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1

Heterogeneous nucleation of hydroxyapatite on protein: structural effect of silk sericin  

PubMed Central

Acidic proteins play an important role during mineral formation in biological systems, but the mechanism of mineral formation is far from understood. In this paper, we report on the relationship between the structure of a protein and hydroxyapatite deposition under biomimetic conditions. Sericin, a type of silk protein, was adopted as a suitable protein for studying structural effect on hydroxyapatite deposition, since it forms a hydroxyapatite layer on its surface in a metastable calcium phosphate solution, and its structure has been reported. Sericin effectively induced hydroxyapatite nucleation when it has high molecular weight and a ? sheet structure. This indicates that the specific structure of a protein can effectively induce heterogeneous nucleation of hydroxyapatite in a biomimetic solution, i.e. a metastable calcium phosphate solution. This finding is useful in understanding biomineralization, as well as for the design of organic polymers that can effectively induce hydroxyapatite nucleation. PMID:16849195

Takeuchi, Akari; Ohtsuki, Chikara; Miyazaki, Toshiki; Kamitakahara, Masanobu; Ogata, Shin-ichi; Yamazaki, Masao; Furutani, Yoshiaki; Kinoshita, Hisao; Tanihara, Masao

2005-01-01

2

Exploring natural silk protein sericin for regenerative medicine: an injectable, photoluminescent, cell-adhesive 3D hydrogel.  

PubMed

Sericin, a major component of silk, has a long history of being discarded as a waste during silk processing. The value of sericin for tissue engineering is underestimated and its potential application in regenerative medicine has just begun to be explored. Here we report the successful fabrication and characterization of a covalently-crosslinked 3D pure sericin hydrogel for delivery of cells and drugs. This hydrogel is injectable, permitting its implantation through minimally invasive approaches. Notably, this hydrogel is found to exhibit photoluminescence, enabling bioimaging and in vivo tracking. Moreover, this hydrogel system possesses excellent cell-adhesive capability, effectively promoting cell attachment, proliferation and long-term survival of various types of cells. Further, the sericin hydrogel releases bioactive reagents in a sustained manner. Additionally, this hydrogel demonstrates good elasticity, high porosity, and pH-dependent degradation dynamics, which are advantageous for this sericin hydrogel to serve as a delivery vehicle for cells and therapeutic drugs. With all these unique features, it is expected that this sericin hydrogel will have wide utility in the areas of tissue engineering and regenerative medicine. PMID:25412301

Wang, Zheng; Zhang, Yeshun; Zhang, Jinxiang; Huang, Lei; Liu, Jia; Li, Yongkui; Zhang, Guozheng; Kundu, Subhas C; Wang, Lin

2014-01-01

3

One-step synthesis of natural silk sericin-based microcapsules with bionic structures.  

PubMed

Different techniques are being developed for fabricating microcapsules; it is still a challenge to fabricate them in an efficient and environment-friendly process. Here, a one-step green route to synthesize silk protein sericin-based microcapsules without any assistance of organic solvents is reported. By carefully changing the concentration of calcium ions accompanied with stirring, the morphology of the microcapsules can easily be regulated to form either discoidal, biconcave, cocoon-like, or tubular structures. The chelation of Ca(2+) and shearing force from agitation may induce the conformational transformation of sericin, which possibly results in the formation of microcapsules through the self-assembly of the protein subsequently. The as-prepared cocoon-like microcapsules exhibit pH-dependent stability. A potential application of microcapsules being fabricated from natural water-soluble silk protein sericin for controlled bioactive molecules loading and release system by a pH-triggered manner is quite feasible. PMID:25168858

Liu, Zhaogang; Cai, Yurong; Jia, Yaru; Liu, Lin; Kong, Xiangdong; Kundu, Subhas C; Yao, Juming

2014-10-01

4

Tuning molecular weights of Bombyx mori (B. mori) silk sericin to modify its assembly structures and materials formation.  

PubMed

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 into biomaterials from two forms of B. mori silk sericin with different molecular weights, one is a low (12.0 kDa) molecular sericin (LS) form and another is a high (66.0 kDa) molecular weight sericin (HS) form. Circular dichroism (CD) spectra showed that LS in hexafluoroacetone (HFA) solvent adopted a predominantly random coil conformation, whereas HS tended to form a ?-sheet structure along with a large content of random coils. In addition, LS and HS in HFA solvent were found to form cylinder-like smaller nanoparticles and larger irregular aggregates before electrospinning, respectively. As a result, biomaterials based on microparticles and nanofibers were successfully fabricated by electrospinning of LS and HS dissolved in HFA, respectively. The cell viability and differentiation assay indicated that nanofibers and microparticles improved cell adhesion, growth, and differentiation, proving that the scaffolds electrospun from sericin are biocompatible regardless of its molecular weight. The microparticles, not common in electrospinning of silk proteins reported previously, were found to promote the osteogenic differentiation of mesenchymal stem cells in comparison to the nanofibers. This study suggested that molecular weight of sericin mediates its secondary structure and assembly structure, which in turn leads to a control of final morphology of the electrospun materials. The microparticles and nanofibers of sericin can be potentially used as building blocks for fabricating the scaffolds for tissue engineering. PMID:25050697

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

2014-08-27

5

Formulation and characterization of silk sericin-PVA scaffold crosslinked with genipin.  

PubMed

A porous-three-dimensional scaffold shows several advantages in terms of tissue engineering since it can provide a framework for cells to attach, proliferate and form an extracellular matrix. Sericin, a by-product from the silk industry, can form a three-dimensional scaffold with PVA after freeze-drying but has a fragile structure. Glycerin (as a plasticizer) and genipin (a crosslinking agent) are necessary to make a strong and stable matrix. Our objective was to investigate the properties of a three-dimensional silk sericin and PVA scaffold with and without glycerin and genipin at various concentrations. SEM showed that adding glycerin into scaffold gave better uniformity and porosity. Smaller pore sizes and better uniformity were found as the concentration of genipin in the scaffold increased. The results of FTIR indicated that glycerin retained a high moisture content and had a major effect at 3286 cm(-1), indicating the presence of water molecule in the matrix structure. Adding genipin into the scaffold resulted in a higher degree of crosslinking or fewer free ?-amino groups, as shown by the decrease in the stretching (=C-H) peak and absorption peaks around 1370-1650 cm(-1), respectively. The sericin/PVA scaffold had a low water sorption capacity, but adding glycerin significantly increased this property. Genipin further enhanced the moisture absorption capacity of the scaffold and extended the time taken to reach equilibrium. After immersing the sericin/PVA scaffold into purified water, the scaffold completely dissolved within an hour, whereas the scaffolds containing glycerin or glycerin with 0.1% genipin swelled 8 and 11 times, respectively, compared with the initial stage after 6h of immersion. In terms of mechanical properties, the sericin/PVA/glycerin scaffold exhibited a similar compressive strength to the scaffold with a high genipin concentration, whereas a low concentration of genipin softened and reduced the compressive strength of the scaffold. A small amount of sericin was released from the scaffold and a higher concentration of genipin, resulting in less protein leaching compared to non-crosslinked sericin/PVA. The fraction of protein released from the sericin/PVA/glycerin scaffold was about 4%, with values of about 1 and 0.04% in the case of scaffolds with 0.01 and 0.1% genipin, respectively. All results indicated that the composition of the scaffolds had a significant effect on their physical properties, and that can easily be tuned to obtain scaffolds suitable for biological applications. PMID:20804781

Aramwit, Pornanong; Siritientong, Tippawan; Kanokpanont, Sorada; Srichana, Teerapol

2010-12-01

6

pH responsive poly amino-acid hydrogels formed via silk sericin templating.  

PubMed

Poly(amino acid) hydrogels have attracted a great deal of attention as biodegradable biomaterials that can limit products of synthetic polymer degradation. Here we report on a stimuli-responsive, porous, composite biomaterial based on the protein templating of the poly(amino acid) hydrogel from poly(aspartic acid) with the silk protein sericin. This low-cost, biocompatible and biodegradable hydrogel demonstrates a greatly increased porosity and improvement in volumetric swelling over networks formed from pure poly(aspartic acid). The swelling capacity measured over a range of pH values surrounding physiological pH 7.0 demonstrates a linear profile, in which hydrogel volume and mass increase to a maximum, with an increase as a function of higher sericin content. In comparison to pure poly(aspartic acid), this demonstrates a nearly 3-fold increase in retention volume at basic pH. The increase in swelling is also demonstrated by the increase in porosity and internal micro-architecture of the hydrogel networks. The biomaterial is then shown to perform well as a scaffold for cells with high mechanical strength and integrity. This protein- and homo poly(amino acid)-based super-swelling hydrogel has applications in drug delivery and tissue engineering as an economical and environmentally friendly biomaterial, in addition to ensuring the species incorporated maintain their biocompatibility during processing. PMID:25073107

Kurland, Nicholas E; Ragland, Robert B; Zhang, Aolin; Moustafa, Mahmoud E; Kundu, Subhas C; Yadavalli, Vamsi K

2014-09-01

7

Self-assembled silk sericin/poloxamer nanoparticles as nanocarriers of hydrophobic and hydrophilic drugs for targeted delivery  

NASA Astrophysics Data System (ADS)

In recent times self-assembled micellar nanoparticles have been successfully employed in tissue engineering for targeted drug delivery applications. In this review, silk sericin protein from non-mulberry Antheraea mylitta tropical tasar silk cocoons was blended with pluronic F-127 and F-87 in the presence of solvents to achieve self-assembled micellar nanostructures capable of carrying both hydrophilic (FITC-inulin) and hydrophobic (anticancer drug paclitaxel) drugs. The fabricated nanoparticles were subsequently characterized for their size distribution, drug loading capability, cellular uptake and cytotoxicity. Nanoparticle sizes ranged between 100 and 110 nm in diameter as confirmed by dynamic light scattering. Rapid uptake of these particles into cells was observed in in vitro cellular uptake studies using breast cancer MCF-7 cells. In vitro cytotoxicity assay using paclitaxel-loaded nanoparticles against breast cancer cells showed promising results comparable to free paclitaxel drugs. Drug-encapsulated nanoparticle-induced apoptosis in MCF-7 cells was confirmed by FACS and confocal microscopic studies using Annexin V staining. Up-regulation of pro-apoptotic protein Bax, down-regulation of anti-apoptotic protein Bcl-2 and cleavage of regulatory protein PARP through Western blot analysis suggested further drug-induced apoptosis in cells. This study projects silk sericin protein as an alternative natural biomaterial for fabrication of self-assembled nanoparticles in the presence of poloxamer for successful delivery of both hydrophobic and hydrophilic drugs to target sites.

Mandal, Biman B.; Kundu, S. C.

2009-09-01

8

Biomedical Applications of Mulberry Silk and its Proteins: A Review  

NASA Astrophysics Data System (ADS)

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.

Nivedita, S.; Sivaprasad, V.

2014-04-01

9

Isolation and processing of silk proteins for biomedical applications.  

PubMed

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

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

2014-09-01

10

Study of the physical properties of whey protein: sericin protein-blended edible films  

Microsoft Academic Search

Effects of the sericin protein on the physical properties of the whey protein isolate film including the mechanical strength,\\u000a water vapor permeability, light transmission, moisture content, solubility and swelling were investigated in the present work.\\u000a The mechanical strength of the whey protein film could be reinforced with 0.1% sericin caused by the hydrogen bonding between\\u000a the sericin and whey protein

Jing Wang; Jiejing Shang; Fazheng Ren; Xiaojing Leng

2010-01-01

11

Production and properties of electrosprayed sericin nanopowder  

NASA Astrophysics Data System (ADS)

Sericin is a proteinous substrate that envelops fibroin (silk) fiber, and its recovery provides significant economical and social benefits. Sericin is an antibacterial agent that resists oxidation and absorbs moisture and UV light. In powder form, sericin has a wide range of applications in food, cosmetics and drug delivery. Asides from other techniques of producing powder, such as precipitation and spray drying, electrospraying can yield solid nanoparticles, particularly in the submicron range. Here, we report the production of sericin nanopowder by electrospraying. Sericin sponge was recovered from Bombyx mori cocoons through a high-temperature, high-pressure process, followed by centrifugation and freeze drying of the sericin solution. The electrospraying solution was prepared by dissolving the sericin sponge in dimethyl sulfoxide. We demonstrate that electrospraying is capable of producing sericin nanopowder with an average particle size of 25 nm, which is by far smaller than the particles produced by other techniques. The electrosprayed sericin nanopowder consists of small crystallites and exhibits a high moisture absorbance.

Hazeri, Najmeh; Tavanai, Hossein; Moradi, Ali Reza

2012-06-01

12

Antioxidant activities of two sericin proteins extracted from cocoon of silkworm (Bombyx mori) measured by DPPH, chemiluminescence, ORAC and ESR methods  

PubMed Central

Recent efforts have focused on the use of sericin proteins extracted from cocoons of silkworm as a healthy food source for human consumption. In this study, we focused on the antioxidative properties of sericin proteins. The antioxidative properties were measured in sericin proteins extracted from the shell of the cocoon, designated hereafter as white sericin protein and yellow-green sericin protein, as well as bread without sericin protein and bread to which white sericin powder had been added using four measurement methods: 1,1-Diphenyl-2-picrylhydrazyl (DPPH), chemiluminescence, oxygen radical absorbance capacity (ORAC) and electron spin resonance (ESR). High antioxidative properties of sericin proteins were indicated by all four methods. A comparison of the two types of sericin proteins revealed that yellow-green sericin protein exhibited high antioxidative properties as indicated by the DPPH, chemiluminescence and ORAC methods. By contrast, a higher antioxidative property was determined in white sericin protein by the ESR method. Consequently, our findings confirmed that sericin proteins have antioxidative properties against multiple radicals. In addition, the antioxidative property of bread was enhanced by the addition of sericin powder to the bread. Therefore, findings of this study suggest that sericin proteins may be efficiently used as beneficial food for human health. PMID:24748975

TAKECHI, TAYORI; WADA, RITSUKO; FUKUDA, TSUBASA; HARADA, KAZUKI; TAKAMURA, HITOSHI

2014-01-01

13

Silk-based biomaterials  

Microsoft Academic Search

Silk from the silkworm, Bombyx mori, has been used as biomedical suture material for centuries. The unique mechanical properties of these fibers provided important clinical repair options for many applications. During the past 20 years, some biocompatibility problems have been reported for silkworm silk; however, contamination from residual sericin (glue-like proteins) was the likely cause. More recent studies with well-defined

Gregory H. Altman; Frank Diaz; Caroline Jakuba; Tara Calabro; Rebecca L. Horan; Jingsong Chen; Helen Lu; John Richmond; David L. Kaplan

2003-01-01

14

Structure and expression of the silk adhesive protein Ser2 in Bombyx mori Barbara Kludkiewicz a,b  

E-print Network

in revised form 27 November 2009 Accepted 30 November 2009 Keywords: Silkworm Sericin Cocoon Silk gland rights reserved. 1. Introduction The silk produced by caterpillars such as the silkworm, Bombyx mori

Â?urovec, Michal

15

Recombinant DNA production of spider silk proteins  

PubMed Central

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 recombinant spider silks. PMID:24119078

Tokareva, Olena; Michalczechen-Lacerda, Valquiria A; Rech, Elibio L; Kaplan, David L

2013-01-01

16

Micro-scale Processing of Silk Protein  

E-print Network

fiber based applications for silk protein such as microspheres for drug delivery and the molding of nano- andnano- to micro-size orifices can now easily be purchased, and have been used to extrude silk fibers (

Breslauer, David Nate

2010-01-01

17

Single Honeybee Silk Protein Mimics Properties of Multi-Protein Silk  

Microsoft Academic Search

Honeybee silk is composed of four fibrous proteins that, unlike other silks, are readily synthesized at full-length and high yield. The four silk genes have been conserved for over 150 million years in all investigated bee, ant and hornet species, implying a distinct functional role for each protein. However, the amino acid composition and molecular architecture of the proteins are

Tara D. Sutherland; Jeffrey S. Church; Xiao Hu; Mickey G. Huson; David L. Kaplan; Sarah Weisman; Markus Buehler

2011-01-01

18

Microbial production of spider silk proteins.  

PubMed

The remarkable properties of spider dragline silk and related protein polymers will find many applications if the materials can be produced economically. We have demonstrated the production of high molecular weight spider dragline silk analog proteins encoded by synthetic genes in several microbial systems, including Escherichia coli and Pichia pastoris. In E. coli, proteins of up to 1000 amino acids in length could be produced efficiently, but the yield and homogeneity of higher molecular weight silk proteins were found to be limited by truncated synthesis, probably as a result of ribosome termination errors. No such phenomenon was observed in the yeast P. pastoris, where higher molecular weight silk proteins could be produced without heterogeneity due to truncated synthesis. Spider dragline silk analog proteins could be secreted by P. pastoris when fused to both the signal sequence and N-terminal pro-sequence of the Saccharomyces cerevisiae alpha-mating factor gene. PMID:11763501

Fahnestock, S R; Yao, Z; Bedzyk, L A

2000-08-01

19

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

PubMed Central

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 the ability to form three-dimensional porous scaffolds from the silk fibroin, but did not affect macrophage activation or ?-sheet content in the materials formed. Methods are also provided to determine the content of residual sericin in silk fibroin solutions and to assess changes in silk fibroin molecular weight. Amino acid composition analysis was used to detect sericin residuals in silk solutions with a detection limit between 1.0% and 10% wt/wt, while fluorescence spectroscopy was used to reproducibly distinguish between silk samples with different molecular weights. Both methods are simple and require minimal sample volume, providing useful quality control tools for silk fibroin preparation processes. PMID:21695778

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

2012-01-01

20

Sequence Requirements of Spider Silk Elastic Proteins.  

National Technical Information Service (NTIS)

Sequence for flagelliform silk proteins from Nephila, Argiope, and Araneus species have been obtained. These sequences show significant sequence differences although the changes are not predicted to affect the secondary structure of the protein and theref...

R. V. Lewis

2001-01-01

21

Single honeybee silk protein mimics properties of multi-protein silk.  

PubMed

Honeybee silk is composed of four fibrous proteins that, unlike other silks, are readily synthesized at full-length and high yield. The four silk genes have been conserved for over 150 million years in all investigated bee, ant and hornet species, implying a distinct functional role for each protein. However, the amino acid composition and molecular architecture of the proteins are similar, suggesting functional redundancy. In this study we compare materials generated from a single honeybee silk protein to materials containing all four recombinant proteins or to natural honeybee silk. We analyse solution conformation by dynamic light scattering and circular dichroism, solid state structure by Fourier Transform Infrared spectroscopy and Raman spectroscopy, and fiber tensile properties by stress-strain analysis. The results demonstrate that fibers artificially generated from a single recombinant silk protein can reproduce the structural and mechanical properties of the natural silk. The importance of the four protein complex found in natural silk may lie in biological silk storage or hierarchical self-assembly. The finding that the functional properties of the mature material can be achieved with a single protein greatly simplifies the route to production for artificial honeybee silk. PMID:21311767

Sutherland, Tara D; Church, Jeffrey S; Hu, Xiao; Huson, Mickey G; Kaplan, David L; Weisman, Sarah

2011-01-01

22

Single Honeybee Silk Protein Mimics Properties of Multi-Protein Silk  

PubMed Central

Honeybee silk is composed of four fibrous proteins that, unlike other silks, are readily synthesized at full-length and high yield. The four silk genes have been conserved for over 150 million years in all investigated bee, ant and hornet species, implying a distinct functional role for each protein. However, the amino acid composition and molecular architecture of the proteins are similar, suggesting functional redundancy. In this study we compare materials generated from a single honeybee silk protein to materials containing all four recombinant proteins or to natural honeybee silk. We analyse solution conformation by dynamic light scattering and circular dichroism, solid state structure by Fourier Transform Infrared spectroscopy and Raman spectroscopy, and fiber tensile properties by stress-strain analysis. The results demonstrate that fibers artificially generated from a single recombinant silk protein can reproduce the structural and mechanical properties of the natural silk. The importance of the four protein complex found in natural silk may lie in biological silk storage or hierarchical self-assembly. The finding that the functional properties of the mature material can be achieved with a single protein greatly simplifies the route to production for artificial honeybee silk. PMID:21311767

Sutherland, Tara D.; Church, Jeffrey S.; Hu, Xiao; Huson, Mickey G.; Kaplan, David L.; Weisman, Sarah

2011-01-01

23

New silk protein: modification of silk protein by gene engineering for production of biomaterials.  

PubMed

The interest in silk fibroin morphology and structure have increased due to its attractiveness for bio-related applications. Silk fibers have been used as sutures for a long time in the surgical field, due to the biocompatibility of silk fibroin fibers with human living tissue. In addition, it has been demonstrated that silk can be used as a substrate for enzyme immobilization in biosensors. A more complete understanding of silk structure would provide the possibility to further exploit silk fibroin for a wide range of new uses, such as the production of oxygen-permeable membranes and biocompatible materials. Silk fibroin-based membranes could be utilized as soft tissue compatible polymers. Baculovirus-mediated transgenesis of the silkworm allows specific alterations in a target sequence. Homologous recombination of a foreign gene downstream from a powerful promoter, such as the fibroin promoter, would allow the constitutive production of a useful protein in the silkworm and the modification of the character of silk protein. A chimeric protein consisted of fibroin and green fluorescent protein was expressed under the control of fibroin in the posterior silk gland and the gene product was spun into the cocoon layer. This technique, gene targeting, will lead to the modification and enhancement of physicochemical properties of silk protein. PMID:11763506

Mori, H; Tsukada, M

2000-08-01

24

Structure of a Protein Superfiber: Spider Dragline Silk  

Microsoft Academic Search

Spider major ampullate (dragline) silk is an extracellular fibrous protein with unique characteristics of strength and elasticity. The silk fiber has been proposed to consist of pseudocrystalline regions of antiparallel beta-sheet interspersed with elastic amorphous segments. The repetitive sequence of a fibroin protein from major ampullate silk of the spider Nephila clavipes was determined from a partial cDNA clone. The

Ming Xu; Randolph V. Lewis

1990-01-01

25

Silk protein aggregation kinetics revealed by Rheo-IR.  

PubMed

The remarkable mechanical properties of silk fibres stem from a multi-scale hierarchical structure created when an aqueous protein "melt" is converted to an insoluble solid via flow. To directly relate a silk protein's structure and function in response to flow, we present the first application of a Rheo-IR platform, which couples cone and plate rheology with attenuated total reflectance infrared spectroscopy. This technique provides a new window into silk processing by linking shear thinning to an increase in molecular alignment, with shear thickening affecting changes in the silk protein's secondary structure. Additionally, compared to other static characterization methods for silk, Rheo-IR proved particularly useful at revealing the intrinsic difference between natural (native) and reconstituted silk feedstocks. Hence Rheo-IR offers important novel insights into natural silk processing. This has intrinsic academic merit, but it might also be useful when designing reconstituted silk analogues alongside other polymeric systems, whether natural or synthetic. PMID:24200713

Boulet-Audet, Maxime; Terry, Ann E; Vollrath, Fritz; Holland, Chris

2014-02-01

26

Variation in Protein Intake Induces Variation in Spider Silk Expression  

PubMed Central

Background It is energetically expensive to synthesize certain amino acids. The proteins (spidroins) of spider major ampullate (MA) silk, MaSp1 and MaSp2, differ in amino acid composition. Glutamine and proline are prevalent in MaSp2 and are expensive to synthesize. Since most orb web spiders express high proline silk they might preferentially attain the amino acids needed for silk from food and shift toward expressing more MaSp1 in their MA silk when starved. Methodology/Principal Findings We fed three spiders; Argiope aetherea, Cyrtophora moluccensis and Leucauge blanda, high protein, low protein or no protein solutions. A. aetherea and L. blanda MA silks are high in proline, while C. moluccesnsis MA silks are low in proline. After 10 days of feeding we determined the amino acid compositions and mechanical properties of each species' MA silk and compared them between species and treatments with pre-treatment samples, accounting for ancestry. We found that the proline and glutamine of A. aetherea and L. blanda silks were affected by protein intake; significantly decreasing under the low and no protein intake treatments. Glutmaine composition in C. moluccensis silk was likewise affected by protein intake. However, the composition of proline in their MA silk was not significantly affected by protein intake. Conclusions Our results suggest that protein limitation induces a shift toward different silk proteins with lower glutamine and/or proline content. Contradictions to the MaSp model lie in the findings that C. moluccensis MA silks did not experience a significant reduction in proline and A. aetherea did not experience a significant reduction in serine on low/no protein. The mechanical properties of the silks could not be explained by a MaSp1 expressional shift. Factors other than MaSp expression, such as the expression of spidroin-like orthologues, may impact on silk amino acid composition and spinning and glandular processes may impact mechanics. PMID:22363691

Blamires, Sean J.; Wu, Chun-Lin; Tso, I-Min

2012-01-01

27

Silk Coatings on PLGA and Alginate Microspheres for Protein Delivery  

PubMed Central

Bombyx mori silk fibroin self-assembles on surfaces to form ultrathin nanoscale coatings based on our prior studies using layer-by-layer deposition techniques driven by hydrophobic interactions between silk fibroin protein molecules. In the present study, polylactic-co-glycolic acid (PLGA) and alginate microspheres were used as substrates and coated with silk fibroin. The coatings were visualized by confocal laser scanning microscopy using fluorescein-labeled silk fibroin. On PLGA microspheres the coating was ~1 ?m and discontinuous, reflecting the porous surface of these microspheres determined by SEM. In contrast, on alginate microspheres the coating was ~10 ?m thick and continuous. The silk fibroin penetrated into the alginate gel matrix. The silk coating on the PLGA microspheres delayed PLGA degradation. The silk coating on the alginate microspheres survived ethylenediamine tetraacetic acid (EDTA) treatment used to remove the Ca+2-cross-links in the alginate gels to solubilize the alginate. This suggests that alginate microspheres can be used as templates to form silk microcapsules. Horseradish peroxidase (HRP) and tetramethylrhodamine-conjugated bovine serum albumin (BSA) as model protein drugs were encapsulated in the PLGA and alginate microspheres with and without the silk fibroin coatings. Drug release was significantly retarded by the silk coatings when compared to uncoated microsphere controls, and was retarded further by methanol-treated silk coating when compared to silk water-based coatings on alginate microspheres. Silk coatings on PLGA and alginate microspheres provide mechanically stable shells as well as a diffusion barrier to the encapsulated protein drugs. This coating technique has potential for biosensor and drug delivery applications due to the aqueous process employed, the ability to control coating thickness and crystalline content, and the biocompatibility of the silk fibroin protein used in the process. PMID:17583788

Wang, Xiaoqin; Wenk, Esther; Hu, Xiao; Castro, Guillermo R.; Meinel, Lorenz; Wang, Xianyan; Li, Chunmei; Merkle, Hans; Kaplan, David L.

2009-01-01

28

Stabilization of viruses by encapsulation in silk proteins.  

PubMed

Viruses are important for a range of modern day applications. However, their utility is limited by their susceptibility to temperature degradation. In this study, we report a simple system to compare the ability of different dried protein films to stabilize viruses against exposure to elevated temperatures. Films from each of three different silks, silkworm, honeybee silk and hornet silk, stabilized entrapped viruses at 37 °C better than films of albumin from bovine serum (BSA) and all four proteins provided substantially more stabilization than no protein controls. A comparison of the molecular structure of the silks and BSA films showed no correlation between the ability of the proteins to stabilize the virus and the secondary structure of the protein in the films. The mechanism of stabilization is discussed and a hypothesis is suggested to explain the superior performance of the silk proteins. PMID:25229876

Sutherland, Tara D; Sriskantha, Alagacone; Church, Jeffrey S; Strive, Tanja; Trueman, Holly E; Kameda, Tsunenori

2014-10-22

29

Tubuliform silk protein: A protein with unique molecular characteristics and mechanical properties in the spider silk fibroin family  

Microsoft Academic Search

Orb–web weavers can produce up to six different types of silk and a glue for various functions. Tubuliform silk is unique\\u000a among them due to its distinct amino acid composition, specific time of production, and atypical mechanical properties. To\\u000a study the protein composing this silk, tubuliform gland cDNA libraries were constructed from three orb–weaving spiders Argiope\\u000a aurantia, Araneus gemmoides, and

M. Tian; R. V. Lewis

2006-01-01

30

Production of synthetic spider dragline silk protein in Pichia pastoris  

Microsoft Academic Search

The methylotrophic yeast Pichia pastoris was tested as a host for the production of long, repetitive protein polymers. Synthetic genes for a designed analog of a\\u000a spider dragline silk protein were readily expressed at high levels under control of the methanol-inducible AOX1 promoter. Transformants containing multiple gene copies produced elevated levels of silk protein, but of a variety of altered

S. R. Fahnestock; L. A. Bedzyk

1997-01-01

31

Solution structure of eggcase silk protein and its implications for silk fiber formation  

PubMed Central

Spider silks are renowned for their excellent mechanical properties and biomimetic and industrial potentials. They are formed from the natural refolding of water-soluble fibroins with ?-helical and random coil structures in silk glands into insoluble fibers with mainly ?-structures. The structures of the fibroins at atomic resolution and silk formation mechanism remain largely unknown. Here, we report the 3D structures of individual domains of a ?366-kDa eggcase silk protein that consists of 20 identical type 1 repetitive domains, one type 2 repetitive domain, and conserved nonrepetitive N- and C-terminal domains. The structures of the individual domains in solution were determined by using NMR techniques. The domain interactions were investigated by NMR and dynamic light-scattering techniques. The formation of micelles and macroscopic fibers from the domains was examined by electron microscopy. We find that either of the terminal domains covalently linked with at least one repetitive domain spontaneously forms micelle-like structures and can be further transformed into fibers at ?37 °C and a protein concentration of >0.1 wt%. Our biophysical and biochemical experiments indicate that the less hydrophilic terminal domains initiate the assembly of the proteins and form the outer layer of the micelles whereas the more hydrophilic repetitive domains are embedded inside to ensure the formation of the micelle-like structures that are the essential intermediates in silk formation. Our results establish the roles of individual silk protein domains in fiber formation and provide the basis for designing miniature fibroins for producing artificial silks. PMID:19458259

Lin, Zhi; Huang, Weidong; Zhang, Jingfeng; Fan, Jing-Song; Yang, Daiwen

2009-01-01

32

Coatings and films made of silk proteins.  

PubMed

Silks are a class of proteinaceous materials produced by arthropods for various purposes. Spider dragline silk is known for its outstanding mechanical properties, and it shows high biocompatibility, good biodegradability, and a lack of immunogenicity and allergenicity. The silk produced by the mulberry silkworm B. mori has been used as a textile fiber and in medical devices for a long time. Here, recent progress in the processing of different silk materials into highly tailored isotropic and anisotropic coatings for biomedical applications such as tissue engineering, cell adhesion, and implant coatings as well as for optics and biosensors is reviewed. PMID:25004395

Borkner, Christian B; Elsner, Martina B; Scheibel, Thomas

2014-09-24

33

Nanostructure and molecular mechanics of spider dragline silk protein assemblies  

PubMed Central

Spider silk is a self-assembling biopolymer that outperforms most known materials in terms of its mechanical performance, despite its underlying weak chemical bonding based on H-bonds. While experimental studies have shown that the molecular structure of silk proteins has a direct influence on the stiffness, toughness and failure strength of silk, no molecular-level analysis of the nanostructure and associated mechanical properties of silk assemblies have been reported. Here, we report atomic-level structures of MaSp1 and MaSp2 proteins from the Nephila clavipes spider dragline silk sequence, obtained using replica exchange molecular dynamics, and subject these structures to mechanical loading for a detailed nanomechanical analysis. The structural analysis reveals that poly-alanine regions in silk predominantly form distinct and orderly beta-sheet crystal domains, while disorderly regions are formed by glycine-rich repeats that consist of 31-helix type structures and beta-turns. Our structural predictions are validated against experimental data based on dihedral angle pair calculations presented in Ramachandran plots, alpha-carbon atomic distances, as well as secondary structure content. Mechanical shearing simulations on selected structures illustrate that the nanoscale behaviour of silk protein assemblies is controlled by the distinctly different secondary structure content and hydrogen bonding in the crystalline and semi-amorphous regions. Both structural and mechanical characterization results show excellent agreement with available experimental evidence. Our findings set the stage for extensive atomistic investigations of silk, which may contribute towards an improved understanding of the source of the strength and toughness of this biological superfibre. PMID:20519206

Keten, Sinan; Buehler, Markus J.

2010-01-01

34

Tubuliform silk protein: A protein with unique molecular characteristics and mechanical properties in the spider silk fibroin family  

NASA Astrophysics Data System (ADS)

Orb-web weavers can produce up to six different types of silk and a glue for various functions. Tubuliform silk is unique among them due to its distinct amino acid composition, specific time of production, and atypical mechanical properties. To study the protein composing this silk, tubuliform gland cDNA libraries were constructed from three orb-weaving spiders Argiope aurantia, Araneus gemmoides, and Nephila clavipes. Amino acid composition comparison between the predicted tubuliform silk protein sequence (TuSp1) and the corresponding gland protein confirms that TuSp1 is the major component in tubuliform gland in three spiders. Sequence analysis suggests that TuSp1 shares no significant similarity with its paralogues, while it has conserved sequence motifs with the most primitive spider, Euagrus chisoseus silk protein. The presence of large side-chain amino acids in TuSp1 sequence is consistent with the frustrated ?-sheet crystalline structure of tubuliform silk observed in transmission electron microscopy. Repeat unit comparison within species as well as among three spiders exhibits high sequence conservation. Parsimony analysis based on carboxy terminal sequence shows that Argiope and Araneus are more closely related than either is to Nephila which is consistent with phylogenetic analysis based on morphological evidence.

Tian, M.; Lewis, R. V.

2006-02-01

35

Solution behavior of synthetic silk peptides and modified recombinant silk proteins  

NASA Astrophysics Data System (ADS)

Spider dragline silk from Nephila clavipes possesses impressive mechanical properties derived in part from repetitive primary sequence containing polyalanine regions that self-assemble into crystalline ?-sheets. In the present study, we have sought to understand more details of redox responses related to conformational transitions of modified silk peptides and a recombinant protein containing encoded methionine triggers. Regardless of the position of the methionine trigger relative to the polyalanine domain, chemical oxidation was rapid and slight increases in the ?-helical structure and decreases in the ?-sheet and random coil content were observed by CD and FTIR in the assembled silk-like peptides and the recombinant protein. CD results indicated that the decrease in ?-sheet and random coil conformations, coupled with the increase in helical content during oxidation, occurred during the first 30 min of the reaction. No further conformational changes occurred after this time and the response was independent of methionine trigger location relative to the penta-alanine domain. These results were confirmed with fluorescence studies. The design, processing and utility of these modified redox triggered silk-like peptides and proteins suggest a range of potential utility, from biomaterials to engineered surface coatings with chemically alterable secondary structure and, thus, properties.

Foo, C. Wong Po; Bini, E.; Huang, J.; Lee, S. Y.; Kaplan, D. L.

2006-02-01

36

Biomaterials Derived from Silk-Tropoelastin Protein Systems  

PubMed Central

A structural protein blend system based on silkworm silk fibroin and recombinant human tropoelastin is described. Silk fibroin, a semicrystalline fibrous protein with beta-sheet crystals provides mechanical strength and controllable biodegradation, while tropoelastin, a noncrystallizable elastic protein provides elasticity. Differential scanning calorimetry (DSC) and temperature modulated DSC (TMDSC) indicated that silk becomes miscible with tropoelastin at different blend ratios, without macrophase separation. Fourier transform infrared spectroscopy (FTIR) revealed secondary structural changes of the blend system (beta-sheet content) before and after methanol treatment. Atomic Force Microscopy (AFM) nano-indentation demonstrated that blending silk and tropoelastin at different ratios resulted in modification of mechanical features, with resilience from ~68% to ~97%, and elastic modulus between 2~9Mpa, depending on the ratio of the two polymers. Some of these values are close to those of native aortic elastin or elastin-like polypeptides. Significantly, during blending and drying silk-tropoelastin form micro- and nano-scale porous morphologies which promote human mesenchymal stem cell attachment and proliferation. These blends offer a new protein biomaterial system for cell support and tailored biomaterial properties to match mechanical needs. PMID:20674969

Hu, Xiao; Wang, Xiuli; Rnjak, Jelena; Weiss, Anthony S.; Kaplan, David L.

2010-01-01

37

Production of synthetic spider dragline silk protein in Pichia pastoris.  

PubMed

The methylotrophic yeast Pichia pastoris was tested as a host for the production of long, repetitive protein polymers. Synthetic genes for a designed analog of a spider dragline silk protein were readily expressed at high levels under control of the methanol-inducible AOX1 promoter. Transformants containing multiple gene copies produced elevated levels of silk protein, but of a variety of altered sizes as a result of gene rearrangements at the time of transformation. Genes up to 3000 codons in length or longer could be expressed with no evidence of the prevalent truncated synthesis observed for similar genes in Escherichia coli, though genes longer than 1600 codons were expressed less efficiently than shorter genes. Silk-producing P. pastoris strains were stable without selection for at least 100 doublings. PMID:9035408

Fahnestock, S R; Bedzyk, L A

1997-01-01

38

Genetic engineering of fibrous proteins: spider dragline silk and collagen  

Microsoft Academic Search

Various strategies have been employed to genetically engineer fibrous proteins. Two examples, the subject of this review, include spider dragline silk from Nephila clavipes and collagen. These proteins are highlighted because of their unique mechanical and biological properties related to controlled release, biomaterials and tissue engineering. Cloning and expression of native genes and synthetic artificial variants of the consensus sequence

Cheryl Wong Po Foo; David L Kaplan

2002-01-01

39

Interfacial rheological properties of recombinant spider-silk proteins Cyrille Vzya  

E-print Network

Interfacial rheological properties of recombinant spider-silk proteins Cyrille Vézya E27 Lehrstuhl to characterize the adsorption kinetics and the rheology of spider-silk films formed at an oil water interface. The high surface activity of the engineered spider-silk proteins results in a fast formation of highly

Bausch, Andreas

40

Molecular characterization and evolutionary study of spider tubuliform (eggcase) silk protein.  

PubMed

As a result of hundreds of millions of years of evolution, orb-web-weaving spiders have developed the use of seven different silks produced by different abdominal glands for various functions. Tubuliform silk (eggcase silk) is unique among these spider silks due to its high serine and very low glycine content. In addition, tubuliform silk is the only silk produced just during a short period of time, the reproductive season, in the spider's life. To understand the molecular characteristics of the proteins composing this silk, we constructed tubuliform-gland-specific cDNA libraries from three different spider families, Nephila clavipes, Argiope aurantia, and Araneus gemmoides. Sequencing of tubuliform silk cDNAs reveals the repetitive architecture of its coding sequence and novel amino acid motifs. The inferred protein, tubuliform spidroin 1 (TuSp1), contains highly homogenized repeats in all three spiders. Amino acid composition comparison of the predicted tubuliform silk protein sequence to tubuliform silk indicates that TuSp1 is the major component of tubuliform silk. Repeat unit alignment of TuSp1 among three spider species shows high sequence conservation among tubuliform silk protein orthologue groups. Sequence comparison among TuSp1 repetitive units within species suggests intragenic concerted evolution, presumably through gene conversion and unequal crossover events. Comparative analysis demonstrates that TuSp1 represents a new orthologue in the spider silk gene family. PMID:15924419

Tian, Maozhen; Lewis, Randolph V

2005-06-01

41

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

NASA Astrophysics Data System (ADS)

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

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

2006-02-01

42

Biosynthesis and Applications of Silk?like and Collagen?like Proteins  

Microsoft Academic Search

Genetic engineering provides a useful strategy to generate native and designed fibrous proteins, such as silks and collagens, for fundamental and applied studies. Recent studies on biosynthesized spider and silkworm silks, and human collagens, are reviewed with a focus on genes, expression systems and biomaterial applications. Genetically engineered silks and collagens can be exploited for fundamental biophysical studies of the

Jia Huang; C. Wong Po Foo; David L. Kaplan

2007-01-01

43

Recombinant spider silk particles for controlled delivery of protein drugs  

Microsoft Academic Search

The engineered and recombinant spider silk protein eADF4(C16) has been shown to be a promising biomaterial for the use as drug delivery system. In previous studies, eADF4(C16) particles were loaded with low molecular weight drugs exhibiting a positive net-charge and sufficient hydrophobicity. Here, we demonstrate that also macromolecular drugs like proteins can be loaded on eADF4(C16) particles. Using lysozyme as

Markus Hofer; Gerhard Winter; Julia Myschik

44

Protein and amino acid composition of silks from the cob weaver, Latrodectus hesperus (black widow).  

PubMed

The silks from the cob weaving spider, Latrodectus hesperus (black widow), have been examined with the goal of expanding our understanding of the relationship between the protein structure and mechanical performance of these unique biomaterials. The scaffolding, dragline and inner egg case silks each appear to be distinct fibers based on mole percent amino acid composition and polypeptide composition. Further, we find that the amino acid composition of dragline and egg case silk are similar to the analogous silks produced by orb weaving spiders, while scaffolding silk may represent a novel silk. The black widow silks are comprised of multiple high molecular weight polypeptides, however, the egg case and scaffolding silks also contain some smaller polypeptides. PMID:10342753

Casem, M L; Turner, D; Houchin, K

1999-01-01

45

Evidence for diet effects on the composition of silk proteins produced by spiders.  

PubMed

Silks are highly expressed, secreted proteins that represent a substantial metabolic cost to the insects and spiders that produce them. Female spiders in the superfamily Araneoidea (the orb-spinning spiders and their close relatives) spin six different kinds of silk (three fibroins and three fibrous protein glues) that differ in amino acid content and protein structure. In addition to this diversity in silks produced by different glands, we found that individual spiders of the same species can spin dragline silks (drawn from the spider's ampullate gland) that vary in content as well. Freely foraging ARGIOPE: argentata (Araneae: Araneoidea), collected from 13 Caribbean islands, produced dragline silk that showed an inverse relationship between the amount of serine and glycine they contained. X-ray microdiffraction of the silks localized these differences to the amorphous regions of the protein that are thought to lend silks their elasticity. The crystalline regions of the proteins, which lend silks their strength, were unaffected. Laboratory experiments with ARGIOPE: keyserlingi suggested that variation in silk composition reflects the type of prey the spiders were fed but not the total amount of prey they received. Hence, it may be that the amino acid content (and perhaps the mechanical properties) of dragline silk spun by ARGIOPE: directly reflect the spiders' diet. The ability to vary silk composition and, possibly, function is particularly important for organisms that disperse broadly, such as Argiope, and that occupy diverse habitats with diverse populations of prey. PMID:11110907

Craig, C L; Riekel, C; Herberstein, M E; Weber, R S; Kaplan, D; Pierce, N E

2000-12-01

46

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

PubMed Central

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

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

2014-01-01

47

Engineering aqueous fiber assembly into silk-elastin-like protein polymers.  

PubMed

Self-assembled peptide/protein nanofibers are valuable 1D building blocks for creating complex structures with designed properties and functions. It is reported that the self-assembly of silk-elastin-like protein polymers into nanofibers or globular aggregates in aqueous solutions can be modulated by tuning the temperature of the protein solutions, the size of the silk blocks, and the charge of the elastin blocks. A core-sheath model is proposed for nanofiber formation, with the silk blocks in the cores and the hydrated elastin blocks in the sheaths. The folding of the silk blocks into stable cores--affected by the size of the silk blocks and the charge of the elastin blocks--plays a critical role in the assembly of silk-elastin nanofibers. Furthermore, enhanced hydrophobic interactions between the elastin blocks at elevated temperatures greatly influence the nanoscale features of silk-elastin nanofibers. PMID:24798978

Zeng, Like; Jiang, Linan; Teng, Weibing; Cappello, Joseph; Zohar, Yitshak; Wu, Xiaoyi

2014-07-01

48

Recombinant Dragline Silk-Like Proteins--Expression and Purification  

PubMed Central

Spider dragline silk is a proteinaceous fiber with impressive physical characteristics making it attractive for use in advanced materials. The fiber is composed of two proteins (spidroins MaSp1 and MaSp2), each of which contains a large central repeat array flanked by non-repetitive N- and C-terminal domains. The repeat arrays appear to be largely responsible for the tensile properties of the fiber, suggesting that the N- and C-terminal domains may be involved in self-assembly. We recently isolated the MaSp1 and MaSp2 N-terminal domains from Nephila clavipes and have incorporated these into mini-silk genes for expression in transgenic systems. Current efforts involve the development of expression vectors that will allow purification using a removable affinity tag for scalable protein purification. PMID:23914141

Gaines, William A.; Marcotte, William R.

2011-01-01

49

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

NASA Astrophysics Data System (ADS)

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

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

2009-03-01

50

Stability of silk and collagen protein materials in space.  

PubMed

Collagen and silk materials, in neat forms and as silica composites, were flown for 18 months on the International Space Station [Materials International Space Station Experiment (MISSE)-6] to assess the impact of space radiation on structure and function. As natural biomaterials, the impact of the space environment on films of these proteins was investigated to understand fundamental changes in structure and function related to the future utility in materials and medicine in space environments. About 15% of the film surfaces were etched by heavy ionizing particles such as atomic oxygen, the major component of the low-Earth orbit space environment. Unexpectedly, more than 80% of the silk and collagen materials were chemically crosslinked by space radiation. These findings are critical for designing next-generation biocompatible materials for contact with living systems in space environments, where the effects of heavy ionizing particles and other cosmic radiation need to be considered. PMID:24305951

Hu, Xiao; Raja, Waseem K; An, Bo; Tokareva, Olena; Cebe, Peggy; Kaplan, David L

2013-01-01

51

Hypotheses that correlate the sequence, structure, and mechanical properties of spider silk proteins.  

PubMed

Several types of silks and silk protein coding genes have been characterized from orb-web weaving spiders. When the protein sequences of major ampullate, minor ampullate, and flagelliform silks from Nephila clavipes are compared, they can be summarized as sets of shared amino acid motifs. Four of these motifs and their likely secondary structures are described. Each structural element, termed a module, is then associated with its impact on the mechanical properties of a silk fiber. In particular, correlations are drawn between an alanine-rich 'crystalline module' and tensile strength and between a proline-containing 'elasticity module' and extensibility. PMID:10342774

Hayashi, C Y; Shipley, N H; Lewis, R V

1999-01-01

52

A Novel Model System for Design of Biomaterials Based on Recombinant Analogs of Spider Silk Proteins  

Microsoft Academic Search

Spider dragline silk possesses impressive mechanical and biochemical properties. It is synthesized by a couple of major ampullate\\u000a glands in spiders and comprises of two major structural proteins—spidroins 1 and 2. The relationship between structure and\\u000a mechanical properties of spider silk is not well understood. Here, we modeled the complete process of the spider silk assembly\\u000a using two new recombinant

Vladimir G. Bogush; Olga S. Sokolova; Lyubov I. Davydova; Dmitri V. Klinov; Konstantin V. Sidoruk; Natalya G. Esipova; Tatyana V. Neretina; Igor A. Orchanskyi; Vsevolod Yu Makeev; Vladimir G. Tumanyan; Konstantin V. Shaitan; Vladimir G. Debabov; Mikhail P. Kirpichnikov

2009-01-01

53

Nanoparticle self-assembly by a highly stable recombinant spider wrapping silk protein subunit.  

PubMed

Artificial spider silk proteins may form fibers with exceptional strength and elasticity. Wrapping silk, or aciniform silk, is the toughest of the spider silks, and has a very different protein composition than other spider silks. Here, we present the characterization of an aciniform protein (AcSp1) subunit named W1, consisting of one AcSp1 199 residue repeat unit from Argiope trifasciata. The structural integrity of recombinant W1 is demonstrated in a variety of buffer conditions and time points. Furthermore, we show that W1 has a high thermal stability with reversible denaturation at ?71°C and forms self-assembled nanoparticle in near-physiological conditions. W1 therefore represents a highly stable and structurally robust module for protein-based nanoparticle formation. PMID:23994530

Xu, Lingling; Tremblay, Marie-Laurence; Orrell, Kathleen E; Leclerc, Jérémie; Meng, Qing; Liu, Xiang-Qin; Rainey, Jan K

2013-10-01

54

Purification and cytotoxcicity of tag-free bioengineered spider silk proteins3  

PubMed Central

Bioengineered spider silk-like proteins can serve as biomaterials for various biomedical applications. These proteins can be assembled in several morphological forms such as films, microcapsules, spheres, fibers, gels and scaffolds. However, crucial points for recombinant spider silks for human use are toxicity and immunogenicity. To assess this issue two bioengineered spider silk proteins composed of different numbers of repetitive motifs of the consensus repeats from spidroin-1 from Nephila clavipes (15X and 6X) were cloned and expressed in E. coli. The proteins were free of tag-sequence and were purified using two methods based on (i) thermal and (ii) organic acid resistance of the spider silks. The soluble spider silk proteins were not cytotoxic and did not activate macrophages over a wide range of concentrations, except when present at the highest concentration. Films made of the different silk variants supported the growth of the cells. Based on these data, and since the biodegradation rate of silk is very slow, the bioengineered spider silks are presumed safe biomaterials for biomedical applications. PMID:22865581

Dams-Kozlowska, Hanna; Majer, Agnieszka; Tomasiewicz, Paulina; Lozinska, Jolanta; Kaplan, David L.; Mackiewicz, Andrzej

2012-01-01

55

Analysis of the conserved N-terminal domains in major ampullate spider silk proteins.  

PubMed

Major ampullate silk, also known as dragline silk, is one of the strongest biomaterials known. This silk is composed of two proteins, major ampullate spidroin 1 (MaSp1) and major ampullate spidroin 2 (MaSp2). Only partial cDNA sequences have been obtained for these proteins, and these sequences are toward the C-terminus. Thus, the N-terminal domains have never been characterized for either protein. Here we report the sequence of the N-terminal region of major ampullate silk proteins from three spider species: Argiope trifasciata, Latrodectus geometricus, and Nephila inaurata madagascariensis. The amino acid sequences are inferred from genomic DNA clones. Northern blotting experiments suggest that the predicted 5' end of the transcripts are present in fibroin mRNA. The presence of more than one Met codon in the N-terminal region indicates the possibility of translation of both a long and a short isoform. The size of the short isoform is consistent with the published, cDNA based, N-terminal sequence found in flagelliform silk. Analyses comparing the level of identity of all known spider silk N-termini show that the N-terminus is the most conserved part of silk proteins. Two DNA sequence motifs identified upstream of the putative transcription start site are potential silk fibroin promoter elements. PMID:16283740

Motriuk-Smith, Dagmara; Smith, Alyson; Hayashi, Cheryl Y; Lewis, Randolph V

2005-01-01

56

Sericin-carboxymethyl cellulose porous matrices as cellular wound dressing material.  

PubMed

In this study, porous three-dimensional (3D) hydrogel matrices are fabricated composed of silk cocoon protein sericin of non-mulberry silkworm Antheraea mylitta and carboxymethyl cellulose. The matrices are prepared via freeze-drying technique followed by dual cross-linking with glutaraldehyde and aluminum chloride. The microstructure of the hydrogel matrices is assessed using scanning electron microscopy and biophysical characterization are carried out using Fourier transform infrared spectroscopy and X-ray diffraction. The transforming growth factor ?1 release from the cross-linked matrices as a growth factor is evaluated by immunosorbent assay. Live dead assay and 3-[4,5-dimethylthiazolyl-2]-2,5-diphenyl tetrazolium bromide assay show no cytotoxicity of blended matrices toward human keratinocytes. The matrices support the cell attachment and proliferation of human keratinocytes as observed through scanning electron microscope and confocal images. Gelatin zymography demonstrates the low levels of matrix metalloproteinase 2 (MMP-2) and insignificant amount of MMP-9 in the culture media of cell seeded matrices. Low inflammatory response of the matrices is indicated through tumor necrosis factor alpha release assay. The results indicate that the fabricated matrices constitute 3D cell-interactive environment for tissue engineering applications and its potential use as a future cellular biological wound dressing material. PMID:23853114

Nayak, Sunita; Kundu, S C

2014-06-01

57

Analysis of tissue-specific region in sericin 1 gene promoter of Bombyx mori  

SciTech Connect

The gene encoding sericin 1 (Ser1) of silkworm (Bombyx mori) is specifically expressed in the middle silk gland cells. To identify element involved in this transcription-dependent spatial restriction, truncation of the 5' terminal from the sericin 1 (Ser1) promoter is studied in vivo. A 209 bp DNA sequence upstream of the transcriptional start site (-586 to -378) is found to be responsible for promoting tissue-specific transcription. Analysis of this 209 bp region by overlapping deletion studies showed that a 25 bp region (-500 to -476) suppresses the ectopic expression of the Ser1 promoter. An unknown factor abundant in fat body nuclear extracts is shown to bind to this 25 bp fragment. These results suggest that this 25 bp region and the unknown factor are necessary for determining the tissue-specificity of the Ser1 promoter.

Liu Yan [College of Biomedical Engineering and Instrument Science, Zhejiang University, Hangzhou 310027 (China); Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031 (China); Yu Lian [College of Biomedical Engineering and Instrument Science, Zhejiang University, Hangzhou 310027 (China); Zhejiang Province Key Laboratory of Preventive Veterinary Medicine, Institute of Preventive Veterinary Medicine, Zhejiang University, Hangzhou 310029 (China); Guo Xiuyang [Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031 (China); Guo Tingqing [Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031 (China); Wang Shengpeng [Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031 (China); Lu Changde [Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031 (China)]. E-mail: cdlu@sibs.ac.cn

2006-03-31

58

Quantitative Correlation Between the Protein Primary Sequences and Secondary Structures in Spider Dragline Silks  

PubMed Central

Synthetic spider silk holds great potential for use in various applications spanning medical uses to ultra lightweight armor, however producing synthetic fibers with mechanical properties comparable to natural spider silk has eluded the scientific community. Natural dragline spider silks are commonly made from proteins that contain highly repetitive amino acid motifs, adopting an array of secondary structures. Before further advances can be made in the production of synthetic fibers based on spider silk proteins, it is imperative to know the percentage of each amino acid in the protein that forms a specific secondary structure. Linking these percentages to the primary amino acid sequence of the protein will establish a structural foundation for synthetic silk. In this study, Nuclear Magnetic Resonance (NMR) techniques are used to quantify the percentage of Ala, Gly, and Ser that form both ?-sheet and helical secondary structures. The fraction of these three amino acids and their secondary structure are quantitatively correlated to the primary amino acid sequence for the proteins that comprise major and minor ampullate silk from the Nephila clavipes spider providing a blueprint for synthetic spider silks. PMID:20000730

Jenkins, Janelle E.; Creager, Melinda S.; Lewis, Randolph V.; Holland, Gregory P.; Yarger, Jeffery L.

2009-01-01

59

Dragline silk: a fiber assembled with low-molecular-weight cysteine-rich proteins.  

PubMed

Dragline silk has been proposed to contain two main protein constituents, MaSp1 and MaSp2. However, the mechanical properties of synthetic spider silks spun from recombinant MaSp1 and MaSp2 proteins have yet to approach natural fibers, implying the natural spinning dope is missing critical factors. Here we report the discovery of novel molecular constituents within the spinning dope that are extruded into dragline silk. Protein studies of the liquid spinning dope from the major ampullate gland, coupled with the analysis of dragline silk fibers using mass spectrometry, demonstrate the presence of a new family of low-molecular-weight cysteine-rich proteins (CRPs) that colocalize with the MA fibroins. Expression of the CRP family members is linked to dragline silk production, specifically MaSp1 and MaSp2 mRNA synthesis. Biochemical data support that CRP molecules are secreted into the spinning dope and assembled into macromolecular complexes via disulfide bond linkages. Sequence analysis supports that CRP molecules share similarities to members that belong to the cystine slipknot superfamily, suggesting that these factors may have evolved to increase fiber toughness by serving as molecular hubs that dissipate large amounts of energy under stress. Collectively, our findings provide molecular details about the components of dragline silk, providing new insight that will advance materials development of synthetic spider silk for industrial applications. PMID:25259849

Pham, Thanh; Chuang, Tyler; Lin, Albert; Joo, Hyun; Tsai, Jerry; Crawford, Taylor; Zhao, Liang; Williams, Caroline; Hsia, Yang; Vierra, Craig

2014-11-10

60

Sequence-structure correlations in the MaSp1 protein of N. clavipes dragline silk  

E-print Network

Silk is a hierarchically structured protein fiber with exceptional tensile strength and extensibility, making it one of the toughest and most versatile biocompatible materials. While experimental studies have shown that ...

Bratzel, Graham Hayden

2011-01-01

61

TNF-? blocker effect of naringenin-loaded sericin microparticles that are potentially useful in the treatment of psoriasis.  

PubMed

This study aims to evaluate the effect of combined use of the racemic flavanone Naringenin (NRG) and the protein sericin as TNF-? blockers. Sericin (SMs) and (R/S) NRG-loaded Sericin (SNRGMs) microparticles were prepared by spray-drying, characterized in terms of morphology and particle size distribution, and encapsulation efficiency was determined. Concerning morphology and particle size distribution of microparticles, results indicated that they were not affected by the presence of NRG. The encapsulation efficiency was almost quantitative (93%), thus proving that sericin can be advantageously loaded with (R/S) NRG. Biological evaluation of (R/S) NRG, SMs and SNRGMs was then performed in lipopolysaccharide (LPS)-stimulated human peripheral blood mononuclear cells (hPBMC). SNRGMs resulted cytotoxic at the higher dose used (200 ?g/mL) and the effect was greater than (R/S) NRG alone. Moreover, even if sericin alone was not effective in suppressing LPS-induced serum TNF-? levels, SNRGMs loaded with 9.3% of (R/S) NRG were significantly more potent than (R/S) NRG alone. In summary, this study provides the proof of concept that sericin-based microspheres loaded with TNF-?-blockers could contribute to the down regulation of the cytokine and represents the starting point for the development of new topical formulations for the treatment of middle-stage psoriasis. PMID:25101847

Chlapanidas, Theodora; Perteghella, Sara; Leoni, Flavio; Faragò, Silvio; Marazzi, Mario; Rossi, Daniela; Martino, Emanuela; Gaggeri, Raffaella; Collina, Simona

2014-01-01

62

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

PubMed

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

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

2013-03-01

63

Recombinant minimalist spider wrapping silk proteins capable of native-like fiber formation.  

PubMed

Spider silks are desirable biomaterials characterized by high tensile strength, elasticity, and biocompatibility. Spiders produce different types of silks for different uses, although dragline silks have been the predominant focus of previous studies. Spider wrapping silk, made of the aciniform protein (AcSp1), has high toughness because of its combination of high elasticity and tensile strength. AcSp1 in Argiope trifasciata contains a 200-aa sequence motif that is repeated at least 14 times. Here, we produced in E. coli recombinant proteins consisting of only one to four of the 200-aa AcSp1 repeats, designated W(1) to W(4). We observed that purified W(2), W(3) and W(4) proteins could be induced to form silk-like fibers by shear forces in a physiological buffer. The fibers formed by W(4) were ?3.4 µm in diameter and up to 10 cm long. They showed an average tensile strength of 115 MPa, elasticity of 37%, and toughness of 34 J cm(-3). The smaller W(2) protein formed fewer fibers and required a higher protein concentration to form fibers, whereas the smallest W(1) protein did not form silk-like fibers, indicating that a minimum of two of the 200-aa repeats was required for fiber formation. Microscopic examinations revealed structural features indicating an assembly of the proteins into spherical structures, fibrils, and silk-like fibers. CD and Raman spectral analysis of protein secondary structures suggested a transition from predominantly ?-helical in solution to increasingly ?-sheet in fibers. PMID:23209681

Xu, Lingling; Rainey, Jan K; Meng, Qing; Liu, Xiang-Qin

2012-01-01

64

Recombinant Minimalist Spider Wrapping Silk Proteins Capable of Native-Like Fiber Formation  

PubMed Central

Spider silks are desirable biomaterials characterized by high tensile strength, elasticity, and biocompatibility. Spiders produce different types of silks for different uses, although dragline silks have been the predominant focus of previous studies. Spider wrapping silk, made of the aciniform protein (AcSp1), has high toughness because of its combination of high elasticity and tensile strength. AcSp1 in Argiope trifasciata contains a 200-aa sequence motif that is repeated at least 14 times. Here, we produced in E. coli recombinant proteins consisting of only one to four of the 200-aa AcSp1 repeats, designated W1 to W4. We observed that purified W2, W3 and W4 proteins could be induced to form silk-like fibers by shear forces in a physiological buffer. The fibers formed by W4 were ?3.4 µm in diameter and up to 10 cm long. They showed an average tensile strength of 115 MPa, elasticity of 37%, and toughness of 34 J cm?3. The smaller W2 protein formed fewer fibers and required a higher protein concentration to form fibers, whereas the smallest W1 protein did not form silk-like fibers, indicating that a minimum of two of the 200-aa repeats was required for fiber formation. Microscopic examinations revealed structural features indicating an assembly of the proteins into spherical structures, fibrils, and silk-like fibers. CD and Raman spectral analysis of protein secondary structures suggested a transition from predominantly ?-helical in solution to increasingly ?-sheet in fibers. PMID:23209681

Xu, Lingling; Rainey, Jan K.; Meng, Qing; Liu, Xiang-Qin

2012-01-01

65

Nanostructure and molecular mechanics of spider dragline silk protein assemblies  

E-print Network

Spider silk is a self-assembling biopolymer that outperforms most known materials in terms of its mechanical performance, despite its underlying weak chemical bonding based on H-bonds. While experimental studies have shown ...

Keten, Sinan

66

Facts and myths of antibacterial properties of silk.  

PubMed

Silk cocoons provide protection to silkworm from biotic and abiotic hazards during the immobile pupal phase of the lifecycle of silkworms. Protection is particularly important for the wild silk cocoons reared in an open and harsh environment. To understand whether some of the cocoon components resist growth of microorganisms, in vitro studies were performed using gram negative bacteria Escherichia coli (E. coli) to investigate antibacterial properties of silk fiber, silk gum, and calcium oxalate crystals embedded inside some cocoons. The results show that the previously reported antibacterial properties of silk cocoons are actually due to residues of chemicals used to isolate/purify cocoon elements, and properly isolated silk fiber, gum, and embedded crystals free from such residues do not have inherent resistance to E. coli. This study removes the uncertainty created by previous studies over the presence of antibacterial properties of silk cocoons, particularly the silk gum and sericin. © 2013 Wiley Periodicals, Inc. Biopolymers 101: 237-245, 2014. PMID:23784754

Kaur, Jasjeet; Rajkhowa, Rangam; Afrin, Tarannum; Tsuzuki, Takuya; Wang, Xungai

2014-03-01

67

A comparison of the composition of silk proteins produced by spiders and insects  

Microsoft Academic Search

Proteins that are highly expressed and composed of amino acids that are costly to synthesize are likely to place a greater drain on an organism’s energy resources than proteins that are composed of ingested amino acids or ones that are metabolically simple to produce. Silks are highly expressed proteins produced by all spiders and many insects. We compared the metabolic

Catherine L. Craig; Michael Hsu; David Kaplan; N. E. Pierce

1999-01-01

68

A protocol for the production of recombinant spider silk-like proteins for artificial fiber spinning  

PubMed Central

The extreme strength and elasticity of spider silks originate from the modular nature of their repetitive proteins. To exploit such materials and mimic spider silks, comprehensive strategies to produce and spin recombinant fibrous proteins are necessary. This protocol describes silk gene design and cloning, protein expression in bacteria, recombinant protein purification and fiber formation. With an improved gene construction and cloning scheme, this technique is adaptable for the production of any repetitive fibrous proteins, and ensures the exact reproduction of native repeat sequences, analogs or chimeric versions. The proteins are solubilized in 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP) at 25–30% (wt/vol) for extrusion into fibers. This protocol, routinely used to spin single micrometer-size fibers from several recombinant silk-like proteins from different spider species, is a powerful tool to generate protein libraries with corresponding fibers for structure–function relationship investigations in protein-based biomaterials. This protocol may be completed in 40 d. PMID:19229199

Teule, Florence; Cooper, Alyssa R; Furin, William A; Bittencourt, Daniela; Rech, Elibio L; Brooks, Amanda; Lewis, Randolph V

2009-01-01

69

Molecular Evolution of Lepidopteran Silk Proteins: Insights from the Ghost Moth, Hepialus californicus  

E-print Network

Ó The Author(s) 2010. This article is published with open access at Springerlink.com Abstract Silk production has independently evolved in numerous arthropod lineages, such as Lepidoptera, the moths and butterflies. Lepidopteran larvae (caterpillars) synthesize silk proteins in modified salivary glands and spin silk fibers into protective tunnels, escape lines, and pupation cocoons. Molecular sequence data for these proteins are necessary to determine critical features of their function and evolution. To this end, we constructed an expression library from the silk glands of the ghost moth, Hepialus californicus, and characterized light chain fibroin and heavy chain fibroin gene transcripts. The predicted H. californicus silk fibroins share many elements with other lepidopteran and trichopteran fibroins, such as conserved placements of cysteine, aromatic, and polar amino acid residues. Further comparative analyses were performed to determine site-specific signatures of selection and to assess whether fibroin genes are informative as phylogenetic markers. We found that purifying selection has constrained mutation within the fibroins and that light chain fibroin is a promising molecular marker. Thus, by characterizing the H. californicus fibroins, we identified key functional amino acids and gained insight into the evolutionary processes that have shaped these adaptive molecules.

Matthew A. Collin; Kazuei Mita; Frantisek Sehnal; Cheryl Y. Hayashi; M. A. Collin; C. Y. Hayashi; K. Mita; F. Sehnal; Lepidoptera Trichoptera Silk Fibroin

70

Snmp-1, a Novel Membrane Protein of Olfactory Neurons of the Silk Moth Antheraea polyphemus with Homology to the CD36 Family of  

E-print Network

Snmp-1, a Novel Membrane Protein of Olfactory Neurons of the Silk Moth Antheraea polyphemus of the wild silk moth Antheraea polyphemus. We have purified and cloned a prominent 67-kDa protein which we of olfactory neuron receptor membranes of the wild silk moth Antheraea polyphemus. The morphology of the A

Vogt, Richard G.

71

High yield recombinant silk-like protein production in transgenic plants through protein targeting.  

PubMed

DP1B is a synthetic analogue of spider dragline silk protein. It can be spun to form silk fiber. Previously, it had been expressed in transgenic plants, showing the general feasibility of the plant-based DP1B production. However, success of such a plant-based platform requires a great increase of DP1B productivity in plant cells to reduce production cost. This report describes a protein targeting approach to accumulate DP1B in apoplast, ER lumen, and vacuole in Arabidopsis cells, by utilizing appropriate combinations of sporamin-targeting determinant peptides and ER retention peptide. The approach has dramatically enhanced DP1B accumulation, resulting in high production yield. The accumulation can be as high as 8.5 and 6.7% total soluble protein in leaf tissue by targeting to apoplast and ER lumen, respectively, or as high as 18 and 8.2% total soluble protein in seeds by targeting to ER lumen and vacuole, respectively. However, the vacuole targeting in leaves and the apoplast targeting in seeds have failed to accumulate full length DP1B molecules or any DP1B at all, respectively, suggesting that they may not be suitable for applications in leaf tissues and seeds. Data in this study recommend a combination of seed-specific expression and ER-targeting as one of the best strategies for yield enhancement of plant-based DP1B production. PMID:16145839

Yang, Jianjun; Barr, Leslie A; Fahnestock, Stephen R; Liu, Zhan-Bin

2005-06-01

72

Tunable self-assembly of genetically engineered silk--elastin-like protein polymers.  

PubMed

Silk--elastin-like protein polymers (SELPs), consisting of the repeating units of silk and elastin blocks, combine a set of outstanding physical and biological properties of silk and elastin. Because of the unique properties, SELPs have been widely fabricated into various materials for the applications in drug delivery and tissue engineering. However, little is known about the fundamental self-assembly characteristics of these remarkable polymers. Here we propose a two-step self-assembly process of SELPs in aqueous solution for the first time and report the importance of the ratio of silk-to-elastin blocks in a SELP's repeating unit on the assembly of the SELP. Through precise tuning of the ratio of silk to elastin, various structures including nanoparticles, hydrogels, and nanofibers could be generated either reversibly or irreversibly. This assembly process might provide opportunities to generate innovative smart materials for biosensors, tissue engineering, and drug delivery. Furthermore, the newly developed SELPs in this study may be potentially useful as biomaterials for controlled drug delivery and biomedical engineering. PMID:21955178

Xia, Xiao-Xia; Xu, Qiaobing; Hu, Xiao; Qin, Guokui; Kaplan, David L

2011-11-14

73

IDENTIFICATION, CHARACTERIZATION AND ANTI-FUNGAL ACTVITIES OF SILK PROTEINS IN ASPERGILLUS FLAVUS  

E-print Network

challenged by A. flavus 3) by mapping the proteome of silk proteins in a A. flavus resistant inbred and 4 and susceptible corn inbreds showed significant activity in the resistant line compared to the susceptible line (p RESISTANT AND SUSCEPTIBLE MAIZE INBREDS By Bela Peethambaran A Dissertation Submitted to the Faculty

Ray, David

74

Study of Protein Conformation and Orientation in Silkworm and Spider Silk Fibers Using Raman Microspectroscopy  

E-print Network

Study of Protein Conformation and Orientation in Silkworm and Spider Silk Fibers Using Raman mori and Samia cynthia ricini silkworms, and from the spider Nephila edulis. It is shown that.19 ( 0.02, respectively, even though the two types of silkworm fibroins strongly differ in their primary

Pezolet, Michel

75

Native-sized recombinant spider silk protein produced in metabolically engineered Escherichia coli results in a strong fiber  

PubMed Central

Spider dragline silk is a remarkably strong fiber that makes it attractive for numerous applications. Much has thus been done to make similar fibers by biomimic spinning of recombinant dragline silk proteins. However, success is limited in part due to the inability to successfully express native-sized recombinant silk proteins (250–320 kDa). Here we show that a 284.9 kDa recombinant protein of the spider Nephila clavipes is produced and spun into a fiber displaying mechanical properties comparable to those of the native silk. The native-sized protein, predominantly rich in glycine (44.9%), was favorably expressed in metabolically engineered Escherichia coli within which the glycyl-tRNA pool was elevated. We also found that the recombinant proteins of lower molecular weight versions yielded inferior fiber properties. The results provide insight into evolution of silk protein size related to mechanical performance, and also clarify why spinning lower molecular weight proteins does not recapitulate the properties of native fibers. Furthermore, the silk expression, purification, and spinning platform established here should be useful for sustainable production of natural quality dragline silk, potentially enabling broader applications. PMID:20660779

Xia, Xiao-Xia; Qian, Zhi-Gang; Ki, Chang Seok; Park, Young Hwan; Kaplan, David L.; Lee, Sang Yup

2010-01-01

76

Formulation of poorly water-soluble substances using self-assembling spider silk protein  

Microsoft Academic Search

In nature self-assembly of soluble monomeric proteins to defined supramolecular structures is an essential process in the formation of morphologically distinct biological materials like cells, tissues, diatoms or mollusk shells. Using these proteins, complex structures in the nanometer and micrometer scale can be generated that are hardly obtainable by other methods. The engineered ADF4(C16) spider silk protein is mimicking the

Burghard Liebmann; Daniel Hümmerich; Thomas Scheibel; Marcus Fehr

2008-01-01

77

Silk protein based hybrid photonic-plasmonic crystal.  

PubMed

We propose a biocompatible hybrid photonic platform incorporating a 3D silk inverse opal (SIO) crystal and a 2D plasmonic crystal formed on the top surface of the SIO. This hybrid photonic-plasmonic crystal (HPPC) structure simultaneously exhibits both an extraordinary transmission and a pseudo-photonic band-gap in its transmission spectrum. We demonstrate the use of the HPPC as a refractive index (RI) sensor. By performing a multispectral analysis to analyze the RI value, a sensitivity of 200,000 nm·?%T/RIU (refractive index unit) is achieved. PMID:23571980

Kim, Sunghwan; Mitropoulos, Alexander N; Spitzberg, Joshua D; Kaplan, David L; Omenetto, Fiorenzo G

2013-04-01

78

Synthetic spider dragline silk proteins and their production in Escherichia coli  

Microsoft Academic Search

Synthetic genes were designed to encode analogs of the two proteins of Nephila clavipes dragline silk, spidroins 1 and 2. The genes were constructed of tandem repeats of relatively long (more than 300?bp) DNA\\u000a sequences assembled from synthetic oligonucleotides, and encoded proteins of high molecular mass (65–163?kDa). Both analogs\\u000a were produced efficiently in Escherichia coli. The yield and homogeneity of

S. R. Fahnestock; S. L. Irwin

1997-01-01

79

Air filter devices including nonwoven meshes of electrospun recombinant spider silk proteins.  

PubMed

Based on the natural sequence of Araneus diadematus Fibroin 4 (ADF4), the recombinant spider silk protein eADF4(C16) has been engineered. This highly repetitive protein has a molecular weight of 48kDa and is soluble in different solvents (hexafluoroisopropanol (HFIP), formic acid and aqueous buffers). eADF4(C16) provides a high potential for various technical applications when processed into morphologies such as films, capsules, particles, hydrogels, coatings, fibers and nonwoven meshes. Due to their chemical stability and controlled morphology, the latter can be used to improve filter materials. In this protocol, we present a procedure to enhance the efficiency of different air filter devices, by deposition of nonwoven meshes of electrospun recombinant spider silk proteins. Electrospinning of eADF4(C16) dissolved in HFIP results in smooth fibers. Variation of the protein concentration (5-25% w/v) results in different fiber diameters (80-1,100 nm) and thus pore sizes of the nonwoven mesh. Post-treatment of eADF4(C16) electrospun from HFIP is necessary since the protein displays a predominantly ?-helical secondary structure in freshly spun fibers, and therefore the fibers are water soluble. Subsequent treatment with ethanol vapor induces formation of water resistant, stable ?-sheet structures, preserving the morphology of the silk fibers and meshes. Secondary structure analysis was performed using Fourier transform infrared spectroscopy (FTIR) and subsequent Fourier self-deconvolution (FSD). The primary goal was to improve the filter efficiency of existing filter substrates by adding silk nonwoven layers on top. To evaluate the influence of electrospinning duration and thus nonwoven layer thickness on the filter efficiency, we performed air permeability tests in combination with particle deposition measurements. The experiments were carried out according to standard protocols. PMID:23685883

Lang, Gregor; Jokisch, Stephan; Scheibel, Thomas

2013-01-01

80

Dielectric breakdown strength of regenerated silk fibroin films as a function of protein conformation.  

PubMed

Derived from Bombyx mori cocoons, regenerated silk fibroin (RSF) exhibits excellent biocompatibility, high toughness, and tailorable biodegradability. Additionally, RSF materials are flexible, optically clear, easily patterned with nanoscale features, and may be doped with a variety bioactive species. This unique combination of properties has led to increased interest in the use of RSF in sustainable and biocompatible electronic devices. In order to explore the applicability of this biopolymer to the development of future bioelectronics, the dielectric breakdown strength (Ebd) of RSF thin films was quantified as a function of protein conformation. The application of processing conditions that increased ?-sheet content (as determined by FTIR analysis) and produced films in the silk II structure resulted in RSF materials with improved Ebd with values reaching up to 400 V/?m. PMID:23987229

Dickerson, Matthew B; Fillery, Scott P; Koerner, Hilmar; Singh, Kristi M; Martinick, Katie; Drummy, Lawrence F; Durstock, Michael F; Vaia, Richard A; Omenetto, Fiorenzo G; Kaplan, David L; Naik, Rajesh R

2013-10-14

81

Molecular cloning and expression of the C-terminus of spider flagelliform silk protein from Araneus ventricosus.  

PubMed

A cDNA coding for the C-terminus of spider flagelliform silk protein (AvFlag) was cloned from Araneus ventricosus. Analysis of the cDNA sequence shows that the C-terminus of AvFlag consists of 167 amino acids of a repetitive region and 87 amino acids of a C-terminal non-repetitive region. The peptide motifs found in spider flagelliform silk proteins, GPGGX and GGX,were conserved in the repetitive region of AvFlag. Phylogenetic analysis further confirmed that AvFlag belongs to the spider flagelliform silk proteins. The AvFlag cDNA was expressed as a 28 kDa polypeptide in baculovirus-infected insect cells. As a new expression approach for spider silk protein,the combination of polyhedrin and AvFlag creates a polyhedrin AvFlag fusion protein (61 kDa) that is produced as recombinant polyhedra; this provides a basis for the source of spider silk proteins for various applications. PMID:17762143

Lee, Kwang Sik; Kim, Bo Yeon; Je, Yeon Ho; Woo, Soo Dong; Sohn, Hung Dae; Jin, Byung Rae

2007-06-01

82

Protein Secondary Structure and Orientation in Silk as Revealed by Raman Spectromicroscopy  

PubMed Central

Taking advantage of recent advances in polarized Raman microspectroscopy, and based on a rational decomposition of the amide I band, the conformation and orientation of proteins have been determined for cocoon silks of the silkworms Bombyx mori and Samia cynthia ricini and dragline silks of the spiders Nephila clavipes and Nephila edulis. This study distinguished between band components due to ?-sheets, ?-turns, 31-helices, and unordered structure for the four fibers. For B. mori, the ?-sheet content is 50%, which matches the proportion of residues that form the GAGAGS fibroin motifs. For the Nephila dragline and S. c. ricini cocoon, the ?-sheet content (36–37% and 45%, respectively) is higher than the proportion of residues that belong to polyalanine blocks (18% and 42%, respectively), showing that adjacent GGA motifs are incorporated into the ?-sheets. Nephila spidroins contain fewer ?-sheets and more flexible secondary structures than silkworm fibroins. The amorphous polypeptide chains are preferentially aligned parallel to the fiber direction, although their level of orientation is much lower than that of ?-sheets. Overall, the results show that the four silks exhibit a common molecular organization, with mixtures of different amounts of ?-sheets and flexible structures, which are organized with specific orientation levels. PMID:17277183

Lefevre, Thierry; Rousseau, Marie-Eve; Pezolet, Michel

2007-01-01

83

Protein secondary structure and orientation in silk as revealed by Raman spectromicroscopy.  

PubMed

Taking advantage of recent advances in polarized Raman microspectroscopy, and based on a rational decomposition of the amide I band, the conformation and orientation of proteins have been determined for cocoon silks of the silkworms Bombyx mori and Samia cynthia ricini and dragline silks of the spiders Nephila clavipes and Nephila edulis. This study distinguished between band components due to beta-sheets, beta-turns, 3(1)-helices, and unordered structure for the four fibers. For B. mori, the beta-sheet content is 50%, which matches the proportion of residues that form the GAGAGS fibroin motifs. For the Nephila dragline and S. c. ricini cocoon, the beta-sheet content (36-37% and 45%, respectively) is higher than the proportion of residues that belong to polyalanine blocks (18% and 42%, respectively), showing that adjacent GGA motifs are incorporated into the beta-sheets. Nephila spidroins contain fewer beta-sheets and more flexible secondary structures than silkworm fibroins. The amorphous polypeptide chains are preferentially aligned parallel to the fiber direction, although their level of orientation is much lower than that of beta-sheets. Overall, the results show that the four silks exhibit a common molecular organization, with mixtures of different amounts of beta-sheets and flexible structures, which are organized with specific orientation levels. PMID:17277183

Lefèvre, Thierry; Rousseau, Marie-Eve; Pézolet, Michel

2007-04-15

84

Synthetic spider dragline silk proteins and their production in Escherichia coli.  

PubMed

Synthetic genes were designed to encode analogs of the two proteins of Nephila clavipes dragline silk, spidroins 1 and 2. The genes were constructed of tandem repeats of relatively long (more than 300 bp) DNA sequences assembled from synthetic oligonucleotides, and encoded proteins of high molecular mass (65-163 kDa). Both analogs were produced efficiently in Escherichia coli. The yield and homogeneity of the products of longer genes were limited by premature termination of synthesis, probably as a result of processivity errors in protein synthesis. Average termination rates were determined to be 1 in 1100 codons to 1 in 300 codons, depending on the length and synonymous codon choices of the gene. Both analog proteins could be induced to form stable aqueous solutions without denaturants. Circular dichroism spectra of the purified proteins in dilute solution resembled spectra of redissolved natural dragline silk in reflecting a largely disordered structure in water and more ordered structures in mixed solvents with methanol and trifluoroethanol. PMID:9035407

Fahnestock, S R; Irwin, S L

1997-01-01

85

Silk as a Biomaterial  

PubMed Central

Silks are fibrous proteins with remarkable mechanical properties produced in fiber form by silkworms and spiders. Silk fibers in the form of sutures have been used for centuries. Recently regenerated silk solutions have been used to form a variety of biomaterials, such as gels, sponges and films, for medical applications. Silks can be chemically modified through amino acid side chains to alter surface properties or to immobilize cellular growth factors. Molecular engineering of silk sequences has been used to modify silks with specific features, such as cell recognition or mineralization. The degradability of silk biomaterials can be related to the mode of processing and the corresponding content of beta sheet crystallinity. Several primary cells and cell lines have been successfully grown on different silk biomaterials to demonstrate a range of biological outcomes. Silk biomaterials are biocompatible when studied in vitro and in vivo. Silk scaffolds have been successfully used in wound healing and in tissue engineering of bone, cartilage, tendon and ligament tissues. PMID:19543442

Vepari, Charu

2009-01-01

86

Autoclaving as a chemical-free process to stabilize recombinant silk-elastinlike protein polymer nanofibers  

NASA Astrophysics Data System (ADS)

We report here that autoclaving is a chemical-free, physical crosslinking strategy capable of stabilizing electrospun recombinant silk-elastinlike protein (SELP) polymer nanofibers. Fourier transform infrared spectroscopy showed that the autoclaving of SELP nanofibers induced a conformational conversion of ?-turns and unordered structures to ordered ?-sheets. Tensile stress-strain analysis of the autoclaved SELP nanofibrous scaffolds in phosphate buffered saline at 37 °C revealed a Young's modulus of 1.02 ± 0.28 MPa, an ultimate tensile strength of 0.34 ± 0.04 MPa, and a strain at failure of 29% ± 3%.

Qiu, Weiguo; Cappello, Joseph; Wu, Xiaoyi

2011-06-01

87

A novel marine silk  

NASA Astrophysics Data System (ADS)

The discovery of a novel silk production system in a marine amphipod provides insights into the wider potential of natural silks. The tube-building corophioid amphipod Crassicorophium bonellii produces from its legs fibrous, adhesive underwater threads that combine barnacle cement biology with aspects of spider silk thread extrusion spinning. We characterised the filamentous silk as a mixture of mucopolysaccharides and protein deriving from glands representing two distinct types. The carbohydrate and protein silk secretion is dominated by complex ?-sheet structures and a high content of charged amino acid residues. The filamentous secretion product exits the gland through a pore near the tip of the secretory leg after having moved through a duct, which subdivides into several small ductules all terminating in a spindle-shaped chamber. This chamber communicates with the exterior and may be considered the silk reservoir and processing/mixing space, in which the silk is mechanically and potentially chemically altered and becomes fibrous. We assert that further study of this probably independently evolved, marine arthropod silk processing and secretion system can provide not only important insights into the more complex arachnid and insect silks but also into crustacean adhesion cements.

Kronenberger, Katrin; Dicko, Cedric; Vollrath, Fritz

2012-01-01

88

A novel marine silk.  

PubMed

The discovery of a novel silk production system in a marine amphipod provides insights into the wider potential of natural silks. The tube-building corophioid amphipod Crassicorophium bonellii produces from its legs fibrous, adhesive underwater threads that combine barnacle cement biology with aspects of spider silk thread extrusion spinning. We characterised the filamentous silk as a mixture of mucopolysaccharides and protein deriving from glands representing two distinct types. The carbohydrate and protein silk secretion is dominated by complex ?-sheet structures and a high content of charged amino acid residues. The filamentous secretion product exits the gland through a pore near the tip of the secretory leg after having moved through a duct, which subdivides into several small ductules all terminating in a spindle-shaped chamber. This chamber communicates with the exterior and may be considered the silk reservoir and processing/mixing space, in which the silk is mechanically and potentially chemically altered and becomes fibrous. We assert that further study of this probably independently evolved, marine arthropod silk processing and secretion system can provide not only important insights into the more complex arachnid and insect silks but also into crustacean adhesion cements. PMID:22057952

Kronenberger, Katrin; Dicko, Cedric; Vollrath, Fritz

2012-01-01

89

Molecular cloning and expression of the C-terminus of spider flagelliform silk protein from Araneus ventricosus  

Microsoft Academic Search

A cDNA coding for the C-terminus of spider flagelliform silk protein (AvFlag) was cloned from Araneus ventricosus. Analysis of the cDNA sequence shows that the C-terminus of AvFlag consists of 167 amino acids of a repetitive region and\\u000a 87 amino acids of a C-terminal non-repetitive region. The peptide motifs found in spider flagelliform silk proteins, GPGGX\\u000a and GGX, were conserved

Kwang Sik Lee; Bo Yeon Kim; Yeon Ho Je; Soo Dong Woo; Hung Dae Sohn; Byung Rae Jin

2007-01-01

90

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

NASA Astrophysics Data System (ADS)

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.

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

2012-02-01

91

Identification and Characterization of Multiple Spidroin 1 Genes Encoding Major Ampullate Silk Proteins in Nephila clavipes  

PubMed Central

Spider dragline silk is primarily composed of proteins called major ampullate spidroins (MaSp) that consist of a large repeat array flanked by non-repetitive N- and C-terminal domains. Until recently, there has been little evidence for more than one gene encoding each of the two major spidroin silk proteins, MaSp1 and MaSp2. Here, we report the deduced N-terminal domain sequences for two distinct MaSp1 genes from Nephila clavipes (MaSp1A and MaSp1B) and for MaSp2. All three MaSp genes are co-expressed in the major ampullate gland. A search of the GenBank database also revealed two distinct MaSp1 C-terminal domain sequences. Sequencing confirmed that both MaSp1 genes are present in all seven Nephila clavipes spiders examined. The presence of nucleotide polymorphisms in these genes confirmed that MaSp1A and MaSp1B are distinct genetic loci and not merely alleles of the same gene. We have experimentally determined the transcription start sites for all three MaSp genes and established preliminary pairing between the two MaSp1 N- and C-terminal domains. Phylogenetic analysis of these new sequences and other published MaSp N- and C-terminal domain sequences illustrated that duplications of MaSp genes may be widespread among spider species. PMID:18828837

Gaines, William A.; Marcotte, William R.

2010-01-01

92

Structure-property relationship of regenerated spider silk protein nano/microfibrous scaffold fabricated by electrospinning.  

PubMed

The regenerated Araneus ventricosus spider dragline silk protein fibrous scaffold with moderate strength and flexibility was fabricated by electrospinning and post treatment with 90 vol % acetone. The effect of collection method on the morphology of regenerated spider silk protein (RSSP) fibrous scaffold, the effects of the post treatment solvents and their concentrations on the molecular conformation, crystallinity and mechanical properties were studied. The results show that the morphology was affected by the solvent used in the coagulation bath. The molecular conformation, crystallinity and mechanical property of this scaffold were strongly affected by the kind of post treatment solvent and slightly influenced by its concentration when it was higher than 50 vol %. The degradation rate of this scaffold was very slow and resulting in little pH change of the degradation medium within 5 months. PC 12 cells were cultured on the electrospun RSSP fibrous scaffold and in its extraction fluid to examine the changes of PC 12 cells after different times of culture. The results show that the electrospun RSSP fibrous scaffold had good biocompatibility with PC 12 cells. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 3828-3837, 2014. PMID:24288266

Yu, Qiaozhen; Xu, Shuiling; Zhang, Hong; Gu, Li; Xu, Yepei; Ko, Frank

2014-11-01

93

Biomimetic production of silk-like recombinant squid sucker ring teeth proteins.  

PubMed

The sucker ring teeth (SRT) of Humboldt squid exhibit mechanical properties that rival those of robust engineered synthetic polymers. Remarkably, these properties are achieved without a mineral phase or covalent cross-links. Instead, SRT are exclusively made of silk-like proteins called "suckerins", which assemble into nanoconfined ?-sheet reinforced supramolecular networks. In this study, three streamlined strategies for full-length recombinant suckerin protein production and purification were developed. Recombinant suckerin exhibited high solubility and colloidal stability in aqueous-based solvents. In addition, the colloidal suspensions exhibited a concentration-dependent conformational switch, from random coil to ?-sheet enriched structures. Our results demonstrate that recombinant suckerin can be produced in a facile manner in E. coli and processed from mild aqueous solutions into materials enriched in ?-sheets. We suggest that recombinant suckerin-based materials offer potential for a range of biomedical and engineering applications. PMID:25068184

Ding, Dawei; Guerette, Paul A; Hoon, Shawn; Kong, Kiat Whye; Cornvik, Tobias; Nilsson, Martina; Kumar, Akshita; Lescar, Julien; Miserez, Ali

2014-09-01

94

Silk-elastinlike protein polymer hydrogels for localized adenoviral gene therapy of head and neck tumors.  

PubMed

Vector dissemination, transient gene expression, and rapid clearance are major obstacles to successful human gene therapy. In this study, we investigated the effect of silk-elastinlike protein polymer (SELP) hydrogels on biodistribution and anticancer efficacy of adenoviral gene therapy in a head and neck cancer model. Transcriptional activities of adenovirus carrying beta-galactosidase (Ad-LacZ) and luciferase (Ad-Luc) reporter genes were evaluated in (nu/nu) mice with head and neck cancer as a function of polymer concentration. Antitumor efficacy of thymidine kinase encoding adenovirus (Ad-Tk) and ganciclovir (GSV) combination was also evaluated. SELP (4 wt %) matrices localized viral release, minimized dissemination to liver, and enhanced reporter gene expression levels by 4-8-fold compared to virus alone. SELP- Ad-Tk with GSV reduced tumor volume significantly compared to the virus alone. SELPs provide a means for temporal and spatial control of viral gene delivery to head and neck tumors. PMID:19722557

Greish, Khaled; Araki, Koji; Li, Daqing; O'Malley, Bert W; Dandu, Ramesh; Frandsen, Jordan; Cappello, Joseph; Ghandehari, Hamidreza

2009-08-10

95

Silk-Elastinlike Protein Polymer Hydrogels for Localized Adenoviral Gene Therapy of Head and Neck Tumors  

PubMed Central

Vector dissemination, transient gene expression and rapid clearance are major obstacles to successful human gene therapy. In this study, we investigated the effect of silk-elastinlike protein polymer (SELP) hydrogels, on biodistribution and anticancer efficacy of adenoviral gene therapy in a head and neck cancer model. Transcriptional activities of adenovirus carrying ?-galactosidase (Ad-LacZ) and luciferase (Ad-Luc) reporter genes were evaluated in (nu/nu) mice with head and neck cancer, as a function of polymer concentration. Antitumor efficacy of thymidine kinase encoding adenovirus (Ad-Tk) and ganciclovir (GSV) combination was also evaluated. 4 wt% SELP matrices localized viral release, minimized dissemination to liver and enhanced reporter gene expression levels by 4–8 fold compared to virus alone. SELP- Ad-Tk with GSV reduced tumor volume significantly compared to the virus alone. SELPs provide a means for temporal and spatial control of viral gene delivery to head and neck tumors. PMID:19722557

Greish, Khaled; Araki, Koji; Li, Daqing; O'Malley, Bert W.; Dandu, Ramesh; Frandsen, Jordan; Cappello, Joseph; Ghandehari, Hamidreza

2009-01-01

96

Spider Silk Fibers Spun from Soluble Recombinant Silk Produced in Mammalian Cells  

Microsoft Academic Search

Spider silks are protein-based ``biopolymer'' filaments or threads secreted by specialized epithelial cells as concentrated soluble precursors of highly repetitive primary sequences. Spider dragline silk is a flexible, lightweight fiber of extraordinary strength and toughness comparable to that of synthetic high-performance fibers. We sought to ``biomimic'' the process of spider silk production by expressing in mammalian cells the dragline silk

Anthoula Lazaris; Steven Arcidiacono; Yue Huang; Jiang-Feng Zhou; François Duguay; Nathalie Chretien; Elizabeth A. Welsh; Jason W. Soares; Costas N. Karatzas

2002-01-01

97

Genetically programmable thermoresponsive plasmonic gold/silk-elastin protein core/shell nanoparticles.  

PubMed

The design and development of future molecular photonic/electronic systems pose the challenge of integrating functional molecular building blocks in a controlled, tunable, and reproducible manner. The modular nature and fidelity of the biosynthesis method provides a unique chemistry approach to one-pot synthesis of environmental factor-responsive chimeric proteins capable of energy conversion between the desired forms. In this work, facile tuning of dynamic thermal response in plasmonic nanoparticles was facilitated by genetic engineering of the structure, size, and self-assembly of the shell silk-elastin-like protein polymers (SELPs). Recombinant DNA techniques were implemented to synthesize a new family of SELPs, S4E8Gs, with amino acid repeats of [(GVGVP)4(GGGVP)(GVGVP)3(GAGAGS)4] and tunable molecular weight. The temperature-reversible conformational switching between the hydrophilic random coils and the hydrophobic ?-turns in the elastin blocks were programmed to between 50 and 60 °C by site-specific glycine mutation, as confirmed by variable-temperature proton NMR and circular dichroism (CD) spectroscopy, to trigger the nanoparticle aggregation. The dynamic self-aggregation/disaggregation of the Au-SELPs nanoparticles was regulated in size and pattern by the ?-sheet-forming, thermally stable silk blocks, as revealed by transmission electron microscopy (TEM) and dynamic light scattering (DLS). The thermally reversible, shell dimension dependent, interparticle plasmon coupling was investigated by both variable-temperature UV-vis spectroscopy and finite-difference time-domain (FDTD)-based simulations. Good agreement between the calculated and measured spectra sheds light on design and synthesis of responsive plasmonic nanostructures by independently tuning the refractive index and size of the SELPs through genetic engineering. PMID:24712906

Lin, Yinan; Xia, Xiaoxia; Wang, Ming; Wang, Qianrui; An, Bo; Tao, Hu; Xu, Qiaobing; Omenetto, Fiorenzo; Kaplan, David L

2014-04-22

98

Silk inverse opals  

NASA Astrophysics Data System (ADS)

Periodic nanostructures provide the facility to control and manipulate the flow of light through their lattices. Three-dimensional photonic crystals enable the controlled design of structural colour, which can be varied by infiltrating the structure with different (typically liquid) fillers. Here, we report three-dimensional photonic crystals composed entirely of a purified natural protein (silk fibroin). The biocompatibility of this protein, as well as its favourable material properties and ease of biological functionalization, present opportunities for otherwise unattainable device applications such as bioresorbable integration of structural colour within living tissue or lattice functionalization by means of organic and inorganic material doping. We present a silk inverse opal that demonstrates a pseudo-photonic bandgap in the visible spectrum and show its associated structural colour beneath biological tissue. We also leverage silk's facile dopability to manufacture a gold nanoparticle silk inverse opal and demonstrate patterned heating mediated by enhancement of nanoparticle absorption at the band-edge frequency of the photonic crystal.

Kim, Sunghwan; Mitropoulos, Alexander N.; Spitzberg, Joshua D.; Tao, Hu; Kaplan, David L.; Omenetto, Fiorenzo G.

2012-12-01

99

Recombinant spider silk particles as drug delivery vehicles  

Microsoft Academic Search

Spider silk has been in the focus of research mainly due to the superior mechanical characteristics of silk fibers. However, it has been previously shown that spider silk proteins can also adopt other morphologies such as submicroparticles. This study examines the applicability of such particles as drug carriers. Particle characterization revealed that particles made of the engineered spider silk protein

Andreas Lammel; Martin Schwab; Markus Hofer; Gerhard Winter; Thomas Scheibel

2011-01-01

100

Hydrophobic drug-triggered self-assembly of nanoparticles from silk-elastin-like protein polymers for drug delivery.  

PubMed

Silk-elastin-like protein polymers (SELPs) combine the mechanical and biological properties of silk and elastin. These properties have led to the development of various SELP-based materials for drug delivery. However, SELPs have rarely been developed into nanoparticles, partially due to the complicated fabrication procedures, nor assessed for potential as an anticancer drug delivery system. We have recently constructed a series of SELPs (SE8Y, S2E8Y, and S4E8Y) with various ratios of silk to elastin blocks and described their capacity to form micellar-like nanoparticles upon thermal triggering. In this study, we demonstrate that doxorubicin, a hydrophobic antitumor drug, can efficiently trigger the self-assembly of SE8Y (SELPs with silk to elastin ratio of 1:8) into uniform micellar-like nanoparticles. The drug can be loaded in the SE8Y nanoparticles with an efficiency around 6.5% (65 ng doxorubicin/?g SE8Y), S2E8Y with 6%, and S4E8Y with 4%, respectively. In vitro studies with HeLa cell lines demonstrate that the protein polymers are not cytotoxic (IC50 > 200 ?g/mL), while the doxorubicin-loaded SE8Y nanoparticles showed a 1.8-fold higher cytotoxicity than the free drug. Confocal laser scanning microscopy (CLSM) and flow cytometry indicate significant uptake of the SE8Y nanoparticles by the cells and suggest internalization of the nanoparticles through endocytosis. This study provides an all-aqueous, facile method to prepare nanoscale, drug-loaded SELPs packages with potential for tumor cell treatments. PMID:24527851

Xia, Xiao-Xia; Wang, Ming; Lin, Yinan; Xu, Qiaobing; Kaplan, David L

2014-03-10

101

Silk from Crickets: A New Twist on Spinning  

Microsoft Academic Search

Raspy crickets (Orthoptera: Gryllacrididae) are unique among the orthopterans in producing silk, which is used to build shelters. This work studied the material composition and the fabrication of cricket silk for the first time. We examined silk-webs produced in captivity, which comprised cylindrical fibers and flat films. Spectra obtained from micro-Raman experiments indicated that the silk is composed of protein,

Andrew A. Walker; Sarah Weisman; Jeffrey S. Church; David J. Merritt; Stephen T. Mudie; Tara D. Sutherland

2012-01-01

102

Mechanical Improvements to Reinforced Porous Silk Scaffolds  

PubMed Central

Load bearing porous biodegradable scaffolds are required to engineer functional tissues such as bone. Mechanical improvements to porogen leached scaffolds prepared from silk proteins were systematically studied through the addition of silk particles in combination with silk solution concentration, exploiting interfacial compatibility between the two components. Solvent solutions of silk up to 32 w/v% were successfully prepared in hexafluoroisopropanaol (HFIP) for the study. The mechanical properties of the reinforced silk scaffolds correlated to the material density and matched by a power law relationship, independent of the ratio of silk particles to matrix. These results were similar to the relationships previously shown for cancellous bone. The mechanism behind the increased mechanical properties was a densification effect, and not the effect of including stiffer silk particles into the softer silk continuous matrix. A continuous interface between the silk matrix and the silk particles, as well as homogeneous distribution of the silk particles within the matrix were observed. Furthermore, we note that the roughness of the pore walls was controllable by varying the ratio of particles matrix, providing a route to control topography. The rate of proteolytic hydrolysis of the scaffolds decreased with increase in mass of silk used in the matrix and with increasing silk particle content. PMID:21793193

Gil, Eun Seok; Kluge, Jonathan A.; Rockwood, Danielle N.; Rajkhowa, Rangam; Wang, Lijing; Wang, Xungai; Kaplan, David L

2012-01-01

103

Molecular spring: from spider silk to silkworm silk  

E-print Network

In this letter, we adopt a new approach combining theoretical modeling with silk stretching measurements to explore the mystery of the structures between silkworm and spider silks, leading to the differences in mechanical response against stretching. Hereby the typical stress-strain profiles are reproduced by implementing the newly discovered and verified "$\\beta$-sheet splitting" mechanism, which primarily varies the secondary structure of protein macromolecules; our modeling and simulation results show good accordance with the experimental measurements. Hence, it can be concluded that the post-yielding mechanical behaviors of both kinds of silks are resulted from the splitting of crystallines while the high extensibility of spider dragline is attributed to the tiny $\\beta$-sheets solely existed in spider silk fibrils. This research reveals for the first time the structural factors leading to the significant difference between spider and silkworm silks in mechanical response to the stretching force. Addition...

Wu, Xiang; Du, Ning; Xu, Gang-Qin; Li, Bao-Wen

2009-01-01

104

Study of protein conformation and orientation in silkworm and spider silk fibers using Raman microspectroscopy.  

PubMed

Raman microspectroscopy has been used for the first time to determine quantitatively the orientation of the beta-sheets in silk monofilaments from Bombyx mori and Samia cynthia ricini silkworms, and from the spider Nephila edulis. It is shown that, for systems with uniaxial symmetry such as silk, it is possible to determine the order parameters P2 and P4 of the orientation distribution function from intensity ratios of polarized Raman spectra. The equations allowing the calculation of P2 and P4 using polarized Raman microspectroscopy for a vibration with a cylindrical Raman tensor were first derived and then applied to the amide I band that is mostly due to the C=O stretching vibration of the peptide groups. The shape of the Raman tensor for the amide I vibration of the beta-sheets was determined from an isotropic film of Bombyx mori silk treated with methanol. For both the Bombyx mori and Samia cynthia ricini fibroin fibers, the values of P2 and P4 obtained are equal to -0.36 +/- 0.03 and 0.19 +/- 0.02, respectively, even though the two types of silkworm fibroins strongly differ in their primary sequences. For the Nephila edulis dragline silk, values of P2 and P4 of -0.32 +/- 0.02 and 0.13 +/- 0.02 were obtained, respectively. These results clearly indicate that the carbonyl groups are highly oriented perpendicular to the fiber axis and that the beta-sheets are oriented parallel to the fiber axis, in agreement with previous X-ray and NMR results. The most probable distribution of orientation was also calculated from the values of P2 and P4 using the information entropy theory. For the three types of silk, the beta-sheets are highly oriented parallel to the fiber axis. The orientation distributions of the beta-sheets are nearly Gaussian functions with a width of 32 degrees and 40 degrees for the silkworm fibroins and the spider dragline silk, respectively. In addition to these results, the comparison of the Raman spectra recorded for the different silk samples and the polarization dependence of several bands has allowed to clarify some important band assignments. PMID:15530039

Rousseau, Marie-Eve; Lefèvre, Thierry; Beaulieu, Lilyane; Asakura, Tetsuo; Pézolet, Michel

2004-01-01

105

Enhanced function of pancreatic islets co-encapsulated with ECM proteins and mesenchymal stromal cells in a silk hydrogel  

PubMed Central

Pancreatic islet encapsulation within biosynthetic materials has had limited clinical success due to loss of islet function and cell death. As an alternative encapsulation material, a silk-based scaffold was developed to reestablish the islet microenvironment lost during cell isolation. Islets were encapsulated with ECM proteins (laminin and collagen IV) and mesenchymal stromal cells (MSCs), known to have immunomodulatory properties or to enhance islet cell graft survival and function. After a 7 day in vitro encapsulation, islets remained viable and maintained insulin secretion in response to glucose stimulation. Islets encapsulated with collagen IV, or laminin had increased insulin secretion at day 2 and day 7, respectively. A 3.2-fold synergistic improvement in islet insulin secretion was observed when islets were co-encapsulated with MSCs and ECM proteins. Furthermore, encapsulated islets had increased gene expression of functional genes; insulin I, insulin II, glucagon, somatostatin, and PDX-1, and lower expression of the de-differentiation genes cytokeratin 19 and vimentin compared to non-encapsulated cells. This work demonstrates that encapsulation in silk with both MSCs and ECM proteins enhances islet function and with further development may have potential as a suitable platform for islet delivery in vivo. PMID:22766242

Davis, Nicolynn E.; Beenken-Rothkopf, Liese N.; Mirsoian, Annie; Kojic, Nikola; Kaplan, David; Barron, Annelise E.; Fontaine, Magali J.

2012-01-01

106

Silk-I is a Hydrate  

NASA Astrophysics Data System (ADS)

In order to study the thermodynamics of supramolecular self-assembly phenomena in crystallizable segments of Bombyx mori silkworm silk fibroin, the osmotic stress method was applied. Precise control of the protein and LiBr concentration enables us to manipulate the phases of the silk fibroin, thus the method provided a means for the direct investigation of microscopic and thermodynamic details of these intermolecular interactions in aqueous media. Under the experimental osmotic pressure range of 0.2-7.6MPa applied by poly(ethylene glycol), it is apparent that as osmotic pressure increases, silk fibroin molecules are crowded together to form a water-soluble crystalline mesophase (silk-I), and then with further increase in osmotic pressure become anti-parallel beta-sheet structure (silk-II). A partial ternary phase diagram of water-silk fibroin-LiBr was constructed based on X-ray results. The microscopic data clearly indicate that silk-I is a hydrated structure since water of hydration is found to be lost in the conversion to silk-II. 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. The phase diagram suggests that processes, such as natural, regenerated or biomimetic silk fibroin spinning, that convert random coil fibroin to silk-II are likely to require a silk-I intermediate structure. This intermediate is necessary for the pre-alignment of the fibroin molecules and to shed water. We tried to utilize hydrated silk-I mesophase to pre-structure the silk fibroin by applying osmotic pressure, for the wet-spinning of silk fibroin.

Sohn, Sungkyun; Strey, Helmut H.; Gido, Samuel P.

2004-03-01

107

Silk Fibroin Electrogelation Mechanisms  

PubMed Central

A silk fibroin gel system (e-gel), formed with weak electric fields has potential utility in medical materials and devices. The mechanism of silk e-gel formation was studied to gain additional insight into the process and control of the material properties. Silk fibroin nanoparticles with sizes of several ten nanometers, composed of metastable conformations, were involved in e-gel formation. Under electric fields, the nanoparticles rapidly assembled into larger nano- or microspheres with size ranges from tens nanometers to several microns. Repulsive forces from the negative surface charge of the acidic groups on the protein were screened by the local decrease in solution pH in the vicinity of the positive electrode. By controlling the formation and content of silk fibroin nanoparticles e-gel could be formed even from low concentration silk fibroin solutions (1%). When e-gel was reversed to the solution state, the aggregated nano- and microspheres dispersed into solution, a significant observation related to future applications for this process, such as for drug delivery. PMID:21345387

Lu, Qiang; Huang, Yongli; Li, Mingzhong; Zuo, Baoqi; Lu, Shenzhou; Wang, Jiannan; Zhu, Hesun; Kaplan, David L.

2012-01-01

108

Snmp-1, a novel membrane protein of olfactory neurons of the silk moth Antheraea polyphemus with homology to the CD36 family of membrane proteins.  

PubMed

While olfactory neurons of silk moths are well known for their exquisite sensitivity to sex pheromone odorants, molecular mechanisms underlying this sensitivity are poorly understood. In searching for proteins that might support olfactory mechanisms, we characterized the protein profile of olfactory neuron receptor membranes of the wild silk moth Antheraea polyphemus. We have purified and cloned a prominent 67-kDa protein which we have named Snmp-1 (sensory neuron membrane protein-1). Northern blot analysis suggests that Snmp-1 is uniquely expressed in antennal tissue; in situ hybridization and immunocytochemical analyses show that Snmp-1 is expressed in olfactory neurons and that the protein is localized to the cilia, dendrites, and somata but not the axons. Snmp-1 mRNA expression increases significantly 1-2 days before the end of adult development, coincident with the functional maturation of the olfactory system. Sequence analysis suggests Snmp-1 is homologous with the CD36 protein family, a phylogenetically diverse family of receptor-like membrane proteins. CD36 family proteins are characterized as having two transmembrane domains and interacting with proteinaceous ligands; Snmp-1 is the first member of this family identified in nervous tissue. These findings argue that Snmp-1 has an important role in olfaction; possible roles of Snmp-1 in odorant detection are discussed. PMID:9169446

Rogers, M E; Sun, M; Lerner, M R; Vogt, R G

1997-06-01

109

Silk-elastinlike protein polymers improve the efficacy of adenovirus thymidine kinase enzyme prodrug therapy of head and neck tumors  

PubMed Central

Background Adenoviral directed enzyme prodrug therapy is a promising approach for head and neck cancer gene therapy. Challenges with this approach however are transient gene expression and dissemination of viruses to distant organs. Methods We used recombinant silk-elastinlike protein copolymer (SELP) matrices for intratumoral delivery of adenoviruses containing both thymidine kinase-1, and luciferase genes in a nude mice model of JHU-022 head and neck tumor. Hydrogels made from two SELP analogues (47K and 815K) with similar silk to elastinlike block ratios but different block lengths were studied for intratumoral viral delivery. Tumor bearing mice were followed up for tumor progression and luciferase gene expression concomitantly for five weeks. Polymer’s safety was evaluated through body weight change, blood count, liver and kidney functions in addition to gross and microscopic histological examination. Results SELP 815K analogues efficiently controlled the duration and extent of transfection in tumors for up to 5 weeks with no detectable spread to the liver. About five-fold greater reduction in tumor volume was obtained with matrix-mediated delivery compared to intra-tumoral injection of adenoviruses in saline. SELP matrix proved safe in all injected mice compared to control group. Conclusion SELP- controlled gene delivery approach could potentially improve the anticancer activity of virus-mediated gene therapy while limiting viral spread to normal organs. PMID:20603862

Greish, Khaled; Frandsen, Jordan; Scharff, Stephanie; Gustafson, Joshua; Cappello, Joseph; Li, Daqing; O'Malley, Bert W.; Ghandehari, Hamidreza

2010-01-01

110

Silk: Caterpillars  

NSDL National Science Digital Library

If you listen closely, you can hear the sounds of a critical element in the production of silk--silkworm caterpillars dining on mulberry leaves. This episode of the two-minute radio show Pulse of the Planet focuses on how silkworms are raised. In the episode, available here in MP3 audio and text formats, a guest biochemist describes the silk farming process, noting the silkworms' dependence on large quantities of human-supplied mulberry leaves. The host points out how much silkworms grow in length and mass as evidence for why so many leaves are needed. Copyright 2005 Eisenhower National Clearinghouse

Planet, Pulse O.

2005-11-23

111

Invited review nonmulberry silk biopolymers.  

PubMed

The silk produced by silkworms are biopolymers and can be classified into two types--mulberry and nonmulberry. Mulberry silk of silkworm Bombyx mori has been extensively explored and used for century old textiles and sutures. But for the last few decades it is being extensively exploited for biomedical applications. However, the transformation of nonmulberry silk from being a textile commodity to biomaterials is relatively new. Within a very short period of time, the combination of load bearing capability and tensile strength of nonmulberry silk has been equally envisioned for bone, cartilage, adipose, and other tissue regeneration. Adding to its advantage is its diverse morphology, including macro to nano architectures with controllable degradation and biocompatibility yields novel natural material systems in vitro. Its follow on applications involve sustained release of model compounds and anticancer drugs. Its 3D cancer models provide compatible microenvironment systems for better understanding of the cancer progression mechanism and screening of anticancer compounds. Diversely designed nonmulberry matrices thus provide an array of new cutting age technologies, which is unattainable with the current synthetic materials that lack biodegradability and biocompatibility. Scientific exploration of nonmulberry silk in tissue engineering, regenerative medicine, and biotechnological applications promises advancement of sericulture industries in India and China, largest nonmulberry silk producers of the world. This review discusses the prospective biomedical applications of nonmulberry silk proteins as natural biomaterials. PMID:22241173

Kundu, S C; Kundu, Banani; Talukdar, Sarmistha; Bano, Subia; Nayak, Sunita; Kundu, Joydip; Mandal, Biman B; Bhardwaj, Nandana; Botlagunta, Mahendran; Dash, Biraja C; Acharya, Chitrangada; Ghosh, Ananta K

2012-06-01

112

Silk from Crickets: A New Twist on Spinning  

PubMed Central

Raspy crickets (Orthoptera: Gryllacrididae) are unique among the orthopterans in producing silk, which is used to build shelters. This work studied the material composition and the fabrication of cricket silk for the first time. We examined silk-webs produced in captivity, which comprised cylindrical fibers and flat films. Spectra obtained from micro-Raman experiments indicated that the silk is composed of protein, primarily in a beta-sheet conformation, and that fibers and films are almost identical in terms of amino acid composition and secondary structure. The primary sequences of four silk proteins were identified through a mass spectrometry/cDNA library approach. The most abundant silk protein was large in size (300 and 220 kDa variants), rich in alanine, glycine and serine, and contained repetitive sequence motifs; these are features which are shared with several known beta-sheet forming silk proteins. Convergent evolution at the molecular level contrasts with development by crickets of a novel mechanism for silk fabrication. After secretion of cricket silk proteins by the labial glands they are fabricated into mature silk by the labium-hypopharynx, which is modified to allow the controlled formation of either fibers or films. Protein folding into beta-sheet structure during silk fabrication is not driven by shear forces, as is reported for other silks. PMID:22355311

Walker, Andrew A.; Weisman, Sarah; Church, Jeffrey S.; Merritt, David J.; Mudie, Stephen T.; Sutherland, Tara D.

2012-01-01

113

Antheraea pernyi Silk Fiber: A Potential Resource for Artificially Biospinning Spider Dragline Silk  

PubMed Central

The outstanding properties of spider dragline silk are likely to be determined by a combination of the primary sequences and the secondary structure of the silk proteins. Antheraea pernyi silk has more similar sequences to spider dragline silk than the silk from its domestic counterpart, Bombyx mori. This makes it much potential as a resource for biospinning spider dragline silk. This paper further verified its possibility as the resource from the mechanical properties and the structures of the A. pernyi silks prepared by forcible reeling. It is surprising that the stress-strain curves of the A. pernyi fibers show similar sigmoidal shape to those of spider dragline silk. Under a controlled reeling speed of 95?mm/s, the breaking energy was 1.04 × 105?J/kg, the tensile strength was 639 MPa and the initial modulus was 9.9 GPa. It should be noted that this breaking energy of the A. pernyi silk approaches that of spider dragline silk. The tensile properties, the optical orientation and the ?-sheet structure contents of the silk fibers are remarkably increased by raising the spinning speeds up to 95?mm/s. PMID:20454537

Zhang, Yaopeng; Yang, Hongxia; Shao, Huili; Hu, Xuechao

2010-01-01

114

Mechanism of silk processing in insects and spiders  

Microsoft Academic Search

Silk spinning by insects and spiders leads to the formation of fibres that exhibit high strength and toughness. The lack of understanding of the protein processing in silk glands has prevented the recapitulation of these properties in vitro from reconstituted or genetically engineered silks. Here we report the identification of emulsion formation and micellar structures from aqueous solutions of reconstituted

Hyoung-Joon Jin; David L. Kaplan

2003-01-01

115

Post-secretion processing influences spider silk performance  

PubMed Central

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 ?-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

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

2012-01-01

116

Mechanical Response of Silk Crystalline Units from Force-Distribution Analysis  

E-print Network

crystals, opening up the road to predict full fiber mechanics. INTRODUCTION Silk proteins build upMechanical Response of Silk Crystalline Units from Force-Distribution Analysis Senbo Xiao, Wolfram of silk fibers is thought to be caused by embedded crystalline units acting as cross links of silk

Gräter, Frauke

117

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

PubMed Central

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. PMID:23824061

Cao, Ting-Ting; Wang, Yuan-Jing; Zhang, Yu-Qing

2013-01-01

118

Mixed protein blends composed of gelatin and Bombyx mori silk fibroin: effects of solvent-induced crystallization and composition.  

PubMed

Novel protein blends have been prepared by mixing gelatin (G) with Bombyx mori silk fibroin (SF) and using aqueous methanol (MeOH) to post-induce SF crystallization. When co-cast from solution, amorphous blends of these polymers appear homogeneous, as discerned from visual observation, microscopy, and Fourier-transform infrared (FTIR) spectroscopy. Upon subsequent exposure to aqueous MeOH, SF undergoes a conformational change from random coil to beta sheet. This transformation occurs in pure SF, as well as in each of the G/SF blends, according to X-ray diffractometry and thermal calorimetry. The influence of MeOH-induced SF crystallization on structure and property development has been ascertained in terms of preparation history and blend composition. Thermal gravimetric analysis reveals that the presence of beta sheets in SF and G/SF blends improves thermal stability, while extensional rheometry confirms that SF crystallization enhances the tensile properties of the blends. By preserving a support scaffold above the G helix-to-coil transition temperature, the formation of crystalline SF networks in G/SF blends can be used to stabilize G-based hydrogels for biomaterial and pharmaceutical purposes. The present study not only examines the properties of G/SF blends before and after SF crystallization, but also establishes the foundation for future research into thermally responsive G/SF bioconjugates. PMID:16529407

Gil, Eun S; Frankowski, David J; Bowman, Michelle K; Gozen, Arif O; Hudson, Samuel M; Spontak, Richard J

2006-03-01

119

Silk film biomaterials for ocular surface repair  

NASA Astrophysics Data System (ADS)

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

Lawrence, Brian David

120

Design, expression and solid-state NMR characterization of silk-like materials constructed from sequences of spider silk, Samia cynthia ricini and Bombyx mori silk fibroins.  

PubMed

Silk has a long history of use in medicine as sutures. To address the requirements of a mechanically robust and biocompatible material, basic research to clarify the role of repeated sequences in silk fibroin in its structures and properties seems important as well as the development of a processing technique suitable for the preparation of fibers with excellent mechanical properties. In this study, three silk-like protein analogs were constructed from two regions selected from among the crystalline region of Bombyx mori silk fibroin, (GAGSGA)(2), the crystalline region of Samia cynthia ricini silk fibroin, (Ala)(12), the crystalline region of spider dragline silk fibroin, (Ala)(6), and the Gly-rich region of spider silk fibroin, (GGA)(4). The silk-like protein analog constructed from the crystalline regions of the spider dragline silk and B. mori silk fibroins, (A(6)SCS)(8), that constructed from the crystalline regions of the S. c.ricini and B. mori silk fibroins, (A(12)SGS)(4), that constructed from and the crystalline region of S. c.ricini silk fibroin and the glycine-rich region of spider dragline silk fibroin, (A(12)SGS)(4),were expressed their molecular weights being about 36.0 kDa, 17.0 kDa and 17.5 kDa, respectively in E. coli by means of genetic engineering technologies. (A(12)SCS)(4) and (A(12)SGS)(4 )undergo a structural transition from alpha-helix to beta-sheet on a change in the solvent treatment from trifluoroacetic acid (TFA) to formic acid (FA). However, (A(6)SCS)(8) takes on the beta-sheet structure predominantly on TFA treatment and FA treatment. Structural analysis was performed on model peptides selected from spider dragline and S. c.ricini silks by means of (13)C CP/MAS NMR. PMID:16002994

Yang, Mingying; Asakura, Tetsuo

2005-06-01

121

Functionalized Silk Biomaterials for Wound Healing  

PubMed Central

Silk protein-biomaterial wound dressings with epidermal growth factor (EGF) and silver sulfadiazine were studied with a cutaneous excisional mouse wound model. Three different material designs (silk films, lamellar porous silk films, electrospun silk nanofibers) and two different drug functionalization techniques (drug coatings or drug loading into the materials) were studied to compare wound healing responses. Changes in wound size and histological assessments of wound tissues over time confirmed that functionalized silk biomaterial wound dressings increased wound healing rate, including reepithelialization, dermis proliferation, collagen synthesis, epidermal differentiation into hair follicles and sebaceous glands, and reduced scar formation, when compared to air-permeable Tegaderm™ tape (3M) (? control) and a commercially sold wound dressing (Tegaderm™ Hydrocolloid dressing) (+ control). All silk biomaterials studied were effective for wound healing, while the porous features of the silk biomaterials (lamellar porous films and electrospun nanofibers) and the incorporation of EGF/silver sulfadiazine, via drug loading or coating, provided the most rapid wound healing responses. This systematic approach to evaluate functionalized silk biomaterial wound dressings demonstrates a useful strategy to select formulations for further study towards new treatment options for chronic wounds. PMID:23184644

Gil, Eun Seok; Panilaitis, Bruce; Bellas, Evangelia

2013-01-01

122

Therapeutic effects of sericin on diabetic keratopathy in Otsuka Long-Evans Tokushima Fatty rats.  

PubMed

An Otsuka Long-Evans Tokushima Fatty (OLETF) rat provides a useful model for studies to develop corneal wound healing drugs for use in diabetic keratopathy resulting from type 2 diabetes mellitus. We investigated the effects of sericin on corneal wound healing in OLETF rats. Corneal wounds were prepared by removal of the corneal epithelium and documented using a TRC-50X. Sericin was instilled into the eyes of rats five times a day following corneal abrasion. The plasma levels of glucose, triglycerides, cholesterol and insulin in 38 wk old OLETF rats were significantly higher than in normal control rats (LETO rats), and the rate of corneal wound healing in OLETF rats was slower than in normal rat, probably due to the suppression of cell migration and proliferation caused by high plasma glucose levels. The corneal wounds of OLETF rats instilled with saline showed almost complete healing 72 h after corneal epithelial abrasion. On the other hand, the instillation of sericin has a potent effect in promoting wound healing and wound size reduction in OLETF rats and the wounds showed almost complete healing at 48 h after abrasion. The sericin may be an effective and safe drug to promote corneal wound healing in diabetic keratopathy. PMID:24379918

Nagai, Noriaki; Ito, Yoshimasa

2013-12-15

123

Continuous production of flexible fibers from transgenically produced honeybee silk proteins.  

PubMed

Flexible and solvent stable fibers are produced after concentrated recombinant honeybee protein solutions are extruded into a methanol bath, dried, drawn in aqueous methanol, then covalently cross-linked using dry heat. Proteins in solution are predominantly coiled coil. Significant levels of non-orientated ß-sheets form during drying or after coagulation in aqueous methanol. Drawing generally aligns the coiled coil component parallel with the fibre axis and ß-sheet component perpendicular to the fiber axis. The fibres are readily handled, stable in the strong protein denaturants, urea and guanidinium, and suitable for a range of applications such as weaving and knitting. PMID:23881528

Poole, Jacinta; Church, Jeffrey S; Woodhead, Andrea L; Huson, Mickey G; Sriskantha, Alagacone; Kyratzis, Ilias L; Sutherland, Tara D

2013-10-01

124

In vivo biological responses to silk proteins functionalized with bone sialoprotein.  

PubMed

Recombinant 6mer?+?BSP protein, combining six repeats of the consensus sequence for Nephila clavipes dragline (6mer) and bone sialoprotein sequence (BSP), shows good support for cell viability and induces the nucleation of hydroxyapatite and tricalcium phosphate during osteoblast in vitro culture. The present study is conducted to characterize this bioengineered protein-based biomaterial further for in vivo behavior related to biocompatibility. 6mer?+?BSP protein films are implanted in subcutaneous pouches in the back of mice and responses are evaluated by flow cytometry and histology. The results show no major differences between the inflammatory responses induced by 6mer?+?BSP films and the responses observed for the controls. Thus, this new chimeric protein could represent an alternative for bone regeneration applications. PMID:23359587

Gomes, Sílvia; Gallego-Llamas, Jabier; Leonor, Isabel B; Mano, João F; Reis, Rui L; Kaplan, David L

2013-04-01

125

Brown recluse spider's nanometer scale ribbons of stiff extensible silk.  

PubMed

The silk of the recluse spider features a ribbon-like morphology unlike any other spider silk or synthetically spun polymer fiber. These protein ribbons represent free-standing polymer films with a thickness of about 50 nm. Stress-strain characterization of individual fibers via atomic force microscopy reveals that these ribbons, only a few molecular layers of protein thin, rival the mechanical performance of the best silks. PMID:24352987

Schniepp, Hannes C; Koebley, Sean R; Vollrath, Fritz

2013-12-23

126

Designing Spider Silk Genes for Materials Applications.  

National Technical Information Service (NTIS)

This project is designed to create new proteins based on naturally occurring spider silk proteins with the goal of controlling elasticity and tensile strength in fibers spun from the proteins. The new genes have been constructed in year 1, the proteins ha...

R. V. Lewis

2006-01-01

127

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

PubMed Central

Silk fibroin protein-based micro- and nanospheres provide new options for drug delivery due to their biocompatibility, biodegradability and their tunable drug loading and release properties. In the present study, we report a new aqueous-based preparation method for silk spheres with controllable sphere size and shape. The preparation was based on phase separation between silk fibroin and polyvinyl alcohol (PVA) at a weight ratio of 1/1 and 1/4. Water-insoluble silk spheres were easily obtained from the blend in a three step process: (1) air-drying the blend solution into a film, (2) film dissolution in water and (3) removal of residual PVA by subsequent centrifugation. In both cases, the spheres had approximately 30% beta-sheet content and less than 5% residual PVA. Spindle-shaped silk particles, as opposed to the spherical particles formed above, were obtained by stretching the blend films before dissolving in water. Compared to the 1/1 ratio sample, the silk spheres prepared from the 1/4 ratio sample showed a more homogeneous size distribution ranging from 300 nm up to 20 ?m. Further studies showed that sphere size and polydispersity could be controlled either by changing the concentration of silk and PVA or by applying ultrasonication on the blend solution. Drug loading was achieved by mixing model drugs in the original silk solution. The distribution and loading efficiency of the drug molecules in silk spheres depended on their hydrophobicity and charge, resulting in different drug release profiles. The entire fabrication procedure could be completed within one day. The only chemical used in the preparation except water was PVA, an FDA-approved ingredient in drug formulations. Silk micro- and nanospheres reported have potential as drug delivery carriers in a variety of biomedical applications. PMID:19945157

Wang, Xiaoqin; Yucel, Tuna; Lu, Qiang; Hu, Xiao; Kaplan, David L.

2009-01-01

128

Mechanism of silk processing in insects and spiders  

NASA Astrophysics Data System (ADS)

Silk spinning by insects and spiders leads to the formation of fibres that exhibit high strength and toughness. The lack of understanding of the protein processing in silk glands has prevented the recapitulation of these properties in vitro from reconstituted or genetically engineered silks. Here we report the identification of emulsion formation and micellar structures from aqueous solutions of reconstituted silkworm silk fibroin as a first step in the process to control water and protein-protein interactions. The sizes (100-200nm diameter) of these structures could be predicted from hydrophobicity plots of silk protein primary sequence. These micelles subsequently aggregated into larger `globules' and gel-like states as the concentration of silk fibroin increased, while maintaining solubility owing to the hydrophilic regions of the protein interspersed among the larger hydrophobic regions. Upon physical shearing or stretching structural transitions, increased birefringence and morphological alignment were demonstrated, indicating that this process mimics the behaviour of similar native silk proteins in vivo. Final morphological features of these silk materials are similar to those observed in native silkworm fibres.

Jin, Hyoung-Joon; Kaplan, David L.

2003-08-01

129

Silk-based delivery systems of bioactive molecules  

PubMed Central

Silks are biodegradable, biocompatible, self-assemblying proteins that can also be tailored via genetic engineering to contain specific chemical features, offering utility for drug and gene delivery. Silkworm silk has been used in biomedical sutures for decades and has recently achieved Food and Drug Administration approval for expanded biomaterials device utility. With the diversity and control of size, structure and chemistry, modified or recombinant silk proteins can be designed and utilized in various biomedical application, such as for the delivery of bioactive molecules. This review focuses on the biosynthesis and applications of silk-based multi-block copolymer systems and related silk protein drug delivery systems. The utility of these systems for the delivery of small molecule drugs, proteins and genes are reviewed. PMID:20298729

Numata, Keiji; Kaplan, David L

2010-01-01

130

Electrospinning of chitosan/sericin/PVA nanofibers incorporated with in situ synthesis of nano silver.  

PubMed

Here, chitosan/sericin/poly(vinyl alcohol) as a biodegradable nanofibrous membrane was prepared through electrospinning with and without silver nitrate. The influences of spinning conditions including volume ratio of chitosan and sericin, voltage and spinning distance at constant feed rate on the fiber morphology and size distribution were examined by SEM and Image J software. The FT-IR spectrum and EDAX were used to indicate the chemical structure of nanofibrous membrane. In addition, the effect of AgNO3 on the nanofibers diameter and its antibacterial activity was investigated. The optimum conditions obtained with chitosan:sericin (50:50, v/v), 22kV voltage, 10cm spinning distance at 0.25mL/h feed rate to prepare nanofibers with small diameter and narrow size distribution without beads. The mean diameter of nanofibers was about 180nm while introducing AgNO3 led to smaller nanofibers diameter about 95nm. Moreover, the presence of AgNO3 produced an excellent antibacterial activity against Escherchia coli. PMID:25256480

Hadipour-Goudarzi, Elmira; Montazer, Majid; Latifi, Masoud; Aghaji, Ali Akbar Ghare

2014-11-26

131

Water-Insoluble Silk Films with Silk I Structure  

PubMed Central

Water-insoluble regenerated silk materials are normally achieved by increasing ?-sheet content (silk II). In the present study, water-insoluble silk films were prepared by controlling very slow drying of B. mori silk solutions, resulting in the formation of stable films with dominating silk I instead of silk II structure. Wide angle x-ray scattering (WAXS) indicated that the silk films stabilized by slow drying were mainly composed of silk I rather than silk II, while water- and methanol-annealed silk films had a higher silk II content. The silk films prepared through slow drying had a globule-like structure in the core with nano-filaments. The core region was composed of silk I and silk II, and these regions are surrounded by hydrophilic nano-filaments containing random, turns, and ?-helix secondary structures. The insoluble silk films prepared by slow drying had unique thermal, mechanical and degradative properties. DSC results revealed that silk I crystals had stable thermal properties up to 250°C, without crystallization above the Tg, but degraded in lower temperature than silk II structure. Compared with water- and methanol-annealed films, the films prepared through slow drying achieved better mechanical ductility and more rapid enzymatic degradation, reflective of the differences in secondary structure achieved via differences in post processing of the cast silk films. Importantly, the silk I structure, a key intermediate secondary structure for the formation of mechanically robust natural silk fibers, was successfully generated in the present approach of very slow drying, mimicking the natural process. The results also point to a new mode to generate new types of silk biomaterials, where mechanical properties can be enhanced, and degradation rates increased, yet water insolubility is maintained along with low beta sheet content. PMID:19874919

Lu, Qiang; Hu, Xiao; Wang, Xiaoqin; Kluge, Jonathan A.; Lu, Shenzhou; Cebe, Peggy; Kaplan, David L.

2009-01-01

132

Water-insoluble Silk Films with Silk I Structure  

SciTech Connect

Water-insoluble regenerated silk materials are normally produced by increasing the {beta}-sheet content (silk II). In the present study water-insoluble silk films were prepared by controlling the very slow drying of Bombyx mori silk solutions, resulting in the formation of stable films with a predominant silk I instead of silk II structure. Wide angle X-ray scattering indicated that the silk films stabilized by slow drying were mainly composed of silk I rather than silk II, while water- and methanol-annealed silk films had a higher silk II content. The silk films prepared by slow drying had a globule-like structure at the core surrounded by nano-filaments. The core region was composed of silk I and silk II, surrounded by hydrophilic nano-filaments containing random turns and {alpha}-helix secondary structures. The insoluble silk films prepared by slow drying had unique thermal, mechanical and degradative properties. Differential scanning calorimetry results revealed that silk I crystals had stable thermal properties up to 250 C, without crystallization above the T{sub g}, but degraded at lower temperatures than silk II structure. Compared with water- and methanol-annealed films the films prepared by slow drying had better mechanical ductility and were more rapidly enzymatically degraded, reflecting the differences in secondary structure achieved via differences in post processing of the cast silk films. Importantly, the silk I structure, a key intermediate secondary structure for the formation of mechanically robust natural silk fibers, was successfully generated by the present approach of very slow drying, mimicking the natural process. The results also point to a new mode of generating new types of silk biomaterials with enhanced mechanical properties and increased degradation rates, while maintaining water insolubility, along with a low {beta}-sheet content.

Lu, Q.; Hu, X; Wang, X; Kluge, J; Lu, S; Cebe, P; Kaplan, D

2010-01-01

133

Sequential origin in the high performance properties of orb spider dragline silk  

PubMed Central

Major ampullate (MA) dragline silk supports spider orb webs, combining strength and extensibility in the toughest biomaterial. MA silk evolved ~376?MYA and identifying how evolutionary changes in proteins influenced silk mechanics is crucial for biomimetics, but is hindered by high spinning plasticity. We use supercontraction to remove that variation and characterize MA silk across the spider phylogeny. We show that mechanical performance is conserved within, but divergent among, major lineages, evolving in correlation with discrete changes in proteins. Early MA silk tensile strength improved rapidly with the origin of GGX amino acid motifs and increased repetitiveness. Tensile strength then maximized in basal entelegyne spiders, ~230?MYA. Toughness subsequently improved through increased extensibility within orb spiders, coupled with the origin of a novel protein (MaSp2). Key changes in MA silk proteins therefore correlate with the sequential evolution high performance orb spider silk and could aid design of biomimetic fibers. PMID:23110251

Blackledge, Todd A.; Perez-Rigueiro, Jose; Plaza, Gustavo R.; Perea, Belen; Navarro, Andres; Guinea, Gustavo V.; Elices, Manuel

2012-01-01

134

Sequential origin in the high performance properties of orb spider dragline silk.  

PubMed

Major ampullate (MA) dragline silk supports spider orb webs, combining strength and extensibility in the toughest biomaterial. MA silk evolved ~376?MYA and identifying how evolutionary changes in proteins influenced silk mechanics is crucial for biomimetics, but is hindered by high spinning plasticity. We use supercontraction to remove that variation and characterize MA silk across the spider phylogeny. We show that mechanical performance is conserved within, but divergent among, major lineages, evolving in correlation with discrete changes in proteins. Early MA silk tensile strength improved rapidly with the origin of GGX amino acid motifs and increased repetitiveness. Tensile strength then maximized in basal entelegyne spiders, ~230?MYA. Toughness subsequently improved through increased extensibility within orb spiders, coupled with the origin of a novel protein (MaSp2). Key changes in MA silk proteins therefore correlate with the sequential evolution high performance orb spider silk and could aid design of biomimetic fibers. PMID:23110251

Blackledge, Todd A; Pérez-Rigueiro, José; Plaza, Gustavo R; Perea, Belén; Navarro, Andrés; Guinea, Gustavo V; Elices, Manuel

2012-01-01

135

Early Events in the Evolution of Spider Silk Genes  

PubMed Central

Silk spinning is essential to spider ecology and has had a key role in the expansive diversification of spiders. Silk is composed primarily of proteins called spidroins, which are encoded by a multi-gene family. Spidroins have been studied extensively in the derived clade, Orbiculariae (orb-weavers), from the suborder Araneomorphae (‘true spiders’). Orbicularians produce a suite of different silks, and underlying this repertoire is a history of duplication and spidroin gene divergence. A second class of silk proteins, Egg Case Proteins (ECPs), is known only from the orbicularian species, Lactrodectus hesperus (Western black widow). In L. hesperus, ECPs bond with tubuliform spidroins to form egg case silk fibers. Because most of the phylogenetic diversity of spiders has not been sampled for their silk genes, there is limited understanding of spidroin gene family history and the prevalence of ECPs. Silk genes have not been reported from the suborder Mesothelae (segmented spiders), which diverged from all other spiders >380 million years ago, and sampling from Mygalomorphae (tarantulas, trapdoor spiders) and basal araneomorph lineages is sparse. In comparison to orbicularians, mesotheles and mygalomorphs have a simpler silk biology and thus are hypothesized to have less diversity of silk genes. Here, we present cDNAs synthesized from the silk glands of six mygalomorph species, a mesothele, and a non-orbicularian araneomorph, and uncover a surprisingly rich silk gene diversity. In particular, we find ECP homologs in the mesothele, suggesting that ECPs were present in the common ancestor of extant spiders, and originally were not specialized to complex with tubuliform spidroins. Furthermore, gene-tree/species-tree reconciliation analysis reveals that numerous spidroin gene duplications occurred after the split between Mesothelae and Opisthothelae (Mygalomorphae plus Araneomorphae). We use the spidroin gene tree to reconstruct the evolution of amino acid compositions of spidroins that perform different ecological functions. PMID:22761664

Starrett, James; Garb, Jessica E.; Kuelbs, Amanda; Azubuike, Ugochi O.; Hayashi, Cheryl Y.

2012-01-01

136

Safety Evaluation of Silk Protein Film (A Novel Wound Healing Agent) in Terms of Acute Dermal Toxicity, Acute Dermal Irritation and Skin Sensitization  

PubMed Central

Acute dermal toxicity study was conducted in rats. The parameters studied were body weight, serum biochemistry and gross pathology. The animals were also observed for clinical signs and mortality after the application of test film. The dermal irritation potential of silk protein film was examined using Draize test. In the initial test, three test patches were applied sequentially for 3 min, 1 and 4 hours, respectively, and skin reaction was graded. The irritant or negative response was confirmed using two additional animals, each with one patch, for an exposure period of 4 hours. The responses were scored at 1, 24, 48 and 72 hours after the patch removal. Skin sensitization study was conducted according to Buehler test in guinea pigs, in which on day 0, 7 and 14, the animals were exposed to test material for 6 hours (Induction phase) and on day 28, the animals were exposed for a period of 24 hours (Challenge phase). The skin was observed and recorded at 24 and 48 hours after the patch removal. In acute dermal toxicity study, the rats dermally treated with silk film did not show any abnormal clinical signs and the body weight, biochemical parameters and gross pathological observations were not significantly different from the control group. In acute dermal irritation study, the treated rabbits showed no signs of erythema, edema and eschar, and the scoring was given as “0” for all time points of observations according to Draize scoring system. In skin sensitization study, there were no skin reactions 24 and 48 hours after the removal of challenge patch, which was scored “0” based on Magnusson/Kligman grading scale. PMID:21430915

Padol, Amol R.; Jayakumar, K.; Shridhar, N. B.; Narayana Swamy, H. D.; Narayana Swamy, M.; Mohan, K.

2011-01-01

137

An experimental confirmation of thermal transitions in native and regenerated spider silks.  

PubMed

Biological structures such as spider silks are formed by proteins. The physical properties of such proteins are determined by environmental conditions such as temperature and humidity. In this paper, we confirm the thermal transitions that take place in spider silks using differential scanning calorimetry and study how the interaction of spider silk proteins with water affects the onset temperatures for these thermal processes. Native fibres and regenerated films of dragline silk and egg sac silk from Argiope argentata spiders were used to study thermal transitions of protein based structures. For the first time, differential scanning calorimetry (DSC) tests were carried out with spider silk samples of relatively large mass (10mg). Previous attempts of DSC tests applied to spider silk samples failed to detect thermal transitions in a conclusive way. The tests reported here, however, show thermal transitions on both natural and regenerated samples that are in agreement with results from dynamic mechanical analysis (DMA) tests reported in the literature. The water content on spider silks seems to lower the temperatures at which such thermal transitions take place. The results also confirm that the amorphous regions of native and regenerated spider silk and silk worm silk give rise to similar thermal transitions. PMID:23827592

Torres, Fernando G; Troncoso, Omar P; Torres, Carlos; Cabrejos, Wilson

2013-04-01

138

Silk material modification and microfabrication strategies for applications in biosensors and drug delivery  

NASA Astrophysics Data System (ADS)

Silk has proven to be a promising biomaterial for the development of a novel generation of biomedical devices due to the material's intrinsic properties of biocompatibility and biodegradability. To further the development of silk-based devices, augmented functionality can be provided to silk by means of microfabrication and material functionalization. In this dissertation, we set out to explore possibilities of silk-based biomedical device development with particular attention to different fabrication strategies that can be leveraged for this purpose, taking inspiration from conventional lithography, contact imprinting and chemical modification of the silk biomaterial. In particular, we have produced a novel silk-based drug delivery device, in the form of microneedles. For this purpose we have developed micromolding strategies which allow the fabrication of high aspect ratio silk structures. Furthermore, we have produced a THz split ring resonator based sensor device on silk films. To realize this device, we have developed a convenient fabrication method, allowing transfer of previously microfabricated metal and oxide microstructures to the silk film surface. This method has proven useful for hard masking and patterning silk films with reactive ion etching. Moreover, we have explored patterning semiconductor and glass substrates with silk films. For this purpose we have modified a standard microfabrication method -lift off- to be amendable to silk. Furthermore, combining silk based fabrication and material functionalization has lead to microfluidics pH sensing applications. Finally we have explored the possibility of utilizing silk for injectable and biodegradable glucose sensors. For this purpose, we have doped silk hydrogels with a fluorescence based protein biosensor. The advanced silk fabrication and material synthesis strategies and the resulting novel devices presented here could potentially lead to a new class of biomedical applications.

Tsioris, Konstantinos

139

Unraveled mechanism in silk engineering: Fast reeling induced silk toughening  

NASA Astrophysics Data System (ADS)

We theoretically and experimentally study the mechanical response of silkworm and spider silks against stretching and the relationship with the underlying structural factors. It is found that the typical stress-strain profiles are predicted in good agreement with experimental measurements by implementing the "?-sheet splitting" mechanism we discovered and verified, primarily varying the secondary structure of protein macromolecules. The functions of experimentally observed structural factors responding to the external stress have been clearly addressed, and optimization of the microscopic structures to enhance the mechanical strength will be pointed out, beneficial to their biomedical and textile applications.

Wu, Xiang; Liu, Xiang-Yang; Du, Ning; Xu, Gangqin; Li, Baowen

2009-08-01

140

Biomechanics of Spider Silks.  

National Technical Information Service (NTIS)

This project aims to gain a fuller understanding of the relationship between processing, structure and mechanical properties of selected spider and silkworm silks. This goal will be pursued by inducing spiders and silkworms to produce threads under a rang...

F. Vollrath

2006-01-01

141

Increased molecular mobility in humid silk fibers under tensile stress  

NASA Astrophysics Data System (ADS)

Silk fibers are semicrystalline nanocomposite protein fibers with an extraordinary mechanical toughness that changes with humidity. Diffusive or overdamped motion on a molecular level is absent in dry silkworm silk, but present in humid silk at ambient temperature. This microscopic diffusion distinctly depends on the externally applied macroscopic tensile force. Quasielastic and inelastic neutron-scattering data as a function of humidity and of tensile strain on humid silk fibers support the model that both the adsorbed water and parts of the amorphous polymers participate in diffusive motion and are affected by the tensile force. It is notable that the quasielastic linewidth of humid silk at 100% relative humidity increases significantly with the applied force. The effect of the tensile force is discussed in terms of an increasing alignment of the polymer chains in the amorphous fraction with increasing tensile stress which changes the geometrical restrictions of the diffusive motions.

Seydel, Tilo; Knoll, Wiebke; Greving, Imke; Dicko, Cedric; Koza, Michael M.; Krasnov, Igor; Müller, Martin

2011-01-01

142

Hornet silk: thermophysical properties  

Microsoft Academic Search

Thermoelectric measurements performed on strips of hornet silk have shown marked rise in the electric charge upon increase of temperature between 20°C and 33°C. The increase was dependent on the relative humidity (RH), occurring only at RH above 90%. In tests of heat capacity, hornet silk was found to possess a specific heat of over 2J\\/gK. within a temperature range

V. Pertsis

2002-01-01

143

Silk film biomaterials for cornea tissue engineering  

PubMed Central

Biomaterials for corneal tissue engineering must demonstrate several critical features for potential utility in vivo, including transparency, mechanical integrity, biocompatibility and slow biodegradation. Silk film biomaterials were designed and characterized to meet these functional requirements. Silk protein films were used in a biomimetic approach to replicate corneal stromal tissue architecture. The films were 2 ?m thick to emulate corneal collagen lamellae dimensions, and were surface patterned to guide cell alignment. To enhance trans-lamellar diffusion of nutrients and to promote cell-cell interaction, pores with 0.5 to 5.0 ?m diameters were introduced into the silk films. Human and rabbit corneal fibroblast proliferation, alignment and corneal extracellular matrix expression on these films in both 2D and 3D cultures was demonstrated. The mechanical properties, optical clarity and surface patterned features of these films, combined with their ability to support corneal cell functions suggest this new biomaterial system offers important potential benefits for corneal tissue regeneration. PMID:19059642

Lawrence, Brian D.; Marchant, Jeffrey K.; Pindrus, Mariya; Omenetto, Fiorenzo; Kaplan, David L.

2009-01-01

144

Molecular dynamics analysis of supercontraction in spider dragline silk  

E-print Network

Spider dragline silk is a material that has evolved over millions of years to develop finely tuned mechanical properties. It is a protein-based fiber, used as the main structural component in spider webs and as a lifeline ...

Batty, Laura

2013-01-01

145

Sequence of Spider Aciniform and Piriform Silks.  

National Technical Information Service (NTIS)

The aciniform glands from Argiope Trifasciata were used to construct a cDNA library. The library was probed with various DNA probes based on known spider silk protein sequences. A large 9kb clone was identified and partial sequencing has been done. The C-...

R. V. Lewis

2001-01-01

146

Liquid crystalline spinning of spider silk  

Microsoft Academic Search

Spider silk has outstanding mechanical properties despite being spun at close to ambient temperatures and pressures using water as the solvent. The spider achieves this feat of benign fibre processing by judiciously controlling the folding and crystallization of the main protein constituents, and by adding auxiliary compounds, to create a composite material of defined hierarchical structure. Because the 'spinning dope'

Fritz Vollrath; David P. Knight

2001-01-01

147

Solid-State NMR Comparison of Various Spiders' Dragline Silk Fiber  

PubMed Central

Major ampullate (dragline) spider silk is a coveted biopolymer due to its combination of strength and extensibility. The dragline silk of different spiders have distinct mechanical properties that can be qualitatively correlated to the protein sequence. This study uses amino acid analysis and carbon-13 solid-state NMR to compare the molecular composition, structure and dynamics of major ampullate dragline silk of four orb-web spider species (Nephila clavipes, Araneus gemmoides, Argiope aurantia and Argiope argentata) and one cobweb species (Latrodectus hesperus). The mobility of the protein backbone and amino acid side chains in water exposed silk fibers is shown to correlate to the proline content. This implies that regions of major ampullate spidroin 2 protein, which is the only dragline silk protein with any significant proline content, become significantly hydrated in dragline spider silk. PMID:20593757

Creager, Melinda S.; Jenkins, Janelle E; Thagard-Yeaman, Leigh A.; Brooks, Amanda E.; Jones, Justin A.; Lewis, Randolph V.; Holland, Gregory P.; Yarger, Jeffery L.

2010-01-01

148

Solid-state NMR comparison of various spiders' dragline silk fiber.  

PubMed

Major ampullate (dragline) spider silk is a coveted biopolymer due to its combination of strength and extensibility. The dragline silk of different spiders have distinct mechanical properties that can be qualitatively correlated to the protein sequence. This study uses amino acid analysis and carbon-13 solid-state NMR to compare the molecular composition, structure, and dynamics of major ampullate dragline silk of four orb-web spider species ( Nephila clavipes , Araneus gemmoides , Argiope aurantia , and Argiope argentata ) and one cobweb species ( Latrodectus hesperus ). The mobility of the protein backbone and amino acid side chains in water exposed silk fibers is shown to correlate to the proline content. This implies that regions of major ampullate spidroin 2 protein, which is the only dragline silk protein with any significant proline content, become significantly hydrated in dragline spider silk. PMID:20593757

Creager, Melinda S; Jenkins, Janelle E; Thagard-Yeaman, Leigh A; Brooks, Amanda E; Jones, Justin A; Lewis, Randolph V; Holland, Gregory P; Yarger, Jeffery L

2010-08-01

149

Blueprint for a High-Performance Biomaterial: Full-Length Spider Dragline Silk Genes  

PubMed Central

Spider dragline (major ampullate) silk outperforms virtually all other natural and manmade materials in terms of tensile strength and toughness. For this reason, the mass-production of artificial spider silks through transgenic technologies has been a major goal of biomimetics research. Although all known arthropod silk proteins are extremely large (>200 kiloDaltons), recombinant spider silks have been designed from short and incomplete cDNAs, the only available sequences. Here we describe the first full-length spider silk gene sequences and their flanking regions. These genes encode the MaSp1 and MaSp2 proteins that compose the black widow's high-performance dragline silk. Each gene includes a single enormous exon (>9000 base pairs) that translates into a highly repetitive polypeptide. Patterns of variation among sequence repeats at the amino acid and nucleotide levels indicate that the interaction of selection, intergenic recombination, and intragenic recombination governs the evolution of these highly unusual, modular proteins. Phylogenetic footprinting revealed putative regulatory elements in non-coding flanking sequences. Conservation of both upstream and downstream flanking sequences was especially striking between the two paralogous black widow major ampullate silk genes. Because these genes are co-expressed within the same silk gland, there may have been selection for similarity in regulatory regions. Our new data provide complete templates for synthesis of recombinant silk proteins that significantly improve the degree to which artificial silks mimic natural spider dragline fibers. PMID:17565367

Ayoub, Nadia A.; Garb, Jessica E.; Tinghitella, Robin M.; Collin, Matthew A.; Hayashi, Cheryl Y.

2007-01-01

150

Unravelling the biodiversity of nanoscale signatures of spider silk fibres  

NASA Astrophysics Data System (ADS)

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.

Silva, Luciano P.; Rech, Elibio L.

2013-12-01

151

Segmented nanofibers of spider dragline silk: Atomic force microscopy and single-molecule force spectroscopy  

PubMed Central

Despite its remarkable materials properties, the structure of spider dragline silk has remained unsolved. Results from two probe microscopy techniques provide new insights into the structure of spider dragline silk. A soluble synthetic protein from dragline silk spontaneously forms nanofibers, as observed by atomic force microscopy. These nanofibers have a segmented substructure. The segment length and amino acid sequence are consistent with a slab-like shape for individual silk protein molecules. The height and width of nanofiber segments suggest a stacking pattern of slab-like molecules in each nanofiber segment. This stacking pattern produces nano-crystals in an amorphous matrix, as observed previously by NMR and x-ray diffraction of spider dragline silk. The possible importance of nanofiber formation to native silk production is discussed. Force spectra for single molecules of the silk protein demonstrate that this protein unfolds through a number of rupture events, indicating a modular substructure within single silk protein molecules. A minimal unfolding module size is estimated to be around 14 nm, which corresponds to the extended length of a single repeated module, 38 amino acids long. The structure of this spider silk protein is distinctly different from the structures of other proteins that have been analyzed by single-molecule force spectroscopy, and the force spectra show correspondingly novel features. PMID:11959907

Oroudjev, E.; Soares, J.; Arcidiacono, S.; Thompson, J. B.; Fossey, S. A.; Hansma, H. G.

2002-01-01

152

Photovoltaic Thai Silk Spinning Machine  

Microsoft Academic Search

This research presents the design and development of a photovoltaic Thai silk spinning machine for silk clothes product for One Tambol One Product (OTOP) in Thailand. This machine is based on PIC microcontroller for control speed of motor and battery charger. The results of this research, the machine helps Thai villagers for silk spinning by hand and makes little products

Denpong Sirikul; Narong Mungkung; Ratchadawan Nimnual

153

Effect of beta-sheet crystals on the thermal and rheological behavior of protein-based hydrogels derived from gelatin and silk fibroin.  

PubMed

Novel protein-based hydrogels have been prepared by blending gelatin (G) with amorphous Bombyx mori silk fibroin (SF) and subsequently promoting the formation of beta-sheet crystals in SF upon exposure to methanol or methanol/water solutions. Differential scanning calorimetry of the resultant hydrogels confirms the presence and thermoreversibility of the G helix-coil transition between ambient and body temperature at high G concentrations. At low G concentrations, this transition is shifted to higher temperatures and becomes progressively less pronounced. Complementary dynamic rheological measurements reveal solid-liquid cross-over at the G helix-coil transition temperature typically between 30 and 36 degrees C in blends prior to the formation of beta-sheet crystals. Introducing the beta-sheet conformation in SF stabilizes the hydrogel network and extends the solid-like behavior of the hydrogels to elevated temperatures beyond body temperature with as little as 10 wt.-% SF. The temperature-dependent elastic modulus across the G helix-coil transition is reversible, indicating that the conformational change in G can be used in stabilized G/SF hydrogels to induce thermally triggered encapsulant release. PMID:16080165

Gil, Eun S; Spontak, Richard J; Hudson, Samuel M

2005-08-12

154

Gumfooted lines in black widow cobwebs and the mechanical properties of spider capture silk.  

PubMed

Orb-weaving spiders produce webs using two types of silk that have radically different mechanical properties. The dragline silk used to construct the supporting frame and radii of the web is stiff and as strong as steel, while the capture spiral is much weaker but more than ten times as extensible. This remarkable divergence in mechanical properties has been attributed to the aqueous glue that coats the capture spiral, which is thought to decrease capture spiral stiffness and increase its extensibility. However, discerning the effect of the aqueous glue on fiber performance is complicated because dragline silk and the capture spiral are assembled from different proteins, which may also affect mechanical performance. Here, we use the sticky gumfooted lines of black widow cobwebs to test the effect of the addition of aqueous glue on the mechanical properties of dragline silk. We also surveyed orb-webs spun by a broad range of species for bundles of looped silk. Such bundles, termed windlasses, have been thought to increase capture spiral extensibility by "paying out" additional lengths of silk. Our results suggest that neither plasticization of silk by aqueous glue nor excess silk in windlasses can by themselves account for the remarkable extensibility of orb-weaver capture silk compared to other spider silks. This argues that the unique amino acid motifs of the flagelliform fibroins that constitute the core of the capture spiral play an essential role in capture silk's extreme extensibility. PMID:16351953

Blackledge, Todd A; Summers, Adam P; Hayashi, Cheryl Y

2005-01-01

155

Piriform Spider Silk Sequences Reveal Unique Repetitive Elements  

PubMed Central

Orb-weaving spider silk fibers are assembled from very large, highly repetitive proteins. The repeated segments contain, in turn, short, simple repetitive amino acid motifs that account for the physical and mechanical properties of the assembled fiber. Of the six orb-weaver silk fibroins, the piriform silk that makes the attachment discs, which lashes the joints of the web and attaches dragline silk to surfaces has not been previously characterized. Piriform silk protein cDNAs were isolated from phage libraries of three species, A. trifasciata, N. clavipes, and N. cruentata. The deduced amino acid sequences from these genes revealed two new repetitive motifs: an alternating proline motif where every other amino acid is proline, and a glutamine-rich motif of 6 to 8 amino acids. Similar to other spider silk proteins, the repeated segments are large (>200 amino acids) and highly homogenized within a species. There is also substantial sequence similarity across the genes from the three species with particular conservation of the repetitive motifs. Northern blot analysis revealed that the messenger RNA is larger than 11kb and is expressed exclusively in the piriform glands of the spider. Phylogenetic analysis of the C-terminal regions of the new proteins with published spidroins robustly shows that the pirifom sequences form an ortholog group. PMID:20954740

Perry, David J.; Bittencourt, Daniela; Siltberg-Liberles, Jessica; Rech, Elibio L.; Lewis, Randolph V.

2010-01-01

156

Evidence for antimicrobial activity associated with common house spider silk  

PubMed Central

Background Spider silk is one of the most versatile materials in nature with great strength and flexibility. Native and synthetically produced silk has been used in a wide range of applications including the construction of artificial tendons and as substrates for human cell growth. In the literature there are anecdotal reports that suggest that native spider silk may also have antimicrobial properties. Findings In this study we compared the growth of a Gram positive and a Gram negative bacterium in the presence and absence of silk produced by the common house spider Tegenaria domestica. We demonstrate that native web silk of Tegenaria domestica can inhibit the growth of the Gram positive bacterium, Bacillus subtilis. No significant inhibition of growth was detected against the Gram negative bacterium, Escherichia coli. The antimicrobial effect against B. subtilis appears to be short lived thus the active agent potentially acts in a bacteriostatic rather than bactericidal manner. Treatment of the silk with Proteinase K appears to reduce the ability to inhibit bacterial growth. This is consistent with the active agent including a protein element that is denatured or cleaved by treatment. Tegenaria silk does not appear to inhibit the growth of mammalian cells in vitro thus there is the potential for therapeutic applications. PMID:22731829

2012-01-01

157

The Digital Silk Road  

Microsoft Academic Search

Existing and proposed mechanisms for digital money all require large overhead to transfer money between parties. This overhead makes them unsuitable for extremely low cost activities, such as delivering and routing packets. The digital silk road is a proposed money system with extremely low transaction cost built into the communication protocols. The money introduced by this system is much more

Norman Hardy; Eric Dean Tribble

1993-01-01

158

Hierarchical Chain Model of Spider Capture Silk Elasticity  

E-print Network

Spider capture silk is a biomaterial with both high strength and high elasticity, but the structural design principle underlying these remarkable properties is still unknown. It was revealed recently by atomic force microscopy that, an exponential force--extension relationship holds both for capture silk mesostructures and for intact capture silk fibers [N. Becker et al., Nature Materials 2, 278 (2003)]. In this Letter a simple hierarchical chain model was proposed to understand and reproduce this striking observation. In the hierarchical chain model, a polymer is composed of many structural motifs which organize into structural modules and supra-modules in a hierarchical manner. Each module in this hierarchy has its own characteristic force. The repetitive patterns in the amino acid sequence of the major flagelliform protein of spider capture silk is in support of this model.

Haijun Zhou; Yang Zhang

2004-12-20

159

Materials: Surprising strength of silkworm silk  

NASA Astrophysics Data System (ADS)

Commercial silkworm silk is presumed to be much weaker and less extensible than spider dragline silk, which has been hailed as a 'super-fibre'. But we show here that the mechanical properties of silkworm silks can approach those of spider dragline silk when reeled under controlled conditions. We suggest that silkworms might be able to produce threads that compare well with spider silk by changing their spinning habits, rather than by having their silk genes altered.

Shao, Zhengzhong; Vollrath, Fritz

2002-08-01

160

Cross-linking in the silks of bees, ants and hornets.  

PubMed

Silk production is integral to the construction of nests or cocoons for many Aculeata, stinging Hymenopterans such as ants, bees and wasps. Here we report the sequences of new aculeate silk proteins and compare cross-linking among nine native silks from three bee species (Apis mellifera, Bombus terrestris and Megachile rotundata), three ant species (Myrmecia forficata, Oecophylla smaragdina and Harpegnathos saltator) and three hornets (Vespa analis, Vespa simillima and Vespa mandarinia). The well studied silks of spiders and silkworms are comprised of large proteins that are cross-linked and stabilized predominantly by intra and intermolecular beta sheet structure. In contrast, the aculeate silks are comprised of relatively small proteins that contain central coiled coil domains and comparatively reduced amounts of beta sheet structure. The hornet silks, which have the most beta sheet structure and the greatest amount of amino acid sequence outside the coiled-coil domains, dissolve in concentrated LiBr solution and appear to be stabilized predominantly by beta sheet structure like the classic silks. In contrast, the ant and bee silks, which have less beta sheet and less sequence outside the coiled-coil domains, could not be dissolved in LiBr and appear to be predominantly stabilized by covalent cross-linking. The iso-peptide cross-linker, ?-(?-glutamyl)-lysine that is produced by transglutaminase enzymes, was demonstrated to be present in all silks by mass spectrometry, but at greater levels in silks of ants and bees. The bee silks and ant cocoons, but not the Oecophylla nest silks, appeared to be further stabilized by tanning reactions. PMID:24607851

Campbell, Peter M; Trueman, Holly E; Zhang, Qiang; Kojima, Katsura; Kameda, Tsunenori; Sutherland, Tara D

2014-05-01

161

Electrodeposited silk coatings for functionalized implant applications  

NASA Astrophysics Data System (ADS)

The mechanical and morphological properties of titanium as well as its biocompatibility and osteoinductive characteristics have made it the material of choice for dental implant systems. Although the success rate of titanium implants exceeds 90% in healthy individuals, a large subset of the population has one or more risk factors that inhibit implant integration. Treatments and coatings have been developed to improve clinical outcomes via introduction of appropriate surface topography, texture and roughness or incorporation of bioactive molecules. It is essential that the coatings and associated deposition techniques are controllable and reproducible. Currently, methods of depositing functional coatings are dictated by numerous parameters (temperature, particle size distribution, pH and voltage), which result in variable coating thickness, strength, porosity and weight, and hinder or preclude biomolecule incorporation. Silk is a highly versatile protein with a unique combination of mechanical and physical properties, including tunable degradation, biocompatibility, drug stabilizing capabilities and mechanical properties. Most recently an electrogelation technique was developed which allows for the deposition of gels which dry seamlessly over the contoured topography of the conductive substrate. In this work we examine the potential use of silk electrogels as mechanically robust implant coatings capable of sequestering and releasing therapeutic agents. Electrodeposition of silk electrogels formed in uniform electric fields was characterized with respect to field intensity and deposition time. Gel formation kinetics were used to derive functions which allowed for the prediction of coating deposition over a range of process and solution parameters. Silk electrogel growth orientation was shown to be influenced by the applied electric field. Coatings were reproducible and tunable via intrinsic silk solution properties and extrinsic process parameters. Adhesion was modulated over a 10-fold range and implant insertion into bone mimics demonstrated that the coatings were able to withstand delamination forces experienced during these mock implantations. Antibiotic release from coated implant studs inhibited bacterial growth and dexamethasone release was shown to stimulate calcium deposition in mesenchymal stem cells.

Elia, Roberto

162

The expression pattern of four odorant-binding proteins in male and female silk moths, Bombyx mori.  

PubMed

Four recombinant odorant-binding proteins of Bombyx mori, pheromone-binding protein (PBP), general odorant-binding protein 1 (GOBP1), general odorant-binding protein 2 (GOBP2) and antennal binding protein X (ABPX), were expressed in E. coli and used to raise polyclonal antisera. Immunoblots of antennal homogenates showed that these antisera were specific. In Western blot analysis and immunocytochemical labelling experiments, the sera against recombinant PBP and GOBP2 of B. mori gave identical results as sera against native PBP and GOBP2 of Antheraea polyphemus, respectively, thus confirming earlier results obtained with the latter. Labelling consecutive cross sections of various sensillum types with all four antisera revealed different labelling patterns in male and female sensilla (s.) trichodea and s. basiconica. Long s. trichodea in males and females represented uniform labelling types, whereas for short s. trichodea, s. intermedia, and s. basiconica a great variety of labelling patterns was observed, some being more common than others. Long s. trichodea, which in males are uniformly tuned to the pheromone components bombykol and bombykal, all strongly expressed PBP; labelling with antisera against the other three odorant-binding proteins hardly was above background, only in some hairs GOBP1 was expressed somewhat more strongly. Long s. trichodea of females, which respond specifically to linalool and benzoic acid, showed a different labelling pattern. Here, we observed strong labelling with antibodies against GOBP2 and medium labelling with anti-GOBP1, sometimes with anti-ABPX. S. basiconica in both sexes most commonly co-expressed GOBP1 and GOBP2, but other patterns were occasionally found, with some of them showing PBP expression, also in females. The great variety of labelling types in short s. trichodea, s. intermedia, and s. basiconica suggests a similar variety of functional subtypes as observed in plant odour-sensitive sensilla of other moth species. PMID:16374716

Maida, R; Mameli, M; Müller, B; Krieger, J; Steinbrecht, R A

2005-03-01

163

Novel aquatic silk genes Simulium (Psilozia) vittatum (Zett) Diptera: Simuliidae.  

PubMed

The silks of arthropods have an elementary role in the natural history of the organisms that spin them, yet they are coded by rapidly evolving genes leading some authors to speculate that silk proteins are non-homologous proteins co-opted multiple times independently for similar functions. However, some general structural patterns are emerging. In this work we identified three major silk gland proteins using a combined biochemical, proteomic, next-generation sequencing and bioinformatic approach. Biochemical characterization determined that they were phosphorylated with multiple isoforms and potentially differential phosphorylation. Structural characterization showed that their structure was more similar to silk proteins from distantly related aquatic Trichopteran species than more closely related terrestrial or aquatic Diptera. Overall, our approach is easily transferable to any non-model species and if used across a larger number of aquatic species, we will be able to better understand the processes involved in linking the secondary structure of silk proteins with their function between in an organisms and its habitat. PMID:24446544

Papanicolaou, Alexie; Woo, Angelica; Brei, Brianna; Ma, Danjun; Masedunskas, Andrius; Gray, Elmer; Guishan Xiao, Gary; Cho, Soochin; Brockhouse, Charles

2013-12-01

164

1H, 13C and 15N NMR assignments of the aciniform spidroin (AcSp1) repetitive domain of Argiope trifasciata wrapping silk.  

PubMed

Spider silk is one of nature's most remarkable biomaterials due to extraordinary strength and toughness not found in today's synthetic materials. Of the seven types of silk, wrapping silk (AcSp1) is the most extensible of the types of silks and has no sequence similarity to the other types. Here we report the chemical shifts for the AcSp1 199 amino acid protein repeat unit and its anticipated secondary structure based on secondary chemical shifts. PMID:21989955

Xu, Lingling; Tremblay, Marie-Laurence; Meng, Qing; Liu, Xiang-Qin; Rainey, Jan K

2012-10-01

165

Characterisation of phenol oxidase and peroxidase from maize silk.  

PubMed

Silk of some maize genotypes contains a high level of phenolics that undergo enzymatic oxidation to form quinones, which condense among themselves or with proteins to form brown pigments. Two phenolic oxidizing enzymes, peroxidase (POD; EC 1.11.1.7) and polyphenol oxidase (PPO; EC 1.10.3.1), from maize (Zea mays L.) silk were characterised with respect to their preferred substrate, different isoforms and specific effectors. One browning silk sample with high, and two non-browning samples with low phenolic content were investigated. Although POD oxidizes a wide range of phenolic substrates in vitro, its activity rate was independent of silk phenolic content. PPO activity, detected with o-diphenolic substrates, was abundant only in browning silk, and low or absent in non-browning silk. Pollination increased POD but not PPO activity. Isoelectric-focusing (IEF) and specific staining for POD and PPO showed a high degree of polymorphism that varied with silk origin. The IEF pattern of POD revealed a number of anionic and several cationic isoenzymes, with the most pronounced having neutral pI 7 and a basic isoform with pI 10. Detected isoforms of PPO were anionic, except for one neutral form found only in browning silk, and occupied positions different from those of POD. Different inhibitory effects of NaN(3), EDTA, KCN, and L-cysteine, as well as different impacts of a variety of cations on the oxidation of chlorogenic acid, mediated by PPO or POD, were detected. The findings are discussed in terms of a possible roles of these enzymes in defence and pollination. PMID:20522176

Sukalovi?, V Hadzi-Taskovi?; Veljovi?-Jovanovi?, S; Maksimovi?, J Dragisi?; Maksimovi?, V; Paji?, Z

2010-05-01

166

Permeability of silk microcapsules made by the interfacial adsorption of Kevin D. Hermanson,a  

E-print Network

- menical is that of the thin-shelled vesicle. In applications such as drug delivery, flavor encapsulation and functionalizability. Recently, the assembly of recombinant spider-silk proteins at an emulsion interface has been used be removed from the emulsion and inserted into a single-phase solution. The final spider-silk shell has

Bausch, Andreas

167

The effects of corn silk on glycaemic metabolism  

PubMed Central

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 ?-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, p < 0.01, respectively), while the level of insulin secretionn was markedly elevated in alloxa-induced hyperglycemic mice (p < 0.05). The alloxan-damaged pancreatic ?-cells of the mice were partly recovered gradually after the mice were administered with corn silk extract 15 days later. Also, the body weight of the alloxan-induced hyperglycemic mice was increased gradually. However, ascension of blood glucose induced by adrenalin and gluconeogenesis induced by L-alanine were not inhibited by corn silk extract treatment (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 ?-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. PMID:19930631

2009-01-01

168

Silk constructs for delivery of muskuloskeletal therapeutics  

PubMed Central

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 exciting for biomedical applications. his pattern along with the versatility of this biopolymer have been translated into progress for musculoskeletal applications. We review the use and potential of silk fibroin for systemic and localized delivery of therapeutics in diseases affecting the musculoskeletal system. We also present future directions for this biopolymer as well as the necessary research and development steps for their achievement. PMID:22522139

Meinel, Lorenz; Kaplan, David L.

2012-01-01

169

Unraveled mechanism in silk engineering: Fast reeling induced silk Xiang Wu,1,2  

E-print Network

response of silkworm and spider silks against stretching and the relationship with the underlying spider silk can stop a Boeing 747 in flight.5 On the other hand, the silkworm silk market has flourished for the last several cen- turies, and the high production of silkworm silk dominates silk engineering

Li, Baowen

170

Silk Fibroin under Osmotic Stress  

NASA Astrophysics Data System (ADS)

The osmotic stress method was applied to study the thermodynamics of supramolecular self-assembly phenomena in crystallizable segments of Bombyx mori silkworm silk fibroin. Controlling compositions and phases of silk fibroin solution, the method provided a means for the direct investigation of microscopic and thermodynamic details of these intermolecular interactions in aqueous media. It is apparent that as osmotic pressure increases, silk fibroin molecules get pressurized to align together to form a water-soluble crystalline mesophase (Silk-I), and then gradually become anti-parallel b-sheet structure (Silk-II) at higher osmotic pressure. This behavior becomes more sensitive as the salt concentration decreases. A partial ternary phase diagram of Water-Silk fibroin-LiBr was constructed based on the results. This phase diagram can be utilized to help design a new route for wet spinning of re-generated silk fibroin. Precise control of compositions and corresponding crystalline structure of a silk fibroin solution may enable us to simulate the natural Bombyx mori silkworm spinning process.

Sohn, Sungkyun; Strey, Helmut H.; Gido, Samuel P.

2003-03-01

171

Tuning Chemical and Physical Crosslinks in Silk Electrogels for Morphological Analysis and Mechanical Reinforcement  

PubMed Central

Electrochemically controlled, reversible assembly of biopolymers into hydrogel structures is a promising technique for on-demand cell or drug encapsulation and release systems. An electrochemically sol-gel transition has been demonstrated in regenerated Bombyx mori silk fibroin, offering a controllable way to generate biocompatible and reversible adhesives and other biomedical materials. Despite the involvement of an electrochemically triggered electrophoretic migration of the silk molecules, the mechanism of the reversible electrogelation remains unclear. It is, however, known that the freshly prepared silk electrogels (e-gels) adopt a predominantly random coil conformation, indicating a lack of crosslinking as well as thermal, mechanical and morphological stabilities. In the present work, the tuning of covalent and physical ?-sheet crosslinks in silk hydrogels was studied for programming the structural properties. Scanning electron microscopy (SEM) revealed delicate morphology, including locally aligned fibrillar structures, in silk e-gels, preserved by combining glutaraldehyde-crosslinking and ethanol dehydration. Fourier transform infrared (FTIR) spectroscopic analysis of either electrogelled, vortex-induced or spontaneously formed silk hydrogels showed that the secondary structure of silk e-gels was tunable between non ?-sheet dominated and ?-sheet dominated states. Dynamic oscillatory rheology confirmed the mechanical reinforcement of silk e-gels provided by controlled chemical and physical crosslinks. The selective incorporation of either chemical or physical or both crosslinks into the electrochemically-responsive, originally unstructured silk e-gel should help in the design for electrochemically-responsive protein polymers. PMID:23859710

Lin, Yinan; Xia, Xiaoxia; Shang, Ke; Elia, Roberto; Huang, Wenwen; Cebe, Peggy; Leisk, Gary; Omenetto, Fiorenzo; Kaplan, David L.

2013-01-01

172

Properties and potential medical applications of silk fibers produced by Rothischildia lebeau.  

PubMed

Rothischildia lebeau which belongs to the Saturniidae family of silk-producing insects secretes protein fibers with properties between that of the Bombyx mori and the common wild silks. Traditionally, wild silks produced by insects such as Antheraea mylitta are considerably coarser and have inferior tensile properties than the domesticated and most commonly used silk produced by B. mori. Recently, it has been demonstrated that some of the wild silks have unique properties and preferable for medical applications. Wild silks are comparatively easier to rear, produce larger cocoons, and could have unique properties. In this research, the structure and properties of the silk fibers produced by R. lebeau were studied and the potential of using the fibers for medical applications was investigated. Fibers produced by R. lebeau had average tensile strength of 3.3?g/den, similar to that of wild silks but lower than that of the B. mori silk. R. lebeau fibers were biocompatible and showed potential to be useful for tissue engineering and other medical applications. PMID:23594071

Reddy, Narendra; Jiang, Qiuran; Yang, Yiqi

2013-01-01

173

Materials: Surprising strength of silkworm silk  

Microsoft Academic Search

Commercial silkworm silk is presumed to be much weaker and less extensible than spider dragline silk, which has been hailed as a 'super-fibre'. But we show here that the mechanical properties of silkworm silks can approach those of spider dragline silk when reeled under controlled conditions. We suggest that silkworms might be able to produce threads that compare well with

Zhengzhong Shao; Fritz Vollrath

2002-01-01

174

Preparation of hexagonal GeO2 particles with particle size and crystallinity controlled by peptides, silk and silk-peptide chimeras.  

PubMed

We demonstrate the use of silk based proteins to control the particle/crystallite size during GeO2 formation, using a bio-mimetic approach at circumneutral pH and ambient temperature. Multicrystalline GeO2 was prepared from germanium tetraethoxide (TEOG) in the presence of different silk-based proteins: Bombyx mori silk (native silk) and two chimeric proteins prepared by linking a germania binding peptide (Ge28: HATGTHGLSLSH) with Bombyx mori silk via chemical coupling at different peptide loadings (silk-Ge28 10% and silk-Ge28 50%). The mineralisation activity of the silk-based proteins was compared with that of peptide Ge28 as a control system. GeO2 mineralisation was investigated in water and in citric acid/bis-tris propane buffer at pH 6. Morphology, particle size, crystallinity, water and organic content of the materials obtained were analysed to study the effect of added biomolecules and mineralisation environment on material properties. In the presence of silk additives well-defined cube-shape hybrid materials composed of hexagonal germania and up to ca. 5 wt% organic content were obtained. The cubic particles ranged from 0.4 to 1.4 ?m in size and were composed of crystalline domains in the range 35-106 nm depending on the additive used and synthesis conditions. The organic material incorporated in the mineral did not appear to affect the unit cell dimensions. The silk and chimeric proteins in water promote material formation and crystal growth, possibly via an effective ion-channelling mechanism, however further studies are needed to assert to what extent the presence of the silk impacts on nucleation and growth stages. The germania binding peptide alone did not have any significant effect on reaction rate, yield or the material's properties compared to the blank. Interestingly, the peptide content in the silk chimeras tested did not affect mineralisation. The presence of buffer inhibited mineral condensation rate and yield. The use of silk-based biomolecules allows control of crystallite/particle size of hybrid materials opening up opportunities for bio-inspired approaches to be applied for the synthesis of functional germania based devices and materials. PMID:25300352

Boix, Estefania; Puddu, Valeria; Perry, Carole C

2014-10-22

175

Structure Formation in Bombyx mori Silk Foams  

NASA Astrophysics Data System (ADS)

The structural evolution of silk stored within the lumen of the silk gland of the silkworm (Silk I) to the highly crystalline spun fiber (Silk II) is not well understood. Conclusive experimental elucidation of Silk I crystal structure has still not been achieved despite nearly fifty years of study. Via a foaming technique, a new insight has been provided into the structural formation of Bombyx mori silk on a microscopic scale in aqueous systems. These formations include an unoriented state, a mesophase, and near perfect single crystals. A similarity between the semicrystalline foam structureand the previously observed interfacial silk (Silk III) was found. Under different conditions, a Silk I-like structure was present in the silk foams as near perfect single crystals.

He, Shi-Juang; Valluzzi, Regina; Gido, Samuel P.

1998-03-01

176

Synthetic spider silk production on a laboratory scale.  

PubMed

As society progresses and resources become scarcer, it is becoming increasingly important to cultivate new technologies that engineer next generation biomaterials with high performance properties. The development of these new structural materials must be rapid, cost-efficient and involve processing methodologies and products that are environmentally friendly and sustainable. Spiders spin a multitude of different fiber types with diverse mechanical properties, offering a rich source of next generation engineering materials for biomimicry that rival the best manmade and natural materials. Since the collection of large quantities of natural spider silk is impractical, synthetic silk production has the ability to provide scientists with access to an unlimited supply of threads. Therefore, if the spinning process can be streamlined and perfected, artificial spider fibers have the potential use for a broad range of applications ranging from body armor, surgical sutures, ropes and cables, tires, strings for musical instruments, and composites for aviation and aerospace technology. In order to advance the synthetic silk production process and to yield fibers that display low variance in their material properties from spin to spin, we developed a wet-spinning protocol that integrates expression of recombinant spider silk proteins in bacteria, purification and concentration of the proteins, followed by fiber extrusion and a mechanical post-spin treatment. This is the first visual representation that reveals a step-by-step process to spin and analyze artificial silk fibers on a laboratory scale. It also provides details to minimize the introduction of variability among fibers spun from the same spinning dope. Collectively, these methods will propel the process of artificial silk production, leading to higher quality fibers that surpass natural spider silks. PMID:22847722

Hsia, Yang; Gnesa, Eric; Pacheco, Ryan; Kohler, Kristin; Jeffery, Felicia; Vierra, Craig

2012-01-01

177

Accelerated in vitro Degradation of Optically Clear Low ?-sheet Silk Films by Enzyme-Mediated Pretreatment  

PubMed Central

Purpose To design patterned, transparent silk films with fast degradation rates for the purpose of tissue engineering corneal stroma, Methods ?-sheet (crystalline) content of silk films was decreased significantly by using a short water annealing time. Additionally, a protocol combining short water annealing time with enzymatic pretreatment of silk films with protease XIV was developed. Results Low ?-sheet content (17–18%) and enzymatic pre-treatment provided film stability in aqueous environments and accelerated degradation of the silk films in the presence of human corneal fibroblasts in vitro. The results demonstrate a direct relationship between reduced ?-sheet content and enzymatic pre-treatment and overall degradation rate of the protein films. Conclusions The novel protocol developed here provides new approaches to modulate the regeneration rate of silk biomaterials for corneal tissue regeneration needs. Translational relevance Patterned silk protein films possess desirable characteristics for corneal tissue engineering, including optical transparency, biocompatibility, cell alignment and tunable mechanical properties, but current fabrication protocols do not provide adequate degradation rates to match the regeneration properties of the human cornea. This novel processing protocol makes silk films more suitable for the construction of human corneal stroma tissue and a promising way to tune silk film degradation properties to match corneal tissue regeneration. PMID:23579493

Shang, Ke; Rnjak-Kovacina, Jelena; Lin, Yinan; Hayden, Rebecca S.; Hu, Tao; Kaplan, David L.

2013-01-01

178

Accelerated In Vitro Degradation of Optically Clear Low ?-Sheet Silk Films by Enzyme-Mediated Pretreatment  

PubMed Central

Purpose: To design patterned, transparent silk films with fast degradation rates for the purpose of tissue engineering corneal stroma. Methods: ?-sheet (crystalline) content of silk films was decreased significantly by using a short water annealing time. Additionally, a protocol combining short water annealing time with enzymatic pretreatment of silk films with protease XIV was developed. Results: Low ?-sheet content (17%–18%) and enzymatic pretreatment provided film stability in aqueous environments and accelerated degradation of the silk films in the presence of human corneal fibroblasts in vitro. The results demonstrate a direct relationship between reduced ?-sheet content and enzymatic pretreatment, and overall degradation rate of the protein films. Conclusions: The novel protocol developed here provides new approaches to modulate the regeneration rate of silk biomaterials for corneal tissue regeneration needs. Translational Relevance: Patterned silk protein films possess desirable characteristics for corneal tissue engineering, including optical transparency, biocompatibility, cell alignment, and tunable mechanical properties, but current fabrication protocols do not provide adequate degradation rates to match the regeneration properties of the human cornea. This novel processing protocol makes silk films more suitable for the construction of human corneal stroma tissue and a promising way to tune silk film degradation properties to match corneal tissue regeneration. PMID:24049717

Shang, Ke; Rnjak-Kovacina, Jelena; Lin, Yinan; Hayden, Rebecca S.; Tao, Hu; Kaplan, David L.

2013-01-01

179

Changes in growth and cell wall extensibility of maize silks following pollination.  

PubMed

In response to pollination maize silks undergo an accelerated process of senescence which involves an inhibition of elongation. To gain insight into the mechanism underlying this growth response, the relationships among silk elongation kinetics, cell wall biophysical properties, pollen tube growth, and expansin protein abundance were investigated. The inhibition of silk elongation became apparent beyond 12 h after pollination. Pollinated walls were less responsive in assays of extension induced by pollen beta-expansin. Expansin protein abundance and endogenous expansin activity were not considerably reduced after pollination. Silk wall plastic compliance was significantly reduced 6 h post-pollination and beyond, suggesting that the wall undergoes structural modifications leading to its rigidification in response to pollination. The reduction in the plastic compliance occurred locally and progressively, shortly after pollen tubes traversed through a region of silk. Though numerous pollen grains germinated and initiated pollen tubes at the silk tip, the density of pollen tubes gradually declined along the length of the silk and only 1-2 reached the ovary even 24 h after pollination. These results support the notion that pollination-induced cell wall rigidification plays multiple roles in maize reproduction, including inhibition of silk growth and prevention of polyspermy. PMID:20656797

Kapu, Nuwan U Sella; Cosgrove, Daniel J

2010-09-01

180

Changes in growth and cell wall extensibility of maize silks following pollination  

PubMed Central

In response to pollination maize silks undergo an accelerated process of senescence which involves an inhibition of elongation. To gain insight into the mechanism underlying this growth response, the relationships among silk elongation kinetics, cell wall biophysical properties, pollen tube growth, and expansin protein abundance were investigated. The inhibition of silk elongation became apparent beyond 12?h after pollination. Pollinated walls were less responsive in assays of extension induced by pollen ?-expansin. Expansin protein abundance and endogenous expansin activity were not considerably reduced after pollination. Silk wall plastic compliance was significantly reduced 6?h post-pollination and beyond, suggesting that the wall undergoes structural modifications leading to its rigidification in response to pollination. The reduction in the plastic compliance occurred locally and progressively, shortly after pollen tubes traversed through a region of silk. Though numerous pollen grains germinated and initiated pollen tubes at the silk tip, the density of pollen tubes gradually declined along the length of the silk and only 1–2 reached the ovary even 24?h after pollination. These results support the notion that pollination-induced cell wall rigidification plays multiple roles in maize reproduction, including inhibition of silk growth and prevention of polyspermy. PMID:20656797

Sella Kapu, Nuwan U.; Cosgrove, Daniel J.

2010-01-01

181

Nutrient deprivation induces property variations in spider gluey silk.  

PubMed

Understanding the mechanisms facilitating property variability in biological adhesives may promote biomimetic innovations. Spider gluey silks such as the spiral threads in orb webs and the gumfoot threads in cobwebs, both of which comprise of an axial thread coated by glue, are biological adhesives that have variable physical and chemical properties. Studies show that the physical and chemical properties of orb web gluey threads change when spiders are deprived of food. It is, however, unknown whether gumfoot threads undergo similar property variations when under nutritional stress. Here we tested whether protein deprivation induces similar variations in spiral and gumfoot thread morphology and stickiness. We manipulated protein intake for the orb web spider Nephila clavipes and the cobweb spider Latrodectus hesperus and measured the diameter, glue droplet volume, number of droplets per mm, axial thread width, thread stickiness and adhesive energy of their gluey silks. We found that the gluey silks of both species were stickier when the spiders were deprived of protein than when the spiders were fed protein. In N. clavipes a concomitant increase in glue droplet volume was found. Load-extension curves showed that protein deprivation induced glue property variations independent of the axial thread extensions in both species. We predicted that changes in salt composition of the glues were primarily responsible for the changes in stickiness of the silks, although changes in axial thread properties might also contribute. We, additionally, showed that N. clavipes' glue changes color under protein deprivation, probably as a consequence of changes to its biochemical composition. PMID:24523902

Blamires, Sean J; Sahni, Vasav; Dhinojwala, Ali; Blackledge, Todd A; Tso, I-Min

2014-01-01

182

Micromolar biosensing of nitric oxide using myoglobin immobilized in a synthetic silk film.  

PubMed

In this work we investigate the use of coiled-coil silk proteins, produced in recombinant Escherichia coli, as a new material for immobilizing biosensors. Myoglobin was embedded in transparent honeybee silk protein films. Immobilized myoglobin maintained a high affinity for nitric oxide (KD NO=52 µM) and good sensitivity with a limit of detection of 5 µM. The immobilized myoglobin-silk protein film was stable and could be stored as a dry film at room temperature for at least 60 days. The effect of immobilization on the structure of myoglobin was fully investigated using UV/visible, Fourier Transform Infrared and Raman spectroscopy, which indicated a weakening in the strength of the iron-histidine bond. This study demonstrates that recombinant coiled-coil silk proteins provide a safe and environmentally friendly alternative to sol-gels for stabilizing heme proteins for use as optical biosensors. PMID:25014754

Rapson, Trevor D; Church, Jeffrey S; Trueman, Holly E; Dacres, Helen; Sutherland, Tara D; Trowell, Stephen C

2014-12-15

183

Uncovering spider silk nanocrystalline variations that facilitate wind-induced mechanical property changes.  

PubMed

Spider major ampullate (MA) silk varies in mechanical properties when spun in different environments. Amino acid compositional changes induced by variations in MaSp1 and MaSp2 expression, and various biochemical and physiological glandular processes induce silk property variability. Quantifying the contributions of these mechanisms on silk variability may facilitate the development of silk biomimetics. Wind is a medium that induces variations in MA silk mechanics. We exposed the spider Cyclosa mulmeinensis to wind and measured the amino acid composition, tensile mechanics, and crystalline structure of its MA silk using HPLC, tensile tests, and X-ray diffraction. We found the mechanical properties of MA silks from spiders exposed to wind to differ from unexposed spiders. The amino acid compositions did not differ, but X-ray diffraction found a lower crystal density and greater ?-sheet alignment relative to the fiber axis in the silks of spiders exposed to wind. We found no evidence that the mechanical property variations were a product of profound changes to the alignment of the protein within the amorphous region. We conclude that variations in the density and alignment of the crystalline ?-sheets, probably accompanied by some alignment change in the amorphous region as a result of "stretching" during spinning of the silk, probably explains the mechanical property variations that we found across treatment subgroups. As C. mulmeinensis MA silk increases both in strength and elasticity when the spiders are exposed to wind, bioengineers may consider it as a model for the development of high-performance silk biomimetics. PMID:23947397

Blamires, Sean J; Wu, Chao-Chia; Wu, Chung-Lin; Sheu, Hwo-Shuenn; Tso, I-Min

2013-10-14

184

Physical characterization of functionalized spider silk: electronic and sensing properties  

NASA Astrophysics Data System (ADS)

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

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

2011-10-01

185

Cell culture's spider silk road.  

PubMed

A number of synthetic and natural materials have been tried in cell culture and tissue engineering applications in recent years. Now Jeffrey Perkel takes a look at one new culture component that might surprise you-spider silk. PMID:24924388

Perkel, Jeffrey

2014-06-01

186

Identification and dynamics of polyglycine II nanocrystals in Argiope trifasciata flagelliform silk.  

PubMed

Spider silks combine a significant number of desirable characteristics in one material, including large tensile strength and strain at breaking, biocompatibility, and the possibility of tailoring their properties. Major ampullate gland silk (MAS) is the most studied silk and their properties are explained by a double lattice of hydrogen bonds and elastomeric protein chains linked to polyalanine ?-nanocrystals. However, many basic details regarding the relationship between composition, microstructure and properties in silks are still lacking. Here we show that this relationship can be traced in flagelliform silk (Flag) spun by Argiope trifasciata spiders after identifying a phase consisting of polyglycine II nanocrystals. The presence of this phase is consistent with the dominant presence of the -GGX- and -GPG- motifs in its sequence. In contrast to the passive role assigned to polyalanine nanocrystals in MAS, polyglycine II nanocrystals can undergo growing/collapse processes that contribute to increase toughness and justify the ability of Flag to supercontract. PMID:24162473

Perea, G B; Riekel, C; Guinea, G V; Madurga, R; Daza, R; Burghammer, M; Hayashi, C; Elices, M; Plaza, G R; Pérez-Rigueiro, J

2013-01-01

187

Identification and dynamics of polyglycine II nanocrystals in Argiope trifasciata flagelliform silk  

PubMed Central

Spider silks combine a significant number of desirable characteristics in one material, including large tensile strength and strain at breaking, biocompatibility, and the possibility of tailoring their properties. Major ampullate gland silk (MAS) is the most studied silk and their properties are explained by a double lattice of hydrogen bonds and elastomeric protein chains linked to polyalanine ?-nanocrystals. However, many basic details regarding the relationship between composition, microstructure and properties in silks are still lacking. Here we show that this relationship can be traced in flagelliform silk (Flag) spun by Argiope trifasciata spiders after identifying a phase consisting of polyglycine II nanocrystals. The presence of this phase is consistent with the dominant presence of the –GGX– and –GPG– motifs in its sequence. In contrast to the passive role assigned to polyalanine nanocrystals in MAS, polyglycine II nanocrystals can undergo growing/collapse processes that contribute to increase toughness and justify the ability of Flag to supercontract. PMID:24162473

Perea, G. B.; Riekel, C.; Guinea, G. V.; Madurga, R.; Daza, R.; Burghammer, M.; Hayashi, C.; Elices, M.; Plaza, G. R.; Perez-Rigueiro, J.

2013-01-01

188

Controlled hierarchical assembly of spider silk-DNA chimeras into ribbons and raft-like morphologies.  

PubMed

Spider silk-DNA conjugates comprising the recombinant spider silk protein eADF4(C16) and short oligonucleotides were arranged in a linear antiparallel and parallel as well as in a branched manner via designed complementarity of the DNA moieties. After cross-? fibril self-assembly, temperature-induced annealing of the DNA moieties triggered fibril association into ribbons, composed of aligned nanofibrils, and rafts composed of ribbons ordered into sharply bordered, squared fibrous microstructures. The formation of the superstructures was clearly dependent on the individual silk-DNA conjugate. A combination of 5'-conjugated silk moieties via complementary nucleic acids enhanced fibril association, whereas mixing complementary 5'- and 3'-silk conjugates inhibited the formation of higher-order structures. PMID:24924514

Humenik, Martin; Drechsler, Markus; Scheibel, Thomas

2014-07-01

189

In Situ Raman Spectroscopic Study of Al-Infiltrated Spider Dragline Silk under Tensile Deformation.  

PubMed

Natural materials consisting of protein structures impregnated with a tiny amount of metals often exhibit impressive mechanical behavior, which represents a new design paradigm for the development of biomimetic materials. Here, we produced Al-infiltrated silks by applying a modified Al2O3 atomic layer deposition process to the dragline silk of the Nephila pilipes spider, which showed unusual mechanical properties. The deformation behavior of the molecular structure of the Al-infiltrated silk was investigated by performing in situ Raman spectroscopy, where Raman shifts were measured concurrently with macroscopic mechanical deformations. For identifying the role of the infiltrated Al atoms, the study was performed in parallel with untreated silk, and the results were compared. Our experimental results revealed that superior mechanical properties of the Al-infiltrated silk are likely to be caused by the alterations of the sizes of the ?-sheet crystals and their distribution. PMID:25203848

Lee, Seung-Mo; Pippel, Eckhard; Moutanabbir, Oussama; Kim, Jae-Hyun; Lee, Hak-Joo; Knez, Mato

2014-10-01

190

Sporicidal/bactericidal textiles via the chlorination of silk.  

PubMed

Bacterial spores, such as those of the Bacillus genus, are extremely resilient, being able to germinate into metabolically active cells after withstanding harsh environmental conditions or aggressive chemical treatments. The toughness of the bacterial spore in combination with the use of spores, such as those of Bacillus anthracis, as a biological warfare agent necessitates the development of new antimicrobial textiles. In this work, a route to the production of fabrics that kill bacterial spores and cells within minutes of exposure is described. Utilizing this facile process, unmodified silk cloth is reacted with a diluted bleach solution, rinsed with water, and dried. The chlorination of silk was explored under basic (pH 11) and slightly acidic (pH 5) conditions. Chloramine-silk textiles prepared in acidified bleach solutions were found to have superior breaking strength and higher oxidative Cl contents than those prepared under caustic conditions. Silk cloth chlorinated for ?1 h at pH 5 was determined to induce >99.99996% reduction in the colony forming units of Escherichia coli, as well as Bacillus thuringiensis Al Hakam (B. anthracis simulant) spores and cells within 10 min of contact. The processing conditions presented for silk fabric in this study are highly expeditionary, allowing for the on-site production of protein-based antimicrobial materials from a variety of agriculturally produced feed-stocks. PMID:22352921

Dickerson, Matthew B; Lyon, Wanda; Gruner, William E; Mirau, Peter A; Slocik, Joseph M; Naik, Rajesh R

2012-03-01

191

Silk properties determined by gland-specific expression of a spider fibroin gene family.  

PubMed

Spiders produce a variety of silks that range from Lycra-like elastic fibers to Kevlar-like superfibers. A gene family from the spider Araneus diadematus was found to encode silk-forming proteins (fibroins) with different proportions of amorphous glycine-rich domains and crystal domains built from poly(alanine) and poly(glycine-alanine) repeat motifs. Spiders produce silks of different composition by gland-specific expression of this gene family, which allows for a range of mechanical properties according to the crystal-forming potential of the constituent fibroins. These principles of fiber property control may be important in the development of genetically engineered structural proteins. PMID:8600519

Guerette, P A; Ginzinger, D G; Weber, B H; Gosline, J M

1996-04-01

192

Silk materials--a road to sustainable high technology.  

PubMed

This review addresses the use of silk protein as a sustainable material in optics and photonics, electronics and optoelectronic applications. These options represent additional developments for this technology platform that compound the broad utility and impact of this material for medical needs that have been recently described in the literature. The favorable properties of the material certainly make a favorable case for the use of silk, yet serve as a broad inspiration to further develop biological foundries for both the synthesis and processing of Nature's materials for technological applications. PMID:22553118

Tao, Hu; Kaplan, David L; Omenetto, Fiorenzo G

2012-06-01

193

Physical Characterization of Functionalized Silk Material for Electronic Application and Devices  

NASA Astrophysics Data System (ADS)

Naturally harvested spider silk fibers are investigated for their physical properties under ambient, humidified, iodine-doped, pyrolized, sputtered gold and carbon nanotube coated conditions. The functional properties include: humidity activated conductivity; enhanced flexibility and carbon yield of pyrolized iodized silk fibers; full metallic conductivity and flexibility of micron-sized gold-sputtered silk fibers; and high strain sensitivity of carbon nanotube coated silk fibers. Magic angle spinning nuclear magnetic resonance (MAS-NMR) and Fourier transform infrared spectroscopy (FTIR) are used to explore the nature of ambient and functionalized spider silk fiber, and significant changes in amino acid-protein backbone signature are correlated with gold sputtering, and iodine-doped conditions. The application of gold-sputtered neat spider silk fibers for making four terminal flexible, clean, ohmic contacts to organic superconductor samples and carbon nanotube coated silk fibers for heart pulse monitoring sensor are demonstrated. The role of silk thin film in organic thin film transistor will be briefly discussed.

Steven, Eden; Jobiliong, Eric; Park, Jin Gyu; Paravastu, Anant; Davidson, Michael; Baird, Michelle; Alamo, Rufina; Kaner, Papatya; Brooks, James; Siegrist, Theo

2012-02-01

194

Total X-Ray Scattering of Spider Dragline Silk  

NASA Astrophysics Data System (ADS)

Total x-ray scattering measurements of spider dragline silk fibers from Nephila clavipes, Argiope aurantia, and Latrodectus hesperus all yield similar structure factors, with only small variations between the different species. Wide-angle x-ray scattering from fibers orientated perpendicular to the beam shows a high degree of anisotropy, and differential pair distribution functions obtained by integrating over wedges of the equatorial and meridian planes indicate that, on average, the majority (95%) of the atom-atom correlations do not extend beyond 1 nm. Futhermore, the atom-atom correlations between 1 and 3 nm are not associated with the most intense diffraction peaks at Q=1-2Å-1. Disordered molecular orientations along the fiber axis are consistent with proteins in similar structural arrangements to those in the equatorial plane, which may be associated with the silk’s greater flexibility in this direction.

Benmore, C. J.; Izdebski, T.; Yarger, J. L.

2012-04-01

195

Structure to function: Spider silk and human collagen  

Microsoft Academic Search

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

Olena S. Rabotyagova

2008-01-01

196

Molecular and mechanical characterization of aciniform silk: uniformity of iterated sequence modules in a novel member of the spider silk fibroin gene family.  

PubMed

Araneoid spiders use specialized abdominal glands to produce up to seven different protein-based silks/glues that have diverse physical properties. The fibroin sequences that encode aciniform fibers (wrapping silk) and the mechanical properties of these fibers have not been characterized previously. To gain a better understanding of the molecular radiation of spider silk fibroin genes, cDNA libraries derived from aciniform glands of the banded garden spider, Argiope trifasciata, were constructed, and unique silk transcripts were sequenced. There was evidence for a single silk fibroin gene that was expressed in the aciniform glands, and the inferred amino acid composition of the novel fibroin closely matched the amino acid contents of these glands. The inferred protein, aciniform spidroin 1 (AcSp1), is composed of highly homogenized repeats that are 200 amino acids in length. The long stretches of poly-alanine and glycine-alanine subrepeats, which are thought to account for the crystalline regions of minor ampullate and major ampullate fibers, are very poorly represented in AcSp1. The AcSp1 repeat unit is iterated minimally 14 times and does not display substantial sequence similarity to any previously described genes or proteins. Database searches, however, showed that the nonrepetitive carboxy-terminus contains stretches of matches to known spider fibroin sequences, suggesting that the AcSp1 gene is a highly divergent member of the spider silk gene family. In phylogenetic analyses of carboxy-terminal sequences from araneid spiders, the aciniform sequence did not group strongly with clusters of fibroins from the flagelliform, minor ampullate, or major ampullate silk glands. Comparisons of stress/strain curves for major ampullate, minor ampullate, and aciniform silks from Argiope trifasciata showed significant differences in ultimate strength, extensibility, and toughness. Remarkably, the toughness of aciniform silk was 50% greater than the highest values typically recorded for major ampullate silk. These differences in performance, in combination with the radical divergence at the sequence level among fibroin paralogs, suggest a possible linkage between silk fibroin sequences and performance that should be explored in future structural/functional studies of aciniform silk. PMID:15240839

Hayashi, Cheryl Y; Blackledge, Todd A; Lewis, Randolph V

2004-10-01

197

Processing of ?-glucosidase-silk fibroin nanoparticle bioconjugates and their characteristics.  

PubMed

Silk fibroin derived from Bombyx mori is a biomacromolecular protein with excellent biocompatibility. The aim of this work was to develop silk fibroin nanoparticles (SFNs) derived from the fibrous protein, which is a novel vector for enzyme modification in food processing. Silk fibroin was dissolved in highly concentrated CaCl2 and subjected to lengthy desalting in water. The resulting liquid silk, which contained water-soluble polypeptides with molecular mass ranging from 10 to 200 kDa, and ?-glucosidase were added rapidly into acetone. The ?-glucosidase molecules were embedded into silk fibroin nanoparticles, forming ?-glucosidase-silk fibroin nanoparticles (?G-SFNs) with a diameter of 50-150 nm. The enzyme activity of the ?G-SFN bioconjugates was determined with p-nitrophenyl-?-D-glucoside as the substrate, and the optimum conditions for the preparation of ?G-SFNs were investigated. The enzyme activity recovery of ?G-SFNs was 59.2 % compared to the free enzyme (specific activity was 1 U mg(-1)). The kinetic parameters of the ?G-SFNs and the free ?-glucosidase were the same. The ?G-SFNs had good operational stability and could be used repeatedly. These results confirmed that silk protein nanoparticles were good carriers as bioconjugates for the modification of enzymes with potential value for research and development. The method used in this study has potential applications in food processing and the production of flavour agents. PMID:24671567

Cao, Ting-Ting; Zhou, Zhen-Zhen; Zhang, Yu-Qing

2014-05-01

198

Polarized light microscopy, variability in spider silk diameters, and the mechanical characterization of spider silk  

Microsoft Academic Search

Spider silks possess a remarkable combination of high tensile strength and exten- sibility that makes them among the toughest materials known. Despite the potential exploi- tation of these properties in biotechnology, very few silks have ever been characterized mechanically. This is due in part to the difficulty of measuring the thin diameters of silk fibers. The largest silk fibers are

Todd A. Blackledge; Richard A. Cardullo; Cheryl Y. Hayashi

199

Structure to function: Spider silk and human collagen  

NASA Astrophysics Data System (ADS)

Nature has the ability to assemble a variety of simple molecules into complex functional structures with diverse properties. Collagens, silks and muscles fibers are some examples of fibrous proteins with self-assembling properties. One of the great challenges facing Science is to mimic these designs in Nature to find a way to construct molecules that are capable of organizing into functional supra-structures by self-assembly. In order to do so, a construction kit consisting of molecular building blocks along with a complete understanding on how to form functional materials is required. In this current research, the focus is on spider silk and collagen as fibrous protein-based biopolymers that can shed light on how to generate nanostructures through the complex process of self-assembly. Spider silk in fiber form offers a unique combination of high elasticity, toughness, and mechanical strength, along with biological compatibility and biodegrability. Spider silk is an example of a natural block copolymer, in which hydrophobic and hydrophilic blocks are linked together generating polymers that organize into functional materials with extraordinary properties. Since silks resemble synthetic block copolymer systems, we adopted the principles of block copolymer design from the synthetic polymer literature to build block copolymers based on spider silk sequences. Moreover, we consider spider silk to be an important model with which to study the relationships between structure and properties in our system. Thus, the first part of this work was dedicated to a novel family of spider silk block copolymers, where we generated a new family of functional spider silk-like block copolymers through recombinant DNA technology. To provide fundamental insight into relationships between peptide primary sequence, block composition, and block length and observed morphological and structural features, we used these bioengineered spider silk block copolymers to study secondary structure, morphological features and assembly. Aside from fundamental perspectives, we anticipate that these results will provide a blueprint for the design of precise materials for a range of potential applications such as controlled release devices, functional coatings, components of tissue regeneration materials and environmentally friendly polymers in future studies. In the second part of this work, human collagen type I was studied as another representative of the family of fibrous proteins. Collagen type I is the most abundant extracellular matrix protein in the human body, providing the basis for tissue structure and directing cellular functions. Collagen has a complex structural hierarchy, organized at different length scales, including the characteristic triple helical feature. In the present study we assessed the relationship between collagen structure (native vs. denatured) and sensitivity to UV radiation with a focus on changes in the primary structure, conformation, microstructure and material properties. Free radical reactions are involved in collagen degradation and a mechanism for UV-induced collagen degradation related to structure was proposed. The results from this study demonstrated the role of collagen supramolecular organization (triple helix) in the context of the effects of electromagnetic radiation on extracellular matrices. Owing to the fact that both silks and collagens are proteins that have found widespread interest for biomaterial related needs, we anticipate that the current studies will serve as a foundation for future biomaterial designs with controlled properties. Furthermore, fundamental insight into self-assembly and environmentally-2mediated degradation, will build a foundation for fundamental understanding of the remodeling and functions of these types of fibrous proteins in vivo and in vitro. This type of insight is essential for many areas of scientific inquiry, from drug delivery, to scaffolds for tissue engineering, and to the stability of materials in space.

Rabotyagova, Olena S.

200

Nanolayer Biomaterial Coatings of Silk Fibroin for Controlled Release  

PubMed Central

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 control of ?-sheet crystal content and the multilayer structure of the silk coatings correlated with the release properties of the incorporated compounds. In particular, higher crystallinity and a thicker silk capping layer suppressed the initial burst of release and prolonged the duration of release. These novel coatings and deposition approach provide a unique option to regulate structure and morphology, and thus release kinetics. The results also suggest these systems as a promising framework for surface engineering of biomaterials and medical devices to regulate the release of drugs, when considered with the all-aqueous process involved, the conformal nature of the coatings, the robust material properties of silk fibroin, and the degradability and biocompatibility of this family of protein. PMID:17628161

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

2009-01-01

201

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

PubMed Central

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(NO3)2-methanol, Ca(NO3)2-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 (?-form, type II ?-turn), while the other treatments produced more silk II (?-form, anti-parallel ?-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. PMID:22676291

2012-01-01

202

Untangling spider silk evolution with spidroin terminal domains  

PubMed Central

Background Spidroins are a unique family of large, structural proteins that make up the bulk of spider silk fibers. Due to the highly variable nature of their repetitive sequences, spidroin evolutionary relationships have principally been determined from their non-repetitive carboxy (C)-terminal domains, though they offer limited character data. The few known spidroin amino (N)-terminal domains have been difficult to obtain, but potentially contain critical phylogenetic information for reconstructing the diversification of spider silks. Here we used silk gland expression data (ESTs) from highly divergent species to evaluate the functional significance and phylogenetic utility of spidroin N-terminal domains. Results We report 11 additional spidroin N-termini found by sequencing ~1,900 silk gland cDNAs from nine spider species that shared a common ancestor > 240 million years ago. In contrast to their hyper-variable repetitive regions, spidroin N-terminal domains have retained striking similarities in sequence identity, predicted secondary structure, and hydrophobicity. Through separate and combined phylogenetic analyses of N-terminal domains and their corresponding C-termini, we find that combined analysis produces the most resolved trees and that N-termini contribute more support and less conflict than the C-termini. These analyses show that paralogs largely group by silk gland type, except for the major ampullate spidroins. Moreover, spidroin structural motifs associated with superior tensile strength arose early in the history of this gene family, whereas a motif conferring greater extensibility convergently evolved in two distantly related paralogs. Conclusions A non-repetitive N-terminal domain appears to be a universal attribute of spidroin proteins, likely retained from the origin of spider silk production. Since this time, spidroin N-termini have maintained several features, consistent with this domain playing a key role in silk assembly. Phylogenetic analyses of the conserved N- and C-terminal domains illustrate dramatic radiation of the spidroin gene family, involving extensive duplications, shifts in expression patterns and extreme diversification of repetitive structural sequences that endow spider silks with an unparalleled range of mechanical properties. PMID:20696068

2010-01-01

203

Silk Optics Mark Cronin-Golomb  

E-print Network

Technologies · Acquired by Allergan 2010 ­ Seri #12;1/27/14 3 Silk Materials ­ A Road to Sustainable High http://onlinelibrary.wiley.com/doi/10.1002/adma.201104477/full#fig2 Silk Materials ­ A Road1/27/14 1 Silk Optics Mark Cronin-Golomb Department of Biomedical Engineering Tufts University

Barthelat, Francois

204

Nephila clavipes Flagelliform silk-like GGX motifs contribute to extensibility and spacer motifs contribute to strength in synthetic spider silk fibers.  

PubMed

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 the Nephila clavipes flagelliform Flag protein that produces this type of fiber. The as-spun fibers were processed in 80% aqueous isopropanol using a standardized process for all four fiber types, which produced improved mechanical properties. Mechanical testing of the recombinant proteins determined that the GGX motif contributes extensibility and the spacer motif contributes strength to the recombinant fibers. Recombinant protein fibers containing the spacer motif were stronger than the proteins constructed without the spacer that contained only the GGX motif or the combination of the GGX and GPGGX motifs. The mechanical and structural X-ray diffraction analysis of the recombinant fibers provide data that suggests a functional role of the spacer motif that produces tensile strength, though the spacer motif is not clearly defined structurally. These results indicate that the spacer is likely a primary contributor of strength, with the GGX motif supplying mobility to the protein network of native N. clavipes flagelliform silk fibers. PMID:23646825

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

2013-06-10

205

Discrimination of cultivated silk and wild silk by conventional instrumental analyses.  

PubMed

In Japan, recent trends have seen wild silk preferred over cultivated silk because of its texture. Some cases of fraud have occurred where cultivated silk garments are sold as wild silk. Samples from these cases, morphological observation using light microscope and polarized microscope have been conducted in forensic science laboratories. Sometimes scanning electron microscopy was also carried out. However, the morphology of silk shows quite wide variation, which makes it difficult to discriminate wild and cultivated silks by this method. In this report, silk discrimination was investigated using conventional instrumental analyses commonly available in forensic laboratories, such as Fourier-transfer infrared spectrometry (FT-IR), pyrolysis-gas chromatography/mass spectrometry (pyr-GC/MS) and differential thermal analysis (DTA). By FT-IR, cultivated and wild silk gave similar infrared spectra, but wild silk had a characteristic peak at 965 cm(-1) from the deformation vibration of the carbon-carbon double bond of the indole ring. Comparison of the pyrograms of cultivated and wild silk showed that wild silk had large indole and skatole peaks that cultivated silk did not, and these peaks might arise from tryptophan. The results of thermogravimetry/DTA showed that the endothermic peak was about 40 °C higher for wild silk than for cultivated silk. Using a combination of these results, cultivated and wild silk could be discriminated by common forensic instrumental techniques. PMID:23742990

Matsuyama, Yuji; Nagatani, Yoshiaki; Goto, Toshiyuki; Suzuki, Shinichi

2013-09-10

206

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

PubMed

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

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

207

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

NASA Astrophysics Data System (ADS)

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

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

2005-06-01

208

Rate-dependent behavior of the amorphous phase of spider dragline silk.  

PubMed

The time-dependent stress-strain behavior of spider dragline silk was already observed decades ago, and has been attributed to the disordered sequences in silk proteins, which compose the soft amorphous matrix. However, the actual molecular origin and magnitude of internal friction within the amorphous matrix has remained inaccessible, because experimentally decomposing the mechanical response of the amorphous matrix from the embedded crystalline units is challenging. Here, we used atomistic molecular dynamics simulations to obtain friction forces for the relative sliding of peptide chains of Araneus diadematus spider silk within bundles of these chains as a representative unit of the amorphous matrix in silk fibers. We computed the friction coefficient and coefficient of viscosity of the amorphous phase to be in the order of 10(-6) Ns/m and 10(4) Ns/m(2), respectively, by extrapolating our simulation data to the viscous limit. Finally, we used a finite element method for the amorphous phase, solely based on parameters derived from molecular dynamics simulations including the newly determined coefficient of viscosity. With this model the time scales of stress relaxation, creep, and hysteresis were assessed, and found to be in line with the macroscopic time-dependent response of silk fibers. Our results suggest the amorphous phase to be the primary source of viscosity in silk and open up the avenue for finite element method studies of silk fiber mechanics including viscous effects. PMID:24896131

Patil, Sandeep P; Markert, Bernd; Gräter, Frauke

2014-06-01

209

Identification and synthesis of novel biomaterials based on spider structural silk fibers  

NASA Astrophysics Data System (ADS)

The diversity in function and mechanical behavior of spider silks, and the ability to produce these silks recombinantly, have tremendous potential in creating a new class of biomimetic materials. Here we investigate the structural and mechanical properties of pyriform silks from the golden orb-weaver, Nephila clavipes. Nanoscale indentation measurements using atomic force microscopy on natural pyriform silk suggests that this biomaterial has high toughness that may be suitable for dissipating high amounts of mechanical energy. We also observed the occurrence of highly organized nanocrystals within the pyriform silk fibers that may contribute to the remarkable energy dissipation capability of these silks. It has been demonstrated that poly-(Gly-Ala) and poly-Ala stretches within the internal block repeat modules of dragline silk fibroins form nanocrystals, and these nanocrystalline structures may be responsible for the high extensibility of the dragline silks. In contrast, amino acid sequence analysis shows that PySp2 does not contain the same motifs. In the absence of poly-(Gly-Ala) and poly-Ala repeats, we hypothesized that PySp2 contains new protein motifs sufficient to polymerize into functional structures. To investigate the functional contributions of these novel motifs during pyriform fiber formation, we expressed different recombinant PySp2 fibroins with various segments spanning its block repeat units. We demonstrate that PySp2 recombinant proteins with the Pro-rich sub-block domain (PXP motifs, where X= sub-set of the amino acids A, L, or R) and/or the Ser + Gln + Ala-rich sub-block domain (QQSSVAQS motifs) are sufficient for artificial fiber formation. Moreover, we show that recombinant PySp2 proteins that contain a single block repeat unit can self-assemble into foam-like nanostructures. Collectively, our findings support the use of PySp2 recombinant proteins for a wide range of biomimetic materials with morphologies ranging from fibers to porous structures.

Hsia, Yang; Gnesa, Eric; Tang, Simon; Jeffery, Felicia; Geurts, Paul; Zhao, Liang; Franz, Andreas; Vierra, Craig

2011-11-01

210

Helicoidal multi-lamellar features of RGD-functionalized silk biomaterials for corneal tissue engineering  

Microsoft Academic Search

RGD-coupled silk protein-biomaterial lamellar systems were prepared and studied with human cornea fibroblasts (hCFs) to match functional requirements. A strategy for corneal tissue engineering was pursued to replicate the structural hierarchy of human corneal stroma within thin stacks of lamellae-like tissues, in this case constructed from scaffolds constructed with RGD-coupled, patterned, porous, mechanically robust and transparent silk films. The influence

Eun Seok Gil; Biman B. Mandal; Sang-Hyug Park; Jeffrey K. Marchant; Fiorenzo G. Omenetto; David L. Kaplan

2010-01-01

211

Judaism and the Silk Route.  

ERIC Educational Resources Information Center

Demonstrates that the Judeans traveled along the Ancient Silk Route. Discusses the Iranian influence on the formation of Jewish religious ideas. Considers the development of Jewish trade networks, focusing on the Radanites (Jewish traders), the Jewish presence in the Far East, and the survival of Judaism in central Asia. (CMK)

Foltz, Richard

1998-01-01

212

Silk-based nanocomplexes with tumor-homing peptides for tumor-specific gene delivery.  

PubMed

Nanoscale complexes of recombinant silk molecules containing THPs with DNA are designed as less cytotoxic and highly target-specific gene carriers. Genetically engineered silk proteins containing poly(L-lysine) domains to interact with pDNA and the THP to bind to specific tumorigenic cells for target-specific pDNA delivery are prepared, followed by in vitro transfection into MDA-MB-435 melanoma cells, highly metastatic human breast tumor MDA-MB-231?cells, and non-tumorigenic MCF-10A breast epithelial cells. The silk/poly(L-lysine) block copolymer containing Lyp1 (ML-Lyp1) shows significant differences from silk/poly(L-lysine) block copolymer containing F3 (ML-F3) in cytotoxicity to MCF10A cells. ML-F3 is the most promising candidate for target delivery into tumorigenic cells. PMID:22052706

Numata, Keiji; Mieszawska-Czajkowska, Aneta J; Kvenvold, Laura A; Kaplan, David L

2012-01-01

213

Fabrication and Biocompatibility of Electrospun Silk Biocomposites  

PubMed Central

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. PMID:24957869

Wei, Kai; Kim, Byoung-Suhk; Kim, Ick-Soo

2011-01-01

214

Secrets of the Silk Road  

NSDL National Science Digital Library

Secrets of the Silk Road is another worthy addition to the growing body of Web-based information about the Silk Road (see, for example, The International Dunhuang Project, founded in 1998 and mentioned in several Scout Reports), launched by the Bowers Museum of Santa Ana, California and presented by University of Pennsylvania Museum of Archaeology and Anthropology. This site includes artifacts documenting the history of the Silk Road in the vast Tarim Basin in the Far Western Xinjiang Uyghur Autonomous Region of China. Three well-preserved mummies from the Tarim Basin have been brought to the United States for the exhibition, along with lavish grave goods. Visitors to the website can view images of "The Beauty of Xiaohe," a female mummy from 1800-1500 BCE and other treasures ranging from a Chrysanthemum Shaped Dessert from the 5th-3rd century BCE, to gold objects such as mask from 5th-6th century CE or the Gold Plaque with Lion Design, 5th-3rd Century BCE.

215

A hybrid silk/RADA-based fibrous scaffold with triple hierarchy for ligament regeneration.  

PubMed

While silk-based microfibrous scaffolds possess excellent mechanical properties and have been used for ligament tissue-engineering applications, the microenvironment in these scaffolds is not biomimetic. We hypothesized that coating a hybrid silk scaffold with an extracellular matrix (ECM)-like network of self-assembling peptide nanofibers would provide a biomimetic three-dimensional nanofibrous microenvironment and enhance ligament tissue regeneration after bone marrow-derived mesenchymal stem cell (BMSC)-seeding. A novel scaffold possessing a triple structural hierarchy comprising macrofibrous knitted silk fibers, a silk microsponge, and a peptide nanofiber mesh was developed by coating self-assembled RADA16 peptide nanofibers on a silk microfiber-reinforced-sponge scaffold. Compared with the uncoated control, RADA-coated scaffolds showed enhanced BMSC proliferation, metabolism, and fibroblastic differentiation during the 3 weeks of culture. BMSC-seeded RADA-coated scaffolds showed an increasing temporal expression of key fibroblastic ECM proteins (collagen type I and III, tenascin-C), with a significantly higher tenascin-C expression compared with the controls. BMSC-seeded RADA-coated scaffolds also showed a temporal increase in total collagen and glycosaminoglycan production (the amount produced being higher than in control scaffolds) during 3 weeks of culture, and possessed 7% higher maximum tensile load compared with the BMSC-seeded control scaffolds. The results indicate that the BMSC-seeded RADA-coated hybrid silk scaffold system has the potential for use in ligament tissue-engineering applications. PMID:22429111

Chen, Kelei; Sahoo, Sambit; He, Pengfei; Ng, Kian Siang; Toh, Siew Lok; Goh, James C H

2012-07-01

216

Reproducibility of the tensile properties of spider (Argiope trifasciata) silk obtained by forced silking.  

PubMed

A modified forced silking procedure was developed to allow an accurate study of the tensile properties of spider (Argiope trifasciata) silk, especially the characterization of the variability of the tensile properties of forcibly silked fibers. The procedure involves an immobilization technique that does not require anesthetization of the spider, a mode of collection that allows immediate access to any silk sample with a minimum manipulation, and a technique to measure the diameters of the spider silk fibers systematically. The forcibly silked fibers obtained by this procedure show reproducible tensile properties in terms of force-displacement curves as well as stress-strain curves. Furthermore, reproducibility also extends to forcibly silked fibers obtained from different spiders when stress-strain is considered. PMID:15612009

Guinea, Gustavo V; Elices, Manuel; Real, José Ignacio; Gutiérrez, Sara; Pérez-Rigueiro, José

2005-01-01

217

Tunable Silk: Using Microfluidics to Fabricate Silk Fibers with Controllable Properties  

PubMed Central

Despite widespread use of silk, it remains a significant challenge to fabricate fibers with properties similar to native silk. It has recently been recognized that the key to tuning silk fiber properties lies in controlling internal structure of assembled ?-sheets. We report an advance in the precise control of silk fiber formation with control of properties via microfluidic solution spinning. We use an experimental approach combined with modeling to accurately predict and independently tune fiber properties including Young’s modulus and diameter to customize fibers. This is the first reported microfluidic approach capable of fabricating functional fibers with predictable properties and provides new insight into the structural transformations responsible for the unique properties of silk. Unlike bulk processes, our method facilitates the rapid and inexpensive fabrication of fibers from small volumes (50 ?L) that can be characterized to investigate sequence-structure-property relationships to optimize recombinant silk technology to match and exceed natural silk properties. PMID:21438624

Kinahan, Michelle E.; Filippidi, Emmanouela; Koster, Sarah; Hu, Xiao; Evans, Heather M.; Pfohl, Thomas; Kaplan, David L.; Wong, Joyce

2011-01-01

218

Complex gene expression in the dragline silk producing glands of the Western black widow (Latrodectus hesperus)  

PubMed Central

Background Orb-web and cob-web weaving spiders spin dragline silk fibers that are among the strongest materials known. Draglines are primarily composed of MaSp1 and MaSp2, two spidroins (spider fibrous proteins) expressed in the major ampullate (MA) silk glands. Prior genetic studies of dragline silk have focused mostly on determining the sequence of these spidroins, leaving other genetic aspects of silk synthesis largely uncharacterized. Results Here, we used deep sequencing to profile gene expression patterns in the Western black widow, Latrodectus hesperus. We sequenced millions of 3?-anchored “tags” of cDNAs derived either from MA glands or control tissue (cephalothorax) mRNAs, then associated the tags with genes by compiling a reference database from our newly constructed normalized L. hesperus cDNA library and published L. hesperus sequences. We were able to determine transcript abundance and alternative polyadenylation of each of three loci encoding MaSp1. The ratio of MaSp1:MaSp2 transcripts varied between individuals, but on average was similar to the estimated ratio of MaSp1:MaSp2 in dragline fibers. We also identified transcription of TuSp1 in MA glands, another spidroin family member that encodes the primary component of egg-sac silk, synthesized in tubuliform glands. In addition to the spidroin paralogs, we identified 30 genes that are more abundantly represented in MA glands than cephalothoraxes and represent new candidates for involvement in spider silk synthesis. Conclusions Modulating expression rates of MaSp1 variants as well as MaSp2 and TuSp1 could lead to differences in mechanical properties of dragline fibers. Many of the newly identified candidate genes likely encode secreted proteins, suggesting they could be incorporated into dragline fibers or assist in protein processing and fiber assembly. Our results demonstrate previously unrecognized transcript complexity in spider silk glands. PMID:24295234

2013-01-01

219

Total x-ray scattering of spider dragline silk.  

PubMed

Total x-ray scattering measurements of spider dragline silk fibers from Nephila clavipes, Argiope aurantia, and Latrodectus hesperus all yield similar structure factors, with only small variations between the different species. Wide-angle x-ray scattering from fibers orientated perpendicular to the beam shows a high degree of anisotropy, and differential pair distribution functions obtained by integrating over wedges of the equatorial and meridian planes indicate that, on average, the majority (95%) of the atom-atom correlations do not extend beyond 1 nm. Futhermore, the atom-atom correlations between 1 and 3 nm are not associated with the most intense diffraction peaks at Q=1-2 Å(-1). Disordered molecular orientations along the fiber axis are consistent with proteins in similar structural arrangements to those in the equatorial plane, which may be associated with the silk's greater flexibility in this direction. PMID:22680907

Benmore, C J; Izdebski, T; Yarger, J L

2012-04-27

220

Development and characterization of silk fibroin coated quantum dots  

NASA Astrophysics Data System (ADS)

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.

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

2008-02-01

221

Seljuk Muqarnas along the Silk Road  

Microsoft Academic Search

The film Seljuk Muqarnas along the Silk Road gives an overview of muqarnas, stalactite vaults, in Seljuk style architecture (1038-1194). The muqarnas are located in portals and niches of car- avansaraies, madrassas and mosques. Starting with the Sultan Han near Kayseri we follow the Silk Road westward till Konya and finally show the Arslanhane Camii in Ankara. Video recordings alternate

Silvia Harmsen; Daniel Jungblut; Susanne Kromker

222

A comparison of convergently evolved insect silks that share ?-sheet molecular structure.  

PubMed

Raspy crickets produce silk webs that are used to build shelters. These webs have been found to consist of both fiber and film components. Raman spectra obtained from both components were found to be very similar for a given species. The protein structure of the fibers and films produced by both species was predominately ?-sheet with lesser amounts of ?-turns, unordered and ?-helical protein also detected. The orientation of the ?-sheet backbone in the fiber was determined to be parallel to the fiber axis. Compared to cocoon and dragline silk the orientation distribution exhibits a significant randomly orientated protein component. Amino acid analysis confirmed the presence of glycine, serine, and alanine in both species, which are known to form antiparallel ?-sheet structures. Both species, although at significantly different concentrations, where found to contain proline. This amino acid is uncommon in insect silks, and likely involved in increasing fiber elasticity. © 2013 Wiley Periodicals, Inc. Biopolymers 101: 630-639, 2014. PMID:24170682

Church, Jeffrey S; Woodhead, Andrea L; Walker, Andrew A; Sutherland, Tara D

2014-06-01

223

Size-dependent mechanical properties of beta-structures in protein materials  

E-print Network

Protein materials such as spider silk can be exceptionally strong, and they can stretch tremendously before failure. Notably, silks are made entirely of proteins, which owe their structure and stability to weak molecular ...

Keten, Sinan

2010-01-01

224

On the strength of ?-sheet crystallites of Bombyx mori silk fibroin.  

PubMed

Silk fibroin, a natural multi-domain protein, has attracted great attention due to its superior mechanical properties such as ultra-high strength and stretchability, biocompatibility, as well as its versatile biodegradability and processability. It is mainly composed of ?-sheet crystallites and amorphous domains. Although its strength is well known to be controlled by the dissociation of protein chains from ?-sheet crystallites, the way that water as the solvent affects its strength and the reason that its theoretically predicted strength is several times higher than experimental measurement remain unclear. We perform all-atom molecular dynamics simulations on a ?-sheet crystallite of Bombyx mori silk. We find that water solvent reduces the number and strength of hydrogen bonds between ?-chains, and thus greatly weakens the strength of silk fibroin. By dissociating protein chains at different locations from the crystallite, we also find that the pulling strength for the interior chains is several times higher than that for the surface/corner chains, with the former being consistent with the theoretically predicted value, while the latter on par with the experimental value. It is shown that the weakest rupture strength controls the failure strength of silk fibre. Hence, this work sheds light on the role of water in the strength of silk fibroin and also provides clues on the origin of the strength difference between theory and experiment. PMID:24789564

Cheng, Yuan; Koh, Leng-Duei; Li, Dechang; Ji, Baohua; Han, Ming-Yong; Zhang, Yong-Wei

2014-07-01

225

A novel methodology to explore the viscoelasticity of spider major ampullate silk.  

PubMed

Even before material science was a recognized discipline, the amazing mechanical properties of spider silk were documented and became the object of much study. In addition to the exceptional material properties of spider silk and the reported low immunogenicity, its concatenated amino acid motif arrangement facilitates a distinct possibility of manipulating the silk to create a designer biomaterial for medical applications. Crystalline protein regions imbedded in a mobile protein matrix give it a distinct set of viscoelastic abilities. Consequently, elasticity cannot be simply quantified by only measuring extensibility. To understand how the sequence of the major ampullate proteins affects elasticity, the hysteresis of single fibers from two different species, Argiope aurantia and Nephila clavipes, were examined using cyclic loading and unloading. The yield point that discriminates a transition from elastic extension to a plastic extension was analyzed by examining three different properties: Young's modulus, energy recovery and slack in the fiber after recovery. Young's modulus remained relatively constant regardless of the cycle. However, the energy recovered decreased as the slack and cycle number increased. Large standard deviations masked any quantitative differences between species and substantiated the necessity of developing synthetic silk to harness the amazing mechanical properties of spider silk. PMID:20799185

Brooks, A E; Brothers, T J; Creager, M S; Lewis, R V

2007-01-01

226

Sustained volume retention in vivo with adipocyte and lipoaspirate seeded silk scaffolds  

PubMed Central

Current approaches to soft tissue regeneration include the use of fat grafts, natural or synthetic biomaterials as filler materials. Fat grafts and natural biomaterials resorb too quickly to maintain tissue regeneration, while synthetic materials do not degrade or regenerate tissue. Here, we present a simple approach to volume stable filling of soft tissue defects. In this study, we combined lipoaspirate with a silk protein matrix in a subcutaneous rat model. Silk biomaterials can be tailored to fit a variety of needs, and here were processed silk biomaterials into a porous sponge format to allow for tissue ingrowth while remaining mechanically robust. Over an 18 month period, the lipoaspirate seeded silk protein matrix regenerated subcutaneous adipose tissue while maintaining the original implanted volume. A silk protein matrix alone was not sufficient to regenerate adipose tissue, but yielded a fibrous tissue, although implanted volume was maintained. This work presents a significant improvement to the standard approaches to filling soft tissue defects by matching biomaterial degradation and tissue regeneration profiles. PMID:23374707

Bellas, Evangelia; Panilaitis, Bruce J.B.; Glettig, Dean L.; Kirker-Head, Carl A.; Yoo, James J.; Marra, Kacey G.; Rubin, J. Peter; Kaplan, David L.

2013-01-01

227

Silk-Road Scholarship Program Background The Silk-Road Scholarship Program is jointly funded by the SBS Cultural Foundation  

E-print Network

Silk-Road Scholarship Program Background The Silk-Road Scholarship Program is jointly funded fee and living expenses. Students from countries located in the area of the Silk-Road may apply. We are currently enrolling or who graduated from the eligible universities of the Silk-Road Scholarship Program

Choi, Sunghyun

228

Evaluation of Gel Spun Silk-Based Biomaterials in a Murine Model of Bladder Augmentation  

PubMed Central

Currently, gastrointestinal segments are considered the gold standard for bladder reconstructive procedures. However, significant complications including chronic urinary tract infection, metabolic abnormalities, urinary stone formation, bowel dysfunction, and secondary malignancies are associated with this approach. Biomaterials derived from silk fibroin may represent a superior alternative due their robust mechanical properties, biodegradable features, and processing plasticity. In the present study, we evaluated the efficacy of a gel spun silk-based matrix for bladder augmentation in a murine model. Over the course of 70 d implantation period, H&E and Masson’s trichrome (MTS) analysis revealed that silk matrices were capable of supporting both urothelial and smooth muscle regeneration at the defect site. Prominent uroplakin and contractile protein expression (?-actin, calponin, and SM22?) was evident by immunohistochemical analysis demonstrating maturation of the reconstituted bladder wall compartments. Gel spun silk matrices also elicited a minimal acute inflammatory reaction following 70 d of bladder integration, in contrast to parallel assessments of small intestinal submucosa (SIS) and polyglycolic acid (PGA) matrices which routinely promoted evidence of fibrosis and chronic inflammatory responses. Voided stain on paper analysis revealed that silk augmented animals displayed similar voiding patterns in comparison to non surgical controls by 42 d of implantation. In addition, cystometric evaluations of augmented bladders at 70 d post-op demonstrated that silk scaffolds supported significant increases in bladder capacity, voided volume, and flow rate while maintaining similar degrees of compliance relative to the control group. These results provide evidence for the utility of gel spun silk-based matrices for functional bladder tissue engineering applications. PMID:20951426

Mauney, Joshua R.; Cannon, Glenn M.; Lovett, Michael L.; Gong, Edward M.; DiVizio, Dolores; Kaplan, David L.; Adam, Rosalyn M.; Estrada, Carlos R.

2013-01-01

229

Dating silk by capillary electrophoresis mass spectrometry.  

PubMed

A new capillary electrophoresis mass spectrometry (CE-MS) technique is introduced for age estimation of silk textiles based on amino acid racemization rates. With an L to D conversion half-life of ~2500 years for silk (B. mori) aspartic acid, the technique is capable of dating silk textiles ranging in age from several decades to a few-thousand-years-old. Analysis required only ~100 ?g or less of silk fiber. Except for a 2 h acid hydrolysis at 110 °C, no other sample preparation is required. The CE-MS analysis takes ~20 min, consumes only nanoliters of the amino acid mixture, and provides both amino acid composition profiles and D/L ratios for ~11 amino acids. PMID:21913691

Moini, Mehdi; Klauenberg, Kathryn; Ballard, Mary

2011-10-01

230

In vitro chondrogenic differentiation of human adipose-derived stem cells with silk scaffolds  

PubMed Central

Human adipose-derived stem cells have shown chondrogenic differentiation potential in cartilage tissue engineering in combination with natural and synthetic biomaterials. In the present study, we hypothesized that porous aqueous-derived silk protein scaffolds would be suitable for chondrogenic differentiation of human adipose-derived stem cells. Human adipose-derived stem cells were cultured up to 6 weeks, and cell proliferation and chondrogenic differentiation were investigated and compared with those in conventional micromass culture. Cell proliferation, glycosaminoglycan, and collagen levels in aqueous-derived silk scaffolds were significantly higher than in micromass culture. Transcript levels of SOX9 and type II collagen were also upregulated in the cell–silk constructs at 6 weeks. Histological examination revealed that the pores of the silk scaffolds were filled with cells uniformly distributed. In addition, chondrocyte-specific lacunae formation was evident and distributed in the both groups. The results suggest the biodegradable and biocompatible three-dimensional aqueous-derived silk scaffolds provided an improved environment for chondrogenic differentiation compared to micromass culture. PMID:23316274

Kim, Hyeon Joo; Park, Sang-Hyug; Durham, Jennah; Gimble, Jeffrey M; Kaplan, David L

2012-01-01

231

Processing and characterisation of a novel electropolymerized silk fibroin hydrogel membrane  

PubMed Central

Silk fibroin can be made into various forms of biocompatible medical materials, including hydrogel due to its excellent properties. Here, we report a novel method for the preparation of electropolymerized silk fibroin hydrogel membrane (ESFHM), which is formed on a nanoporous film as a barrier using a homemade device at a higher DC voltage. Regenerated silk fibroin solution in Tris buffer (pH 6.55–7.55) was added into a reservoir with a negative charge, and the silk molecules migrated toward the positive charge at 80VDC, resulting in the formation of the ESFHM on the barrier film. Barrier film with a MWCO of 10?kDa is favourable to the formation of the ESFHM. Semi-transparent ESFHM with a swelling ratio of 1056.4% predominantly consisted of a mixture of ?-sheets and ?-helix crystalline structures. SEM studies revealed that the ESFHM consisted of a 3D mesh structure woven by a chain of silk fibroin nanoparticles with a size of approximately 30 nanometres, similar to a pearl necklace. In vitro studies indicated that the ESFHM was degradable and was sufficient for cell adhesion and growth. Thus, ESFHM is a promising candidate for loading bioactive protein and appropriate cells, as artificial skin or for use in transplantation. PMID:25154713

Wang, Hai-Yan; Zhang, Yu-Qing

2014-01-01

232

Processing and characterisation of a novel electropolymerized silk fibroin hydrogel membrane.  

PubMed

Silk fibroin can be made into various forms of biocompatible medical materials, including hydrogel due to its excellent properties. Here, we report a novel method for the preparation of electropolymerized silk fibroin hydrogel membrane (ESFHM), which is formed on a nanoporous film as a barrier using a homemade device at a higher DC voltage. Regenerated silk fibroin solution in Tris buffer (pH 6.55-7.55) was added into a reservoir with a negative charge, and the silk molecules migrated toward the positive charge at 80VDC, resulting in the formation of the ESFHM on the barrier film. Barrier film with a MWCO of 10?kDa is favourable to the formation of the ESFHM. Semi-transparent ESFHM with a swelling ratio of 1056.4% predominantly consisted of a mixture of ?-sheets and ?-helix crystalline structures. SEM studies revealed that the ESFHM consisted of a 3D mesh structure woven by a chain of silk fibroin nanoparticles with a size of approximately 30 nanometres, similar to a pearl necklace. In vitro studies indicated that the ESFHM was degradable and was sufficient for cell adhesion and growth. Thus, ESFHM is a promising candidate for loading bioactive protein and appropriate cells, as artificial skin or for use in transplantation. PMID:25154713

Wang, Hai-Yan; Zhang, Yu-Qing

2014-01-01

233

Thermal characterization of social vespid silk  

Microsoft Academic Search

In this paper we try to establish a link between the microclimate in the wasp nest and the structure and thermal stability\\u000a of vespid silk. We suggest that there are at least two types of water that is absorbed by the silk of Oriental hornets, namely,\\u000a surface water and intrinsic structural water. The release of both types of water was

Diana Golodnitsky; Avi Ulus; Jacob S. Ishay

2003-01-01

234

Spinning an elastic ribbon of spider silk.  

PubMed

The Sicarid spider Loxosceles laeta spins broad but very thin ribbons of elastic silk that it uses to form a retreat and to capture prey. A structural investigation into this spider's silk and spinning apparatus shows that these ribbons are spun from a gland homologous to the major ampullate gland of orb web spiders. The Loxosceles gland is constructed from the same basic parts (separate transverse zones in the gland, a duct and spigot) as other spider silk glands but construction details are highly specialized. These differences are thought to relate to different ways of spinning silk in the two groups of spiders. Loxosceles uses conventional die extrusion, feeding a liquid dope (spinning solution) to the slit-like die to form a flat ribbon, while orb web spiders use an extrusion process in which the silk dope is processed in an elongated duct to produce a cylindrical thread. This is achieved by the combination of an initial internal draw down, well inside the duct, and a final draw down, after the silk has left the spigot. The spinning mechanism in Loxosceles may be more ancestral. PMID:11911779

Knight, David P; Vollrath, Fritz

2002-02-28

235

Spinning an elastic ribbon of spider silk.  

PubMed Central

The Sicarid spider Loxosceles laeta spins broad but very thin ribbons of elastic silk that it uses to form a retreat and to capture prey. A structural investigation into this spider's silk and spinning apparatus shows that these ribbons are spun from a gland homologous to the major ampullate gland of orb web spiders. The Loxosceles gland is constructed from the same basic parts (separate transverse zones in the gland, a duct and spigot) as other spider silk glands but construction details are highly specialized. These differences are thought to relate to different ways of spinning silk in the two groups of spiders. Loxosceles uses conventional die extrusion, feeding a liquid dope (spinning solution) to the slit-like die to form a flat ribbon, while orb web spiders use an extrusion process in which the silk dope is processed in an elongated duct to produce a cylindrical thread. This is achieved by the combination of an initial internal draw down, well inside the duct, and a final draw down, after the silk has left the spigot. The spinning mechanism in Loxosceles may be more ancestral. PMID:11911779

Knight, David P; Vollrath, Fritz

2002-01-01

236

The mechanical design of spider silks: from fibroin sequence to mechanical function.  

PubMed

Spiders produce a variety of silks, and the cloning of genes for silk fibroins reveals a clear link between protein sequence and structure-property relationships. The fibroins produced in the spider's major ampullate (MA) gland, which forms the dragline and web frame, contain multiple repeats of motifs that include an 8-10 residue long poly-alanine block and a 24-35 residue long glycine-rich block. When fibroins are spun into fibres, the poly-alanine blocks form (&bgr;)-sheet crystals that crosslink the fibroins into a polymer network with great stiffness, strength and toughness. As illustrated by a comparison of MA silks from Araneus diadematus and Nephila clavipes, variation in fibroin sequence and properties between spider species provides the opportunity to investigate the design of these remarkable biomaterials. PMID:10562512

Gosline, J M; Guerette, P A; Ortlepp, C S; Savage, K N

1999-12-01

237

Inducing ?-Sheets Formation in Synthetic Spider Silk Fibers by Aqueous Post-Spin Stretching  

PubMed Central

As a promising biomaterial with numerous potential applications, various types of synthetic spider silk fibers have been produced and studied in an effort to produce manmade fibers with mechanical and physical properties comparable to those of native spider silk. In this study, two recombinant proteins based on Nephila clavipes Major ampullate Spidroin 1 (MaSp1) consensus repeat sequence were expressed and spun into fibers. Mechanical test results showed that fiber spun from the higher molecular weight protein had better overall mechanical properties (70 KD versus 46 KD), whereas postspin stretch treatment in water helped increase fiber tensile strength significantly. Carbon-13 solid-state NMR studies of those fibers further revealed that the postspin stretch in water promoted protein molecule rearrangement and the formation of ?-sheets in the polyalanine region of the silk. The rearrangement correlated with improved fiber mechanical properties and indicated that postspin stretch is key to helping the spider silk proteins in the fiber form correct secondary structures, leading to better quality fibers. PMID:21574576

Hinman, Michael B.; Holland, Gregory P.; Yarger, Jeffery L.; Lewis, Randolph V.

2012-01-01

238

Inducing ?-sheets formation in synthetic spider silk fibers by aqueous post-spin stretching.  

PubMed

As a promising biomaterial with numerous potential applications, various types of synthetic spider silk fibers have been produced and studied in an effort to produce man-made fibers with mechanical and physical properties comparable to those of native spider silk. In this study, two recombinant proteins based on Nephila clavipes Major ampullate Spidroin 1 (MaSp1) consensus repeat sequence were expressed and spun into fibers. Mechanical test results showed that fiber spun from the higher molecular weight protein had better overall mechanical properties (70 KD versus 46 KD), whereas postspin stretch treatment in water helped increase fiber tensile strength significantly. Carbon-13 solid-state NMR studies of those fibers further revealed that the postspin stretch in water promoted protein molecule rearrangement and the formation of ?-sheets in the polyalanine region of the silk. The rearrangement correlated with improved fiber mechanical properties and indicated that postspin stretch is key to helping the spider silk proteins in the fiber form correct secondary structures, leading to better quality fibers. PMID:21574576

An, Bo; Hinman, Michael B; Holland, Gregory P; Yarger, Jeffery L; Lewis, Randolph V

2011-06-13

239

Preparation of Conducting Fibres from Cellulose and Silk by Polypyrrole Coating  

Microsoft Academic Search

cellulose fibre (e.g., cotton) and a protein fibre (e.g., silk), after under going pre- treatment and activating stages, were coated by conductive polymers such as pyr- role by two methods: Vapour and liquid phases. The value of penetration of con- ductive polymer into fibres by scanning electron microscope (SEM) was determined. Then assessments were made on physical properties and optimization

S. Hossein Hosseini; Ali Pairovi

240

Bioengineered silk gene delivery system for nuclear targeting.  

PubMed

Gene delivery research has gained momentum with the use of lipophilic vectors that mimic viral systems to increase transfection efficiency. Maintaining cell viability with these systems remains a major challenge. Therefore, biocompatible biopolymers that are designed by combining non-immunological viral mimicking components with suitable carrier are explored to address these limitations. In the present study, dragline silk recombinant proteins are modified with DNA condensing units and the proton sponge endosomal escape pathway is utilized for enhanced delivery. Transfection efficiency in a COS-7 cell line is enhanced compared to lipofectamine and polyethyleneimine (PEI), as is cell viability. PMID:24889658

Yigit, Sezin; Tokareva, Olena; Varone, Antonio; Georgakoudi, Irene; Kaplan, D L

2014-09-01

241

Novel silk fibroin/elastin wound dressings.  

PubMed

Silk fibroin (SF) and elastin (EL) scaffolds were successfully produced for the first time for the treatment of burn wounds. The self-assembly properties of SF, together with the excellent chemical and mechanical stability and biocompatibility, were combined with elastin protein to produce scaffolds with the ability to mimic the extracellular matrix (ECM). Porous scaffolds were obtained by lyophilization and were further crosslinked with genipin (GE). Genipin crosslinking induces the conformational transition from random coil to ?-sheet of SF chains, yielding scaffolds with smaller pore size and reduced swelling ratios, degradation and release rates. All results indicated that the composition of the scaffolds had a significant effect on their physical properties, and that can easily be tuned to obtain scaffolds suitable for biological applications. Wound healing was assessed through the use of human full-thickness skin equivalents (EpidermFT). Standardized burn wounds were induced by a cautery and the best re-epithelialization and the fastest wound closure was obtained in wounds treated with 50SF scaffolds; these contain the highest amount of elastin after 6 days of healing in comparison with other dressings and controls. The cytocompatibility demonstrated with human skin fibroblasts together with the healing improvement make these SF/EL scaffolds suitable for wound dressing applications. PMID:22546517

Vasconcelos, Andreia; Gomes, Andreia C; Cavaco-Paulo, Artur

2012-08-01

242

Silk Roads or Steppe Roads? The Silk Roads in World History.  

ERIC Educational Resources Information Center

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)

Christian, David

2000-01-01

243

Polarized light microscopy, variability in spider silk diameters, and the mechanical characterization of spider silk  

E-print Network

is as precise as scanning electron microscopy (SEM) across repeated meas- urements of individual samples of silk spiders. Additional key words: major ampullate, flagelliform, fibers, tensile test, orb web Spider silks. One of two different strategies is usually employed. The first approach is to use scanning electron

Blackledge, Todd

244

In vitro evaluation of bi-layer silk fibroin scaffolds for gastrointestinal tissue engineering  

PubMed Central

Silk fibroin scaffolds were investigated for their ability to support attachment, proliferation, and differentiation of human gastrointestinal epithelial and smooth muscle cell lines in order to ascertain their potential for tissue engineering. A bi-layer silk fibroin matrix composed of a porous silk fibroin foam annealed to a homogeneous silk fibroin film was evaluated in parallel with small intestinal submucosa scaffolds. AlamarBlue analysis revealed that silk fibroin scaffolds supported significantly higher levels of small intestinal smooth muscle cell, colon smooth muscle cell, and esophageal smooth muscle cell attachment in comparison to small intestinal submucosa. Following 7?days of culture, relative numbers of each smooth muscle cell population maintained on both scaffold groups were significantly elevated over respective 1-day levels—indicative of cell proliferation. Real-time reverse transcription polymerase chain reaction and immunohistochemical analyses demonstrated that both silk fibroin and small intestinal submucosa scaffolds were permissive for contractile differentiation of small intestinal smooth muscle cell, colon smooth muscle cell, esophageal smooth muscle cell as determined by significant upregulation of ?-smooth muscle actin and SM22? messenger RNA and protein expression levels following transforming growth factor-?1 stimulation. AlamarBlue analysis demonstrated that both matrix groups supported similar degrees of attachment and proliferation of gastrointestinal epithelial cell lines including colonic T84 cells and esophageal epithelial cells. Following 14?days of culture on both matrices, spontaneous differentiation of T84 cells toward an enterocyte lineage was confirmed by expression of brush border enzymes, lactase, and maltase, as determined by real-time reverse transcription polymerase chain reaction and immunohistochemical analyses. In contrast to small intestinal submucosa scaffolds, silk fibroin scaffolds supported spontaneous differentiation of esophageal epithelial cells toward a suprabasal cell lineage as indicated by significant upregulation of cytokeratin 4 and cytokeratin 13 messenger RNA transcript levels. In addition, esophageal epithelial cells maintained on silk fibroin scaffolds also produced significantly higher involucrin messenger RNA transcript levels in comparison to small intestinal submucosa counterparts, indicating an increased propensity for superficial, squamous cell specification. Collectively, these data provide evidence for the potential of silk fibroin scaffolds for gastrointestinal tissue engineering applications.

Franck, Debra; Chung, Yeun Goo; Coburn, Jeannine; Kaplan, David L; Estrada, Carlos R

2014-01-01

245

Mechanisms of monoclonal antibody stabilization and release from silk biomaterials  

PubMed Central

The availability of stabilization and sustained delivery systems for antibody therapeutics remains a major clinical challenge, despite the growing development of antibodies for a wide range of therapeutic applications due to their specificity and efficacy. A mechanistic understanding of protein-matrix interactions is critical for the development of such systems and is currently lacking as a mode to guide the field. We report mechanistic insight to address this need by using well-defined matrices based on silk gels, in combination with a monoclonal antibody. Variables including antibody loading, matrix density, charge interactions, hydrophobicity and water access were assessed to clarify mechanisms involved in the release of antibody from the biomaterial matrix. The results indicate that antibody release is primarily governed by hydrophobic interactions and hydration resistance, which are controlled by silk matrix chemistry, peptide domain distribution and protein density. Secondary ionic repulsions are also critical in antibody stabilization and release. Matrix modification by free methionine incorporation was found to be an effective strategy for mitigating encapsulation induced antibody oxidation. Additionally, these studies highlight a characterization approach to improve the understanding and development of other protein sustained delivery systems, with broad applicability to the rapidly developing monoclonal antibody field. PMID:23859659

Guziewicz, Nicholas A.; Massetti, Andrew J.; Perez-Ramirez, Bernardo J.; Kaplan, David L.

2013-01-01

246

Biomaterials 24 (2003) 401416 Silk-based biomaterials  

E-print Network

with other commonly used biomaterials such as polylactic acid and collagen. Furthermore, the unique different types of silks. Each of these different silks has a different amino acid composition and exhibits

Lu, Helen H.

247

Scroop, luster, and hand : the science and sensuality of silk  

E-print Network

For five thousand years, silk threads have woven through the fabric of human history. Since its accidental discovery in China all that time ago, silk has played roles, major or minor, in many cultures. In both the East and ...

Boyce, Jennifer E. (Jennifer Elaine)

2005-01-01

248

Chromosome Mapping of Dragline Silk Genes in the Genomes of Widow Spiders (Araneae, Theridiidae)  

PubMed Central

With its incredible strength and toughness, spider dragline silk is widely lauded for its impressive material properties. Dragline silk is composed of two structural proteins, MaSp1 and MaSp2, which are encoded by members of the spidroin gene family. While previous studies have characterized the genes that encode the constituent proteins of spider silks, nothing is known about the physical location of these genes. We determined karyotypes and sex chromosome organization for the widow spiders, Latrodectus hesperus and L. geometricus (Araneae, Theridiidae). We then used fluorescence in situ hybridization to map the genomic locations of the genes for the silk proteins that compose the remarkable spider dragline. These genes included three loci for the MaSp1 protein and the single locus for the MaSp2 protein. In addition, we mapped a MaSp1 pseudogene. All the MaSp1 gene copies and pseudogene localized to a single chromosomal region while MaSp2 was located on a different chromosome of L. hesperus. Using probes derived from L. hesperus, we comparatively mapped all three MaSp1 loci to a single region of a L. geometricus chromosome. As with L. hesperus, MaSp2 was found on a separate L. geometricus chromosome, thus again unlinked to the MaSp1 loci. These results indicate orthology of the corresponding chromosomal regions in the two widow genomes. Moreover, the occurrence of multiple MaSp1 loci in a conserved gene cluster across species suggests that MaSp1 proliferated by tandem duplication in a common ancestor of L. geometricus and L. hesperus. Unequal crossover events during recombination could have given rise to the gene copies and could also maintain sequence similarity among gene copies over time. Further comparative mapping with taxa of increasing divergence from Latrodectus will pinpoint when the MaSp1 duplication events occurred and the phylogenetic distribution of silk gene linkage patterns. PMID:20877726

Zhao, Yonghui; Ayoub, Nadia A.; Hayashi, Cheryl Y.

2010-01-01

249

An investigation of the divergence of major ampullate silk fibers from Nephila clavipes and Argiope aurantia.  

PubMed

The major ampullate fiber of both Nephila clavipes and Argiope aurantia is composed of two different proteins, MaSp1 and MaSp2. Each of these proteins has a highly conserved pattern of silk-associated amino acid motifs. The GPGXX motif is the only source of proline and is unique to MaSp2. On the basis of the percent of proline, Nephila clavipes major ampullate silk was calculated to consist of 19% MaSp2 and 81% MaSp1, while Argiope aurantia was calculated to have a significantly higher MaSp2 content of 59% with MaSp1 comprising the remaining 41%. To investigate the functional implications of the difference in protein composition, major ampullate silk fibers from Nephila clavipes and Argiope aurantia were mechanically tested and compared. Stress-strain curves produced from polynomial regression show that the two significant differences between major ampullate silk fibers from Nephila clavipes and Argiope aurantia are the average peak load stress and Young's modulus, with Argiope higher for both. PMID:16283732

Brooks, Amanda E; Steinkraus, Holly B; Nelson, Shane R; Lewis, Randolph V

2005-01-01

250

Acellular bi-layer silk fibroin scaffolds support tissue regeneration in a rabbit model of onlay urethroplasty.  

PubMed

Acellular scaffolds derived from Bombyx mori silk fibroin were investigated for their ability to support functional tissue regeneration in a rabbit model of urethra repair. A bi-layer silk fibroin matrix was fabricated by a solvent-casting/salt leaching process in combination with silk fibroin film casting to generate porous foams buttressed by homogeneous silk fibroin films. Ventral onlay urethroplasty was performed with silk fibroin grafts (Group 1, N = 4) (Width × Length, 1 × 2 cm(2)) in adult male rabbits for 3 m of implantation. Parallel control groups consisted of animals receiving small intestinal submucosa (SIS) implants (Group 2, N = 4) or urethrotomy alone (Group 3, N = 3). Animals in all groups exhibited 100% survival prior to scheduled euthanasia and achieved voluntary voiding following 7 d of initial catheterization. Retrograde urethrography of each implant group at 3 m post-op revealed wide urethral calibers and preservation of organ continuity similar to pre-operative and urethrotomy controls with no evidence of contrast extravasation, strictures, fistulas, or stone formation. Histological (hematoxylin and eosin and Masson's trichrome), immunohistochemical, and histomorphometric analyses demonstrated that both silk fibroin and SIS scaffolds promoted similar extents of smooth muscle and epithelial tissue regeneration throughout the original defect sites with prominent contractile protein (?-smooth muscle actin and SM22?) and cytokeratin expression, respectively. De novo innervation and vascularization were also evident in all regenerated tissues indicated by synaptophysin-positive neuronal cells and vessels lined with CD31 expressing endothelial cells. Following 3 m post-op, minimal acute inflammatory reactions were elicited by silk fibroin scaffolds characterized by the presence of eosinophil granulocytes while SIS matrices promoted chronic inflammatory responses indicated by mobilization of mononuclear cell infiltrates. The results of this study demonstrate that bi-layer silk fibroin scaffolds represent promising biomaterials for onlay urethroplasty, capable of promoting similar degrees of tissue regeneration in comparison to conventional SIS scaffolds, but with reduced immunogenicity. PMID:24632740

Chung, Yeun Goo; Tu, Duong; Franck, Debra; Gil, Eun Seok; Algarrahi, Khalid; Adam, Rosalyn M; Kaplan, David L; Estrada, Carlos R; Mauney, Joshua R

2014-01-01

251

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

PubMed Central

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 consisted of smooth, compact multi-laminates of parallel-oriented silk fibers while GS2 scaffolds were composed of porous (pore size range, 5–50µm) lamellar-like sheets buttressed by a dense outer layer. Bi-layer FF scaffolds were comprised of porous foams (pore size, ~400 µm) fused on their external face with a homogenous, nonporous silk film. Silk groups and small intestinal submucosa (SIS) matrices were evaluated in a rat model of augmentation cystoplasty for 10 weeks of implantation and compared to cystotomy controls. Gross tissue evaluations revealed the presence of intra-luminal stones in all experimental groups. The incidence and size of urinary calculi was the highest in animals implanted with gel spun silk matrices and SIS with frequencies ?57% and stone diameters of 3–4mm. In contrast, rats augmented with FF scaffolds displayed substantially lower rates (20%) and stone size (2mm), similar to the levels observed in controls (13%, 2mm). Histological (hematoxylin and eosin, Masson's trichrome) and immunohistochemical (IHC) analyses showed comparable extents of smooth muscle regeneration and contractile protein (?-smooth muscle actin and SM22?) expression within defect sites supported by all matrix groups similar to controls. Parallel evaluations demonstrated the formation of a transitional, multi-layered urothelium with prominent uroplakin and p63 protein expression in all experimental groups. De novo innervation and vascularization processes were evident in all regenerated tissues indicated by Fox3-positive neuronal cells and vessels lined with CD31 expressing endothelial cells. In comparison to other biomaterial groups, cystometric analyses at 10 weeks post-op revealed that animals implanted with the FF matrix configuration displayed superior urodynamic characteristics including compliance, functional capacity, as well as spontaneous non voiding contractions consistent with control levels. Our data demonstrate that variations in scaffold processing techniques can influence the in vivo functional performance of silk matrices in bladder reconstructive procedures. PMID:23545287

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

252

Finding inspiration in argiope trifasciata spider silk fibers  

NASA Astrophysics Data System (ADS)

The outstanding mechanical properties of silk fibers from the spider Argiope trifasciata are reviewed in this article, particularly the tensile behavior under controlled humidity and temperature. Samples obtained by forced silking showed a remarkable reproducibility. A novel procedure, wet stretching, developed by the authors, promises to shed light on the spinning of artificial silk fibers.

Elices, Manuel; Guinea, Gustavo V.; Pérez-Rigueiro, José; Plaza, Gustavo R.

2005-02-01

253

Finding inspiration in argiope trifasciata spider silk fibers  

Microsoft Academic Search

The outstanding mechanical properties of silk fibers from the spider Argiope trifasciata are reviewed in this article, particularly\\u000a the tensile behavior under controlled humidity and temperature. Samples obtained by forced silking showed a remarkable reproducibility.\\u000a A novel procedure, wet stretching, developed by the authors, promises to shed light on the spinning of artificial silk fibers.

Manuel Elices; Gustavo V. Guinea; José Pérez-Rigueiro; Gustavo R. Plaza

2005-01-01

254

Biomimetic calcium phosphate coatings on recombinant spider silk fibres  

Microsoft Academic Search

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

Liang Yang; My Hedhammar; Tobias Blom; Klaus Leifer; Jan Johansson; Pamela Habibovic; Clemens A. van Blitterswijk

2010-01-01

255

Amphiphilic Spider Silk-Like Block Copolymers with Tunable Physical Properties and Morphology for Biomedical Applications  

NASA Astrophysics Data System (ADS)

Silk-based materials are important candidates for biomedical applications because of their excellent biocompatibility and biodegradability. To generate silk amphiphilic biopolymers with potential use in guided tissue repair and drug delivery, a novel family of spider silk-like block copolymers was synthesized by recombinant DNA technology. Block copolymer thermal properties, structural conformations, protein-water interactions, and self-assembly morphologies were studied with respect to well controlled protein amino acid sequences. A theoretical model was used to predict the heat capacity of the protein and protein-water complex. Using thermal analysis, two glass transitions were observed: Tg1 is related to conformational changes caused by bound water removal, while Tg2 (>Tg1) is the glass transition of dry protein. Real-time infrared spectroscopy and X-ray diffraction confirmed that different secondary structural changes occur during the two Tg relaxations. Using scanning electron microscopy, fibrillar networks and hollow vesicles are observed, depending on protein block copolymer sequence. This study provides a deeper understanding of the relationship between protein physical properties and amino acid sequence, with implications for design of other protein-based materials.

Huang, Wenwen; Krishnaji, Sreevidhya; Kaplan, David; Cebe, Peggy

2013-03-01

256

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

PubMed

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

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

2014-02-01

257

Preparation of uniaxial multichannel silk fibroin scaffolds for guiding primary neurons.  

PubMed

Physical guidance cues have been exploited to stimulate neuron adhesion and neurite outgrowth. In the present study, three-dimensional (3-D) silk fibroin scaffolds with uniaxial multichannels (42-142 ?m in diameter) were prepared by a directional temperature field freezing technique, followed by lyophilization. By varying the initial silk fibroin concentration, the chemical potential and quantity of free water around cylindrical ice crystals could be controlled to control the cross-section morphology of the scaffold channels. Aligned ridges also formed on the inner surface of the multichannels in parallel to the direction of the channels. In vitro, primary hippocampal neurons were seeded in these 3-D silk fibroin scaffolds with uniaxial multichannels of ?120 ?m in diameter. The morphology of the neurons was multipolar and alignment along the scaffold channels was observed. Cell-cell networks and cell-matrix interactions established by newly formed axons were observed after 7 days in culture. These neurons expressed ?-III-tubulin, nerve filament and microtubule-associated protein, while glial fibrillary acidic protein immunofluorescence was barely above background. The ridges on the inner surface of the channels played a critical role in the adhesion and extension of neurons by providing continuous contact guidance. These new 3-D silk scaffolds with uniaxial multichannels provided a favorable microenvironment for the development of hippocampal neurons by guiding axonal elongation and cell migration. PMID:22465574

Zhang, Qiang; Zhao, Yahong; Yan, Shuqin; Yang, Yumin; Zhao, Huijing; Li, Mingzhong; Lu, Shenzhou; Kaplan, David L

2012-07-01

258

Spider Silk-Based Gene Carriers for Tumor Cell-Specific Delivery  

PubMed Central

The present study demonstrates pDNA complexes of recombinant silk proteins containing poly(L-lysine) and tumor-homing peptides (THPs), which are globular and approximately 150–250 nm in diameter, show significant enhancement of target specificity to tumor cells by additions of F3 and CGKRK THPs. We report herein the preparation and study of novel nano-scale silk-based ionic complexes containing pDNA able to home specifically to tumor cells. Particular focus was on how the THP, F3 (KDEPQRRSARLSAKPAPPKPEPKPKKAPAKK) and CGKRK, enhanced transfection specificity to tumor cells. Genetically engineered silk proteins containing both poly(L-lysine) domains to interact with pDNA and the THP to bind to specific tumor cells for target-specific pDNA delivery were prepared using Escherichia coli, followed by in-vitro and in-vivo transfection experiments into MDA-MB-435 melanoma cells and highly metastatic human breast tumor MDA-MB-231 cells. Non-tumorigenic MCF-10A breast epithelial cells were used as a control cell line for in-vitro tumor-specific delivery studies. These results demonstrate that combination of the bioengineered silk delivery systems and THP can serve as a versatile and useful new platform for non-viral gene delivery. PMID:21739966

Numata, Keiji; Reagan, Michaela R; Goldstein, Robert H; Rosenblatt, Michael; Kaplan, David L

2011-01-01

259

Spider silk-based gene carriers for tumor cell-specific delivery.  

PubMed

The present study demonstrates pDNA complexes of recombinant silk proteins containing poly(L-lysine) and tumor-homing peptides (THPs), which are globular and approximately 150-250 nm in diameter, show significant enhancement of target specificity to tumor cells by additions of F3 and CGKRK THPs. We report herein the preparation and study of novel nanoscale silk-based ionic complexes containing pDNA able to home specifically to tumor cells. Particular focus was on how the THP, F3 (KDEPQRRSARLSAKPAPPKPEPKPKKAPAKK), and CGKRK, enhanced transfection specificity to tumor cells. Genetically engineered silk proteins containing both poly(L-lysine) domains to interact with pDNA and the THP to bind to specific tumor cells for target-specific pDNA delivery were prepared using Escherichia coli, followed by in vitro and in vivo transfection experiments into MDA-MB-435 melanoma cells and highly metastatic human breast tumor MDA-MB-231 cells. Non-tumorigenic MCF-10A breast epithelial cells were used as a control cell line for in vitro tumor-specific delivery studies. These results demonstrate that combination of the bioengineered silk delivery systems and THP can serve as a versatile and useful new platform for nonviral gene delivery. PMID:21739966

Numata, Keiji; Reagan, Michaela R; Goldstein, Robert H; Rosenblatt, Michael; Kaplan, David L

2011-08-17

260

A new class of animal collagen masquerading as an insect silk  

PubMed Central

Collagen is ubiquitous throughout the animal kingdom, where it comprises some 28 diverse molecules that form the extracellular matrix within organisms. In the 1960s, an extracorporeal animal collagen that forms the cocoon of a small group of hymenopteran insects was postulated. Here we categorically demonstrate that the larvae of a sawfly species produce silk from three small collagen proteins. The native proteins do not contain hydroxyproline, a post translational modification normally considered characteristic of animal collagens. The function of the proteins as silks explains their unusual collagen features. Recombinant proteins could be produced in standard bacterial expression systems and assembled into stable collagen molecules, opening the door to manufacture a new class of artificial collagen materials. PMID:24091725

Sutherland, Tara D.; Peng, Yong Y.; Trueman, Holly E.; Weisman, Sarah; Okada, Shoko; Walker, Andrew A.; Sriskantha, Alagacone; White, Jacinta F.; Huson, Mickey G.; Werkmeister, Jerome A.; Glattauer, Veronica; Stoichevska, Violet; Mudie, Stephen T.; Haritos, Victoria S.; Ramshaw, John A. M.

2013-01-01

261

Surface Immobilization of antibody on silk fibroin through conformational transition  

PubMed Central

In recent studies silk fibroin has been explored as a new material platform for biosensors. Based on these developments a procedure for the immobilization of antibodies on silk fibroin substrates was developed as a route to functionalizing these biosensor systems. By controlling the conformational transition of the silk fibroin, a primary antibody was immobilized and enriched at the surface of silk fibroin substrates under mild reaction conditions to maintain antibody function. Compared to chemical crosslinking, the immobilization efficiency in the present approach was increased significantly. This method, achieving high loading of antibody while retaining function, improves the feasibility of silk fibroin as a platform material for biosensor applications. PMID:21382528

Lu, Qiang; Wang, Xiaoqin; Zhu, Hesun; Kaplan, David L.

2011-01-01

262

Analyzing the clustering effects of major ampullate silk mechanical properties - biomed 2009.  

PubMed

Although spider silk displays an amazing combination of strength and extensibility unrivalled by most synthetic biomaterials, its molecular architecture is relatively simplistic. Four primary amino acid block motifs (An, (GA)n, GPGXX, GGX) have been correlated with mechanical functions. Recent genetic engineering to control the mechanical behavior of synthetic silk fibers has verified much of the proposed structure/function relationship; however, the genetically defined exchange between strength and elasticity has proven not to be a direct relationship. Thus, complete control over the mechanical properties of a synthetic spider silk based fiber continues to elude scientists. The yet undefined factor(s) may be an element of the fabrication process. Natural silk production results from a combination of dehydration and protein alignment that occurs during concurrent spin and draw processes. While synthetic fiber production attempts to mimic 1) dehydration with a series of coagulating solvents and 2) protein alignment through the controlled extrusion of a concentration dependent spinning solution, the spinning and drawing processes are separated and occur sequentially. Many studies have been conducted which have examined multiple parameters; however, the spinning conditions which produce consistent mechanical properties, necessary for the progression toward any medical, commercial or military application, have not been identified. Here, we report on mathematical methods based on data from a variety of spinning conditions to characterize different impacting properties as either primary (i.e. a condition which directs or dictates mechanical properties of an individual fiber) or ptimizing (i.e. a condition which increases the engineered properties of the silk). PMID:19369768

Brooks, Amanda E; Brooks, Benjamin D; Creager, Melinda S; Lewis, Randolph V

2009-01-01

263

Porous Silk Scaffolds for Delivery of Growth Factors and Stem Cells to Enhance Bone Regeneration  

PubMed Central

Stem cell-based tissue engineering shows promise for bone regeneration and requires artificial microenvironments to enhance the survival, proliferation and differentiation of the seeded cells. Silk fibroin, as a natural protein polymer, has unique properties for tissue regeneration. The present study aimed to evaluate the influence of porous silk scaffolds on rat bone marrow stem cells (BMSCs) by lenti-GFP tracking both in vitro and in vivo in cranial bone defects. The number of cells seeded within silk scaffolds in rat cranial bone defects increased from 2 days to 2 weeks after implantation, followed by a decrease at eight weeks. Importantly, the implanted cells survived for 8 weeks in vivo and some of the cells might differentiate into endothelial cells and osteoblasts induced by the presence of VEGF and BMP-2 in the scaffolds to promote angiogenesis and osteogenesis. The results demonstrate that porous silk scaffolds provide a suitable niche to maintain long survival and function of the implanted cells for bone regeneration. PMID:25050556

Xu, Ling; Zhang, Xiaochen; Wu, Qianju; Zhang, Xiuli; Kaplan, David L.; Jiang, Xinquan

2014-01-01

264

Silk and Silicon: Technology Paradigms and Education.  

ERIC Educational Resources Information Center

Reviews "The Silk Code" (novel by Paul Levinson, Professor of Communications, New York University) that encourages questions about how to choose and use the full range of technology; and compares and applies its ideas to current practices in educational technology. Topics include divergent views in educational theory; and examples of the use of…

Hokanson, Brad

2001-01-01

265

Constructing Knowledge with Silk Road Visuals  

ERIC Educational Resources Information Center

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…

Bisland, Beverly Milner

2008-01-01

266

Silk-Screening a la Andy.  

ERIC Educational Resources Information Center

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)

Mathes, Len

2000-01-01

267

Opportunities for Intervention in Thailand's Silk Subsector.  

National Technical Information Service (NTIS)

The silk subsector holds enormous promise as a potential engine of income growth for poor village households. But to take advantage of the growth, most producers will have to shift from the low-productivity, low-growth traditional markets to the rapidly g...

S. Haggblade, N. Ritchie

1992-01-01

268

High-resolution NMR characterization of a spider-silk mimetic composed of 15 tandem repeats and a CRGD motif  

PubMed Central

Multidimensional solution NMR spectroscopic techniques have been used to obtain atomic level information about a recombinant spider silk construct in hexafluoro-isopropanol (HFIP). The synthetic 49 kDa silk-like protein mimics authentic silk from Nephila clavipes, with the inclusion of an extracellular matrix recognition motif. 2D 1H-15N HSQC NMR spectroscopy reveals 33 cross peaks, which were assigned to amino acid residues in the semicrystalline repeat units. Signals from the amorphous segments in the primary sequence were weak and broad, suggesting that this region is highly dynamic and undergoing conformational exchange. An analysis of the deviations of the 13C?, 13C?, and 13CO chemical shifts relative to the expected random coil values reveals two highly ?-helical regions from amino acid 12–19 and 26–32, which comprise the polyalanine track and a GGLGSQ sequence. This finding is further supported by ?-value analysis and sequential and medium-range NOE interactions. Pulsed field gradient NMR measurements indicate that the topology of the silk mimetic in HFIP is nonglobular. Moreover, the 3D 15N-NOESY HSQC spectrum exhibits few long-range NOEs. Similar spectral features have been observed for repeat modules in other polypeptides and are characteristic of an elongated conformation. The results provide a residue-specific description of a silk sequence in nonaqueous solution and may be insightful for understanding the fold and topology of highly concentrated, stable silk before spinning. Additionally, the insights obtained may find application in future design and large-scale production and storage of synthetic silks in organic solvents. PMID:19177364

McLachlan, Glendon D; Slocik, Joseph; Mantz, Robert; Kaplan, David; Cahill, Sean; Girvin, Mark; Greenbaum, Steve

2009-01-01

269

Antibiotic-Releasing Silk Biomaterials for Infection Prevention and Treatment  

PubMed Central

Effective treatment of infections in avascular and necrotic tissues can be challenging due to limited penetration into the target tissue and systemic toxicities. Controlled release polymer implants have the potential to achieve the high local concentrations needed while also minimizing systemic exposure. Silk biomaterials possess unique characteristics for antibiotic delivery including biocompatibility, tunable biodegradation, stabilizing effects, water-based processing and diverse material formats. We report on functional release of antibiotics spanning a range of chemical properties from different material formats of silk (films, microspheres, hydrogels, coatings). The release of penicillin and ampicillin from bulk-loaded silk films, drug-loaded silk microspheres suspended in silk hydrogels and bulk-loaded silk hydrogels was investigated and in vivo efficacy of ampicillin-releasing silk hydrogels was demonstrated in a murine infected wound model. Silk sponges with nanofilm coatings were loaded with gentamicin and cefazolin and release was sustained for 5 and 3 days, respectively. The capability of silk antibiotic carriers to sequester, stabilize and then release bioactive antibiotics represents a major advantage over implants and pumps based on liquid drug reservoirs where instability at room or body temperature is limiting. The present studies demonstrate that silk biomaterials represent a novel, customizable antibiotic platform for focal delivery of antibiotics using a range of material formats (injectable to implantable). PMID:23483738

Pritchard, Eleanor M.; Valentin, Thomas; Panilaitis, Bruce; Omenetto, Fiorenzo; Kaplan, David L.

2012-01-01

270

Dear Silk Road FIG student, Welcome to the University of Oregon, and to the Silk Road FIG! I am Patricia Komoda, your FA  

E-print Network

Dear Silk Road FIG student, Welcome to the University of Oregon, and to the Silk Road FIG! I am freshman year, I was in the Silk Road FIG, just as you are now. I am currently a sophomore majoring and Taata. He also loves ice cream metaphors, and tells a great story! The primary focus of the Silk Road

Oregon, University of

271

The Influence of Elasticity and Surface Roughness on Myogenic and Osteogenic-Differentiation of Cells on Silk-Elastin Biomaterials  

PubMed Central

The interactions of C2C12 myoblasts and human bone marrow stem cells (hMSCs) with silk-tropoelastin biomaterials, and the capacity of each to promote attachment, proliferation, and either myogenic- or osteogenic-differentiation were investigated. Temperature-controlled water vapor annealing was used to control beta-sheet crystal formation to generate insoluble silk-tropoelastin biomaterial matrices at defined ratios of the two proteins. These ratios controlled surface roughness and micro/nano-scale topological patterns, and elastic modulus, stiffness, yield stress, and tensile strength. A combination of low surface roughness and high stiffness in the silk-tropoelastin materials promoted proliferation and myogenic-differentiation of C2C12 cells. In contrast, high surface roughness with micro/nano-scale surface patterns was favored by hMSCs. Increasing the content of human tropoelastin in the silk-tropoelastin materials enhanced the proliferation and osteogenic-differentiation of hMSCs. We conclude that the silk-tropoelastin composition facilitates fine tuning of the growth and differentiation of these cells. PMID:21872326

Hu, Xiao; Park, Sang-Hyug; Gil, Eun Seok; Xia, Xiao-Xia; Weiss, Anthony S.; Kaplan, David L.

2011-01-01

272

Silk-Based Nano-Complexes with Tumor Homing Peptides for Tumor-Specific Gene Delivery a  

PubMed Central

Nano-scale complexes of recombinant silk molecules containing tumor homing peptides (THPs) with DNA were designed as less-cytotoxic and highly target-specific gene carriers. We report the preparation and study of these nano-scale silk-based ionic complexes containing pDNA that are able to home specifically to tumorigenic cells due to the presence of THPs, namely, F3 and Lyp1. Genetically engineered silk proteins (MaSp1 monomer) containing poly(L-lysine) domains to interact with pDNA and the THP to bind to specific tumorigenic cells for target-specific pDNA delivery were prepared using Escherichia coli. This process was followed by in-vitro transfection experiments into MDA-MB-435 melanoma cells, highly metastatic human breast tumor MDA-MB-231 cells, and non-tumorigenic MCF-10A breast epithelial cells. The silk-poly(L-lysine) block copolymer containing Lyp1 (ML-Lyp1) showed significant differences from silk-poly(L-lysine) block copolymer containing F3 (ML-F3) in cytotoxicity to MCF10A cells. ML-F3 was the most useful candidate for target delivery into tumorigenic cells. The target specificity of the pDNA complexes to tumorigenic cells was regulated by specific adsorption processes on the cell surface, based on field emission scanning electron microscopy observations. PMID:22052706

Numata, Keiji; Mieszawska-Czajkowska, Aneta J; Kvenvold, Laura A; Kaplan, David L

2013-01-01

273

Inkjet printing of silk nest arrays for cell hosting.  

PubMed

An inkjet printing approach is presented for the facile fabrication of microscopic arrays of biocompatible silk "nests" capable of hosting live cells for prospective biosensors. The patterning of silk fibroin nests were constructed by the layer-by-layer (LbL) assembly of silk polyelectrolytes chemically modified with poly-(l-lysine) and poly-(l-glutamic acid) side chains. The inkjet-printed silk circular regions with a characteristic "nest" shape had diameters of 70-100 ?m and a thickness several hundred nanometers were stabilized by ionic pairing and by the formation of the silk II crystalline secondary structure. These "locked-in" silk nests remained anchored to the substrate during incubation in cell growth media to provide a biotemplated platform for printing-in, immobilization, encapsulation and growth of cells. The process of inkjet-assisted printing is versatile and can be applied on any type of substrate, including rigid and flexible, with scalability and facile formation. PMID:24605757

Suntivich, Rattanon; Drachuk, Irina; Calabrese, Rossella; Kaplan, David L; Tsukruk, Vladimir V

2014-04-14

274

23. SOUTH FACADE, FACING RIVER, HOUSES ON SILK STOCKING ROW, ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

23. SOUTH FACADE, FACING RIVER, HOUSES ON SILK STOCKING ROW, NEWHALEM, 1990. - Skagit Power Development, Skagit River & Newhalem Creek Hydroelectric Project, On Skagit River, Newhalem, Whatcom County, WA

275

Volume constancy during stretching of spider silk.  

PubMed

The characterization of silk properties requires a reliable measurement of stress-strain curves from tensile tests, which calls for a detailed analysis of what is considered the cross section of the sample and how it varies during the experiments. Here, spider silk fibers from the major ampullate gland (MAS) of Argiope trifasciata spiders are tensile tested, and the cross-sectional area is measured under different strained configurations. It has been found that the fiber volume remains practically constant during stretching, and deformation proceeds homogeneously in all the fibers. The conservation of volume is validated independently of the type of fiber and the strain level. This result, applied to compute true stress-strain curves for different MAS fibers, shows that the description of their properties depends noticeably on which set of tensile parameters is chosen (true or engineering), and that engineering values could lead to misinterpretation of experiments that combine results from different strain ranges. PMID:16827584

Guinea, G V; Pérez-Rigueiro, J; Plaza, G R; Elices, M

2006-07-01

276

An investigation into the effect of potassium ions on the folding of silk fibroin studied by generalized two-dimensional NMR-NMR correlation and Raman spectroscopy.  

PubMed

We used generalized two-dimensional NMR-NMR correlation to examine the effect of potassium ions on the conformation transition in silk fibroin to investigate the possibility that the fairly high K+ ion content found in the distal end of silk-secreting ducts in the silkworms could have a bearing on natural formation of the silk fiber. This has enabled us to propose a detailed mechanism for the transition process. Our evidence indicates that increasing the [K+] from 0 to 3.7 mg.g(-1) in the silk fibroin, as is thought to occur as the silk fibroin moves through the secretory pathway to the spigot, produces a sequence of secondary structural changes: helix and/or random coil-->helix-like-->beta-sheet-like-->beta-sheet. The sequence is the same as that produced in silk fibroin films by decreasing the pH of fibroin from 6.8 to 4.8. In addition, we used Raman spectroscopy to study the effect of K+ ions on the Fermi doublet resonance of the tyrosyl phenolic ring at 850 and 830 cm(-1). The intensity ratio I(850)/I(830) at these wave numbers indicated that the hydrogen bonding formed by the tyrosyl phenolic-OH becomes more stable with an increase in the K+ ion concentration as above. Our investigation on the effect of K+ ions on fibroin may help provide a theoretical basis for understanding the natural silk-spinning process and the conditions required for biomimetic spinning. It may also have relevance to the aggregation of other beta-sheet proteins, including prion proteins, neurofibrillary proteins and amyloid plaques. PMID:18081855

Ruan, Qing-Xia; Zhou, Ping; Hu, Bing-Wen; Ji, Dan

2008-01-01

277

Molecular mechanisms of phoxim-induced silk gland damage and TiO2 nanoparticle-attenuated damage in Bombyx mori.  

PubMed

Phoxim is a useful organophosphate (OP) pesticide used in agriculture in China, however, exposure to this pesticide can result in a significant reduction in cocooning in Bombyx mori (B. mori). Titanium dioxide nanoparticles (TiO2 NPs) have been shown to decrease phoxim-induced toxicity in B. mori; however, very little is known about the molecular mechanisms of silk gland damage due to OP exposure and repair of gland damage by TiO2 NP pretreatment. In the present study, exposure to phoxim resulted in a significant reduction in cocooning rate in addition to silk gland damage, whereas TiO2 NP attenuated phoxim-induced gland damage, increased the antioxidant capacity of the gland, and increased cocooning rate in B. mori. Furthermore, digital gene expression data suggested that phoxim exposure led to significant alterations in the expression of 833 genes. In particular, phoxim exposure caused significant down-regulation of Fib-L, Ser2, Ser3, and P25 genes involved in silk protein synthesis, and up-regulation of SFGH, UCH3, and Salhh genes involved in silk protein hydrolysis. A combination of both phoxim and TiO2 NP treatment resulted in marked changes in the expression of 754 genes, while treatment with TiO2 NPs led to significant alterations in the expression of 308 genes. Importantly, pretreatment with TiO2 NPs increased Fib-L, Ser2, Ser3, and P25 expression, and decreased SFGH, UCH3, and Salhh expression in silk protein in the silk gland under phoxim stress. Therefore, Fib-L, Ser2, Ser3, P25, SFGH, UCH3, and Salhh may be potential biomarkers of silk gland toxicity in B. mori caused by phoxim exposure. PMID:24331035

Li, Bing; Yu, Xiaohong; Gui, Suxin; Xie, Yi; Zhao, Xiaoyang; Hong, Jie; Sun, Qingqing; Sang, Xuezi; Sheng, Lei; Cheng, Zhe; Cheng, Jie; Hu, Rengping; Wang, Ling; Shen, Weide; Hong, Fashui

2014-06-01

278

Templated native silk smectic gels  

NASA Technical Reports Server (NTRS)

One aspect of the present invention relates to a method of preparing a fibrous protein smectic hydrogel by way of a solvent templating process, comprising the steps of pouring an aqueous fibrous protein solution into a container comprising a solvent that is not miscible with water; sealing the container and allowing it to age at about room temperature; and collecting the resulting fibrous protein smectic hydrogel and allowing it to dry. Another aspect of the present invention relates to a method of obtaining predominantly one enantiomer from a racemic mixture, comprising the steps of pouring an aqueous fibrous protein solution into a container comprising a solvent that is not miscible with water; sealing the container and allowing it to age at about room temperature; allowing the enantiomers of racemic mixture to diffuse selectively into the smectic hydrogel in solution; removing the smectic hydrogel from the solution; rinsing predominantly one enantiomer from the surface of the smectic hydrogel; and extracting predominantly one enantiomer from the interior of the smectic hydrogel. The present invention also relates to a smectic hydrogel prepared according to an aforementioned method.

Jin, Hyoung-Joon (Inventor); Park, Jae-Hyung (Inventor); Valluzzi, Regina (Inventor)

2009-01-01

279

Animal silks: their structures, properties and artificial production.  

PubMed

This feature article reviews recent progress in the understanding of the hierarchically organized structures, the perfectly balanced mechanical properties and the structure-property relationship of the natural animal silk fibres, as well as the experimental attempts to fabricate man-made silk fibres by means of wet spinning, dry spinning, electrospinning and transgenosis. PMID:19865641

Fu, Chengjie; Shao, Zhengzhong; Fritz, Vollrath

2009-11-21

280

Solid-state NMR relaxation studies of Australian spider silks.  

PubMed

Solid-state NMR techniques were used to study two different types of spider silk from two Australian orb-web spider species, Nephila edulis and Argiope keyserlingi. A comparison of (13)C-T(1) and (1)H-T(1rho) solid-state NMR relaxation data of the Ala Calpha, Ala Cbeta, Gly Calpha, and carbonyl resonances revealed subtle differences between dragline and cocoon silk. (13)C-T(1rho) and (1)H-T(1) relaxation experiments showed significant differences between silks of the two species with possible structural variations. Comparison of our data to previous (13)C-T(1) relaxation studies of silk from Nephila clavipes (A. Simmons et al., Macromolecules, 1994, Vol. 27, pp. 5235-5237) also supports the finding that differences in molecular mobility of dragline silk exist between species. Interspecies differences in silk structure may be due to different functional properties. Relaxation studies performed on wet (supercontracted) and dry silks showed that the degree of hydration affects relaxation properties, and hence changes in molecular mobility are correlated with functional properties of silk. PMID:12115143

Kishore, A I; Herberstein, M E; Craig, C L; Separovic, F

281

Project for a Comprehensive Study of the Silk Roads  

Microsoft Academic Search

From time immemorial the Silk Road, with its hundreds of byways, was considered the highway that linked the worlds of East and West. In the mid dle of the twentieth century, scholarly research revealed the crucial role played by oases in linking the North to the South; these oases, even more than the transversal lines of the Silk Roads, played

Vadim Eliseyev

1995-01-01

282

Geographic Perspectives with Elementary Students: The Silk Road  

ERIC Educational Resources Information Center

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…

Bisland, Beverly Milner

2006-01-01

283

Silk Fiber Mechanics from Multiscale Force Distribution Analysis Murat Cetinkaya,  

E-print Network

strength comparable to steel, toughness greater than that of Kevlar (DuPont, Wil- mington, DE polymeric systems. Hence, it is an efficient tool for the design of artificial silk fibers. INTRODUCTION subunits by acting as comparably stiff cross-linking sites (3). The amorphous subunit of a silk fiber

Gräter, Frauke

284

[Preparation and spectroscopic studies of nanosilver/silk-fibroin composite].  

PubMed

The development of reliable, eco-friendly processes for the synthesis of nanomaterials is an important aspect of nanotechnology today. One approach that shows immense potential is based on the in situ synthesis of noble metal nanocolloids using natural biological material such as silk fibroin. In the present paper, the nano-Ag/silk-fibroin colloids were prepared by in situ reducing AgNO3 with silk-fibroin at room temperature without any reducing agent. UV-Vis absorption, atomic force microscopy, fluorescence spectra and resonance scattering spectra were used to study the preparation process and characterization of the nanocomposite. It was found that the formative Ag nanoparticles were attributed to the tyrosine residues in the chains of silk-fibroin that act as reduction agent in the reduction course of AgNO3, and the Ag nanoparticles were uniformly embedded in silk-fibroin colloid, which possess good dispersity and stability and can be kept for a long period. The strength of fluorescence spectra increased after reaction of silk-fibroin with AgNO3 solution, showing that there was a strong chemical adsorption between silver nanoparticles and silk-fibroin, and a stable complex layer was formed in silver nanoparticles. Meanwhile the existence of silver nanoparticles in silk-fibroin was proved by resonance scattering spectra. PMID:19093576

Ai, Shi-yun; Gao, Ji-gang; Zhu, Lu-sheng; Ma, Zhi-jun; Li, Xiao-chen

2008-09-01

285

Understanding the variability of properties in Antheraea pernyi silk fibres.  

PubMed

Variability is a common feature of natural silk fibres, caused by a range of natural processing conditions. Better understanding of variability will not only be favourable for explaining the enviable mechanical properties of animal silks but will provide valuable information for the design of advanced artificial and biomimetic silk-like materials. In this work, we have investigated the origin of variability in forcibly reeled Antheraea pernyi silks from different individuals using dynamic mechanical thermal analysis (DMTA) combined with the effect of polar solvent penetration. Quasi-static tensile curves in different media have been tested to show the considerable variability of tensile properties between samples from different silkworms. The DMTA profiles (as a function of temperature or humidity) through the glass transition region of different silks as well as dynamic mechanical properties after high temperature and water annealing are analysed in detail to identify the origin of silk variability in terms of molecular structures and interactions, which indicate that different hydrogen bonded structures exist in the amorphous regions and they are notably different for silks from different individuals. Solubility parameter effects of solvents are quantitatively correlated with the different glass transitions values. Furthermore, the overall ordered fraction is shown to be a key parameter to quantify the variability in the different silk fibres, which is consistent with DMTA and FTIR observations. PMID:25030083

Wang, Yu; Guan, Juan; Hawkins, Nick; Porter, David; Shao, Zhengzhong

2014-09-01

286

Introduction Spider silk is an inspiration for biomimetic super fibers  

E-print Network

2452 Introduction Spider silk is an inspiration for biomimetic super fibers because of the high affects the mechanical function of silk fibers (e.g. Gatesy et al., 2001; Guerette et al., 1996; Hayashi and Lewis, 1998; Simmons et al., 1996; Tian and Lewis, 2005). Finally, the mechanical process of spinning

Blackledge, Todd

287

Visual Literacy with Picture Books: The Silk Road  

ERIC Educational Resources Information Center

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. …

Bisland, Beverly Milner Lee

2007-01-01

288

Self-assembly of silk fibroin under osmotic stress  

NASA Astrophysics Data System (ADS)

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

Sohn, Sungkyun

289

Optical surface profiling of orb-web spider capture silks.  

PubMed

Much spider silk research to date has focused on its mechanical properties. However, the webs of many orb-web spiders have evolved for over 136 million years to evade visual detection by insect prey. It is therefore a photonic device in addition to being a mechanical device. Herein we use optical surface profiling of capture silks from the webs of adult female St Andrews cross spiders (Argiope keyserlingi) to successfully measure the geometry of adhesive silk droplets and to show a bowing in the aqueous layer on the spider capture silk between adhesive droplets. Optical surface profiling shows geometric features of the capture silk that have not been previously measured and contributes to understanding the links between the physical form and biological function. The research also demonstrates non-standard use of an optical surface profiler to measure the maximum width of a transparent micro-sized droplet (microlens). PMID:20710068

Kane, D M; Joyce, A M; Staib, G R; Herberstein, M E

2010-09-01

290

Traveling the Silk Road: A Measurement Analysis of a Large Anonymous Online  

E-print Network

Traveling the Silk Road: A Measurement Analysis of a Large Anonymous Online Marketplace Nicolas analysis of Silk Road, an anonymous, international online marketplace that operates as a Tor hidden service a detailed picture of the type of goods sold on Silk Road, and of the revenues made both by sellers and Silk

Sadeh, Norman M.

291

Studies on silk secretion in the Trichoptera (F. Limmephilidae). II. Structure and amino acid composition of the silk.  

PubMed

The ultrastructure and amino acid composition of the secreted silk of two species of trichopteran larvae, Pycnopsyche guttifer (Walk.) and Neophylax concinnus McL., were investigated. The spinnerets of these two animals were also examined by scanning electron microscopy. The silk consists of double-stranded, flat ribbons (1-4 mu wide), composed of bundles of 15-25 A filaments. There are two components of the silk: the fiber proper and a surrounding coat thought to be a silk "gum". Only the outer coat is positive to the EM PATP technique of Thiery (1967), which indicated the presence of neutral sugars. Amino acid analyses of Pycnopsyche silk show that, like other silks, two predominant amino acids are glycine and serine. Arginine, unexpectedly, is the third most abundant and there are a large number of basic and long side-chain amino acids. X-ray diffraction studies of the silk indicate that it has a less crystalline, more amorphous structure than that of other silks. PMID:1277288

Engster, M S

1976-06-11

292

The design of silk fiber composition in moths has been conserved for more than 150 million years.  

PubMed

The silk of caterpillars is secreted in the labial glands, stored as a gel in their lumen, and converted into a solid filament during spinning. Heavy chain fibroin (H-fibroin), light chain fibroin (L-fibroin), and P25 protein constitute the filament core in a few species that have been analyzed. Identification of these proteins in Yponomeuta evonymella, a moth from a family which diverged from the rest of Lepidoptera about 150 million years ago, reveals that the mode of filament construction is highly conserved. It is proposed that association of the three proteins is suited for long storage of hydrated silk dope and its rapid conversion to filament. Interactions underlying these processes depend on conserved spacing of critical amino acid residues that are dispersed through the L-fibroin and P25 and assembled in the short ends of the H-fibroin molecule. Strength, elasticity, and other physical properties of the filament are determined by simple amino acid motifs arranged in repetitive modules that build up most of the H-fibroin. H-Fibroin synergy with L-fibroin and P25 does not interfere with motif diversification by which the filament acquires new properties. Several types of motifs in complex repeats occur in the silks used for larval cobwebs and pupal cocoons. Restriction of silk use to cocoon construction in some lepidopteran families has been accompanied by simplification of H-fibroin repeats. An extreme deviation of the silk structure occurs in the Saturniidae silkmoths, which possess modified H-fibroin and lack L-fibroin and P25. PMID:16755355

Yonemura, Naoyuki; Sehnal, Frantisek

2006-07-01

293

Helicoidal multi-lamellar features of RGD-functionalized silk biomaterials for corneal tissue engineering  

PubMed Central

RGD-coupled silk protein-biomaterial lamellar systems were prepared and studied with human cornea fibroblasts (hCFs) to match functional requirements. A strategy for corneal tissue engineering was pursued to replicate the structural hierarchy of human corneal stroma within thin stacks of lamellae-like tissues, in this case constructed from scaffolds constructed with RGD-coupled, patterned, porous, mechanically robust and transparent silk films. The influence of RGD-coupling on the orientation, proliferation, ECM organization, and gene expression of hCFs was assessed. RGD surface modification enhanced cell attachment, proliferation, alignment and expression of both collagens (type I and V) and proteoglycans (decorin and biglycan). Confocal and histological images of the lamellar systems revealed that the bio-functionalized silk human cornea 3D constructs exhibited integrated corneal stroma tissue with helicoidal multi-lamellar alignment of collagen-rich and proteoglycan-rich extracellular matrix, with transparency of the construct. This biomimetic approach to replicate corneal stromal tissue structural hierarchy and architecture demonstrates a useful strategy for engineering human cornea. Further, this approach can be exploited for other tissue systems due to the pervasive nature of such helicoids in most human tissues. PMID:20801503

Gil, Eun Seok; Mandal, Biman B.; Park, Sang-Hyug; Marchant, Jeffrey K.; Omenetto, Fiorenzo G.; Kaplan, David L.

2010-01-01

294

Facile preparation of paclitaxel loaded silk fibroin nanoparticles for enhanced antitumor efficacy by locoregional drug delivery.  

PubMed

Non-toxic, safe materials and preparation methods are among the most important factors when designing nanoparticles (NPs) for future clinical application. Here we report a novel and facile method encapsulating anticancer drug paclitaxel (PTX) into silk fibroin (SF), a biocompatible and biodegradable natural polymer, without adding any toxic organic solvents, surfactants or other toxic agents. The paclitaxel loaded silk fibroin nanoparticles (PTX-SF-NPs) with a diameter of 130 nm were formed in an aqueous solution at room temperature by self-assembling of SF protein, which demonstrated mainly silk I conformation in the NPs. In cellular uptake experiments, coumarin-6 loaded SF NPs were taken up efficiently by two human gastric cancer cell lines BGC-823 and SGC-7901. In vitro cytotoxicity studies demonstrated that PTX kept its pharmacological activity when incorporating into PTX-SF-NPs, while SF showed no cytotoxicity to cells. The in vivo antitumor effects of PTX-SF-NPs were evaluated on gastric cancer nude mice exnograft model. We found that locoregional delivery of PTX-SF-NPs demonstrated superior antitumor efficacy by delaying tumor growth and reducing tumor weights compared with systemic administration. Furthermore, the organs of mice in NP treated groups didn't show obvious toxicity, indicating the in vivo safety of SF NPs. These results suggest that SF NPs are promising drug delivery carriers, and locoregional delivery of SF NPs could be a potential future clinical cancer treatment regimen. PMID:24274601

Wu, Puyuan; Liu, Qin; Li, Rutian; Wang, Jing; Zhen, Xu; Yue, Guofeng; Wang, Huiyu; Cui, Fangbo; Wu, Fenglei; Yang, Mi; Qian, Xiaoping; Yu, Lixia; Jiang, Xiqun; Liu, Baorui

2013-12-11

295

?-Sheet nanocrystalline domains formed from phosphorylated serine-rich motifs in caddisfly larval silk: a solid state NMR and XRD study.  

PubMed

Adhesive silks spun by aquatic caddisfly (order Trichoptera) larvae are used to build both intricate protective shelters and food harvesting nets underwater. In this study, we use (13)C and (31)P solid-state NMR and wide angle X-ray diffraction (WAXD) as tools to elucidate molecular protein structure of caddisfly larval silk from the species Hesperophylax consimilis . Caddisfly larval silk is a fibroin protein based biopolymer containing mostly repetitive amino acid motifs. NMR and X-ray results provide strong supporting evidence for a structural model in which phosphorylated serine repeats (pSX)4 complex with divalent cations Ca(2+) and Mg(2+) to form rigid nanocrystalline ?-sheet structures in caddisfly silk. (13)C NMR data suggests that both phosphorylated serine and neighboring valine residues exist in a ?-sheet conformation while glycine and leucine residues common in GGX repeats likely reside in random coil conformations. Additionally, (31)P chemical shift anisotropy (CSA) analysis indicates that the phosphates on phosphoserine residues are doubly ionized, and are charge-stabilized by divalent cations. Positively charged arginine side chains also likely play a role in charge stabilization. Finally, WAXD results finds that the silk is at least 7-8% crystalline, with ?-sheet interplane spacings of 3.7 and 4.5 Å. PMID:23452243

Addison, J Bennett; Ashton, Nicholas N; Weber, Warner S; Stewart, Russell J; Holland, Gregory P; Yarger, Jeffery L

2013-04-01

296

?-Sheet Nanocrystalline Domains Formed from Phosphorylated Serine-Rich Motifs in Caddisfly Larval Silk: A Solid State NMR and XRD Study  

PubMed Central

Adhesive silks spun by aquatic caddisfly (order Trichoptera) larvae are used to build both intricate protective shelters and food harvesting nets underwater. In this study, we use 13C and 31P solid-state Nuclear Magnetic Resonance (NMR) and Wide Angle X-ray Diffraction (WAXD) as tools to elucidate molecular protein structure of caddisfly larval silk from the species Hesperophylax consimilis. Caddisfly larval silk is a fibroin protein based biopolymer containing mostly repetitive amino acid motifs. NMR and X-ray results provide strong supporting evidence for a structural model in which phosphorylated serine repeats (pSX)4 complex with divalent cations Ca2+ and Mg2+ to form rigid nanocrystalline ?-sheet structures in caddisfly silk. 13C NMR data suggests that both phosphorylated serine and neighboring valine residues exist in a ?-sheet secondary structure conformation while glycine and leucine residues common in GGX repeats likely reside in random coil conformations. Additionally, 31P chemical shift anisotropy (CSA) analysis indicates that the phosphates on phosphoserine residues are doubly ionized, and are charge-stabilized by divalent cations. Positively charged arginine side chains also likely play a role in charge stabilization. Finally, WAXD results finds that the silk is at least 7–8% crystalline, with ?-sheet inter-plane spacings of 3.7 and 4.5 Å. PMID:23452243

Addison, J. Bennett; Ashton, Nicholas N.; Weber, Warner S.; Stewart, Russell J.; Holland, Gregory P.; Yarger, Jeffery L.

2013-01-01

297

Nanofeatured silk fibroin membranes for dermal wound healing applications.  

PubMed

As an effort to create the next generation of improved skin graft materials, in this study, we modified the surfaces of a previously investigated material, silk fibroin, using a NaOH alkaline treatment to obtain a biologically inspired nanofeatured surface morphology. Such surfaces were characterized for roughness, energy, and chemistry. In addition, keratinocyte (skin-forming cells) adhesion and proliferation on such nanofeatured silk fibroin wound dressings were studied in an initial attempt to determine the promotion of an epidermal cover on the wound bed to form a new epidermal barrier. Dermal fibroblast adhesion and proliferation were also studied to assess the ability of nanostructured silk fibroin to replace damaged dermal tissue in chronic wounds (i.e., for diabetic foot ulcers). Results demonstrated for the first time that keratinocyte and fibroblast cell density was greater on nanofeatured silk fibroin membranes compared with non-treated silk fibroin surfaces. The enhancement in cellular functions was correlated with an increase in silk surface nanotopography, wettability and change in chemistry after NaOH treatment. Due to the present promising results, the newly developed nanofeatured silk fibroin membranes are exciting alternative skin graft materials which should be further studied for various skin patch and wound dressing applications. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 135-144, 2015. PMID:24616219

Karahalilo?lu, Zeynep; Ercan, Batur; Denkba?, Emir B; Webster, Thomas J

2015-01-01

298

Silk-elastinlike recombinant polymers for gene therapy of head and neck cancer: from molecular definition to controlled gene expression  

PubMed Central

Silk-elastinlike protein polymers (SELP’s) are block copolymers of silk-like and elastin-like tandem repeats. With appropriate sequence and composition SELPs can be liquid at room temperature and form hydrogels at body temperature. The influence of polymer structure and concentration on biodegradation in vitro and controlled delivery of adenoviruses carrying reporter genes to head and neck tumors in vivo was evaluated using hydrogels made from three SELP analogs. SELP-815K, with eight silk and fifteen elastin units and a lysine (K) modified elastin, was compared to SELP-415K and SELP-47K. Hydrogels with higher silk content and concentration degraded at a slower rate compared to other analogs. Intratumoral injection of adenoviruses with SELPs enhanced gene expression in tumor tissue up to 10 fold compared to viral injection without polymer. Viruses delivered with SELP-815K at 4 wt% polymer concentration showed the highest level of gene expression. Tumor to liver ratio of expression was up to 55 fold higher for SELP-mediated systems. This study demonstrates the influence of exquisite control over polymer structure using recombinant techniques on spatial and temporal control over adenoviral delivery and establishes the utility of SELP matrices as biodegradable systems for gene therapy of head and neck cancer. PMID:19470397

Gustafson, Joshua; Greish, Khaled; Frandsen, Jordan; Cappello, Joseph; Ghandehari, Hamidreza

2009-01-01

299

Engineered disulfides improve mechanical properties of recombinant spider silk  

PubMed Central

Nature's high-performance polymer, spider silk, is composed of specific proteins, spidroins, which form solid fibers. So far, fibers made from recombinant spidroins have failed in replicating the extraordinary mechanical properties of the native material. A recombinant miniature spidroin consisting of four poly-Ala/Gly-rich tandem repeats and a nonrepetitive C-terminal domain (4RepCT) can be isolated in physiological buffers and undergoes self assembly into macrofibers. Herein, we have made a first attempt to improve the mechanical properties of 4RepCT fibers by selective introduction of AA ? CC mutations and by letting the fibers form under physiologically relevant redox conditions. Introduction of AA ? CC mutations in the first poly-Ala block in the miniature spidroin increases the stiffness and tensile strength without changes in ability to form fibers, or in fiber morphology. These improved mechanical properties correlate with degree of disulfide formation. AA ? CC mutations in the forth poly-Ala block, however, lead to premature aggregation of the protein, possibly due to disulfide bonding with a conserved Cys in the C-terminal domain. Replacement of this Cys with a Ser, lowers thermal stability but does not interfere with dimerization, fiber morphology or tensile strength. These results show that mutagenesis of 4RepCT can reveal spidroin structure-activity relationships and generate recombinant fibers with improved mechanical properties. PMID:19388023

Grip, S; Johansson, J; Hedhammar, M

2009-01-01

300

Silk fibroin derived polypeptide-induced biomineralization of collagen.  

PubMed

Silk fibroin (SF) is extensively investigated in osteoregenerative therapy as it combines extraordinary mechanical properties and directs calcium-phosphate formation. However, the role of the peptidic fractions in inducing the protein mineralization has not been previously decoded. In this study, we investigated the mineralization of fibroin-derived polypeptides (FDPs), which were obtained through the chymotryptic separation of the hydrophobic crystalline (Cp) fractions and of the hydrophilic electronegative amorphous (Cs) fractions. When immersed in simulated body fluid (SBF), only Cs fragments demonstrated the formation of carbonated apatite, providing experimental evidence that the mineralization of SF is dictated exclusively by its electronegative amino-acidic sequences. The potential of Cs to conceptually mimic the role of anionic non-collagenous proteins in biomineralization processes was investigated via their incorporation (up to 10% by weight) in bulk osteoid-like dense collagen (DC) gels. Within 6 h in SBF, apatite was formed in DC-Cs hybrid gels, and by day 7, carbonated hydroxylapatite crystals were extensively formed. This accelerated 3-D mineralization resulted in a nine-fold increase in the compressive modulus of the hydrogel. The tailoring of the mineralization and mechanical properties of hydrogels through hybridization with FDPs could potentially have a significant impact on cell delivery and bone regenerative medicine. PMID:21982293

Marelli, Benedetto; Ghezzi, Chiara E; Alessandrino, Antonio; Barralet, Jake E; Freddi, Giuliano; Nazhat, Showan N

2012-01-01

301

Antennal SNMPs (Sensory Neuron Membrane Proteins) of Lepidoptera Define a Unique Family of  

E-print Network

: SNMP1-Apol is an antennal-specific protein of the wild silk moth Antheraea polyphemus; the proteinDa olfactory-specific protein cloned from an adult antenna-specific cDNA library from the silk moth Antheraea polyphemus (Rogers et al., 1997). SNMP is uniquely expressed in olfactory neurons and is localized

Vogt, Richard G.

302

NEW SILKS ROADS: PROMISES AND PERILS OF THE INTERNET IN THE THAI SILK INDUSTRY  

Microsoft Academic Search

The Internet is often touted as a panacea for perceived deficiencies in economic development. Its space-transcending abilities, which can instantly connect producers with consumers, have the potential to cut out intermediaries and to redistribute economic surplus in a more equitable manner. This dissertation asks whether the promises of the Internet are being realized in the Thai silk industry.\\u000aThe project

Mark Graham

2008-01-01

303

Plasticity in Major Ampullate Silk Production in Relation to Spider Phylogeny and Ecology  

PubMed Central

Spider major ampullate silk is a high-performance biomaterial that has received much attention. However, most studies ignore plasticity in silk properties. A better understanding of silk plasticity could clarify the relative importance of chemical composition versus processing of silk dope for silk properties. It could also provide insight into how control of silk properties relates to spider ecology and silk uses. We compared silk plasticity (defined as variation in the properties of silk spun by a spider under different conditions) between three spider clades in relation to their anatomy and silk biochemistry. We found that silk plasticity exists in RTA clade and orbicularian spiders, two clades that differ in their silk biochemistry. Orbiculariae seem less dependent on external spinning conditions. They probably use a valve in their spinning duct to control friction forces and speed during spinning. Our results suggest that plasticity results from different processing of the silk dope in the spinning duct. Orbicularian spiders seem to display better control of silk properties, perhaps in relation to their more complex spinning duct valve. PMID:21818328

Boutry, Cecilia; Rezac, Milan; Blackledge, Todd Alan

2011-01-01

304

Synthesis and characterization of silk-inspired thermoplastic polyurethane elastomers  

E-print Network

Segmented polyurethane elastomers containing additional ordered structures within the hard or soft domains were developed to mimic the hierarchical structure and superior properties observed in spider silk fibers. The ...

Pollock, Gregory S

2005-01-01

305

21 CFR 878.5030 - Natural nonabsorbable silk surgical suture.  

Code of Federal Regulations, 2012 CFR

...FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GENERAL AND PLASTIC SURGERY DEVICES Surgical Devices § 878.5030 Natural nonabsorbable silk surgical suture. (a)...

2012-04-01

306

21 CFR 878.5030 - Natural nonabsorbable silk surgical suture.  

...FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GENERAL AND PLASTIC SURGERY DEVICES Surgical Devices § 878.5030 Natural nonabsorbable silk surgical suture. (a)...

2014-04-01

307

21 CFR 878.5030 - Natural nonabsorbable silk surgical suture.  

Code of Federal Regulations, 2013 CFR

...FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GENERAL AND PLASTIC SURGERY DEVICES Surgical Devices § 878.5030 Natural nonabsorbable silk surgical suture. (a)...

2013-04-01

308

21 CFR 878.5030 - Natural nonabsorbable silk surgical suture.  

Code of Federal Regulations, 2011 CFR

...FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GENERAL AND PLASTIC SURGERY DEVICES Surgical Devices § 878.5030 Natural nonabsorbable silk surgical suture. (a)...

2011-04-01

309

VIEW ALONG SEVENTEENTH STREET. NOTE THE MATURE SILK OAK TREES ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

VIEW ALONG SEVENTEENTH STREET. NOTE THE MATURE SILK OAK TREES LINING THE STREET, WHICH DO NOT PROVIDE A CANOPY VIEW FACING NORTHWEST. - Hickam Field, Hickam Historic Housing, Honolulu, Honolulu County, HI

310

Silk fibroin and polyethylene glycol-based biocompatible tissue adhesives  

PubMed Central

Tissue sealants have emerged in recent years as strong candidates for hemostasis. A variety of formulations are currently commercially available and though they satisfy many of the markets’ needs there are still key aspects of each that need improvement. Here we present a new class of blends, based on silk fibroin and chemically active polyethylene glycols (PEGs) with strong adhesive properties. These materials are cytocompatible, crosslink within seconds via chemical reaction between thiols and maleimides present on the constituent PEGs and have the potential to further stabilize through ?-sheet formation by silk. Based on the silk concentration in the final formulation, the adhesive properties of these materials are comparable or better than the current leading PEG-based sealant. In addition, the silk-PEG based materials show decreased swelling and longer degradation times. Such properties would make them suitable for applications for which the current sealants are contraindicated. PMID:21681949

Serban, Monica A.; Panilaitis, Bruce; Kaplan, David L.

2012-01-01

311

Immobilized Lentivirus Vector on Chondroitin Sulfate-Hyaluronate Acid-Silk Fibroin Hybrid Scaffold for Tissue-Engineered Ligament-Bone Junction  

PubMed Central

The lack of a fibrocartilage layer between graft and bone remains the leading cause of graft failure after anterior cruciate ligament (ACL) reconstruction. The objective of this study was to develop a gene-modified silk cable-reinforced chondroitin sulfate-hyaluronate acid-silk fibroin (CHS) hybrid scaffold for reconstructing the fibrocartilage layer. The scaffold was fabricated by lyophilizing the CHS mixture with braided silk cables. The scanning electronic microscopy (SEM) showed that microporous CHS sponges were formed around silk cables. Each end of scaffold was modified with lentiviral-mediated transforming growth factor-?3 (TGF-?3) gene. The cells on scaffold were transfected by bonded lentivirus. In vitro culture demonstrated that mesenchymal stem cells (MSCs) on scaffolds proliferated vigorously and produced abundant collagen. The transcription levels of cartilage-specific genes also increased with culture time. After 2 weeks, the MSCs were distributed uniformly throughout scaffold. Deposited collagen was also found to increase. The chondral differentiation of MSCs was verified by expressions of collagen II and TGF-?3 genes in mRNA and protein level. Histology also confirmed the production of cartilage extracellular matrix (ECM) components. The results demonstrated that gene-modified silk cable-reinforced CHS scaffold was capable of supporting cell proliferation and differentiation to reconstruct the cartilage layer of interface. PMID:25019087

Sun, Liguo; Li, Hongguo; Qu, Ling; Zhu, Rui; Fan, Xiangli; Xue, Yingsen; Xie, Zhenghong; Fan, Hongbin

2014-01-01

312

Studies on silk secretion in the trichoptera (F. Limnephilidae)  

Microsoft Academic Search

The ultrastructure and amino acid composition of the secreted silk of two species of trichopteran larvae, Pycnopsyche guttifer (Walk.) and Neophylax concinnus McL., were investigated. The spinnerets of these two animals were also examined by scanning electron microscopy. The silk consists of double-stranded, flat ribbons (1–4 µ wide), composed of bundles of 15–25 Å filaments. There are two components of

Maryann S. Engster

1976-01-01

313

SilkDB: a knowledgebase for silkworm biology and genomics  

Microsoft Academic Search

The Silkworm Knowledgebase (SilkDB) is a web- based repository for the curation, integration and study of silkworm genetic and genomic data. With the recent accomplishment of a 6X draft genome sequence of the domestic silkworm (Bombyx mori), SilkDB provides an integrated representation of the large-scale, genome-wide sequence assembly, cDNAs, clusters of expressed sequence tags (ESTs), transposable elements (TEs), mutants, single

Jing Wang; Qingyou Xia; Ximiao He; Mingtao Dai; Jue Ruan; Jie Chen; Guo Yu; Haifeng Yuan; Yafeng Hu; Ruiqiang Li; Tao Feng; Chen Ye; Cheng Lu; Jun Wang; Songgang Li; Gane Ka-shu Wong; Huanming Yang; Jian Wang; Zhonghuai Xiang; Zeyang Zhou; Jun Yu

2005-01-01

314

Laminar silk scaffolds for aligned tissue fabrication  

PubMed Central

3D biomaterial scaffolds with aligned architecture are of vital importance in tissue regeneration to mimic native tissue hierarchy and hence function. We demonstrate a generic method to produce aligned biomaterial scaffolds using the physics of directional ice freezing. Homogeneously aligned 3D silk scaffold with high porosity and alignment was demonstrated. The method can be adapted to a wide range of polymers and is devoid of any chemical reactions, thus avoiding potential complications associated with by-products and purification procedures. Subsequently, the 3D aligned system was tested for mechanical properties and cellular responses with chondrocytes and bone marrow derived human mesenchymal stem cells, assessing survival, proliferation and differentiation. In vivo tests suggested biocompatibility of the matrices for future tissue engineering applications, specifically in areas where high cellular alignment is needed. PMID:23161731

Mandal, Biman B.; Gil, Eun Seok; Panilaitis, Bruce; Kaplan, David L.

2013-01-01

315

Electricity from the silk cocoon membrane.  

PubMed

Silk cocoon membrane (SCM) is an insect engineered structure. We studied the electrical properties of mulberry (Bombyx mori) and non-mulberry (Tussar, Antheraea mylitta) SCM. When dry, SCM behaves like an insulator. On absorbing moisture, it generates electrical current, which is modulated by temperature. The current flowing across the SCM is possibly ionic and protonic in nature. We exploited the electrical properties of SCM to develop simple energy harvesting devices, which could operate low power electronic systems. Based on our findings, we propose that the temperature and humidity dependent electrical properties of the SCM could find applications in battery technology, bio-sensor, humidity sensor, steam engines and waste heat management. PMID:24961354

Tulachan, Brindan; Meena, Sunil Kumar; Rai, Ratan Kumar; Mallick, Chandrakant; Kusurkar, Tejas Sanjeev; Teotia, Arun Kumar; Sethy, Niroj Kumar; Bhargava, Kalpana; Bhattacharya, Shantanu; Kumar, Ashok; Sharma, Raj Kishore; Sinha, Neeraj; Singh, Sushil Kumar; Das, Mainak

2014-01-01

316

Amorphous Silk Fibroin Membranes for Separation of CO2  

NASA Technical Reports Server (NTRS)

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.

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

2009-01-01

317

Multifunctional Silk Nerve Guides for Axon Outgrowth  

NASA Astrophysics Data System (ADS)

Peripheral nerve regeneration is a critical issue as 2.8% of trauma patients present with this type of injury, estimating a total of 200,000 nerve repair procedures yearly in the United States. While the peripheral nervous system exhibits slow regeneration, at a rate of 0.5 mm -- 9 mm/day following trauma, this regenerative ability is only possible under certain conditions. Clinical repairs have changed slightly in the last 30 years and standard methods of treatment include suturing damaged nerve ends, allografting, and autografting, with the autograft the gold standard of these approaches. Unfortunately, the use of autografts requires a second surgery and there is a shortage of nerves available for grafting. Allografts are a second option however allografts have lower success rates and are accompanied by the need of immunosuppressant drugs. Recently there has been a focus on developing nerve guides as an "off the shelf" approach. Although some natural and synthetic guidance channels have been approved by the FDA, these nerve guides are unfunctionalized and repair only short gaps, less than 3 cm in length. The goal of this project was to identify strategies for functionalizing peripheral nerve conduits for the outgrowth of neuron axons in vitro . To accomplish this, two strategies (bioelectrical and biophysical) were indentified for increasing axon outgrowth and promoting axon guidance. Bioelectrical strategies exploited electrical stimulation for increasing neurite outgrowth. Biophysical strategies tested a range of surface topographies for axon guidance. Novel methods were developed for integrating electrical and biophysical strategies into silk films in 2D. Finally, a functionalized nerve conduit system was developed that integrated all strategies for the purpose of attaching, elongating, and guiding nervous tissue in vitro. Future directions of this work include silk conduit translation into a rat sciatic nerve model in vivo for the purpose of repairing long (> 3 cm) peripheral nerve gaps.

Tupaj, Marie C.

318

Stability and cytocompatibility of silk fibroin-capped gold nanoparticles.  

PubMed

Surface engineering is crucial in the colloidal stability and biocompatibility of nanoparticles (NPs). Protein silk fibroin (SF), which gained interest in biomaterial and regenerative medicine, was used in this study to stabilize gold (Au) NPs. Characterization results from UV-Vis spectroscopy revealed that SF-capped Au NPs (SF-Au NPs) possessed remarkable colloidal stabilities in the pH range of 2 to 11 and salt concentration range of 50mM to 1000mM. In addition, dried particle samples were resuspended after lyophilization without aggregation. The results indicated that the steric hindrance rather than the electrostatic repulsion of SF-Au NPs was essential for colloidal stability. The SF-Au NPs manifested improved cytocompatibility compared with bare Au NPs, which was attributed to the inherent non-cytotoxicity of SF and the good colloidal stability of the NPs. The proposed method was simpler, more efficient, and more cost effective than the conventional modification strategies for Au NPs; thus, SF-Au NPs can be potentially used in biomedical applications. PMID:25175209

Jia, Lan; Guo, Li; Zhu, Jingxin; Ma, Yanlong

2014-10-01

319

Structure and ultrastructure of the silk glands and spinneret of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae).  

PubMed

This study provides comprehensive documentation of silk production in the pest moth Helicoverpa armigera from gland secretion to extrusion of silk thread. The structure of the silk glands, accessory structures and extrusion apparatus are reported. The general schema of the paired silk glands follows that found for Lepidoptera. Morphology of the duct, silk press, muscle attachments and spigot are presented as a three-dimensional reconstruction and the cuticular crescent-shaped profile of the silk press is demonstrated in both open and closed forms with attendant muscle blocks, allowing advances in our knowledge of how the silk press functions to regulate the extrusion of silk. Growth of the spigot across instars is documented showing a distinctive developmental pattern for this extrusion device. Its shape and structure are related to use and load-bearing activity. PMID:18089054

Sorensen, Gregory S; Cribb, Bronwen W; Merritt, David; Johnson, Marie-Louise; Zalucki, Myron P

2006-03-01

320

Reversible Assembly of ?-Sheet Nanocrystals within Caddisfly Silk  

PubMed Central

Nuclear magnetic resonance (NMR) and X-ray diffraction (XRD) experiments reveal the structural importance of divalent cation–phosphate complexes in the formation of ?-sheet nanocrystals from phosphorylated serine-rich regions within aquatic silk from caddisfly larvae of the species Hesperophyla consimilis. Wide angle XRD data on native caddisfly silk show that the silk contains a significant crystalline component with a repetitive orthorhombic unit cell aligned along the fiber axis with dimensions of 5.9 Å × 23.2 Å × 17.3 Å. These nanocrystalline domains depend on multivalent cations, which can be removed through chelation with ethylenediaminetetraacetic acid (EDTA). A comparison of wide angle X-ray diffraction data before and after EDTA treatment reveals that the integrated peak area of reflections corresponding to the nanocrystalline regions decreases by 15–25% while that of the amorphous background reflections increases by 20%, indicating a partial loss of crystallinity. 31P solid-state NMR data on native caddisfly silk also show that the phosphorylated serine-rich motifs transform from a rigid environment to one that is highly mobile and water-solvated after treatment with EDTA. The removal of divalent cations through exchange and chelation has therefore caused a collapse of the ?-sheet structure. However, NMR results show that the rigid phosphorus environment is mostly recovered after the silk is re-treated with calcium. The 31P spin–lattice (T1) relaxation times were measured at 7.6 ± 3.1 and 1 ± 0.5 s for this calcium-recovered sample and the native silk sample, respectively. The shorter 31P T1 relaxation times measured for the native silk sample are attributed to the presence of paramagnetic iron that is stripped away during EDTA chelation treatment and replaced with diamagnetic calcium. PMID:24576204

Addison, J. Bennett; Weber, Warner S.; Mou, Qiushi; Ashton, Nicholas N.; Stewart, Russell J.; Holland, Gregory P.; yarger, Jeffery L.

2014-01-01

321

Tough silk fibers prepared in air using a biomimetic microfluidic chip.  

PubMed

Microfluidic chips with single channel were built to mimic the shear and elongation conditions in the spinning apparatus of spider and silkworm. Silk fibers dry-spun from regenerated silk fibroin (RSF) aqueous solution using the chip could be tougher than degummed natural silk. The artificial silk exhibited a breaking strength up to 614 MPa, a breaking elongation up to 27% and a breaking energy of 101 kJ/kg. PMID:24613677

Luo, Jie; Zhang, Lele; Peng, Qingfa; Sun, Mengjie; Zhang, Yaopeng; Shao, Huili; Hu, Xuechao

2014-05-01

322

Research Journal Highlights The silk road to bio-integrated electronics  

E-print Network

Research Journal Highlights The silk road to bio-integrated electronics Nature Materials, April 19 Mutations confer increased risk of breast and ovarian cancer (Nature Genetics) · 19 Apr The silk road to bio rights reserved. Page 1 of 1The silk road to bio-integrated electronics (Nature Materials): NPG Nature

Rogers, John A.

323

SilkRoad: A Multithreaded Runtime System with Software Distributed Shared Memory for SMP Clusters  

E-print Network

SilkRoad: A Multithreaded Runtime System with Software Distributed Shared Memory for SMP Clusters L for user level shared memory. In this paper, we describe SilkRoad, an extension of distributed Cilk, which im- plementing the Lazy Release Consistency (LRC) mem- ory model. In the SilkRoad runtime system

Wong, Weng Fai

324

Antimicrobial Silk Clothing in the Treatment of Atopic Dermatitis Proves Comparable to Topical Corticosteroid Treatment  

Microsoft Academic Search

Background: Atopic dermatitis (AD) is aggravated by mechanical irritation and bacterial colonization. Objective: This study compared the efficacy of an antimicrobial silk fabric (DermaSilk®) with that of a topical corticosteroid in the treatment of AD. Methods: Fifteen children were enrolled and wore a dress, where the left side was made of DermaSilk and the right side was made of cotton.

G. Senti; L. S. Steinmann; B. Fischer; R. Kurmann; T. Storni; P. Johansen; P. Schmid-Grendelmeier; B. Wüthrich; T. M. Kündig

2006-01-01

325

Hierarchical Chain Model of Spider Capture Silk Elasticity Haijun Zhou1  

E-print Network

Hierarchical Chain Model of Spider Capture Silk Elasticity Haijun Zhou1 and Yang Zhang2 1 Max 2005) Spider capture silk is a biomaterial with both high strength and high elasticity of spider capture silk is in support of this model. DOI: 10.1103/PhysRevLett.94.028104 PACS numbers: 87

Zhang, Yang

326

INTRODUCTION Most of the world's 40,000 species of spiders produce dragline silk  

E-print Network

spider silk an important model for biomimetic research (Gosline et al., 1986; Vadlamudi, 1995; Vollrath the impressive performance of silk is not limited solely to tensile mechanics. Here, we show that spider silk provide inspiration for biomimetic muscle. The cellulose fibrils in wheat awns expand and contract

Agnarsson, Ingi

327

SILK: Scout Paths in the Linux Kernel Andy Bavier Thiemo Voigt Mike Wawrzoniak Larry Peterson  

E-print Network

SILK: Scout Paths in the Linux Kernel Andy Bavier #5; Thiemo Voigt #3; Mike Wawrzoniak #5; Larry Peterson #5; Per Gunningberg z November 20, 2001 Abstract SILK stands for Scout In the Linux Kernel, and is a port of the Scout operating system to run as a Linux kernel mod­ ule. SILK forms a replacement

Voigt, Thiemo

328

The Silk Roads. Mathematical model. A.S.Malkov, Keldysh Institute of Applied Mathematics, Moscow, Russia  

E-print Network

The Silk Roads. Mathematical model. A.S.Malkov, Keldysh Institute of Applied Mathematics, Moscow. The dynamics of the Silk Roads is described by means of formal spatial equations. Historical data conveys the facts that the location of the trade routes known as the Silk Roads altered profoundly enough from epoch

White, Douglas R.

329

A novel property of spider silk: chemical defence against ants  

PubMed Central

Spider webs are made of silk, the properties of which ensure remarkable efficiency at capturing prey. However, remaining on, or near, the web exposes the resident spiders to many potential predators, such as ants. Surprisingly, ants are rarely reported foraging on the webs of orb-weaving spiders, despite the formidable capacity of ants to subdue prey and repel enemies, the diversity and abundance of orb-web spiders, and the nutritional value of the web and resident spider. We explain this paradox by reporting a novel property of the silk produced by the orb-web spider Nephila antipodiana (Walckenaer). These spiders deposit on the silk a pyrrolidine alkaloid (2-pyrrolidinone) that provides protection from ant invasion. Furthermore, the ontogenetic change in the production of 2-pyrrolidinone suggests that this compound represents an adaptive response to the threat of natural enemies, rather than a simple by-product of silk synthesis: while 2-pyrrolidinone occurs on the silk threads produced by adult and large juvenile spiders, it is absent on threads produced by small juvenile spiders, whose threads are sufficiently thin to be inaccessible to ants. PMID:22113027

Zhang, Shichang; Koh, Teck Hui; Seah, Wee Khee; Lai, Yee Hing; Elgar, Mark A.; Li, Daiqin

2012-01-01

330

Self-Assembly of Natural Silk Fibroin under Osmotic Stress  

NASA Astrophysics Data System (ADS)

Osmotic stress method was applied to investigate the supramolecular self-assembly behavior of natural silk fibroin within the gland. As with the experiments on re-generated silk fibroin, poly(ethylene glycol) 8K was used to apply an osmotic stress of 0.2-7.6 MPa to the gland of Bombyx mori silkworm, in vitro. Fibroin samples were extracted from the fibroin-dominant, water-soluble posterior region, and from three different parts in the middle region of the gland. Calcium chloride of 0.01 M was added to each stressing solution to balance the physiological salt content of the sample. Microscopic and thermodynamic details of this self-assembly process along the spinline have been assessed by wide angle X- ray diffraction, optical microscopy, etc. It is apparent that as osmotic stress increases, isotropic silk fibroin molecules in the posterior region are assembled together to form a water-soluble crystalline mesophase known as silk-I. Further increases in osmotic stress induce an anti- parallel beta-sheet structure known as silk-II.

Sohn, Sungkyun; Hata, Tamako; Strey, Helmut H.

2005-03-01

331

Protease inhibitor from insect silk-activities of derivatives expressed in vitro and in transgenic potato.  

PubMed

Several recombinant derivatives of serine protease inhibitor called silk protease inhibitor 2 (SPI2), which is a silk component in Galleria mellonella (Lepidoptera, Insecta), were prepared in the expression vector Pichia pastoris. Both the native and the recombinant protease inhibitors were highly active against subtilisin and proteinase K. The synthetic SPI2 gene with Ala codon in the P1 position was fused with mGFP-5 to facilitate detection of the transgene and its protein product. A construct of the fusion gene with plant regulatory elements (promoter 35S and terminator OCS) was inserted into the binary vector pRD400. The final construct was introduced into Agrobacterium tumefaciens that was then used for genetic transformation of the potato variety Velox. The transgene expression was monitored with the aid of ELISA employing polyclonal antibody against natural SPI2. In vitro tests showed increased resistance to the late blight Phytophthora infestans in several transformed lines. No effect was seen on the growth, mortality, life span or reproduction of Spodoptera littoralis (Lepidoptera, Insecta) caterpillars, while feeding on transformed potato plants expressing the fusion protein, indicating that the transformed potatoes may be harmless to non-target organisms. PMID:23824530

Kodrík, Dalibor; Kludkiewicz, Barbara; Navrátil, Old?ich; Skoková Habuštová, Oxana; Horá?ková, Vendulka; Svobodová, Zde?ka; Vinokurov, Konstantin S; Sehnal, František

2013-09-01

332

The Silk Road, Marco Polo, a Bible and its proteome: a detective story.  

PubMed

Around the end of XIII century (at the time of young Marco Polo's first trip to China at the court of Khubilai Khan in Khan Baliq) a pocket Bible was delivered by a Franciscan friar to the Mogul Emperor, in the framework of the evangelization program of the Far East. Four centuries later, in 1685, this Bible was rediscovered by the Jesuit Philippe Couplet in the house of a rich Chinese in Nanchin and donated to Cosimo III, Grand Duke of Tuscany. This Bible was recently "unearthed" in the Biblioteca Medicea Laurenziana in Florence, wrapped up in a precious yellow silk cloth, in a rather ruined state. After two years of restoration, the Bible will return to China in 2012 for a celebration of its >700years of life and of its remarkable return trip on the Silk Road. On account of the thinness of the parchment (barely 80?m thickness, the size of each foil being 16.5×11cm) it was widely held that the pages were produced from foetal lambskins. On tiny fragments of the margins of a foil, after several unsuccessful attempts at digesting the vellum, we were able to obtain a tryptic peptide mixture, which, upon mass spectrometry analysis, yielded the identity of 8 unique proteins, belonging to the genus Bos taurus, thus confirming the origin of the vellum from calfskins rather than from foetal lambskins. Our results prove that it is possible to obtain reliable protein extraction and IDs from ancient parchment documents. PMID:22504796

Toniolo, Lucia; D'Amato, Alfonsina; Saccenti, Riccardo; Gulotta, Davide; Righetti, Pier Giorgio

2012-06-18

333

The Consolidation Behavior of Silk Hydrogels  

PubMed Central

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 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

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

2010-01-01

334

Enhanced osteoinductivity and osteoconductivity through hydroxyapatite coating of silk-based tissue-engineered ligament scaffold.  

PubMed

Hybrid silk scaffolds combining knitted silk fibers and silk sponge have been recently developed for use as ligament-alone grafts. Incorporating an osteoinductive phase into the ends of a ligament scaffold may potentially generate an integrated "bone-ligament-bone" graft and improve graft osteointegration with host bone. To explore the possible application of hydroxyapatite (HA) coating in the fabrication of osteoinductive ends of silk-based scaffold, HA was coated on the hybrid silk scaffold and the effects to the bone-related cells were evaluated. HA could be coated in a uniform and controlled manner on the silk sponge, using an alternate soaking technology, with the amount deposited being dependent on the number of soaking cycles. HA coating also progressively reduced the hydrophobicity of silk surface (decreasing water contact angle from 87° to 42-76°, after 1-3 soaking cycles), making the HA-coated silk scaffold less favorable for initial cell attachments; but the attached cells showed viability and sustained proliferation on the HA-coated scaffold. As demonstrated by real-time polymerase chain reaction and alkaline phosphatase assay, the osteoinductivity of HA-coated silk scaffolds resulted in the osteogenic differentiation of bone marrow mesenchymal stem cells, and the osteoconductivity of HA-coated silk scaffolds supported osteoblasts growth and maintained the properties of mature osteoblasts. These properties of HA-coating demonstrated its possible application in fabricating osteoinductive ends of the silk-based ligament graft to potentially enhance graft-to-host bone integration. PMID:22949167

He, Pengfei; Sahoo, Sambit; Ng, Kian Siang; Chen, Kelei; Toh, Siew Lok; Goh, James Cho Hong

2013-02-01

335

Investigation of the properties and potential medical applications of natural silk fibers produced by Eupackardia calleta.  

PubMed

Silk has been considered biocompatible and used for medical applications for centuries. However, most of the silk currently used is produced by the domesticated silkworm, Bombyx mori. Recently, it has been demonstrated that silk produced by saturniidae insects such as Antheraea mylitta, Phylisomia ricini, and Antheraea pernyi had unique properties and suitable for medical applications. Therefore, efforts are being made to identify and study the structure and properties of silks produced by wild insects (non B. mori), spiders, and ants to understand their suitability for various medical applications. Eupackardia calleta belongs to the Saturniidae family of insects, but the structure, properties, and potential medical uses of silk fibers produced by E. calleta are not known. In this research, we have characterized the properties of silk fibers produced by E. calleta and the ability of the fibers to support the attachment and proliferation of mouse fibroblast cells. Silk produced by E. calleta had considerably different amino acid composition than B. mori and A. mylitta, P. ricini, and A. pernyi silks. Breaking tenacity of the E. calleta silk fibers at 400 MPa was between that of B. mori silk and A. mylitta, P. ricini, and A. pernyi silks. E. calleta showed good attachment and spreading of mouse fibroblast cells suggesting potential for medical applications. PMID:23565687

Reddy, Narendra; Jiang, Qiuran; Yang, Yiqi

2013-01-01

336

Environmentally friendly surface modification of silk fiber: Chitosan grafting and dyeing  

NASA Astrophysics Data System (ADS)

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.

Davarpanah, Saideh; Mahmoodi, Niyaz Mohammad; Arami, Mokhtar; Bahrami, Hajir; Mazaheri, Firoozmehr

2009-01-01

337

Electrospun silk-elastin-like fibre mats for tissue engineering applications.  

PubMed

Protein-based polymers are present in a wide variety of organisms fulfilling structural and mechanical roles. Advances in protein engineering and recombinant DNA technology allow the design and production of recombinant protein-based polymers (rPBPs) with an absolute control of its composition. Although the application of recombinant proteins as biomaterials is still an emerging technology, the possibilities are limitless and far superior to natural or synthetic materials, as the complexity of the structural design can be fully customized. In this work, we report the electrospinning of two new genetically engineered silk-elastin-like proteins (SELPs) consisting of alternate silk- and elastin-like blocks. Electrospinning was performed with formic acid and aqueous solutions at different concentrations without addition of further agents. The size and morphology of the electrospun structures was characterized by scanning electron microscopy showing its dependence on the concentration and solvent used. Treatment with methanol-saturated air was employed to stabilize the structure and promote water insolubility through a time-dependent conversion of random coils into ?-sheets (FTIR). The resultant methanol-treated electrospun mats were characterized for swelling degree (570-720%), water vapour transmission rate (1083 g/m(2)/day) and mechanical properties (modulus of elasticity ?126 MPa). Furthermore, the methanol-treated SELP fibre mats showed no cytotoxicity and were able to support adhesion and proliferation of normal human skin fibroblasts. Adhesion was characterized by a filopodia-mediated mechanism. These results demonstrate that SELP fibre mats can provide promising solutions for the development of novel biomaterials suitable for tissue engineering applications. PMID:24287397

Machado, Raul; da Costa, André; Sencadas, Vitor; Garcia-Arévalo, Carmen; Costa, Carlos M; Padrão, Jorge; Gomes, Andreia; Lanceros-Méndez, Senentxu; Rodríguez-Cabello, José Carlos; Casal, Margarida

2013-12-01

338

Carbonic anhydrase generates CO2 and H+ that drive spider silk formation via opposite effects on the terminal domains.  

PubMed

Spider silk fibers are produced from soluble proteins (spidroins) under ambient conditions in a complex but poorly understood process. Spidroins are highly repetitive in sequence but capped by nonrepetitive N- and C-terminal domains (NT and CT) that are suggested to regulate fiber conversion in similar manners. By using ion selective microelectrodes we found that the pH gradient in the silk gland is much broader than previously known. Surprisingly, the terminal domains respond in opposite ways when pH is decreased from 7 to 5: Urea denaturation and temperature stability assays show that NT dimers get significantly stabilized and then lock the spidroins into multimers, whereas CT on the other hand is destabilized and unfolds into ThT-positive ?-sheet amyloid fibrils, which can trigger fiber formation. There is a high carbon dioxide pressure (pCO2) in distal parts of the gland, and a CO2 analogue interacts with buried regions in CT as determined by nuclear magnetic resonance (NMR) spectroscopy. Activity staining of histological sections and inhibition experiments reveal that the pH gradient is created by carbonic anhydrase. Carbonic anhydrase activity emerges in the same region of the gland as the opposite effects on NT and CT stability occur. These synchronous events suggest a novel CO2 and proton-dependent lock and trigger mechanism of spider silk formation. PMID:25093327

Andersson, Marlene; Chen, Gefei; Otikovs, Martins; Landreh, Michael; Nordling, Kerstin; Kronqvist, Nina; Westermark, Per; Jörnvall, Hans; Knight, Stefan; Ridderstråle, Yvonne; Holm, Lena; Meng, Qing; Jaudzems, Kristaps; Chesler, Mitchell; Johansson, Jan; Rising, Anna

2014-08-01

339

Osteochondral Tissue Engineering In Vivo: A Comparative Study Using Layered Silk Fibroin Scaffolds from Mulberry and Nonmulberry Silkworms  

PubMed Central

The ability to treat osteochondral defects is a major clinical need. Existing polymer systems cannot address the simultaneous requirements of regenerating bone and cartilage tissues together. The challenge still lies on how to improve the integration of newly formed tissue with the surrounding tissues and the cartilage-bone interface. This study investigated the potential use of different silk fibroin scaffolds: mulberry (Bombyx mori) and non-mulberry (Antheraea mylitta) for osteochondral regeneration in vitro and in vivo. After 4 to 8 weeks of in vitro culture in chondro- or osteo-inductive media, non-mulberry constructs pre-seeded with human bone marrow stromal cells exhibited prominent areas of the neo tissue containing chondrocyte-like cells, whereas mulberry constructs pre-seeded with human bone marrow stromal cells formed bone-like nodules. In vivo investigation demonstrated neo-osteochondral tissue formed on cell-free multi-layer silk scaffolds absorbed with transforming growth factor beta 3 or recombinant human bone morphogenetic protein-2. Good bio-integration was observed between native and neo-tissue within the osteochondrol defect in patellar grooves of Wistar rats. The in vivo neo-matrix formed comprised of a mixture of collagen and glycosaminoglycans except in mulberry silk without growth factors, where a predominantly collagenous matrix was observed. Immunohistochemical assay showed stronger staining of type I and type II collagen in the constructs of mulberry and non-mulberry scaffolds with growth factors. The study opens up a new avenue of using inter-species silk fibroin blended or multi-layered scaffolds of a combination of mulberry and non-mulberry origin for the regeneration of osteochondral defects. PMID:24260335

Saha, Sushmita; Kundu, Banani; Kirkham, Jennifer; Wood, David; Kundu, Subhas C.; Yang, Xuebin B.

2013-01-01

340

Dynamic Culture Conditions to Generate Silk-Based Tissue-Engineered Vascular Grafts  

PubMed Central

Tissue engineering is an alternative approach for the preparation of small-diameter (<6 mm) vascular grafts due to the potential to control thrombosis, anastomotic cellular hyperplasia and matrix production. This control also requires the maintenance of graft patency in vivo, appropriate mechanical properties and the formation of a functional endothelium. As a first step in generating such tissue-engineered vascular grafts (TEVG), our objective was to develop a tissue-engineered construct that mimicked the structure of blood vessels using tubular electrospun silk fibroin scaffolds (ESFS) with suitable mechanical properties. Human coronary artery smooth muscle cells (HCASMCs) and human aortic endothelial cells (HAECs) were sequentially seeded onto the luminal surface of the tubular scaffolds and cultivated under physiological pulsatile flow. The results demonstrated that TEVGs under dynamic flow conditions had better outcome than static culture controls in terms of cell proliferation and alignment, ECM production and cell phenotype based on transcript and protein level assessments. The metabolic activity of HCASMCs present in the TEGs indicated the advantage of dynamic flow over static culture in effective nutrient and oxygen distribution to the cells. A matrigel coating as a basement membrane mimic for ECM significantly improved endothelium coverage and retention under physiological shear forces. The results demonstrate the successful integration of vascular cells into silk electrospun tubular scaffolds as a step toward the development of a TEVG similar to native vessels in terms of vascular cell outcomes and mechanical properties. PMID:19232717

Zhang, Xiaohui; Wang, Xiuli; Keshav, Vinny; Wang, Xiaoqin; Johanas, Jacqueline; Leisk, Gary; Kaplan, David L

2009-01-01

341

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

PubMed

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

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

2014-01-01

342

Focal Infection Treatment using Laser-Mediated Heating of Injectable Silk Hydrogels with Gold Nanoparticles  

PubMed Central

Medical treatment of subcutaneous bacterial abscesses usually involves systemic high-dose antibiotics and incision-drainage of the wound. Such an approach suffers from two main deficiencies: bacterial resistance to antibiotics and pain associated with multiple incision-drainage-wound packing procedures. Furthermore, the efficacy of high-dose systemic antibiotics is limited because of the inability to penetrate into the abscess. To address these obstacles, we present a treatment relying on laser-induced heating of gold nanoparticles embedded in an injectable silk-protein hydrogel. Although bactericidal nanoparticle systems have been previously employed based on silver and nitric oxide, they have limitations regarding customization and safety. The method we propose is safe and uses biocompatible, highly tunable materials: an injectable silk hydrogel and Au nanoparticles, which are effective absorbers at low laser powers such as those provided by hand held devices. We demonstrate that a single 10-minute laser treatment of a subcutaneous infection in mice preserves the general tissue architecture, while achieving a bactericidal effect - even resulting in complete eradication in some cases. The unique materials platform presented here can provide the basis for an alternative treatment of focal infections. PMID:24015118

Kojic, Nikola; Pritchard, Eleanor M.; Tao, Hu; Brenckle, Mark A.; Mondia, Jessica P.; Panilaitis, Bruce; Omenetto, Fiorenzo; Kaplan, David L.

2013-01-01

343

SILK: Higher Level Rules with Defaults and Semantic Scalability  

E-print Network

is the target for KA: "The KR is the deep UI" · Web knowledge interchange (with merging) for scalability://www.mit.edu/~bgrosof/ http://silk.semwebcentral.org ** 3rd International Conference on Web Reasoning and Rules, Chantilly, VA of the world's scientific and similar knowledge on-line · Answer questions, act as personal tutor, with deep

Polz, Martin

344

RECONSTRUCTION OF COMMERCIAL WAYS AND SILK ROAD IN NURATAU AREA  

Microsoft Academic Search

This manuscript is dedicated to history of economical relations and reconstruction of commercial routes on territory of Nuratau from 20 century B.C. till beginning of 20 century. This time interval was conditionally divided onto three periods: before, in time and past of existing the Great Silk Road (GSR). An attempt to reconstruct commercial routes in the region based on analysis

I. T. Muminova

345

Another Way of Knowing: Visualizing the Ancient Silk Routes  

ERIC Educational Resources Information Center

One way that people learn, remember and communicate is visually. We combine past experiences with new visual information to construct meaning. In this study, elementary teachers introduced their students to the peoples and places of the ancient silk routes using illustrations from two children's picture books, "Marco Polo," written by Gian Paolo…

Bisland, Beverly Milner

2010-01-01

346

Teaching the Silk Road: A Journey of Pedagogical Discovery.  

ERIC Educational Resources Information Center

Describes a course for first-year college students that focuses on the Silk Road. Discusses the problems that occurs in such a course, types of resources used, basic strategies and tactics taken, and the focus on mapmaking in the beginning of the course. Includes an annotated bibliography. (CMK)

Andrea, A. J.; Mierse, William

2002-01-01

347

Monday, April 19, 2010 Brain Interfaces Made of Silk  

E-print Network

://www.seas.upenn.edu/~littlab/Site/Brian.html) , professor of neurology and bioengineering at the University of Pennsylvania Medical School. "We wanted transistor arrays built on silk, and tested them in animals--just not in the brain yet. Schwartz says other groups have recognized the importance of multiplexing and signal amplification, but have been working

Rogers, John A.

348

12. NORTH FACADE, OPERATOR'S COTTAGE, SILK STOCKING ROW, NEWHALEM. THE ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

12. NORTH FACADE, OPERATOR'S COTTAGE, SILK STOCKING ROW, NEWHALEM. THE HOUSES WERE CONSTRUCTED SO THAT WHAT APPEARS TO BE THE FRONT OF THE HOUSE FACES THE RIVER AND THE FACADE FACING THE STREET LOOKS LIKE A BACK, 1989. - Skagit Power Development, Skagit River & Newhalem Creek Hydroelectric Project, On Skagit River, Newhalem, Whatcom County, WA

349

11. SOUTH FACADE (FRONT) OF AN OPERATOR'S COTTAGE ON SILK ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

11. SOUTH FACADE (FRONT) OF AN OPERATOR'S COTTAGE ON SILK STOCKING ROW. THESE COTTAGES WERE THE FIRST PERMANENT HOUSING CONSTRUCTED ON THE SKAGIT AND FOR MANY YEARS WERE CONSIDERED TO BE THE BEST. THEY WERE RESERVED FOR POWERHOUSE OPERATORS AND SUPERVISORS AND THEIR FAMILIES, 1989. - Skagit Power Development, Skagit River & Newhalem Creek Hydroelectric Project, On Skagit River, Newhalem, Whatcom County, WA

350

Directly obtaining pristine magnetic silk fibers from silkworm.  

PubMed

By feeding the silkworms with the nano Fe3O4 powder together with mulberry leaves, we directly obtained silkworm spun pristine magnetic silk fiber, MSF. To compare with the normal SF found that this MSF not only has expected magnetic properties, but also has enhanced thermal stability and mechanical properties, e.g. stress and strain. PMID:24269584

Wang, Jun-Ting; Li, Lu-Lu; Feng, Lei; Li, Jin-Fan; Jiang, Lin-Hai; Shen, Qing

2014-02-01

351

Silk fibroin-derived nanoparticles for biomedical applications.  

PubMed

The treatment of disease in the future will be influenced by the ability to produce therapeutic formulations that have high availability at the disease site, sustained and long-term release, with minimal to no toxicity to healthy tissues. Biologically derived delivery systems offer promise in this regard owing to minimization of adverse effects while increasing the efficacy of the entrapped therapeutic. Silk fibroin nanoparticles overcome barriers set by synthetic nondegradable nanoparticles made of silicone, polyethylene glycol and degradable polylactic acid-polyglycolic acid polymers. Silk fibroin-mediated delivery has demonstrated high efficacy in breast cancer cells. While the targeting is associated with the specificity of entrapped therapeutic for the diseased cells, silk fibroin-derived particles enhance intracellular uptake and retention resulting in downmodulation of more than one pathway due to longer availability of the therapeutic. The mechanism of targeting for the nanoparticle is based on the silk fibroin composition, beta-sheet structure and self-assembly into beta-barrels. PMID:20662650

Mathur, Anshu B; Gupta, Vishal

2010-07-01

352

Viscous friction between crystalline and amorphous phase of dragline silk.  

PubMed

The hierarchical structure of spider dragline silk is composed of two major constituents, the amorphous phase and crystalline units, and its mechanical response has been attributed to these prime constituents. Silk mechanics, however, might also be influenced by the resistance against sliding of these two phases relative to each other under load. We here used atomistic molecular dynamics (MD) simulations to obtain friction forces for the relative sliding of the amorphous phase and crystalline units of Araneus diadematus spider silk. We computed the coefficient of viscosity of this interface to be in the order of 10(2) Ns/m(2) by extrapolating our simulation data to the viscous limit. Interestingly, this value is two orders of magnitude smaller than the coefficient of viscosity within the amorphous phase. This suggests that sliding along a planar and homogeneous surface of straight polyalanine chains is much less hindered than within entangled disordered chains. Finally, in a simple finite element model, which is based on parameters determined from MD simulations including the newly deduced coefficient of viscosity, we assessed the frictional behavior between these two components for the experimental range of relative pulling velocities. We found that a perfectly relative horizontal motion has no significant resistance against sliding, however, slightly inclined loading causes measurable resistance. Our analysis paves the way towards a finite element model of silk fibers in which crystalline units can slide, move and rearrange themselves in the fiber during loading. PMID:25119288

Patil, Sandeep P; Xiao, Senbo; Gkagkas, Konstantinos; Markert, Bernd; Gräter, Frauke

2014-01-01

353

SilkRoute: trading between relations and XML  

Microsoft Academic Search

XML is the standard format for data exchange between inter-enterprise applications on the Internet. To facilitate data exchange, industry groups define public document type definitions (DTDs) that specify the format of the XML data to be exchanged between their applications. In this paper, we address the problem of automating the conversion of relational data into XML. We describe SilkRoute, a

Mary F. Fernandez; Wang-Chiew Tan; Dan Suciu

2000-01-01

354

Bladder tissue regeneration using acellular bi-layer silk scaffolds in a large animal model of augmentation cystoplasty.  

PubMed

Acellular scaffolds derived from Bombyx mori silk fibroin were investigated for their ability to support functional tissue regeneration in a porcine model of augmentation cystoplasty. Two bi-layer matrix configurations were fabricated by solvent-casting/salt leaching either alone (Group 1) or in combination with silk film casting (Group 2) to yield porous foams buttressed by heterogeneous surface pore occlusions or homogenous silk films, respectively. Bladder augmentation was performed with each scaffold group (6 × 6 cm(2)) in juvenile Yorkshire swine for 3 m of implantation. Augmented animals exhibited high rates of survival (Group 1: 5/6, 83%; Group 2: 4/4, 100%) and voluntary voiding over the course of the study period. Urodynamic evaluations demonstrated mean increases in bladder capacity over pre-operative levels (Group 1: 277%; Group 2: 153%) which exceeded nonsurgical control gains (144%) encountered due to animal growth. Similarly, elevations in bladder compliance were substantially higher in augmented animals from baseline (Group 1: 357%; Group 2: 147%) in comparison to controls (41%). Gross tissue evaluations revealed that both matrix configurations supported extensive de novo tissue formation throughout the entire original implantation site which exhibited ultimate tensile strength similar to nonsurgical counterparts. Histological and immunohistochemical analyses showed that both implant groups promoted comparable extents of smooth muscle regeneration and contractile protein (?-smooth muscle actin and SM22?) expression within defect sites similar to controls. Parallel evaluations demonstrated the formation of a transitional, multi-layered urothelium with prominent cytokeratin, uroplakin, and p63 protein expression in both matrix groups. De novo innervation and vascularization processes were evident in all regenerated tissues indicated by synaptophysin-positive neuronal cells and vessels lined with CD31 expressing endothelial cells. Ex vivo organ bath studies demonstrated that regenerated tissues supported by both silk matrices displayed contractile responses to carbachol, ?,?-methylene-ATP, KCl, and electrical field stimulation similar to controls. Our data detail the ability of acellular silk scaffolds to support regeneration of innervated, vascularized smooth muscle and urothelial tissues within 3 m with structural, mechanical, and functional properties comparable to native tissue in a porcine model of bladder repair. PMID:23953839

Tu, Duong D; Chung, Yeun Goo; Gil, Eun Seok; Seth, Abhishek; Franck, Debra; Cristofaro, Vivian; Sullivan, Maryrose P; Di Vizio, Dolores; Gomez, Pablo; Adam, Rosalyn M; Kaplan, David L; Estrada, Carlos R; Mauney, Joshua R

2013-11-01

355

Bioprospecting Finds the Toughest Biological Material: Extraordinary Silk from a Giant Riverine Orb Spider  

PubMed Central

Background Combining high strength and elasticity, spider silks are exceptionally tough, i.e., able to absorb massive kinetic energy before breaking. Spider silk is therefore a model polymer for development of high performance biomimetic fibers. There are over 41.000 described species of spiders, most spinning multiple types of silk. Thus we have available some 200.000+ unique silks that may cover an amazing breadth of material properties. To date, however, silks from only a few tens of species have been characterized, most chosen haphazardly as model organisms (Nephila) or simply from researchers' backyards. Are we limited to ‘blindly fishing’ in efforts to discover extraordinary silks? Or, could scientists use ecology to predict which species are likely to spin silks exhibiting exceptional performance properties? Methodology We examined the biomechanical properties of silk produced by the remarkable Malagasy ‘Darwin's bark spider’ (Caerostris darwini), which we predicted would produce exceptional silk based upon its amazing web. The spider constructs its giant orb web (up to 2.8 m2) suspended above streams, rivers, and lakes. It attaches the web to substrates on each riverbank by anchor threads as long as 25 meters. Dragline silk from both Caerostris webs and forcibly pulled silk, exhibits an extraordinary combination of high tensile strength and elasticity previously unknown for spider silk. The toughness of forcibly silked fibers averages 350 MJ/m3, with some samples reaching 520 MJ/m3. Thus, C. darwini silk is more than twice tougher than any previously described silk, and over 10 times better than Kevlar®. Caerostris capture spiral silk is similarly exceptionally tough. Conclusions Caerostris darwini produces the toughest known biomaterial. We hypothesize that this extraordinary toughness coevolved with the unusual ecology and web architecture of these spiders, decreasing the likelihood of bridgelines breaking and collapsing the web into the river. This hypothesis predicts that rapid change in material properties of silk co-occurred with ecological shifts within the genus, and can thus be tested by combining material science, behavioral observations, and phylogenetics. Our findings highlight the potential benefits of natural history–informed bioprospecting to discover silks, as well as other materials, with novel and exceptional properties to serve as models in biomimicry. PMID:20856804

Agnarsson, Ingi; Kuntner, Matjaz; Blackledge, Todd A.

2010-01-01

356

Quantitative proteomic and transcriptomic analyses of molecular mechanisms associated with low silk production in silkworm Bombyx mori.  

PubMed

To investigate the molecular mechanisms underlying the low fibroin production of the ZB silkworm strain, we used both SDS-PAGE-based and gel-free-based proteomic techniques and transcriptomic sequencing technique. Combining the data from two different proteomic techniques was preferable in the characterization of the differences between the ZB silkworm strain and the original Lan10 silkworm strain. The correlation analysis showed that the individual protein and transcript were not corresponded well, however, the differentially changed proteins and transcripts showed similar regulated direction in function at the pathway level. In the ZB strain, numerous ribosomal proteins and transcripts were down-regulated, along with the transcripts of translational related elongation factors and genes of important components of fibroin. The proteasome pathway was significantly enhanced in the ZB strain, indicating that protein degradation began on the third day of fifth instar when fibroin would have been produced in the Lan10 strain normally and plentifully. From proteome and transcriptome levels of the ZB strain, the energy-metabolism-related pathways, oxidative phosphorylation, glycolysis/gluconeogenesis, and citrate cycle were enhanced, suggesting that the energy metabolism was vigorous in the ZB strain, while the silk production was low. This may due to the inefficient energy employment in fibroin synthesis in the ZB strain. These results suggest that the reason for the decreasing of the silk production might be related to the decreased ability of fibroin synthesis, the degradation of proteins, and the inefficiency of the energy exploiting. PMID:24428189

Wang, Shao-Hua; You, Zheng-Ying; Ye, Lu-Peng; Che, Jiaqian; Qian, Qiujie; Nanjo, Yohei; Komatsu, Setsuko; Zhong, Bo-Xiong

2014-02-01

357

Human dental pulp progenitor cell behavior on aqueous and hexafluoroisopropanol (HFIP) based silk scaffolds  

PubMed Central

Silk scaffolds have been successfully used for a variety of tissue engineering applications due to their biocompatibility, diverse physical characteristics, and ability to support cell attachment and proliferation. Our prior characterization of 4-day postnatal rat tooth bud cells grown on hexafluoro-2-propanol (HFIP) silk scaffolds showed that the silk scaffolds not only supported osteodentin formation, but also guided the size and shape of the formed osteodentin. In this study, interactions between human dental pulp cells and HFIP and aqueous based silk scaffolds were studied under both in vitro and in vivo conditions. Silk scaffold porosity and incorporation of RGD and DMP peptides were examined. We found that the degradation of aqueous based silk is much faster than HFIP based silk scaffolds. Also, HFIP based silk scaffolds supported the soft dental pulp formation better than the aqueous based silk scaffolds. No distinct hard tissue regeneration was found in any of the implants, with or without additional cells. We conclude that alternative silk scaffold materials, and hDSC pre-seeding cell treatments or sorting and enrichment methods, need to be considered for successful dental hard tissue regeneration. PMID:21484985

Zhang, Weibo; Ahluwalia, Ivy Pruitt; Literman, Robert; Kaplan, David L.; Yelick, Pamela C.

2011-01-01

358

Interactions between Spider Silk and Cells - NIH/3T3 Fibroblasts Seeded on Miniature Weaving Frames  

PubMed Central

Background Several materials have been used for tissue engineering purposes, since the ideal matrix depends on the desired tissue. Silk biomaterials have come to focus due to their great mechanical properties. As untreated silkworm silk has been found to be quite immunogenic, an alternative could be spider silk. Not only does it own unique mechanical properties, its biocompatibility has been shown already in vivo. In our study, we used native spider dragline silk which is known as the strongest fibre in nature. Methodology/Principal Findings Steel frames were originally designed and manufactured and woven with spider silk, harvesting dragline silk directly out of the animal. After sterilization, scaffolds were seeded with fibroblasts to analyse cell proliferation and adhesion. Analysis of cell morphology and actin filament alignment clearly revealed adherence. Proliferation was measured by cell count as well as determination of relative fluorescence each after 1, 2, 3, and 5 days. Cell counts for native spider silk were also compared with those for trypsin-digested spider silk. Spider silk specimens displayed less proliferation than collagen- and fibronectin-coated cover slips, enzymatic treatment reduced adhesion and proliferation rates tendentially though not significantly. Nevertheless, proliferation could be proven with high significance (p<0.01). Conclusion/Significance Native spider silk does not require any modification to its application as a biomaterial that can rival any artificial material in terms of cell growth promoting properties. We could show adhesion mechanics on intracellular level. Additionally, proliferation kinetics were higher than in enzymatically digested controls, indicating that spider silk does not require modification. Recent findings concerning reduction of cell proliferation after exposure could not be met. As biotechnological production of the hierarchical composition of native spider silk fibres is still a challenge, our study has a pioneer role in researching cellular mechanics on native spider silk fibres. PMID:20711495

Kuhbier, Joern W.; Allmeling, Christina; Reimers, Kerstin; Hillmer, Anja; Kasper, Cornelia; Menger, Bjoern; Brandes, Gudrun; Guggenheim, Merlin; Vogt, Peter M.

2010-01-01

359

How Protein Materials Balance Strength, Robustness And Adaptability  

E-print Network

Proteins form the basis of a wide range of biological materials such as hair, skin, bone, spider silk, or cells, which play an important role in providing key functions to biological systems. The focus of this article is ...

Buehler, Markus J.

360

Orientational order of Australian spider silks as determined by solid-state NMR.  

PubMed

A simple solid-state NMR method was used to study the structure of (13)C- and (15)N-enriched silk from two Australian orb-web spider species, Nephila edulis and Argiope keyserlingi. Carbon-13 and (15)N spectra from alanine- or glycine-labeled oriented dragline silks were acquired with the fiber axis aligned parallel or perpendicular to the magnetic field. The fraction of oriented component was determined from each amino acid, alanine and glycine, using each nucleus independently, and attributed to the ordered crystalline domains in the silk. The relative fraction of ordered alanine was found to be higher than the fraction of ordered glycine, akin to the observation of alanine-rich domains in silk-worm (Bombyx mori) silk. A higher degree of crystallinity was observed in the dragline silk of N. edulis compared with A. keyserlingi, which correlates with the superior mechanical properties of the former. PMID:16463360

Bonev, B; Grieve, S; Herberstein, M E; Kishore, A I; Watts, A; Separovic, F

2006-06-01

361

Sample selection, preparation methods, and the apparent tensile properties of silkworm (B. mori) cocoon silk.  

PubMed

Reported literature values of the tensile properties of natural silk cover a wide range. While much of this inconsistency is the result of variability that is intrinsic to silk, some is also a consequence of differences in the way that silk is prepared for tensile tests. Here we explore how measured mechanical properties of Bombyx mori cocoon silk are affected by two intrinsic factors (the location from which the silk is collected within the cocoon, and the color of the silk), and two extrinsic factors (the storage conditions prior to testing, and different styles of reeling the fiber). We find that extrinsic and therefore controllable factors can affect the properties more than the intrinsic ones studied. Our results suggest that enhanced inter-laboratory collaborations, that lead to standardized sample collection, handling, and storage protocols prior to mechanical testing, would help to decrease unnecessary (and complicating) variation in reported tensile properties. PMID:22057343

Reed, Emily J; Bianchini, Lindsay L; Viney, Christopher

2012-06-01

362

Fabrication of CeO2 nanoparticle-modified silk for UV protection and antibacterial applications.  

PubMed

To endow silk with UV-shielding ability and antibacterial activity, CeO2 nanoparticles were immobilized on silk surface via a dip-coating approach without changing silk structure. Surface density of the nanoparticles could be easily adjusted by controlling the number of dip-coating cycle. Enhanced thermal stability of the modified silk is exhibited in thermogravimetric analysis (TGA) and derivative thermogravimetric analysis (DTG). The excellent UV-protection ability and antibacterial property of the CeO2 nanoparticle-coated silk are demonstrated in UV-vis diffuse reflectance spectroscopy and colony-forming capability test, respectively. Based on the data, it can be concluded that CeO2 nanoparticles could be used as a very promising coating material to modify silk for UV-protection and antibacterial applications. PMID:25203972

Lu, Zhisong; Mao, Cuiping; Meng, Mei; Liu, Sangui; Tian, Yunli; Yu, Ling; Sun, Bai; Li, Chang Ming

2014-12-01

363

Study of the microstructure of silk artifacts recovered from a historic deep-ocean site  

Microsoft Academic Search

A comparative study of the microstructure of three marine silk textiles recovered from the site of the S.S. Central America, reference silk fabric, and three historic silks from the same era as the marine textiles was conducted. The analytical techniques used include optical microscopy, scanning electron microscopy, energy dispersive x-ray spectrometry, x-ray diffraction, infrared microspectroscopy, and differential scanning calorimetry. Results

Rekha Srinivasan

2001-01-01

364

THE PHYSICAL PROPERTIES OF SPIDER'S SILK AND THEIR ROLE IN THE DESIGN OF ORB-WEBS  

Microsoft Academic Search

SUMMARY 1. Determinations were made of several physical properties of the viscid and frame silks of the orb-webs built by the spider Araneus sericatus (Cl.). 2. Both types of silk show a breaking stress of approximately 1 GN\\/m2 and an initial resilience of approximately 0-35. 3. The breaking extension ratio of viscid silk (A = 3*00) is much greater than

MARK DENNY

365

Entrapment of both glucose oxidase and peroxidase in regenerated silk fibroin membrane  

Microsoft Academic Search

Two enzmyes, glucose oxidase and peroxidase, were for the first time simultaneously immobilized in regenerated silk fibroin\\u000a membrane. The structure and morphology of the regenerated silk fibroin membrane containing both glucose oxidase and peroxidase\\u000a were investigated with IR spectra and SEM. The bienzymes do not change the structures of the regenerated silk fibroin in the\\u000a membrane, which has an islands-sea

Yongcheng Liu; Haiying Liu; Jianghong Qian; Jiaqi Deng; Tongyin Yu

1996-01-01

366

Stabilization of vaccines and antibiotics in silk and eliminating the cold chain  

PubMed Central

Sensitive biological compounds, such as vaccines and antibiotics, traditionally require a time-dependent “cold chain” to maximize therapeutic activity. This flawed process results in billions of dollars worth of viable drug loss during shipping and storage, and severely limits distribution to developing nations with limited infrastructure. To address these major limitations, we demonstrate self-standing silk protein biomaterial matrices capable of stabilizing labile vaccines and antibiotics, even at temperatures up to 60?°C over more than 6 months. Initial insight into the mechanistic basis for these findings is provided. Importantly, these findings suggest a transformative approach to the cold chain to revolutionize the way many labile therapeutic drugs are stored and utilized throughout the world. PMID:22778443

Zhang, Jeney; Pritchard, Eleanor; Hu, Xiao; Valentin, Thomas; Panilaitis, Bruce; Omenetto, Fiorenzo G.; Kaplan, David L.

2012-01-01

367

Stretching of supercontracted fibers: a link between spinning and the variability of spider silk.  

PubMed

The spinning of spider silk requires a combination of aqueous environment and stretching, and the aim of this work was to explore the role of stretching silk fibers in an aqueous environment and its effect on the tensile properties of spider silk. In particular, the sensitivity of the spider silk tensile behaviour to wet-stretching could be relevant in the search for a relationship between processing and the variability of the tensile properties. Based on this idea and working with MAS silk from Argiope trifasciata orb-web building spiders, we developed a novel procedure that permits modification of the tensile properties of spider silk: silk fibers were allowed to supercontract and subsequently stretched in water. The ratio between the length after stretching and the initial supercontracted length was used to control the process. Tensile tests performed in air, after drying, demonstrated that this simple procedure allows to predictable reproduction of the stress-strain curves of either naturally spun or forcibly silked fibers. These results suggest that the supercontracted state has a critical biological function during the spinning process of spider silk. PMID:15601874

Guinea, G V; Elices, M; Pérez-Rigueiro, J; Plaza, G R

2005-01-01

368

Diffraction from the beta-sheet crystallites in spider silk  

E-print Network

We analyze the wide angle x-ray scattering from oriented spider silk fibers in terms of a quantitative scattering model, including both structural and statistical parameters of the $\\beta$-sheet crystallites of spider silk in the amorphous matrix. The model is based on kinematic scattering theory and allows for rather general correlations of the positional and orientational degrees of freedom, including the crystallite's size, composition and dimension of the unit cell. The model is evaluated numerically and compared to experimental scattering intensities allowing us to extract the geometric and statistical parameters. We show explicitly that for the experimentally found mosaicity (width of the orientational distribution) inter-crystallite effects are negligible and the data can be analyzed in terms of single crystallite scattering, as is usually assumed in the literature.

Stephan Ulrich; Anja Glišovi?; Tim Salditt; Annette Zippelius

2008-11-25

369

A golden-silk spider spins its web  

NASA Technical Reports Server (NTRS)

On the grounds of Kennedy Space Center, a female Golden-Silk Spider repairs its web. The female can be identified by its brownish-green abdomen with a white spotted irregular pattern. The golden-silk spider repairs the webbing each day, replacing half but never the whole web at one time. Its web may measure two to three feet across. The center shares a boundary with the Merritt Island National Wildlife Refuge, a 92,000-acre refuge that is a habitat for more than 331 species of birds, 31 mammals, 117 fishes, and 65 amphibians and reptiles. The marshes and open water of the refuge provide wintering areas for 23 species of migratory waterfowl, as well as a year-round home for great blue herons, great egrets, wood storks, cormorants, brown pelicans and other species of marsh and shore birds, as well as a variety of insects.

1999-01-01

370

The Preparation of Silk Fibroin Drug-loading Microspheres  

Microsoft Academic Search

This paper deals with the development of a kind of water-in-oil-in-water (w\\/o\\/w) multiple-emulsification method to fabricate the SF drug-loading microspheres. The silk fibroin (SF) microspheres were prepared by using SF as vehicle and dexamethasone sodium phosphate (DSP) as drug model. The surface morphology was observed by SEM, the particle diameter and its distribution were observed with Laser particle sizer. The

Rui-Juan Xie; Hai-Yan Wu; Jian-Mei Xu; Qi-Ming Deng

2008-01-01

371

Finishing of silk with EDTA under thermal treatment  

Microsoft Academic Search

Establishment of ester linkages and relevant cross-linkages in the silk fabric by pad-dry-cure technique was studied using disodium salt of ethylene diamine tetra acetic acid (EDTA) in the presence of sodium dihydrogen phosphate, trisodium citrate, sodium formate and potassium sodium tartrate. Potassium sodium tartrate produced the most balanced effects. Treatment with 8% EDTA and 4.8% potassium sodium tartrate at 30°C

D. Das; A. Mukherjee; P. Bhattacharya; D. Chakrabarty

2012-01-01

372

Finishing of silk with EDTA under thermal treatment  

Microsoft Academic Search

Establishment of ester linkages and relevant cross-linkages in the silk fabric by pad-dry-cure technique was studied using disodium salt of ethylene diamine tetra acetic acid (EDTA) in the presence of sodium dihydrogen phosphate, trisodium citrate, sodium formate and potassium sodium tartrate. Potassium sodium tartrate produced the most balanced effects. Treatment with 8% EDTA and 4.8% potassium sodium tartrate at 30°C

D. Das; A. Mukherjee; P. Bhattacharya; D. Chakrabarty

2011-01-01

373

Annulus fibrosus tissue engineering using lamellar silk scaffolds  

PubMed Central

Degeneration of the intervertebral disc (IVD) represents a significant muscular skeletal disease. Recently, scaffolds composed of synthetic, natural and hybrid biomaterials have been investigated as options to restore the IVD; however, they lack the hallmark lamellar morphological features of annulus fibrosus (AF) tissue. The goal of regenerating disc is to achieve anatomic morphology as well as restoration of mechanical and biological function. In this study, two types of scaffold morphologies formed from silk fibroin were investigated towards the goal of AF tissue restoration. The first design mimics the lamellar features of the IVD that is associated with the AF region. The second is a porous spongy scaffold that serves as a control. Toroidal scaffolds were formed from the lamellar and porous silk material systems to generate structures with an outer diameter of 8 mm, inner diameter of 3.5 mm and a height of 3 mm. The inter-lamellar spacing in the lamellar scaffold was 150~250 ?m and the average pore sizes in the porous scaffolds were 100~250 ?m. The scaffolds were seeded with porcine AF cells and, after growth over defined time frames in vitro, histology, biochemical assays, mechanical testing and gene expression indicated that the lamellar scaffold generated results that were more favorable in terms of ECM expression and tissue function than the porous scaffold for AF tissue. Further, the seeded porcine AF cells supported the native shape of AF tissue in the lamellar silk scaffolds. The lamellar silk scaffolds were effective in the formation of AF-like tissue in vitro. PMID:22311816

Park, Sang-Hyug; Gil, Eun Seok; Mandal, Biman B.; Cho, Hong Sik; Kluge, Jonathan A.; Min, Byoung-Hyun; Kaplan, David L.

2012-01-01

374

Evidence of the Most Stretchable Egg Sac Silk Stalk, of the European Spider of the Year Meta menardi  

E-print Network

Spider silks display generally strong mechanical properties, even if differences between species and within the same species can be observed. While many different types of silks have been tested, the mechanical properties ...

Lepore, Emiliano

375

Reversible hydrogel-solution system of silk with high beta-sheet content.  

PubMed

Silkworm silk has been widely used as a textile fiber, as biomaterials and in optically functional materials due to its extraordinary properties. The ?-sheet-rich natural nanofiber units of about 10-50 nm in diameter are often considered the origin of these properties, yet it remains unclear how silk self-assembles into these hierarchical structures. A new system composed of ?-sheet-rich silk nanofibers about 10-20 nm in diameter is reported here, where these nanofibers formed into "flowing hydrogels" at 0.5-2% solutions and could be transformed back into the solution state at lower concentrations, even with a high ?-sheet content. This is in contrast with other silk processed materials, where significant ?-sheet content negates reversibility between solution and solid states. These fibers are formed by regulating the self-assembly process of silk in aqueous solution, which changes the distribution of negative charges while still supporting ?-sheet formation in the structures. Mechanistically, there appears to be a shift toward negative charges along the outside of the silk nanofibers in our present study, resulting in a higher zeta potential (above -50 mV) than previous silk materials which tend to be below -30 mV. The higher negative charge on silk nanofibers resulted in electrostatic repulsion strong enough to negate further assembly of the nanofibers. Changing silk concentration changed the balance between hydrophobic interactions and electrostatic repulsion of ?-sheet-rich silk nanofibers, resulting in reversible hydrogel-solution transitions. Furthermore, the silk nanofibers could be disassembled into shorter fibers and even nanoparticles upon ultrasonic treatment following the transition from hydrogel to solution due to the increased dispersion of hydrophobic smaller particles, without the loss of ?-sheet content, and with retention of the ability to transition between hydrogel and solution states through reversion to longer nanofibers during self-assembly. These reversible solution-hydrogel transitions were tunable with ultrasonic intensity, time, or temperature. PMID:25056606

Bai, Shumeng; Zhang, Xiuli; Lu, Qiang; Sheng, Weiqin; Liu, Lijie; Dong, Boju; Kaplan, David L; Zhu, Hesun

2014-08-11

376

Evidence of the Most Stretchable Egg Sac Silk Stalk, of the European Spider of the Year Meta menardi  

E-print Network

Evidence of the Most Stretchable Egg Sac Silk Stalk, of the European Spider of the Year Meta properties of stalks of silk taken from the egg sac of the cave spider Meta menardi have not yet been elongation has never been reported for egg sac silk stalks and suggests a huge unrolling microscopic

Buehler, Markus J.

377

Expression analysis of inhibitor of apoptosis and related caspases in the midgut and silk gland of the greater wax moth, Galleria mellonella, during metamorphosis and under starvation.  

PubMed

We cloned a cDNA encoding inhibitor of apoptosis (IAP) from the greater wax moth, Galleria mellonella. The deduced amino acid sequence showed that GmIAP contains two baculoviral IAP repeat (BIR) motifs, followed by a RING finger. The sequence comparison showed that GmIAP had high homology to lepidopteran IAPs and baculoviral IAPs, as well as dipteran IAPs. GmIAP transcript and its protein appeared in both the midgut and the silk gland during metamorphosis and starvation where cell death was detected by TUNEL test. IAP, and capases-1, -3, -4 and -6 appeared as at least two peaks in the midgut and silk gland during metamorphosis. Caspase-1 transcript appeared at the highest level among caspases, while caspase-3 and caspase-6 seemed to be the most relevant caspases to IAP during metamorphosis suggesting that IAP and caspases may be involved in a core apoptosis pathway in the wax moth as in flies and mosquitoes. PMID:22975212

Khoa, Dao Bach; Takeda, Makio

2012-12-01

378

Thermoelectric effect in hornet ( Vespa orientalis) silk and thermoregulation in a hornet's nest  

Microsoft Academic Search

The silk caps of the pupae of the Oriental hornet (Vespa orientalis, Vespinae, Hymenoptera) were measured for spontaneous electric current flow as a function of temperature. The measurements were made in the dark, within a range of biological temperatures. A clear correlation was detected between temperature rise and the increase in electric current forming in the silk. Thus at the

Jacob S. Ishay; Vered Barenholz-Paniry

1995-01-01

379

Biodegradable Material Featuring Embedded Silicon-on-Silk Friday, March 12, 2010  

E-print Network

adverse biological reactions while the silk conforms to tissue and dissolves harmlessly over time, factors single crystalline silicon electronics, where the silicon is in the form of nanomembranes, onto water soluble and biocompatible silk substrates," they add. "Electrical, bending, water dissolution, and http

Rogers, John A.

380

The Effect of Food and Silk Reserve Manipulation On Decoration-building of Argiope Aetheroides  

Microsoft Academic Search

Summary In this study I investigated how decoration-building of Argiope aetheroides is proximately controlled. Since decoration silk is produced from the aciniform glands and these glands' secretion is also used in prey-wrapping, I tested if manipulating the amount of silk in the aciniform glands would lead to changes in the building of decorations. When the aciniform glands of A. aetheroides

I-Min Tso

2004-01-01

381

Traces of Ancient Earthquakes in Medieval Cities Along the Silk Road, Northern Tien Shan and Dzhungaria  

Microsoft Academic Search

The evidence for earthquake destruction at medieval cities along the Silk Road from 800 A.D. to 1500 A.D. was examined using techniques of archaeoseismological mapping of architectural components at three currently excavated sites in southern Kazakhstan: Koylyk (Antonovka), Talgar (Talkhir) and Akyrtash, located on the northern branch of the Silk Road, Kazakhstan. This study revealed the following features of seismic

ANDREY KORJENKOV; KARL BAIPAKOV; CLAUDIA CHANG; YURY PESHKOV; TAMARA SAVELIEVA

2003-01-01

382

Electronic commerce on the new silk road: a cornucopia of research opportunities  

Microsoft Academic Search

The Silk Road was an important trade route between Europe and Asia a thousand years ago. It connected locations in Central and Eastern Europe with those scattered around West, Central and Eastern Asia. In more recent times, it has been neglected commercially, with the development of other ports and routes. In this paper, the potential rejuvenation of the Silk Road

Robert M. Davison; Douglas R. Vogel; Joze Gricar; Roger Harris; Maddalena Sorrentino

2003-01-01

383

Different Matrilineal Contributions to Genetic Structure of Ethnic Groups in the Silk Road Region in China  

Microsoft Academic Search

Previous studies have shown that there were extensive genetic admixtures in the Silk Road region. In the present study, we analyzed 252 mtDNAs of five ethnic groups (Uygur, Uzbek, Kazak, Mongolian, and Hui) from Xinjiang Province, China (through which the Silk Road once ran) together with some reported data from the adjacent regions in Central Asia. In a simple way,

Yong-Gang Yao; Qing-Peng Kong; Cheng-Ye Wang; Chun-Ling Zhu; Ya-Ping Zhang

2004-01-01

384

European Silk Road Tourists' and Their Tour Guides' Perceptions of Product and Service Quality  

Microsoft Academic Search

The development potential of the China Silk Road tour has been recognised by international organisations. In a preliminary survey product and service quality perceptions of 78 European Silk Road tourists and of 20 guides were assessed. The tourists were quite critical of food served outside their hotels, the interest created by the attractions, the friendliness of locals and the price

C. P. Grabowski; Wang Geng

2000-01-01

385

More than a safety line: jump-stabilizing silk of salticids.  

PubMed

Salticids are diurnal hunters known for acute vision, remarkable predatory strategies and jumping ability. Like other jumpers, they strive for stability and smooth landings. Instead of using inertia from swinging appendages or aerodynamic forces by flapping wings as in other organisms, we show that salticids use a different mechanism for in-air stability by using dragline silk, which was previously believed to function solely as a safety line. Analyses from high-speed images of jumps by the salticid Hasarius adansoni demonstrate that despite being subject to rearward pitch at take-off, spiders with dragline silk can change body orientation in the air. Instantaneous drag and silk forces calculated from kinematic data further suggest a comparable contribution to deceleration and energy dissipation, and reveal that adjustments by the spider to the silk force can reverse its body pitch for a predictable and optimal landing. Without silk, upright-landing spiders would slip or even tumble, deferring completion of landing. Thus, for salticids, dragline silk is critical for dynamic stability and prey-capture efficiency. The dynamic functioning of dragline silk revealed in this study can advance the understanding of silk's physiological control over material properties and its significance to spider ecology and evolution, and also provide inspiration for future manoeuvrable robot designs. PMID:23925983

Chen, Yung-Kang; Liao, Chen-Pan; Tsai, Feng-Yueh; Chi, Kai-Jung

2013-10-01

386

More than a safety line: jump-stabilizing silk of salticids  

PubMed Central

Salticids are diurnal hunters known for acute vision, remarkable predatory strategies and jumping ability. Like other jumpers, they strive for stability and smooth landings. Instead of using inertia from swinging appendages or aerodynamic forces by flapping wings as in other organisms, we show that salticids use a different mechanism for in-air stability by using dragline silk, which was previously believed to function solely as a safety line. Analyses from high-speed images of jumps by the salticid Hasarius adansoni demonstrate that despite being subject to rearward pitch at take-off, spiders with dragline silk can change body orientation in the air. Instantaneous drag and silk forces calculated from kinematic data further suggest a comparable contribution to deceleration and energy dissipation, and reveal that adjustments by the spider to the silk force can reverse its body pitch for a predictable and optimal landing. Without silk, upright-landing spiders would slip or even tumble, deferring completion of landing. Thus, for salticids, dragline silk is critical for dynamic stability and prey-capture efficiency. The dynamic functioning of dragline silk revealed in this study can advance the understanding of silk's physiological control over material properties and its significance to spider ecology and evolution, and also provide inspiration for future manoeuvrable robot designs. PMID:23925983

Chen, Yung-Kang; Liao, Chen-Pan; Tsai, Feng-Yueh; Chi, Kai-Jung

2013-01-01

387

The Performance Model of SilkRoad -A Multithreaded DSM System for Clusters  

E-print Network

The Performance Model of SilkRoad - A Multithreaded DSM System for Clusters Liang Peng, Weng (DSM) is a highly desir- able programming model for cluster based computing. Even though a number performance model. In this paper, we propose and analyze the performance model of the soft- ware DSM of Silk

Wong, Weng Fai

388

The Expansion of Silk Textile Export in Northern Italy, 1919-1929  

Microsoft Academic Search

The purpose of this article is to examine the expansion and the aspect of silk textile export in Northern Italy, especially in Como district, during the period of 1919-1929, when the Italian silk textile industry gained an international competitive advantage. The author would like to propound some views to which little attention has been given. Generally speaking, the modernization of

Makiko Hino

2011-01-01

389

Development and evaluation of silk fibroin-based nerve grafts used for peripheral nerve regeneration  

Microsoft Academic Search

Silk fibroin (SF), derived from natural silk long used as a textile material, has recently become an important biomaterial for tissue engineering applications. 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 biomimetic design of the SF-based nerve graft (SF graft) which was

Yumin Yang; Fei Ding; Jian Wu; Wen Hu; Wei Liu; Jie Liu; Xiaosong Gu

2007-01-01

390

Evaluation of the Antibacterial Activity of a Special Silk Textile in the Treatment of Atopic Dermatitis  

Microsoft Academic Search

Background: Increased skin Staphylococcus aureus colonization is frequently found in atopic patients. The reduction of local overinfection decreases skin inflammation and improves the flares. Objective: To evaluate the effectiveness of the antimicrobial activity of a silk fabric (MICROAIR DermaSilk®) coated with alkoxysilane quaternary ammonium with durable antimicrobial properties (AEGIS AEM 5572\\/5) in children affected by atopic dermatitis (AD). Methods: Sixteen

Giampaolo Ricci; Annalisa Patrizi; Paolo Mandrioli; Fernando Specchia; Matelda Medri; Giuseppe Menna; Massimo Masi

2006-01-01

391

Macro/microporous silk fibroin scaffolds with potential for articular cartilage and meniscus tissue engineering applications.  

PubMed

This study describes the developmental physicochemical properties of silk fibroin scaffolds derived from high-concentration aqueous silk fibroin solutions. The silk fibroin scaffolds were prepared with different initial concentrations (8, 10, 12 and 16%, in wt.%) and obtained by combining the salt-leaching and freeze-drying methodologies. The results indicated that the antiparallel ?-pleated sheet (silk-II) conformation was present in the silk fibroin scaffolds. All the scaffolds possessed a macro/microporous structure. Homogeneous porosity distribution was achieved in all the groups of samples. As the silk fibroin concentration increased from 8 to 16%, the mean porosity decreased from 90.8±0.9 to 79.8±0.3% and the mean interconnectivity decreased from 97.4±0.5 to 92.3±1.3%. The mechanical properties of the scaffolds exhibited concentration dependence. The dry state compressive modulus increased from 0.81±0.29 to 15.14±1.70 MPa and the wet state dynamic storage modulus increased by around 20- to 30-fold at each testing frequency when the silk fibroin concentration increased from 8 to 16%. The water uptake ratio decreased with increasing silk fibroin concentration. The scaffolds present favorable stability as their structure integrity, morphology and mechanical properties were maintained after in vitro degradation for 30 days. Based on these results, the scaffolds developed in this study are proposed to be suitable for use in meniscus and cartilage tissue-engineered scaffolding. PMID:22019518

Yan, Le-Ping; Oliveira, Joaquim M; Oliveira, Ana L; Caridade, Sofia G; Mano, João F; Reis, Rui L

2012-01-01

392

Effects of different post-spin stretching conditions on the mechanical properties of synthetic spider silk fibers  

PubMed Central

Spider silk is a biomaterial with impressive mechanical properties, resulting in various potential applications. Recent research has focused on producing synthetic spider silk fibers with the same mechanical properties as the native fibers. For this study, three proteins based on the Argiope aurantia Major ampullate Spidroin 2 consensus repeat sequence were expressed, purified and spun into fibers. A number of post-spin draw conditions were tested to determine the effect of each condition on the mechanical properties of the fiber. In all cases, post-spin stretching improved the mechanical properties of the fibers. Aqueous isopropanol was the most effective solution for increasing extensibility, while other solutions worked best for each fiber type for increasing tensile strength. The strain values of the stretched fibers correlated with the length of the proline-rich protein sequence. Structural analysis, including X-ray diffraction and Raman spectroscopy, showed surprisingly little change in the initial as-spun fibers compared with the post-spin stretched fibers. PMID:24113297

Albertson, Amy E.; Teule, Florence; Weber, Warner; Yarger, Jeffery L.; Lewis, Randolph V.

2014-01-01

393

Effects of different post-spin stretching conditions on the mechanical properties of synthetic spider silk fibers.  

PubMed

Spider silk is a biomaterial with impressive mechanical properties, resulting in various potential applications. Recent research has focused on producing synthetic spider silk fibers with the same mechanical properties as the native fibers. For this study, three proteins based on the Argiope aurantia Major ampullate Spidroin 2 consensus repeat sequence were expressed, purified and spun into fibers. A number of post-spin draw conditions were tested to determine the effect of each condition on the mechanical properties of the fiber. In all cases, post-spin stretching improved the mechanical properties of the fibers. Aqueous isopropanol was the most effective solution for increasing extensibility, while other solutions worked best for each fiber type for increasing tensile strength. The strain values of the stretched fibers correlated with the length of the proline-rich protein sequence. Structural analysis, including X-ray diffraction and Raman spectroscopy, showed surprisingly little change in the initial as-spun fibers compared with the post-spin stretched fibers. PMID:24113297

Albertson, Amy E; Teulé, Florence; Weber, Warner; Yarger, Jeffery L; Lewis, Randolph V

2014-01-01

394

Forced reeling of Bombyx mori silk: separating behavior and processing conditions.  

PubMed

Controlled reeling is a powerful tool to investigate the details of silk processing. However, consistent forced reeling of silkworms is hindered by the significant degree of behaviorally induced variation caused by the animal. This paper proposes silkworm paralysis as a novel method to control the animal and thus in vivo spinning conditions. Using these methods, we achieve low and consistent reeling forces during the collection of over 500 m of individual silk fiber while monitoring filament variability, morphology, and properties. Novel techniques to measure the irregular silk cross-sectional areas lead to the more accurate calculation of the true engineering values and mechanical property variation of individual silk fibers. Combining controlled reeling and accurate thread measurement techniques allows us to present the relative contributions of processing and behavior in the performance envelope of Bombyx mori silk. PMID:24004380

Mortimer, Beth; Holland, Chris; Vollrath, Fritz

2013-10-14

395

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

PubMed Central

Silk-elastin-like polymers (SELPs) are protein-based polymers composed of repetitive amino acid sequence motifs found in silk fibroin (GAGAGS) and mammalian elastin (VPGVG). These polymers are of much interest, both from a fundamental and applied point of view, finding potential application in biomedicine, nanotechnology and as materials. The successful employment of such polymers in such diverse fields, however, requires the ready availability of a variety of different forms with novel enhanced properties and which can be simply prepared in large quantities on an industrial scale. In an attempt to create new polymer designs with improved properties and applicability, we have developed four novel SELPs wherein the elastomer forming sequence poly(VPGVG) is replaced with a plastic-like forming sequence, poly(VPAVG), and combined in varying proportions with the silk motif. Furthermore, we optimised a simplified production procedure for these, making use of an autoinduction medium to reduce process intervention and with the production level obtained being 6-fold higher than previously reported for other SELPs, with volumetric productivities above 150?mg/L. Finally, we took advantage of the known enhanced stability of these polymers in developing an abridged, non-chromatographic downstream processing and purification protocol. A simple acid treatment allowed for cell disruption and the obtention of relative pure SELP in one-step, with ammonium sulphate precipitation being subsequently used to enable improved purity. These simplified production and purification procedures improve process efficiency and reduce costs in the preparation of these novel polymers and enhances their potential for application. PMID:23384239

2013-01-01

396

An Amperometric New Methylene Blue N-Mediating Sensor for Hydrogen Peroxide Based on Regenerated Silk Fibroin as an Immobilization Matrix for Peroxidase  

Microsoft Academic Search

A simple and effective procedure was described for the immobilization of peroxidase in regenerated silk fibroin membrane prepared from waste silk. The membranes of regenerated silk fibroin with or without peroxidase, before or after the ethanol treatment, were characterized by ir spectra. An amperometric H2O2sensor, based on the immobilized peroxidase in regenerated silk fibroin membrane, in the use of new

Jianghong Qian; Yongcheng Liu; Haiying Liu; Tongyin Yu; Jiaqi Deng

1996-01-01

397

Abnormal hemoglobins in the Silk Road region of China  

Microsoft Academic Search

A review is presented of the occurrence of 24 abnormal hemoglobins (13 a-chain variants and 11 ß-chain variants) in populations in the Silk Road area of Northwestern China. Most frequently occurring were Hb D-Punjab [ß21(GH4)Glu?Gln] in Uygurs, Kazaks, and Khalkhas, Hb G-Taipei [ß22(B4)Glu?Gly] in persons of the Han nationality, and Hb G-Coushatta [ß22 (B4)Glu?Ala] in the Uygurs, Kazaks, Hans, and

Hou-jun Li; Xian-ning Zhao; Fan Qin; Hui-wu Li; Li Li; Xiao-jun He; Xiao-su Chang; Zhe-meng Li; Ke-xing Liang; Fa-li Xing; Wen-sun Chang; Ru-zhau Wong; Iu-li Yang; Fen-sheng Li; Tuen-tian Zhang; Ri-jun Tian; Brooke B. Webber; Jerry B. Wilson; Titus H. J. Huisman

1990-01-01

398

Characterization of tyrosine-rich Antheraea pernyi silk fibroin hydrolysate.  

PubMed

Antheraea pernyi silk fibroin (SF) hydrolysate were characterized using UV-VIS spectrometer, amino acid composition and heavy metal contents to explore its potential sources for food or cosmetic additives. The hydrolyzed A. pernyi SF was separated into two parts: (a) SFA, alanine-rich fraction and (b) SFB, tyrosine-rich fraction. SFB exhibited strong absorption peaks at 210 and 280 nm due to the presence of the tyrosine. Heavy metal analysis showed that arsenic and mercury did not detect. Other heavy metals, which includes lead, cadmium, etc., were recorded only a trace amount. Therefore, A. pernyi SF hydrolysate could be safely used as sources of food, cosmetic and pharmaceuticals. PMID:20937302

Lee, Kwang-gill; Kweon, HaeYong; Yeo, Joo-hong; Woo, SoonOk; Han, SangMi; Kim, Jong-Ho

2011-01-01

399

Study of the microstructure of silk artifacts recovered from a historic deep-ocean site  

NASA Astrophysics Data System (ADS)

A comparative study of the microstructure of three marine silk textiles recovered from the site of the S.S. Central America, reference silk fabric, and three historic silks from the same era as the marine textiles was conducted. The analytical techniques used include optical microscopy, scanning electron microscopy, energy dispersive x-ray spectrometry, x-ray diffraction, infrared microspectroscopy, and differential scanning calorimetry. Results indicate that the physical microstructure of two of the marine silk artifacts has been altered significantly. An increase in fiber diameter was observed in these two marine silks and their gross physical structure was considerably damaged. Crystallinity indices obtained from IR and XRD are lower in all of the marine and two of the historic silks in comparison with reference silk. Specimens vary in the extent to which short- and long-range order have been disturbed as a result of age and/or marine exposure. In all marine silks and two historic silks, the glass transition is suppressed and degradation temperature altered. Crystallite size perpendicular to the intersheet- and interchain-directions, and the unit cell dimensions, have not altered appreciably in any of the specimens. The primary effect of marine exposure appears to be conversion of some of the crystalline phase into an ordered-amorphous structure. Additionally, the amorphous phase is partially dissolved or recrystallized to an ordered-amorphous phase. No qualitative alteration of the chemical microstructure of fibroin was observed. Organic and inorganic surface deposits are observed on the marine silks but these are not as extensive on the historic artifacts. The formation of biofilm is observed on both the historic and marine silk, but to varying extents. Applicability of the analytical tools to analyze rare artifacts is addressed. While basic analytical studies are important in understanding the structure and degradation of rare artifacts, once understood, subsequent work should build on this research and employ techniques which require smaller samples. From a conservator's perspective, the results indicate that the marine silks are more susceptible to degradation than are historic silks. Handling and storage treatments should focus on controlling the environment and on treating the artifacts to minimize subsequent degradation.

Srinivasan, Rekha

400

Control of fibroin conformation: Toward the development of a biomimetic spinning process for silk fibers  

NASA Astrophysics Data System (ADS)

Nature has shown that silks are sophisticated structural materials with remarkable mechanical properties; however, they are produced using far milder conditions than high-performance synthetic polymer fibers. While recent advances in molecular biotechnology have taken great strides toward the production of proteinaceous biopolymers, little is known about the processing conditions needed to spin fibers with the correct microstructures and mechanical properties. It is the purpose of this research to gain a fundamental understanding about how processing conditions affect the molecular structure of a model protein biopolymer, Bombyx mori silkworm fibroin, the structural protein of cocoon silk. Fibers of B. mori fibroin were wet spun from 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP) into a methanol coagulation bath. X-ray fiber diffraction and quantitative Raman spectroscopy were used to determine that both naturally- and synthetically-spun fibers contain a high degree of beta-sheet (~50%). Fibers subjected to a post-spinning draw exhibited a preferential molecular alignment parallel to the fiber axis resulting in increased strength, stiffness, and extensibility. Fibers with microstructures and mechanical properties most similar to those of naturally-spun fibers were reproduced in synthetically-spun fibers with a draw ratio of 3.5. The transformation of helical fibroin in HFIP to beta-sheet sheet fibroin in synthetically-spun fibers was determined to be caused by the methanol coagulation bath. The kinetics beta-sheet fibroin crystallization from aqueous solution was investigated by monitoring the sigmoidal progression of gel formation using turbidity and Raman spectroscopy. Gelation kinetics were evaluated by measuring lag time, maximum gelation rate, and optical density to determine the effects of protein concentration, detergent concentration (nucleating agent), headgroup chemistry, ionic strength, pH, and temperature. An optimal molar ratio between SDS and fibroin (100:1) was found to produce gels with minimum lag times and maximum gelation rates. Fluorescence spectroscopy and a two-step denaturation and aggregation model for gelation were used to explore the mechanism of fibroin gelation. Conditions that lead to fibroin expansion (dilute fibroin, low ionic strength, highly charged fibroin, or increased temperature) result in decreased lag times. The results presented in this dissertation should aid in developing biomimetic spinning techniques for proteinaceous, biopolymers through the use of amphiphilic sheet nucleating agents.

Carlson, Kimberly Ann Trabbic

1999-09-01