is currently known about the structure and function of gland thread cells and we speculate about the mechanism that these cells use to produce a mechanically robust fiber that is almost one hundred thousand times longer than it is wide. We propose that a key feature of this mechanism involves the unidirectional rotation of the cell’s nucleus, which would serve to twist disorganized filaments into a coherent thread and impart a torsional stress on the thread that would both facilitate coiling and drive energetic unravelling in seawater.

  16. 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.

  17. 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

  18. Ultrasonography of thyroid gland

    Kim, Chu Wan; Lee, Kwan Seh; Chung, Yong In; Lee, Kyung Hi; Lee, Mun Ho; Koh, Chang Soon; Kim, Byung Kook [Seoul National University College of Medicine, Seoul (Korea, Republic of)

    1985-12-15

    The fundamental requirement for ultrasound in thyroid disease is to differentiate cystic masses from solid ones. But with the advance in scan equipment and technique the more detailed information about the lesion, echogenicity, internal echo pattern and surroundings casts but a faint on the clues to give more specific categorical or histological diagnosis of the lesions. Authors reviewed 186 cases of ultrasonography of thyroid gland performed from March to September, 1979 at Seoul National University Hospital and concluded that; 1. Differentiation of solid mass from cystic one is easily and readily made but a few of solid masses appear as cystic ones in ultrasonogram because of inappropriate scan technique. 2. Categorical or histological diagnosis is still beyond the scope of our knowledge but some clues suggest that only accumulation of data will solve the problem in the future.

  19. Ultrasonography of thyroid gland

    The fundamental requirement for ultrasound in thyroid disease is to differentiate cystic masses from solid ones. But with the advance in scan equipment and technique the more detailed information about the lesion, echogenicity, internal echo pattern and surroundings casts but a faint on the clues to give more specific categorical or histological diagnosis of the lesions. Authors reviewed 186 cases of ultrasonography of thyroid gland performed from March to September, 1979 at Seoul National University Hospital and concluded that; 1. Differentiation of solid mass from cystic one is easily and readily made but a few of solid masses appear as cystic ones in ultrasonogram because of inappropriate scan technique. 2. Categorical or histological diagnosis is still beyond the scope of our knowledge but some clues suggest that only accumulation of data will solve the problem in the future.

  20. Trastuzumab in Treating Patients With Metastatic or Recurrent Salivary Gland Cancer

    2013-02-27

    High-grade Salivary Gland Mucoepidermoid Carcinoma; Recurrent Salivary Gland Cancer; Salivary Gland Acinic Cell Tumor; Salivary Gland Adenocarcinoma; Salivary Gland Poorly Differentiated Carcinoma; Stage IVA Salivary Gland Cancer; Stage IVB Salivary Gland Cancer; Stage IVC Salivary Gland Cancer

  1. Study on different surface modification treatments on poly (D, L-lactic acid) with silk Fibroin and their effects on the culture of osteoblast in vitro

    YAO Kang-de; CAI Kai-yong; LIN Song-bai; YANG Zhi-ming; LI Xiu-qiong; XIE Hui-qi; QING Ting-wu

    2001-01-01

    @@ One of the challenges in the field of tissue engineering is the development of biomaterial/cell interactions. For this purpose, the use of biological materials in tissue engineering requires the application of different chemical or physical procedures for surface modification in order to improve the cell affinity of biomaterials. We have studied the effects of one chemical treatment and one physical treatment: 1-(3-dimethylaminopropyl )-3-ethylcarbodiimide (WSC) and entrapment.

  2. Meibomian gland imaging: A review

    Deanne L. Nicholas

    2015-03-01

    Full Text Available The meibomian glands of the upper and lower eyelids play a valuable role in secreting the lipidlayer of the tear film. Disturbances in meibomian gland function may result in altered secretion andvariations in tear composition which may lead to meibomian gland dysfunction and evaporativedry eye, leading to ocular discomfort. To diagnose and monitor the structural and functionalchanges occurring within the glands of the eyelids, various imaging techniques are available.Some of the methods used to evaluate the tears and therefore the meibum within the tears includeevaporimetry, interferometry, tear osmolarity and meibometry. With these techniques, changesin the lipid layer of the tear film can be quantified and alterations in meibomian gland functionassessed. Meibography is an additional method that can be used; it has the unique feature ofallowing the assessment of meibomian gland morphology during ocular surface disease processes.The aim of this review is to create an improved understanding of the meibomian glands and theways that they may be investigated in order to expand on the treatment methods available.

  3. The Effects of Silk Protein Solution on Quality of Mangosteen

    Full text: Sericin silk protein solution prepared from irradiated silk waste was applied as a wax coating on mangosteen fruits but was found not appropriate. Nonetheless, when the solution was sprayed onto fruits from the setting stage till maturity, it enhanced the fruit quality. Fruits showed a bigger stalk and greener, thicker and larger-angled calyx than untreated ones. They were edible after being kept at 25C for 3 to 4 weeks. Fruits sprayed with the silk protein solution and untreated fruits were separately harvested and packed for export. A half of them were irradiated with gamma radiation for insect eradication and then kept at 10C with 80 percents relative humidity. The unirradiated half was stored similarly. Results showed that rind color of irradiated fruits changed slower than unirradiated ones. However, silk protein solution spraying and irradiation did not affect fruit firmness and total soluble solids of mangosteen flesh. For sensory evaluation, it was found that sprayed and irradiated fruits showed no differences from the untreated ones in their physical properties, color, taste and odor. Fruits sprayed with the silk protein solution without irradiation had the longest shelf life of 6 weeks

  4. Spider web and silk performance landscapes across nutrient space

    Blamires, Sean J.; Tseng, Yi-Hsuan; Wu, Chung-Lin; Toft, Søren; Raubenheimer, David; Tso, I.-Min

    2016-01-01

    Predators have been shown to alter their foraging as a regulatory response to recent feeding history, but it remains unknown whether trap building predators modulate their traps similarly as a regulatory strategy. Here we fed the orb web spider Nephila pilipes either live crickets, dead crickets with webs stimulated by flies, or dead crickets without web stimulation, over 21 days to enforce spiders to differentially extract nutrients from a single prey source. In addition to the nutrients extracted we measured web architectures, silk tensile properties, silk amino acid compositions, and web tension after each feeding round. We then plotted web and silk “performance landscapes” across nutrient space. The landscapes had multiple peaks and troughs for each web and silk performance parameter. The findings suggest that N. pilipes plastically adjusts the chemical and physical properties of their web and silk in accordance with its nutritional history. Our study expands the application of the geometric framework foraging model to include a type of predatory trap. Whether it can be applied to other predatory traps requires further testing. PMID:27216252

  5. 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.

  6. 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 β-sheet structure were determined by ThT fluorescence spectral analysis. The content of β-sheet among the four polypeptides followed the order: eGS > eGV > eGY > eGA.

  7. [Tumor of the Parotid Gland].

    Pötzl, Teresa; Iselin, Sabine; Husner, Alexander

    2016-05-11

    Salivary gland tumors are a rare, histologically heterogeneous group of tumors which constitute approximately 4–6 % of all head and neck neoplasms. In 2/3 of cases they are benign, especially in the parotid gland. We report about a rare tumor of the parotid gland presenting as an extraskeletal chondroma. Histologically there were multiple S 100 protein-positive nests of chondrocytes. The externally completed cytology suspected a pleomorphic adenoma, nevertheless, the final histopathological findings showed another tumor entity. PMID:27167480

  8. 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)

  9. Impact and dynamic mechanical thermal properties of textile silk reinforced epoxy resin composites

    Yang, K.; Guan, J.

    2016-07-01

    Silk fabric reinforced epoxy resin composites (SFRPs) were prepared using simple techniques of hand lay-up, hot-press and vacuum treatment, and a series of volume fractions of silk reinforcements were achieved. The impact properties and dynamic mechanical properties of SFRPs were investigated using a pendulum impact testing method and dynamic mechanical thermal analysis (DMTA). The results suggest that silk reinforcement could greatly enhance the mechanical performances of SFRPs. The impact strength reached a maximum of 71 kJ/m2 for 60%-silk SFRP, which demonstrated a potential of silk composites for defence and impact- resistant materials.

  10. Development of new smart materials and spinning systems inspired by natural silks and their applications

    Cheng, Jie; Lee, Sang-Hoon

    2015-12-01

    Silks produced by spiders and silkworms are charming natural biological materials with highly optimized hierarchical structures and outstanding physicomechanical properties. The superior performance of silks relies on the integration of a unique protein sequence, a distinctive spinning process, and complex hierarchical structures. Silks have been prepared to form a variety of morphologies and are widely used in diverse applications, for example, in the textile industry, as drug delivery vehicles, and as tissue engineering scaffolds. This review presents an overview of the organization of natural silks, in which chemical and physical functions are optimized, as well as a range of new materials inspired by the desire to mimic natural silk structure and synthesis.

  11. The consolidation behavior of silk hydrogels.

    Kluge, Jonathan A; Rosiello, Nicholas C; Leisk, Gary G; Kaplan, David L; Dorfmann, A Luis

    2010-04-01

    Hydrogels have mechanical properties and structural features that are similar to load-bearing soft tissues including intervertebral disc and articular cartilage, and can be implanted for tissue restoration or for local release of therapeutic factors. To help predict their performance, mechanical characterization and mathematical modeling are the available methods for use in tissue engineering and drug delivery settings. In this study, confined compression creep tests were performed on silk hydrogels, over a range of concentrations, to examine the phenomenological behavior of the gels under a physiological loading scenario. Based on the observed behavior, we show that the time-dependent response can be explained by a consolidation mechanism, and modeled using Biot's poroelasticity theory. Two observations are in strong support of this modeling framework, namely, the excellent numerical agreement between increasing load step creep data and the linear Terzaghi theory, and the similar values obtained from numerical simulations and direct measurements of the permeability coefficient. The higher concentration gels (8% and 12% w/v) clearly show a strain-stiffening response to creep loading with increasing loads, while the lower concentration gel (4% w/v) does not. A nonlinear elastic constitutive formulation is employed to account for the stiffening. Furthermore, an empirical formulation is used to represent the deformation-dependent permeability. PMID:20142112

  12. Soft magnetic memory of silk cocoon membrane.

    Roy, Manas; Dubey, Amarish; Singh, Sushil Kumar; Bhargava, Kalpana; Sethy, Niroj Kumar; Philip, Deepu; Sarkar, Sabyasachi; Bajpai, Alok; Das, Mainak

    2016-01-01

    Silk cocoon membrane (SCM), a solid matrix of protein fiber, responds to light, heat and moisture and converts these energies to electrical signals. Essentially it exhibits photo-electric and thermo-electric properties; making it a natural electro-magnetic sensor, which may influence the pupal development. This raises the question: 'is it only electricity?', or 'it also posses some kind of magnetic memory?' This work attempted to explore the magnetic memory of SCM and confirm its soft magnetism. Fe, Co, Ni, Mn, Gd were found in SCM, in traces, through energy dispersive X-ray analysis (EDX), X-ray photoelectron spectroscopy (XPS) and inductively coupled plasma mass spectrometry (ICP-MS). Presence of iron was ascertained by electron paramagnetic resonance (EPR). In addition, EPR-spectra showed the presence of a stable pool of carbon-centric free radical in the cocoon structure. Carbon-centric free radicals behaves as a soft magnet inherently. Magnetic-Hysteresis (M-H) of SCM confirmed its soft magnetism. It can be concluded that the soft bio-magnetic feature of SCM is due to the entrapment of ferromagnetic elements in a stable pool of carbon centric radicals occurring on the super-coiled protein structure. Natural soft magnets like SCM provide us with models for developing eco-friendly, protein-based biological soft magnets. PMID:27374752

  13. 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...

  14. 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.

  15. Improved human tenocyte proliferation and differentiation in vitro by optimized silk degumming

    Wang Xiao; Qiu Yiwei; Carr, Andrew J; Triffitt, James T; Sabokbar, Afsie; Xia Zhidao, E-mail: z.xia@swansea.ac.uk [Botnar Research Centre, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, University of Oxford, Oxford (United Kingdom)

    2011-06-15

    Tendon disorders are common clinical conditions. Tendon tissue engineering provides a new approach for tendon repair by integrating engineered substitutes with their native counterparts. Silk is considered to be a promising candidate for tendon engineering because of its biological and mechanical properties. However, a major concern with using silk for biomedical applications is the immune responses generated by sericin, a glue-like protein that coats the silk fibres. This study improves the existing protocols for silk 'degumming' which removes sericin and enables preparation of silk that is suitable for tendon regeneration. Bombyx mori silks were treated by sequential treatments with different proteases. The efficiency of degumming was determined by measuring weight loss, picric acid and carmine staining and scanning electron microscopy. To evaluate the cellular responses after degumming, the growth and differentiation of human tenocytes on silks were examined. The results showed that sequential protease treatment effectively degummed raw silks. The sequentially degummed silks showed enhanced tenocyte proliferation and upregulated mRNA levels of tendon markers. Thick cell multilayers formed on the treated silks, with cells and collagen fibres penetrating into the spaces in individual silk filaments, resulting in a structure resembling human tendon.

  16. Silk reinforced with graphene or carbon nanotubes spun by spiders

    Lepore, Emiliano; Bonaccorso, Francesco; Bruna, Matteo; Bosia, Federico; Taioli, Simone; Garberoglio, Giovanni; Ferrari, Andrea C.; Pugno, Nicola Maria

    2015-01-01

    Here, we report the production of silk incorporating graphene and carbon nanotubes directly by spider spinning, after spraying spiders with the corresponding aqueous dispersions. We observe a significant increment of the mechanical properties with respect to the pristine silk, in terms of fracture strength, Young's and toughness moduli. We measure a fracture strength up to 5.4 GPa, a Young's modulus up to 47.8 GPa and a toughness modulus up to 2.1 GPa, or 1567 J/g, which, to the best of our k...

  17. 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.

  18. Synthesis and characterization of spider silk calcite composite

    Svetlana Dmitrović

    2016-03-01

    Full Text Available Spider silk poses excellent mechanical properties, tenacity and elasticity and it has been used as a template for calcite mineralization to improve load bearing strength of osteoconductive calcite. The samples were obtained by mimicking biomineralization for five days in order to follow formation and growth of calcite on the surface of spider silk. Crystal phase was detected by XRD and FTIR spectroscopy. Microstructure, crystal size and its morphology were studied by means of FESEM. After two days of processing, pure calcite phase was obtained, and a size of the formed crystals increased with prolongation of biomineralization.

  19. 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

  20. 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