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1

The promotion of osseointegration of titanium surfaces by coating with silk protein sericin.  

PubMed

A promising strategy to influence the osseointegration process around orthopaedic titanium implants is the immobilization of bioactive molecules. This recruits appropriate interaction between the surface and the tissue by directing cells adhesion, proliferation, differentiation and active matrix remodelling. In this study, we aimed to investigate the functionalization of metallic implant titanium with silk protein sericin. Titanium surface was immobilized with non-mulberry Antheraea mylitta sericin using glutaraldehyde as crosslinker. To analyse combinatorial effects the sericin immobilized titanium was further conjugated with integrin binding peptide sequence Arg-Gly-Asp (RGD) using ethyl (dimethylaminopropyl) carbodiimide and N-hydroxysulfosuccinimide as coupling agents. The surface of sericin immobilized titanium was characterized biophysically. Osteoblast-like cells were cultured on sericin and sericin/RGD functionalized titanium and found to be more viable than those on pristine titanium. The enhanced adhesion, proliferation, and differentiation of osteoblast cells were observed. RT-PCR analysis showed that mRNA expressions of bone sialoprotein, osteocalcin and alkaline phosphatase were upregulated in osteoblast cells cultured on sericin and sericin/RGD immobilized titanium substrates. Additionally, no significant amount of pro-inflammatory cytokines TNF-?, IL-1? and nitric oxide production were recorded when macrophages cells and osteoblast-macrophages co culture cells were grown on sericin immobilized titanium. The findings demonstrate that the sericin immobilized titanium surfaces are potentially useful bioactive coated materials for titanium-based medical implants. PMID:23357374

Nayak, Sunita; Dey, Tuli; Naskar, Deboki; Kundu, Subhas C

2013-04-01

2

Potential applications of silk sericin, a natural protein from textile industry by-products.  

PubMed

Silk is composed of two major proteins, fibroin (fibrous protein) and sericin (globular, gumming protein). Fibroin has been used in textile manufacturing and for several biomaterial applications, whereas sericin is considered a waste material in the textile industry. Sericin has recently been found to activate the proliferation of several cell-lines and has also shown various biological activities. Sericin can form a gel by itself; however, after mixing with other polymers and cross-linking it can form a film or a scaffold with good characteristics that can be used in the cosmetic and pharmaceutical industries. Sericin is proven to cause no immunological responses, which has resulted in a more acceptable material for biological applications. PMID:21558082

Aramwit, Pornanong; Siritientong, Tippawan; Srichana, Teerapol

2012-03-01

3

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

PubMed

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

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

2013-10-01

4

Maturation and fertilisation of sheep oocytes cultured in serum-free medium containing silk protein sericin.  

PubMed

Sericin is a water-soluble component of silk and has been used as a biomaterial due to its antibacterial and ultraviolet radiation-resistant properties. This study was designed to evaluate the effect of sericin supplementation in a maturation medium on the meiotic competence and fertilisability of sheep oocytes. Cumulus-oocyte complexes (COCs) were cultured in TCM199 supplemented with sericin at various concentrations of 0 (control), 0.1, 0.25 and 0.5%, either with or without bovine serum albumin (BSA). When the COCs were matured without BSA, the supplementation of 0.1% sericin significantly increased the rates of maturation to metaphase II and the total fertilisation of oocytes compared with the other concentrations of sericin. When the COCs were matured with BSA, the beneficial effects of 0.1% sericin supplementation on the maturation and fertilisation of oocytes were not observed. Our findings indicate that supplementation with 0.1% sericin during maturation culture may improve the nuclear maturation and fertilisability of sheep oocytes. Moreover, it may be possible to replace BSA with sericin in chemically defined media without the risk of disease transmission. PMID:25655418

Yasmin, Cut; Otoi, Takeshige; Setiadi, Mohamad Agus; Karja, Ni Wayan Kurniani

2015-03-01

5

Fabrication of silk sericin nanofibers from a silk sericin-hope cocoon with electrospinning method.  

PubMed

In this study, silk sericin nanofibers from sericin hope-silkworm, whose cocoons consist almost exclusively of sericin were successfully prepared by electrospinning method. Scanning electron microscopy (SEM) was used to observe the morphology of the fibers. The effect of spinning conditions, including the concentration of sericin cocoon solution, acceleration voltage, spinning distance and flow rate on the fiber morphologies and the size distribution of sericin nanofibers were examined. The structure and physical properties were also observed by Fourier transform infrared (FT-IR), differential scanning calorimetry (DSC) and thermogravimetric analysis (TG). The optimum conditions for producing finely thinner fibrous sericin nanofibers without beads were the concentration of sericin solution above 6-8 wt%, acceleration voltage ranging from 25 to 32 kV, spinning distance above 9 cm, and flow rate above 0.06 cm min(-1). The mean diameter of as spun sericin fibers varied from 114 to 430 nm at the different spinning conditions. In the as-spun fibers, silk sericin was present in a random coil conformation, while after methanol treatment, the molecular structure of silk sericin was transformed into a ?-sheet containing structure. Sericin hope nanofiber demonstrated thermal degradation at lower temperature than the sericin hope cocoon, which probably due to the randomly coiled rich structure of the sericin hope nanofiber. PMID:22198656

Zhang, Xianhua; Khan, Md Majibur Rahman; Yamamoto, Toshio; Tsukada, Masuhiro; Morikawa, Hideaki

2012-03-01

6

Anti-inflammatory potential of silk sericin.  

PubMed

Silk sericin was found to suppress the production of pro-inflammatory cytokines, which are related to the inflammatory reaction. The objectives of this study were to investigate the anti-inflammatory effect of sericin in vivo using the carrageenan-induced rat edema model and changes in the histology of tissues. The effects of sericin on the expression of COX-2 and iNOS were also evaluated. Sericin solutions at 0.004-0.080 mg/mL were applied topically to the top of the hind paw and carrageenan (1.0 mg) was injected subcutaneously to the plantar surface of the right hind paw. Our results indicated that sericin significantly reduced the inflammation in rats' paw compared with the negative control (water and acetone) and its effect at 0.080 mg/mL was only slightly lower than that of 1.0% w/v indomethacin. Similar numbers of polymorphonuclear and macrophage cells were found in rats' tissue treated with indomethacin and sericin solution, while the numbers were significantly higher in their absence. The gene expression results by RT-PCR showed that the COX-2 and iNOS genes were down-regulated in samples treated with sericin in a dose dependent manner. These data indicated that the anti-inflammatory properties of sericin may be partly attributable to the suppression of the COX-2 enzyme and nitric oxide production. PMID:23738464

Aramwit, Pornanong; Towiwat, Pasarapa; Srichana, Teerapol

2013-04-01

7

Isolation and bioactivities of a non-sericin component from cocoon shell silk sericin of the silkworm Bombyx mori.  

PubMed

The cocoon shell of the silkworm Bombyx mori consists of silk fibroin fiber (70%) surrounded by a sericin layer made up of sericin (25%) and non-sericin (5%) components. The non-sericin component which consists of carbohydrate, salt, wax, flavonoids and derivatives is often overlooked in applied research into sericin and its hydrolysate. Here, sericin and non-sericin compounds were obtained from the sericin layer of five types of cocoon shell by means of degumming in water followed by extraction and separation in ethanol. These ethanol extracts were found to mainly contain flavonoids and free amino acids possessing scavenging activities of the 2,2-diphenyl -1-picrylhydrazyl (DPPH) free radical and inhibiting activities of tyrosinase, which were much greater than the corresponding activities of the purified sericin proteins. The extracts also strongly inhibited ?-glucosidase while the sericins had no such activity. In particular, the inhibitory activities of the ethanol extract of Daizo cocoons were much greater than those of the other cocoons. The IC(50) values of the Daizo cocoons for DPPH free radicals, tyrosinase, and ?-glucosidase were 170, 27, and 110 ?g mL(-1), respectively. The bioactivities of the non-sericin component were much higher than the activity of sericin alone. In addition, the in vivo test showed preliminarily that the administration of the non-sericin component had effectively resistant activity against streptozocin (STZ) oxidation and that of the purified sericin could also evidently decrease the induction ratio of diabetic mice induced by STZ. Therefore, ethanol extract protocols of the sericin layer of cocoon shells provide a novel stock which, together with sericin protein, has potential uses in functional food, biotechnological and medical applications. PMID:22101964

Wang, Hai-Yan; Wang, Yuan-Jing; Zhou, Li-Xia; Zhu, Lin; Zhang, Yu-Qing

2012-02-01

8

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

9

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

PubMed Central

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

10

Functional conservation and structural diversification of silk sericins in two moth species.  

PubMed

Sericins are hydrophilic structural proteins produced by caterpillars in the middle section of silk glands and layered over fibroin proteins secreted in the posterior section. In the process of spinning, fibroins form strong solid filaments, while sericins seal the pair of filaments into a single fiber and glue the fiber into a cocoon. Galleria mellonella and the previously examined Bombyx mori harbor three sericin genes that encode proteins containing long repetitive regions. Galleria sericin genes are similar to each other and the protein repeats are built from short and extremely serine-rich motifs, while Bombyx sericin genes are diversified and encode proteins with long and complex repeats. Developmental changes in sericin properties are controlled at the level of gene expression and splicing. In Galleria , MG-1 sericin is produced throughout larval life until the wandering stage, while the production of MG-2 and MG-3 reaches a peak during cocoon spinning. PMID:23593923

Zurovec, Michal; Kludkiewicz, Barbara; Fedic, Robert; Sulitkova, Jitka; Mach, Vaclav; Kucerova, Lucie; Sehnal, Frantisek

2013-06-10

11

Effects of silk sericin on the proliferation and apoptosis of colon cancer cells.  

PubMed

Sericin is a silk protein woven from silkworm cocoons (Bombyx mori). In animal model, sericin has been reported to have anti-tumoral action against colon cancer. The mechanisms underlying the activity of sericin against cancer cells are not fully understood. The present study investigated the effects of sericin on human colorectal cancer SW480 cells compared to normal colonic mucosal FHC cells. Since the size of the sericin protein may be important for its activity, two ranges of molecular weight were tested. Sericin was found to decrease SW480 and FHC cell viability. The small sericin had higher anti-proliferative effects than that of the large sericin in both cell types. Increased apoptosis of SW480 cells is associated with increased caspase-3 activity and decreased Bcl-2 expression. The anti-proliferative effect of sericin was accompanied by cell cycle arrest at the S phase. Thus, sericin reduced SW480 cell viability by inducing cell apoptosis via caspase-3 activation and down-regulation of Bcl-2 expression. The present study provides scientific data that support the protective effect of silk sericin against cancer cells of the colon and suggests that this protein may have significant health benefits and could potentially be developed as a dietary supplement for colon cancer prevention. PMID:22688983

Kaewkorn, Waraporn; Limpeanchob, Nanteetip; Tiyaboonchai, Waree; Pongcharoen, Sutatip; Sutheerawattananonda, Manote

2012-01-01

12

Cryopreservation for bovine embryos in serum-free freezing medium containing silk protein sericin.  

PubMed

Because the use of serum in the embryo cryopreservation increases the probability of animal health problems such as bovine spongiform encephalopathy (BSE) and viral infections, this study was conducted to examine the effects of sericin supplementation for serum-free freezing medium on the survival and development of bovine embryos after freezing-thawing and direct transfer to recipients. When in vitro-produced bovine embryos were frozen conventionally in the freezing medium supplemented with various concentrations (0.1%, 0.5%, and 1.0%) of sericin, the percentages of damaged zona pellucida, survival, and development of frozen-thawed embryos were similar to those of embryos frozen in freezing medium supplemented with 0.4% bovine serum albumin (BSA) and 20% fetal bovine serum (FBS) (0.4BSA/20F; control). When in vivo-derived embryos were frozen with 0.4BSA/20F (control), 0.5% sericin +20% FBS (0.5S/20F) or 0.5% sericin (0.5S) and were subsequently transferred directly to recipients, the percentages of recipients with pregnancy and normal calving in the 0.5S/20F group were higher than those in the control group (47.3% vs. 40.1% and 94.6% vs. 87.3%, respectively). Moreover, the percentages of recipients with pregnancy and normal calving (42.2% and 92.4%, respectively) in the 0.5S group were similar with those of other groups. In conclusion, these results indicate that serum-free freezing medium supplemented with sericin is available for the cryopreservation of bovine embryos and that it is beneficial for the elimination of a potential source of biological contamination by serum or BSA. PMID:23850826

Isobe, Tomohiro; Ikebata, Yoshihisa; Onitsuka, Takeshi; Do, Lanh Thi Kim; Sato, Yoko; Taniguchi, Masayasu; Otoi, Takeshige

2013-10-01

13

Dietary silk protein, sericin, improves epidermal hydration with increased levels of filaggrins and free amino acids in NC/Nga mice.  

PubMed

Epidermal hydration is maintained primarily by natural moisturising factors (NMF), of which free amino acids (AA) are major constituents that are generated by filaggrin degradation. To identify dietary sources that may improve skin dryness of atopic dermatitis (AD), we investigated dietary effects of silk proteins, sericin and fibroin, on epidermal levels of hydration, filaggrins and free AA, as well as PPAR?, peptidylarginine deiminase-3 (PAD3) and caspase-14 proteins involved in filaggrin expression and degradation processes. NC/Nga mice, an animal model of AD, were fed a control diet (group CA: atopic control) or diets with 1 % sericin (group S) or fibroin (group F) for 10 weeks. In group S, epidermal levels of hydration, total filaggrins and total free AA, as well as PPAR?, PAD3 and caspase-14, which were reduced in group CA, were increased to higher or similar levels of a normal control group of BALB/c mice (group C). Furthermore, profilaggrin, a precursor with multiple filaggrin repeats, and three repeat intermediates were increased, while two repeat intermediates and filaggrin were decreased in parallel with increased levels of glutamate and serine, major AA of NMF in group S. Despite increased levels of total filaggrins, total free AA, PPAR? and PAD3, epidermal levels of hydration, glutamate, serine and caspase-14 were not increased, but other minor AA of NMF were highly detected in group F. Dietary sericin improves epidermal hydration in parallel with enhancing profilaggrin expression and degradation into free AA that is coupled with elevated levels of PPAR?, PAD3 and caspase-14 proteins. PMID:22244094

Kim, Hyunae; Lim, Yu-ji; Park, Ji-Ho; Cho, Yunhi

2012-11-28

14

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

15

Production of silk sericin/silk fibroin blend nanofibers  

NASA Astrophysics Data System (ADS)

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

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

2011-08-01

16

Silk sericin ameliorates wound healing and its clinical efficacy in burn wounds.  

PubMed

The aim of this study was to evaluate the effect of silk sericin, a protein from silkworm cocoon, on scratch wound healing in vitro. For applicable result in clinical use, we also study the efficacy of sericin added to a standard antimicrobial cream, silver zinc sulfadiazine, for open wound care in the treatment of second-degree burn wounds. In vitro scratch assays show that sericin at concentration 100 ?g/mL can promote the migration of fibroblast L929 cells similar to epidermal growth factor (positive control) at 100 ?g/mL. After 1 day of treatment, the length of scratch in wounds treated with sericin was significantly shorter than the length of negative control wounds (culture medium without sericin). For clinical study, a total of 29 patients with 65 burn wounds which covered no less than 15 % of total body surface area were randomly assigned to either control (wounds treated with silver zinc sulfadiazine cream) or treatment (wounds treated with silver zinc sulfadiazine with added sericin cream) group in this randomized, double-blind, standard-controlled study. The results showed that the average time to reach 70 % re-epithelialization of the burned surface and complete healing in the treatment group was significantly shorter, approximately 5-7 days, than in the control group. Regarding time for complete healing, control wounds took approximately 29.28 ± 9.27 days, while wounds treated with silver zinc sulfadiazine with added sericin cream took approximately 22.42 ± 6.33 days, (p = 0.001). No infection or severe reaction was found in any wounds. This is the first clinical study to show that silk sericin is safe and beneficial for burn wound treatment when it is added to silver sulfadiazine cream. PMID:23748948

Aramwit, Pornanong; Palapinyo, Sirinoot; Srichana, Teerapol; Chottanapund, Suthat; Muangman, Pornprom

2013-09-01

17

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

18

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

NASA Astrophysics Data System (ADS)

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

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

19

Biomimetic Nucleation of Hydroxyapatite Crystals Mediated by Antheraea pernyi Silk Sericin Promotes Osteogenic Differentiation of Human Bone Marrow Derived Mesenchymal Stem Cells  

PubMed Central

Biomacromolecules have been used as templates to grow hydroxyapatite crystals (HAps) by biomineralization to fabricate mineralized materials for potential application in bone tissue engineering. Silk sericin is a protein with features desirable as a biomaterial, such as increased hydrophilicity and biodegradation. Mineralization of the silk sericin from Antheraea pernyi (A. pernyi) silkworm has rarely been reported. Here, for the first time, nucleation of HAps on A. pernyi silk sericin (AS) was attempted through a wet precipitation method and consequently the cell viability and osteogenic differentiation of BMSCs on mineralized AS were investigated. It was found that AS mediated the nucleation of HAps in the form of nanoneedles while self-assembling into ?-sheet conformation, leading to the formation of a biomineralized protein based biomaterial. The cell viability assay of BMSCs showed that the mineralization of AS stimulated cell adhesion and proliferation, showing that the resultant AS biomaterial is biocompatible. The differentiation assay confirmed that the mineralized AS significantly promoted the osteogenic differentiation of BMSCs when compared to nonmineralized AS as well as other types of sericin (B. mori sericin), suggesting that the resultant mineralized AS biomaterial has potential in promoting bone formation. This result represented the first work proving the osteogenic differentiation of BMSCs directed by silk sericin. Therefore, the biomineralization of A. pernyi silk sericin coupled with seeding BMSCs on the resultant mineralized biomaterials is a useful strategy to develop the potential application of this unexplored silk sericin in the field of bone tissue engineering. This study lays the foundation for the use of A. pernyi silk sericin as a potential scaffold for tissue engineering. PMID:24666022

2015-01-01

20

Biomimetic nucleation of hydroxyapatite crystals mediated by Antheraea pernyi silk sericin promotes osteogenic differentiation of human bone marrow derived mesenchymal stem cells.  

PubMed

Biomacromolecules have been used as templates to grow hydroxyapatite crystals (HAps) by biomineralization to fabricate mineralized materials for potential application in bone tissue engineering. Silk sericin is a protein with features desirable as a biomaterial, such as increased hydrophilicity and biodegradation. Mineralization of the silk sericin from Antheraea pernyi (A. pernyi) silkworm has rarely been reported. Here, for the first time, nucleation of HAps on A. pernyi silk sericin (AS) was attempted through a wet precipitation method and consequently the cell viability and osteogenic differentiation of BMSCs on mineralized AS were investigated. It was found that AS mediated the nucleation of HAps in the form of nanoneedles while self-assembling into ?-sheet conformation, leading to the formation of a biomineralized protein based biomaterial. The cell viability assay of BMSCs showed that the mineralization of AS stimulated cell adhesion and proliferation, showing that the resultant AS biomaterial is biocompatible. The differentiation assay confirmed that the mineralized AS significantly promoted the osteogenic differentiation of BMSCs when compared to nonmineralized AS as well as other types of sericin (B. mori sericin), suggesting that the resultant mineralized AS biomaterial has potential in promoting bone formation. This result represented the first work proving the osteogenic differentiation of BMSCs directed by silk sericin. Therefore, the biomineralization of A. pernyi silk sericin coupled with seeding BMSCs on the resultant mineralized biomaterials is a useful strategy to develop the potential application of this unexplored silk sericin in the field of bone tissue engineering. This study lays the foundation for the use of A. pernyi silk sericin as a potential scaffold for tissue engineering. PMID:24666022

Yang, Mingying; Shuai, Yajun; Zhang, Can; Chen, Yuyin; Zhu, Liangjun; Mao, Chuanbin; OuYang, Hongwei

2014-04-14

21

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

PubMed Central

Wound healing is a biological process directed to the restoration of tissue that has suffered an injury. An important phase of wound healing is the generation of a basal epithelium able to wholly replace the epidermis of the wound. A broad range of products derived from fibroin and sericin from Bombyx mori silk are used to stimulate wound healing. However, so far the molecular mechanism underlying this phenomenon has not been elucidated. The aim of this work was to determine the molecular basis underlying wound healing properties of silk proteins using a cell model. For this purpose, we assayed fibroin and sericin in a wound healing scratch assay using MDA-MB-231 and Mv1Lu cells. Both proteins stimulated cell migration. Furthermore, treatment with sericin and fibroin involved key factors of the wound healing process such as upregulation of c-Jun and c-Jun protein phosphorylation. Moreover, fibroin and sericin stimulated the phosphorylation of ERK 1/2 and JNK 1/2 kinases. All these experiments were done in the presence of specific inhibitors for some of the cell signalling pathways referred above. The obtained results revealed that MEK, JNK and PI3K pathways are involved in fibroin and sericin stimulated cells migration. Inhibition of these three kinases prevented c-Jun upregulation and phosphorylation by fibroin or sericin. Fibroin and sericin were tested in the human keratinocyte cell line, HaCaT, with similar results. Altogether, our results showed that fibroin and sericin initiate cell migration by activating the MEK, JNK and PI3K signalling pathways ending in c-Jun activation. PMID:22860103

García-Vizcaíno, Eva María; Alcaraz, Antonia; Cenis, José Luis; Nicolás, Francisco José

2012-01-01

22

Silk sericin/polyacrylamide in situ forming hydrogels for dermal reconstruction.  

PubMed

In situ forming tissue sealants are advantageous due to ease in application, complete coverage of defect site and assured comfort levels to patients. The interconnected three-dimensional hydrophilic networks perfectly manage typical dermal wounds by suitably scaffolding skin fibroblast, diffusing the nutrients, therapeutics and exudates while still maintaining an adequately moist environment. We evaluate the cell homing ability of semi-interpenetrating non-mulberry tropical tasar silk sericin/polyacrylamide hydrophilic network with a keen understanding of its network characteristics and correlation of protein concentration with the performance as cell scaffold. Interconnectivity of porous networks observed through scanning electron micrograph revealed pore sizes ranging from 23 to 52 ?m. The enhanced ?-sheet content with the increasing sericin concentration in far red spectroscopy study supported their corresponding improved compressive strength. These semi-interpenetrating networks were found to possess a maximum fluid uptake of 112% of its weight, hence preventing the accumulation of exudates at the wound area. The present systems appear to possess characteristics like rapid gelation (~5min) at 37 °C, 98% porosity enabling the migration of fibroblasts during healing (observed through confocal and scanning electron micrographs), cell adhesion together with the absence of any cyto-toxic effect suggesting its potential as in situ tissue sealants. The compressive strength up to 61 kPa ensured ease in handling even when wet. The results prove the suitability to use non-mulberry tasar cocoon silk sericin/polyacrylamide semi-interpenetrating network as a reconstructive dermal sealant. PMID:22819495

Kundu, Banani; Kundu, Subhas C

2012-10-01

23

Mineralization and biocompatibility of Antheraea pernyi (A. pernyi) silk sericin film for potential bone tissue engineering.  

PubMed

This study aimed to investigate the mineralization of Antheraea pernyi (A. pernyi) silk sericin. Mineralization of A. pernyi sericin was performed by alternative soaking in calcium and phosphate. The inhibition of precipitation of calcium carbonate and von Kossa staining on A. pernyi sericin were tested, and the corresponding results prove that A. pernyi sericin has Ca binding activity. Scanning electron microscope (SEM) observation shows that spherical crystals could be nucleated on the A. pernyi sericin film. These crystals were confirmed to be hydroxyapatite according to FT-IR and XRD spectra, indicating that A. pernyi sericin is capable of mineralization. In addition, cell adhesion and growth activity assay demonstrate that A. pernyi sericin shows excellent biocompatibility for the growth of MG-63 cells. PMID:24211968

Yang, Mingying; Mandal, Namita; Shuai, Yajun; Zhou, Guanshan; Min, Sijia; Zhu, Liangjun

2014-01-01

24

Accelerated healing of full-thickness wounds by genipin-crosslinked silk sericin/PVA scaffolds.  

PubMed

Silk sericin has recently been studied for its advantageous biological properties, including its ability to promote wound healing. This study developed a delivery system to accelerate the healing of full-thickness wounds. Three-dimensional scaffolds were fabricated from poly(vinyl alcohol) (PVA), glycerin (as a plasticizer) and genipin (as a crosslinking agent), with or without sericin. The physical and biological properties of the genipin-crosslinked sericin/PVA scaffolds were investigated and compared with those of scaffolds without sericin. The genipin-crosslinked sericin/PVA scaffolds exhibited a higher compressive modulus and greater swelling in water than the scaffolds without sericin. Sericin also exhibited controlled release from the scaffolds. The genipin-crosslinked sericin/PVA scaffolds promoted the attachment and proliferation of L929 mouse fibroblasts. After application to full-thickness rat wounds, the wounds treated with genipin-crosslinked sericin/PVA scaffolds showed a significantly greater reduction in wound size, collagen formation and epithelialization compared with the control scaffolds without sericin but lower numbers of macrophages and multinucleated giant cells. These results indicate that the delivery of sericin from the novel genipin-crosslinked scaffolds efficiently healed the wound. Therefore, these genipin-crosslinked sericin/PVA scaffolds represent a promising candidate for the accelerated healing of full-thickness wounds. PMID:23307034

Aramwit, Pornanong; Siritienthong, Tippawan; Srichana, Teerapol; Ratanavaraporn, Juthamas

2013-01-01

25

Effect of silk sericin on morphology and structure of calcium carbonate crystal  

NASA Astrophysics Data System (ADS)

In this paper, silk sericin was employed to regulate the mineralization of calcium carbonate (CaCO3). CaCO3 composite particles were prepared by the precipitation reaction of sodium carbonate with calcium chloride solution in the presence of silk sericin. The as-prepared samples were collected at different reaction time to study the crystallization process of CaCO3 by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA) and X-ray diffraction (XRD). The results showed that silk sericin significantly affected the morphology and crystallographic polymorph of CaCO3. With increasing the reaction time, the crystal phase of CaCO3 transferred from calcite dominated to vaterite dominated mixtures, while the morphology of CaCO3 changed from disk-like calcite crystal to spherical vaterite crystal. These studies showed the potential of silk sericin used as a template molecule to control the growth of inorganic crystal.

Zhao, Rui-Bo; Han, Hua-Feng; Ding, Shao; Li, Ze-Hao; Kong, Xiang-Dong

2013-06-01

26

Inhibitory effects of silk protein, sericin on UVB-induced acute damage and tumor promotion by reducing oxidative stress in the skin of hairless mouse  

Microsoft Academic Search

This study was conducted to assess protective effect of an antioxidant protein, sericin, on UVB-induced acute damage and tumor promotion in mouse skin. In experiment 1, HR-1 hairless mice were treated with 180 mJ\\/cm2 of ultraviolet B light (UVB) once daily for 1 and 7 days. The treatment for 7 days caused red sunburn lesions of the skin. The intensity

Siqin Zhaorigetu; Noriyuki Yanaka; Masahiro Sasaki; Hiromitsu Watanabe; Norihisa Kato

2003-01-01

27

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

E-print Network

Structure and expression of the silk adhesive protein Ser2 in Bombyx mori Barbara Kludkiewicz a in revised form 27 November 2009 Accepted 30 November 2009 Keywords: Silkworm Sericin Cocoon Silk gland Repetitive sequence a b s t r a c t Sericins are soluble silk components encoded in Bombyx mori by three

Â?urovec, Michal

28

Sericin removal from raw Bombyx mori silk scaffolds of high hierarchical order.  

PubMed

Silk fibroin has previously been described as a promising candidate for ligament tissue engineering (TE) approaches. For biocompatibility reasons, silkworm silk requires removal of sericin, which can elicit adverse immune responses in the human body. One disadvantage of the required degumming process is the alteration of the silk fiber structural properties, which can hinder textile engineering of high order hierarchical structures. Therefore, the aim of this study was to find a way to remove sericin from a compact and highly ordered raw silk fiber matrix. The wire rope design of the test model scaffold comprises several levels of geometric hierarchy. Commonly used degumming solutions fail in removing sericin in this wire rope design. Weight loss measurements, picric acid and carmine staining as well as scanning electron microscopy demonstrated that the removal of sericin from the model scaffold of a wire rope design can be achieved through a borate buffer-based system. Furthermore, the borate buffer degummed silks were shown to be nontoxic and did not alter cell proliferation behavior. The possibility to remove sericin after the textile engineering process has taken place eases the production of highly ordered scaffold structures and may expand the use of silk as scaffold material in further TE and regenerative medicine applications. PMID:24066942

Teuschl, Andreas Herbert; van Griensven, Martijn; Redl, Heinz

2014-05-01

29

Effect of Silk Protein Processing on Drug Delivery from Silk Films  

PubMed Central

Sericin removal from the core fibroin protein of silkworm silk is a critical first step in the use of silk for biomaterial-related applications, but degumming can affect silk biomaterial properties, including molecular weight, viscosity, diffusivity and degradation behavior. Increasing the degumming time (10, 30, 60 and 90 min) decreases the average molecular weight of silk protein in solution, silk solution viscosity, and silk film glass transition temperature, and increases the rate of degradation of silk film by protease. Model compounds spanning a range of physical-chemical properties generally showed an inverse relationship between degumming time and release rate through a varied degumming time silk coating. Degumming provides a useful control point to manipulate silk’s material properties. PMID:23349062

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

2013-01-01

30

Effect of silk protein processing on drug delivery from silk films.  

PubMed

Sericin removal from the core fibroin protein of silkworm silk is a critical first step in the use of silk for biomaterial-related applications, but degumming can affect silk biomaterial properties, including molecular weight, viscosity, diffusivity and degradation behavior. Increasing the degumming time (10, 30, 60, and 90?min) decreases the average molecular weight of silk protein in solution, silk solution viscosity, and silk film glass-transition temperature, and increases the rate of degradation of a silk film by protease. Model compounds spanning a range of physical-chemical properties generally show an inverse relationship between degumming time and release rate through a varied degumming time silk coating. Degumming provides a useful control point to manipulate silk's material properties. PMID:23349062

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

2013-03-01

31

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

PubMed

Hox genes are well-known master regulators in developmental morphogenesis along the anteroposterior axis of animals. However, the molecular mechanisms by which Hox proteins regulate their target genes and determine cell fates are not fully understood. The silk gland of Bombyx mori is a tubular tissue divided into several subparts along the anteroposterior axis, and the silk genes are expressed with specific patterns. The sericin-1 gene (ser1) is expressed in the middle silk gland (MSG) with sublocal specificity. Here we show that the Hox protein Antp is a component of the middle silk gland-specific complex, MIC (MSG-intermolt-specific complex), binds to the essential promoter element of ser1, and activates its expression. Ectopic expression of Antp in transgenic silkworms induced the expression of ser1 in the posterior silk gland (PSG), but not in the anterior part of MSG (MSG-A). Correspondingly, a MIC-like complex was formed by the addition of recombinant Antp in extracts from PSG with its cofactors Exd and Hth, but not in extracts from MSG-A. Splicing patterns of ser1 mRNA induced by the ectopic expression of Antp in PSG were almost the same as those in MSG at the fifth instar and altered depending on the induction timing of Antp. Other Hox genes were expressed with sublocal specificity in the silk gland. The Bombyx silk gland might provide a useful system for understanding how Hox proteins select and regulate their target genes. PMID:24333180

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

2014-02-01

32

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

33

Identification of a Novel Type of Silk Protein and Regulation of Its Expression*  

E-print Network

Identification of a Novel Type of Silk Protein and Regulation of Its Expression* (Received The silk of lepidopteran insects has been studied ex- tensively as proteins of two categories: the fibroins, which are produced in the posterior section of silk glands, and the sericins, which are secreted

Â?urovec, Michal

34

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

35

Vibrational spectroscopic study of sulphated silk proteins  

NASA Astrophysics Data System (ADS)

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

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

2007-05-01

36

Preparation of regenerated silk fibroin/silk sericin fibers by coaxial electrospinning.  

PubMed

The coaxial electrospinning using the regenerated silk fibroin (SF) and silk sericin (SS) aqueous solutions as the core and shell spinning dopes, respectively, was carried out to prepare regenerated SF/SS composite fibers with components and core-shell structure similar to the natural silkworm silks. It was found from the scanning electron microscope (SEM) and transmission electron microscope (TEM) results that the core dope (SF aqueous solution) flow rate (Q(c)) and the applied voltage (V) had some effects on the morphology of the composite fiber. With an increase in Q(c), the diameter nonuniformity and eccentricity of the core fiber became serious, while the increasing V played an inverse role. In this work, the suitable Q(c) for the fiber formation with better electrospinnability was about 6 ?L/min, and the corresponding optimum V was 40 kV. Moreover, the results from Raman spectra analysis, modulated differential scanning calorimetry (MDSC), thermogravimetry (TG) measurement and mechanical property test showed that, compared with the pure SF fiber, the coaxially electrospun SF/SS fiber had more ?-sheet conformation, better thermostability and mechanical properties. This was probably because that SS played significant roles in dehydrating SF molecules and inducing the conformational transition of SF to ?-sheet structure. PMID:22935694

Hang, Yichun; Zhang, Yaopeng; Jin, Yuan; Shao, Huili; Hu, Xuechao

2012-12-01

37

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

38

The characteristics of bacterial nanocellulose gel releasing silk sericin for facial treatment.  

PubMed

BackgroundRecently, naturally derived facial masks with beneficial biological properties have received increasing interest. In this study, silk sericin-releasing bacterial nanocellulose gel was developed to be applied as a bioactive mask for facial treatment.ResultsThe silk sericin-releasing bacterial nanocellulose gel produced at a pH of 4.5 had an ultrafine and extremely pure fiber network structure. The mechanical properties and moisture absorption ability of the gel were improved, compared to those of the commercially available paper mask. Silk sericin could be control-released from the gel. A peel test with porcine skin showed that the gel was less adhesive than the commercially available paper mask, which would be removed from the face more easily without pain. The in vitro cytotoxicity test showed that the gel was not toxic to L929 mouse fibroblast and HaCaT human keratinocyte cells. Furthermore, when implanted subcutaneously and evaluated according to ISO10993-6 standard, the gel was not irritant to tissue.ConclusionThe silk sericin-releasing bacterial nanocellulose gel had appropriate physical and biological properties and safety for the facial treatment application. PMID:25487808

Aramwit, Pornanong; Bang, Nipaporn

2014-12-01

39

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

40

Development of ethyl alcohol-precipitated silk sericin/polyvinyl alcohol scaffolds for accelerated healing of full-thickness wounds.  

PubMed

Silk sericin has been recently reported for its advantageous biological properties to promote wound healing. In this study, we established that the ethyl alcohol (EtOH) could be used to precipitate sericin and form the stable sericin/polyvinyl alcohol (PVA) scaffolds without the crosslinking. The sericin/PVA scaffolds were fabricated via freeze-drying and subsequently precipitating in various concentrations of EtOH. The EtOH-precipitated sericin/PVA scaffolds showed denser structure, higher compressive modulus, but lower water swelling ability than the non-precipitated scaffolds. Sericin could be released from the EtOH-precipitated sericin/PVA scaffolds in a sustained manner. After cultured with L929 mouse fibroblasts, the 70 vol% EtOH-precipitated sericin/PVA scaffolds showed the highest potential to promote cell proliferation. After applied to the full-thickness wounds of rats, the 70 vol% EtOH-precipitated sericin/PVA scaffolds showed significantly higher percentage of wound size reduction and higher extent of type III collagen formation and epithelialization, compared with the control scaffolds without sericin. The accelerated wound healing by the 70 vol% EtOH-precipitated sericin/PVA scaffolds was possibly due to (1) the bioactivity of sericin itself to promote wound healing, (2) the sustained release of precipitated sericin from the scaffolds, and (3) the activation and recruitment of wound healing-macrophages by sericin to the wounds. This finding suggested that the EtOH-precipitated sericin/PVA scaffolds were more effective for the wound healing, comparing with the EtOH-precipitated PVA scaffolds without sericin. PMID:23022662

Siritienthong, Tippawan; Ratanavaraporn, Juthamas; Aramwit, Pornanong

2012-12-15

41

Bioengineered silk proteins to control cell and tissue functions.  

PubMed

Silks are defined as protein polymers that are spun into fibers by some lepidoptera larvae such as silkworms, spiders, scorpions, mites, and flies. Silk proteins are usually produced within specialized glands in these animals after biosynthesis in epithelial cells that line the glands, followed by secretion into the lumen of the gland prior to spinning into fibers.The most comprehensively characterized silks are from the domesticated silkworm (Bombyx mori) and from some spiders (Nephila clavipes and Araneus diadematus). Silkworm silk has been used commercially as biomedical sutures for decades and in textile production for centuries. Because of their impressive mechanical properties, silk proteins provide an important set of material options in the fields of controlled drug release, and for biomaterials and scaffolds for tissue engineering. Silkworm silk from B. mori consists primarily of two protein components, fibroin, the structural protein of silk fibers, and sericins, the water-soluble glue-like proteins that bind the fibroin fibers together. Silk fibroin consists of heavy and light chain polypeptides linked by a disulfide bond. Fibroin is the protein of interest for biomedical materials and it has to be purified/extracted from the silkworm cocoon by removal of the sericin. Characteristics of silks, including biodegradability, biocompatibility, controllable degradation rates, and versatility to generate different material formats from gels to fibers and sponges, have attracted interest in the field of biomaterials. Cell culture and tissue formation using silk-based biomaterials have been pursued, where appropriate cell adhesion, proliferation, and differentiation on or in silk biomaterials support the regeneration of tissues. The relative ease with which silk proteins can be processed into a variety of material morphologies, versatile chemical functionalization options, processing in water or solvent, and the related biological features of biocompatibility and enzymatic degradability make these proteins interesting candidates for biomedical applications. PMID:23504416

Preda, Rucsanda C; Leisk, Gary; Omenetto, Fiorenzo; Kaplan, David L

2013-01-01

42

Comparative proteomics reveal diverse functions and dynamic changes of Bombyx mori silk proteins spun from different development stages.  

PubMed

Silkworms (Bombyx mori) produce massive amounts of silk proteins to make cocoons during the final stages of larval development. Although the major components, fibroin and sericin, have been the focus for a long time, few researchers have realized the complexity of the silk proteome. We collected seven kinds of silk fibers spun by silkworm larvae at different developmental stages: the silks spun by new hatched larvae, second instar day 0 larvae, third instar day 0 larvae, fourth instar day 0 larvae, and fourth instar molting larvae, the scaffold silk used to attach the cocoon to the substrate and the cocoon silk. Analysis by liquid chromatography-tandem mass spectrometry identified 500 proteins from the seven silks. In addition to the expected fibroins, sericins, and some known protease inhibitors, we also identified further protease inhibitors, enzymes, proteins of unknown function, and other proteins. Unsurprisingly, our quantitative results showed fibroins and sericins were the most abundant proteins in all seven silks. Except for fibroins and sericins, protease inhibitors, enzymes, and proteins of unknown function were more abundant than other proteins. We found significant change in silk protein compositions through development, being consistent with their different biological functions and complicated formation. PMID:24093152

Dong, Zhaoming; Zhao, Ping; Wang, Chen; Zhang, Yan; Chen, Jianping; Wang, Xin; Lin, Ying; Xia, Qingyou

2013-11-01

43

Silk sericin-alginate-chitosan microcapsules: hepatocytes encapsulation for enhanced cellular functions.  

PubMed

The encapsulation based technology permits long-term delivery of desired therapeutic products in local regions of body without the need of immunosuppressant drugs. In this study microcapsules composed of sericin and alginate micro bead as inner core and with an outer chitosan shell are prepared. This work is proposed for live cell encapsulation for potential therapeutic applications. The sericin protein is obtained from cocoons of non-mulberry silkworm Antheraea mylitta. The sericin-alginate micro beads are prepared via ionotropic gelation under high applied voltage. The beads further coated with chitosan and crosslinked with genipin. The microcapsules developed are nearly spherical in shape with smooth surface morphology. Alamar blue assay and confocal microscopy indicate high cell viability and uniform encapsulated cell distribution within the sericin-alginate-chitosan microcapsules indicating that the microcapsules maintain favourable microenvironment for the cells. The functional analysis of encapsulated cells demonstrates that the glucose consumption, urea secretion rate and intracellular albumin content increased in the microcapsules. The study suggests that the developed sericin-alginate-chitosan microcapsule contributes towards the development of cell encapsulation model. It also offers to generate enriched population of metabolically and functionally active cells for the future therapeutics especially for hepatocytes transplantation in acute liver failure. PMID:24486492

Nayak, Sunita; Dey, Sanchareeka; Kundu, Subhas C

2014-04-01

44

The influence of silkworm species on cellular interactions with novel PVA/silk sericin hydrogels.  

PubMed

Sericin peptides and PVA are chemically modified with methacrylate groups to produce a covalent PVA/sericin hydrogel. Preservation of the sericin bioactivity following methacrylation is confirmed, and PVA/sericin hydrogels are fabricated for both B. mori and A. mylitta sericin. Cell adhesion studies confirm the preservation of sericin bioactivity post incorporation in PVA gels. PVA/A. mylitta gels are observed to facilitate cell adhesion to a significantly greater degree than PVA/B. mori gels. Overall, the incorporation of sericin does not alter the physical properties of the PVA hydrogels but does result in significantly improved cellular interaction, particularly from A. mylitta gels. PMID:22493796

Lim, Khoon S; Kundu, Joydip; Reeves, April; Poole-Warren, Laura A; Kundu, Subhas C; Martens, Penny J

2012-03-01

45

Preliminary characterization of genipin-cross-linked silk sericin/poly(vinyl alcohol) films as two-dimensional wound dressings for the healing of superficial wounds.  

PubMed

The genipin-cross-linked silk sericin/poly(vinyl alcohol) (PVA) films were developed aiming to be applied as two-dimensional wound dressings for the treatment of superficial wounds. The effects of genipin cross-linking concentration on the physical and biological properties of the films were investigated. The genipin-cross-linked silk sericin/PVA films showed the increased surface density, tensile strength, and percentage of elongation, but decreased percentage of light transmission, water vapor transmission rate, and water swelling, compared to the non-cross-linked films. This explained that the cross-linking bonds between genipin and silk sericin would reduce the mobility of molecular chains within the films, resulting in the more rigid molecular structure. Silk sericin was released from the genipin-cross-linked films in a sustained manner. In addition, either L929 mouse fibroblast or HaCat keratinocyte cells showed high percentage of viability when cultured on the silk sericin/PVA films cross-linked with 0.075 and 0.1%?w/v genipin. The in vivo safety test performed according to ISO 10993-6 confirmed that the genipin-cross-linked silk sericin/PVA films were safe for the medical usages. The efficacy of the films for the treatment of superficial skin wounds will be further investigated in vivo and clinically. The genipin-cross-linked silk sericin/PVA films would be promising choices of two-dimensional wound dressings for the treatment of superficial wounds. PMID:24106722

Siritientong, Tippawan; Ratanavaraporn, Juthamas; Srichana, Teerapol; Aramwit, Pornanong

2013-01-01

46

Preliminary Characterization of Genipin-Cross-Linked Silk Sericin/Poly(vinyl alcohol) Films as Two-Dimensional Wound Dressings for the Healing of Superficial Wounds  

PubMed Central

The genipin-cross-linked silk sericin/poly(vinyl alcohol) (PVA) films were developed aiming to be applied as two-dimensional wound dressings for the treatment of superficial wounds. The effects of genipin cross-linking concentration on the physical and biological properties of the films were investigated. The genipin-cross-linked silk sericin/PVA films showed the increased surface density, tensile strength, and percentage of elongation, but decreased percentage of light transmission, water vapor transmission rate, and water swelling, compared to the non-cross-linked films. This explained that the cross-linking bonds between genipin and silk sericin would reduce the mobility of molecular chains within the films, resulting in the more rigid molecular structure. Silk sericin was released from the genipin-cross-linked films in a sustained manner. In addition, either L929 mouse fibroblast or HaCat keratinocyte cells showed high percentage of viability when cultured on the silk sericin/PVA films cross-linked with 0.075 and 0.1%?w/v genipin. The in vivo safety test performed according to ISO 10993-6 confirmed that the genipin-cross-linked silk sericin/PVA films were safe for the medical usages. The efficacy of the films for the treatment of superficial skin wounds will be further investigated in vivo and clinically. The genipin-cross-linked silk sericin/PVA films would be promising choices of two-dimensional wound dressings for the treatment of superficial wounds. PMID:24106722

Siritientong, Tippawan; Ratanavaraporn, Juthamas; Srichana, Teerapol; Aramwit, Pornanong

2013-01-01

47

In vitro development of OPU-derived bovine embryos cultured either individually or in groups with the silk protein sericin and the viability of frozen-thawed embryos after transfer.  

PubMed

The optimization of single-embryo culture conditions is very important, particularly in the in vitro production of bovine embryos using the ovum pick-up (OPU) procedure. The purpose of this study was to examine the development of embryos derived from oocytes obtained by OPU that were cultured either individually or in groups in medium supplemented with or without sericin and to investigate the viability of the frozen-thawed embryos after a direct transfer. When two-cell-stage embryos were cultured either individually or in groups for 7 days in CR1aa medium supplemented with or without 0.5% sericin, the rates of development to blastocysts and freezable blastocysts were significantly lower for the embryos cultured individually without sericin than for the embryos cultured in groups with or without sericin. Moreover, the rate of development to freezable blastocysts of the embryos cultured individually with sericin was significantly higher than that of the embryos cultured without sericin. When the frozen-thawed embryos were transferred directly to recipients, the rates of pregnancy, abortion, stillbirth and normal calving in the recipients were similar among the groups, irrespective of the culture conditions and sericin supplementation. Our findings indicate that supplementation with sericin during embryo culture improves the quality of the embryos cultured individually but not the viability of the frozen-thawed embryos after transfer to recipients. PMID:25488699

Isobe, Tomohiro; Ikebata, Yoshihisa; Do, Lanh Thi Kim; Tanihara, Fuminori; Taniguchi, Masayasu; Otoi, Takeshige

2014-12-01

48

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

49

Green synthesis of silk sericin-capped silver nanoparticles and their potent anti-bacterial activity.  

PubMed

In this study, a 'green chemistry' approach was introduced to synthesize silk sericin (SS)-capped silver nanoparticles (AgNPs) under an alkaline condition (pH 11) using SS as a reducing and stabilizing agent instead of toxic chemicals. The SS-capped AgNPs were successfully synthesized at various concentrations of SS and AgNO3, but the yields were different. A higher yield of SS-capped AgNPs was obtained when the concentrations of SS and AgNO3 were increased. The SS-capped AgNPs showed a round shape and uniform size with diameter at around 48 to 117 nm. The Fourier transform infrared (FT-IR) spectroscopy result proved that the carboxylate groups obtained from alkaline degradation of SS would be a reducing agent for the generation of AgNPs while COO- and NH2?+ groups stabilized the AgNPs and prevented their precipitation or aggregation. Furthermore, the SS-capped AgNPs showed potent anti-bacterial activity against various gram-positive bacteria (minimal inhibitory concentration (MIC) 0.008 mM) and gram-negative bacteria (MIC ranging from 0.001 to 0.004 mM). Therefore, the SS-capped AgNPs would be a safe candidate for anti-bacterial applications. PMID:24533676

Aramwit, Pornanong; Bang, Nipaporn; Ratanavaraporn, Juthamas; Ekgasit, Sanong

2014-01-01

50

Green synthesis of silk sericin-capped silver nanoparticles and their potent anti-bacterial activity  

PubMed Central

In this study, a ‘green chemistry’ approach was introduced to synthesize silk sericin (SS)-capped silver nanoparticles (AgNPs) under an alkaline condition (pH 11) using SS as a reducing and stabilizing agent instead of toxic chemicals. The SS-capped AgNPs were successfully synthesized at various concentrations of SS and AgNO3, but the yields were different. A higher yield of SS-capped AgNPs was obtained when the concentrations of SS and AgNO3 were increased. The SS-capped AgNPs showed a round shape and uniform size with diameter at around 48 to 117 nm. The Fourier transform infrared (FT-IR) spectroscopy result proved that the carboxylate groups obtained from alkaline degradation of SS would be a reducing agent for the generation of AgNPs while COO? and NH2?+ groups stabilized the AgNPs and prevented their precipitation or aggregation. Furthermore, the SS-capped AgNPs showed potent anti-bacterial activity against various gram-positive bacteria (minimal inhibitory concentration (MIC) 0.008 mM) and gram-negative bacteria (MIC ranging from 0.001 to 0.004 mM). Therefore, the SS-capped AgNPs would be a safe candidate for anti-bacterial applications. PMID:24533676

2014-01-01

51

Effect of degumming condition on the solution properties and electrospinnablity of regenerated silk solution.  

PubMed

The application of silk on tissue engineering scaffolds has been studied intensively because silk has an electrospinning technique using a good blood compatibility, excellent cytocompatibility and biodegradability. Silk consists of two polymers, fibroin and sericin. In spite of importance of sericin, most studies were focused on the fibroin only and the effect of residual sericin on the electrospinning performance of silk has not been considered. In this study, regenerated silk with different residual sericin contents was prepared by controlling the degumming conditions. The effects of the degumming conditions on the solution properties and electrospinning performance of silk were examined. The fast protein liquid chromatography (FPLC) measurements confirmed that the molecular weight of the regenerated silk decreased slightly with increasing residual sericin content. More molecular aggregation of silk occurred with increasing sericin content, resulting in an increase in the solution turbidity of formic acid. All silk formic acid solutions exhibited almost Newtonian fluid behavior and the viscosity increased with increasing sericin content. Interestingly, the dope solution viscosity of silk increased remarkably at sericin contents <1% (or degumming ratio >25%) leading to significant improvements in electrospinnability and an increase in the fiber diameter of the silk web. PMID:23295206

Ko, Jae Sang; Yoon, Kyunghwan; Ki, Chang Seok; Kim, Hyun Ju; Bae, Do Gyu; Lee, Ki Hoon; Park, Young Hwan; Um, In Chul

2013-04-01

52

Assembly mechanism of recombinant spider silk proteins  

E-print Network

Assembly mechanism of recombinant spider silk proteins S. Rammensee*, U. Slotta , T. Scheibel October 5, 2007) Spider silk threads are formed by the irreversible aggregation of silk proteins engineered and recombinantly produced spider dragline silk proteins eADF3 (engineered Araneus diadematus

Bausch, Andreas

53

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

54

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, Valquíria A; Rech, Elíbio L; Kaplan, David L

2013-01-01

55

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

56

A novel poly(?-glutamic acid)/silk-sericin hydrogel for wound dressing: Synthesis, characterization and biological evaluation.  

PubMed

A novel multifunctional poly(?-glutamic acid)/silk sericin (?-PGA/SS) hydrogel has been developed and used as wound dressing. The physical and chemical properties of the ?-PGA/SS gels were systemically investigated. Furthermore, these ?-PGA/SS gels have been found to promote the L929 fibroblast cells proliferate, and in the in vivo study, significant stimulatory effects were also observed on granulation and capillary formation on day 9 in H-2-treated wounds, indicating that this new complex hydrogel could maintain a moist healing environment, protect the wound from bacterial infection, absorb excess exudates, and promote cell proliferation to reconstruct damaged tissue. Considering the simple preparation process and excellent biological property, this ?-PGA/SS hydrogel might have a wide range of applications in biomedical and clinical areas. PMID:25579954

Shi, Lu; Yang, Ning; Zhang, Hao; Chen, Li; Tao, Lei; Wei, Yen; Liu, Hui; Luo, Ying

2015-03-01

57

Potential of 2D crosslinked sericin membranes with improved biostability for skin tissue engineering.  

PubMed

Silk sericin protein is a natural, hydrophilic, macromolecular glycoprotein mainly synthesized in the middle silk gland of the silkworm. It constitutes 25-30% of the silk cocoon. Sericin proteins have antioxidant, antimicrobial, UV-resistant properties, promote wound healing and support cell proliferation even in serum-free media. Most of the sericin is discarded as waste in silk processing industries. This study aims at improving the mechanical strength and stability of sericin extracted from the silk cocoons during processing and utilize it as a biocompatible natural biopolymer in biomedical applications. Crosslinked sericin membranes, from the cocoon of non-mulberry tropical silkworm, Antheraea mylitta, were prepared using gluteraldehyde as the crosslinking agent. Physical and structural characteristics of the membranes were analyzed using scanning electron microscopy, atomic force microscopy, Fourier transform infrared spectroscopy and X-ray diffraction along with swelling and degradation studies. The secondary structure of the membrane indicates that crosslinking provides a more integrated structure that significantly improves the stability and mechanical strength of the membranes. In vitro cytocompatibility of the membranes was evaluated by MTT assay and cell cycle analysis of feline fibroblast cells. The adherence, growth and proliferation patterns of cells on membranes were assessed by confocal microscopy, which demonstrated that the latter is non-toxic and supports cell growth. Cell cycle analyses indicate cytocompatibility with normal cell cycle pattern. This study reveals that silk sericin protein can be used as a biocompatible natural biopolymer for various applications in the biomedical field. PMID:22327482

Nayak, Sunita; Talukdar, Sarmistha; Kundu, Subhas C

2012-03-01

58

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

PubMed

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

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

2014-12-01

59

Effect of sericin on preimplantation development of bovine embryos cultured individually.  

PubMed

The silk protein sericin has been identified as a potent antioxidant in mammalian cells. This study was conducted to examine the effects of sericin on preimplantation development and quality of bovine embryos cultured individually. When two-cell-stage embryos were cultured individually for 7 days in CR1aa medium supplemented with 0, 0.1, 0.5, or 1% sericin, rates of total blastocyst formation and development to expanded blastocysts from embryos cultured with 0.5% sericin were higher (P < 0.05) than those from embryos cultured with 0 or 1% sericin. When embryos were cultured individually for 7 days in the CR1aa medium supplemented with 0 or 0.5% sericin under two oxidative stress conditions (50 or 100 ?m H(2)O(2)), the addition of sericin significantly improved the blastocyst formation rate of embryos exposed to 100 ?m H(2)O(2). However, the protective effect of sericin was not observed in development of embryos exposed to 50 ?m H(2)O(2). When embryos were exposed to 100 ?m H(2)O(2) during culture, the DNA fragmentation index of total blastocysts from embryos cultured with 0.5% sericin was lower than blastocysts derived from embryos cultured without sericin (4.4 vs. 6.8%; P < 0.01). In conclusion, the addition of 0.5% sericin to in vitro culture medium improved preimplantation development and quality of bovine embryos cultured individually by preventing oxidative stress. PMID:22626775

Isobe, T; Ikebata, Y; Onitsuka, T; Wittayarat, M; Sato, Y; Taniguchi, M; Otoi, T

2012-09-01

60

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

PubMed

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

2014-05-01

61

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

62

Insect silk contains both a Kunitz-type and a unique Kazal-type proteinase inhibitor  

E-print Network

Insect silk contains both a Kunitz-type and a unique Kazal-type proteinase inhibitor Xavier Nirmala Republic Insect silk is made up of structural fibrous (fibroins) and sticky (sericins) proteins inhibit bacterial and fungal proteinases (subtilisin, proteinase K and pronase). These `silk protei- nase

Â?urovec, Michal

63

Genipin-cross-linked thermosensitive silk sericin/poly(N-isopropylacrylamide) hydrogels for cell proliferation and rapid detachment.  

PubMed

To overcome release of silk sericin (SS) from semi-interpenetrating polymer network (semi-IPN) SS/poly(N-isopropylacrylamide) (PNIPAm) hydrogels, natural biocompatible genipin (GNP) was adopted as cross-linking agent of SS. The GNP/SS/PNIPAm hydrogels with various GNP contents were prepared by radical polymerization. Depending on GNP content, the resultant hydrogels present white, yellow, or dark blue. Required time of color change for GNP/SS mixture solution shortened with increasing GNP ratio. The GNP/SS/PNIPAm hydrogels present good oscillatory shrinking-swelling behavior between 20 and 37°C. The behaviors of L929 cell proliferation, desorption, and transshipment on the surface of hydrogels and tissue culture polystyrene were investigated by 3-(4,5-dimethy thioazol-2-yl)-2,5-di-phenytetrazoliumromide and scanning electron microscopy method. In comparison with pure SS/PNIPAm hydrogels, the introduction of certain GNP can accelerate cell adhesion and proliferation. Due to reversible change between hydrophobicity and hydrophilicity, by lowering temperature to 4°C from 37°C, L929 cells could spontaneously detach from the surface of hydrogels without the need for trypsin or ethylenediaminetetraacetic acid. The detached cells could subsequently be recultured. The prepared hydrogel and detached cells have potential applications in biomedical fields, such as organs or tissue regeneration and cancer or disease therapy. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2013. PMID:23606462

Zhang, Qingsong; Dong, Panpan; Chen, Li; Wang, Xiaozhao; Lu, Si

2013-04-18

64

Preparation of sericin film with different polymers  

Microsoft Academic Search

Silk sericin (SS) is a natural protein derived from silkworm Bombyx mori. It is applicable as an antioxidant in the field of medicines, cosmetics, foods, and food additive SS protein can be blended\\u000a with polymers, to produce materials with improved properties. SS films were prepared using different polymers which are polyvinyl\\u000a alcohol (PVA), polyoxyethylene-polyoxypropylene block copolymer (Plu) and glycerine (G)

Kamol Maikrang; Pornanong Aramwit

65

Thermal crystallization mechanism of silk fibroin protein  

NASA Astrophysics Data System (ADS)

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

Hu, Xiao

66

Dietary sericin enhances epidermal levels of glucosylceramides and ceramides with up-regulating protein expressions of glucosylceramide synthase, ?-glucocerebrosidase and acidic sphingomyelinase in NC/Nga mice.  

PubMed

We have previously reported that dietary sericin improves epidermal dryness with the increased total Ceramide (Cer) in NC/Nga mice, an animal model of atopic dermatitis (AD). In this study, we hypothesized that the increased level of total Cer induced by dietary sericin would be related to the altered metabolism of glucosylceramide (GlcCer) and sphingomyelin (SM), major precursors of Cer generation. NC/Nga mice were fed a control diet (group CA: atopic control) or diets with 1% silk protein, either sericin (group S) or fibroin (group F) for 10 weeks. In the epidermis of group CA, total Cer (including Cer1, 2, 3/4 and 6) and all GlcCer species were reduced; these levels in group S were increased to levels similar to or higher than in the normal control group of BALB/c mice (group C). In addition, the protein expressions, but not mRNA expressions, of GlcCer synthase, ?-glucocerebrosidase, and acidic sphingomyelinase, enzymes for GlcCer synthesis, GlcCer and SM hydrolysis, respectively, were highly increased in group S. The epidermal levels of total Cer (including Cer2, 3/4, and 6) and all GlcCer species and of these enzyme proteins in group F were lower than in group S. Notably, alterations in total SM, SM1, SM3, and SM synthase 1, which were increased in group CA, were not significant between groups S and F. Cer5 and SM2 were not altered among groups. Dietary sericin enhanced the epidermal levels of all GlcCer and most Cer species with up-regulating protein expressions of GlcCer synthase, ?-glucocerebrosidase, and acidic sphingomyelinase. PMID:23244541

Kim, Hyunae; Lee, Jongsun; Cho, Yunhi

2012-12-01

67

A Materiomics Approach to Spider Silk: Protein Molecules to Webs  

E-print Network

A Materiomics Approach to Spider Silk: Protein Molecules to Webs ANNA TARAKANOVA1 and MARKUS J-assembling silk biopolymers have been extensively studied experimentally and in computa- tional investigations length scales in silk, ranging from atomistic models of protein constituents to the spider web

Buehler, Markus J.

68

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

69

Photoprotection by silk cocoons.  

PubMed

A silk cocoon protects a silkworm during its pupal stage from various threats. We systematically investigated the role of fiber, sericin, and embedded crystals in the UV protection of a silk cocoon. Diffuse reflectance and UV absorbance were measured and free radicals generated during exposure to UV radiation were quantified using photoinduced chemiluminescence (PICL). We identified the response to both UV-A and UV-B radiations by silk materials and found that sericin was primarily responsible for UV-A absorption. When sericin was removed, the photoinduced chemiluminescence intensity increased significantly, indicating higher UV-A-induced reactions of cocoons in the absence of sericin. There is progressively higher sericin content toward the outer part of the cocoon shell that allows an effective shield to pupae from UV radiation and resists photodegradation of silk fibers. The study will inspire development of advanced organic photoprotective materials and designing silk-based, free-radical-scavenging antioxidants. PMID:24000973

Kaur, Jasjeet; Rajkhowa, Rangam; Tsuzuki, Takuya; Millington, Keith; Zhang, Jin; Wang, Xungai

2013-10-14

70

Inhibition of the binding of MSG-intermolt-specific complex, MIC, to the sericin-1 gene promoter and sericin-1 gene expression by POU-M1/SGF-3.  

PubMed

The sericin-1 gene encoding a glue protein is expressed in the middle silk gland (MSG) of the silkworm, Bombyx mori. A member of the class III POU domain transcription factors, POU-M1, was cloned as the factor bound to the SC site of the sericin-1 promoter and has been proposed to be a positive transcription factor. In this study, we analyzed the expression pattern of the POU-M1 gene in fourth and fifth instars in comparison with the pattern of the sericin-1 gene. The POU-M1 gene was expressed strongly in the region anterior to the sericin-1-expressing portion of the silk gland at both feeding stages. As the sericin-1-expressing region expands from the posterior to middle portions of the MSG in the fifth instar, the POU-M1-expressing region retreated from the middle to anterior portion. Introduction of the expression vector of POU-M1 into the silk glands by gene gun technology repressed promoter activity of the sericin-1 gene, suggesting that POU-M1 regulates the sericin-1 gene negatively. An in vitro binding assay showed that POU-M1 bound not only to the SC site but also to other promoter elements newly detected in vivo. Another spatiotemporal specific factor MIC binds to these elements, and POU-M1 competed with MIC to bind at the -70 site essential for promoter activity. These results suggest that POU-M1 is involved in restricting the anterior boundary of the sericin-1-expressing region in the silk gland by inhibiting the binding of the transcriptional activator to the promoter elements. PMID:23070540

Kimoto, Mai; Kitagawa, Tsuyuki; Kobayashi, Isao; Nakata, Tomohiro; Kuroiwa, Asato; Takiya, Shigeharu

2012-11-01

71

High-toughness silk produced by a transgenic silkworm expressing spider (Araneus ventricosus) dragline silk protein.  

PubMed

Spider dragline silk is a natural fiber that has excellent tensile properties; however, it is difficult to produce artificially as a long, strong fiber. Here, the spider (Araneus ventricosus) dragline protein gene was cloned and a transgenic silkworm was generated, that expressed the fusion protein of the fibroin heavy chain and spider dragline protein in cocoon silk. The spider silk protein content ranged from 0.37 to 0.61% w/w (1.4-2.4 mol%) native silkworm fibroin. Using a good silk-producing strain, C515, as the transgenic silkworm can make the raw silk from its cocoons for the first time. The tensile characteristics (toughness) of the raw silk improved by 53% after the introduction of spider dragline silk protein; the improvement depended on the quantity of the expressed spider dragline protein. To demonstrate the commercial feasibility for machine reeling, weaving, and sewing, we used the transgenic spider silk to weave a vest and scarf; this was the first application of spider silk fibers from transgenic silkworms. PMID:25162624

Kuwana, Yoshihiko; Sezutsu, Hideki; Nakajima, Ken-ichi; Tamada, Yasushi; Kojima, Katsura

2014-01-01

72

High-Toughness Silk Produced by a Transgenic Silkworm Expressing Spider (Araneus ventricosus) Dragline Silk Protein  

PubMed Central

Spider dragline silk is a natural fiber that has excellent tensile properties; however, it is difficult to produce artificially as a long, strong fiber. Here, the spider (Araneus ventricosus) dragline protein gene was cloned and a transgenic silkworm was generated, that expressed the fusion protein of the fibroin heavy chain and spider dragline protein in cocoon silk. The spider silk protein content ranged from 0.37 to 0.61% w/w (1.4–2.4 mol%) native silkworm fibroin. Using a good silk-producing strain, C515, as the transgenic silkworm can make the raw silk from its cocoons for the first time. The tensile characteristics (toughness) of the raw silk improved by 53% after the introduction of spider dragline silk protein; the improvement depended on the quantity of the expressed spider dragline protein. To demonstrate the commercial feasibility for machine reeling, weaving, and sewing, we used the transgenic spider silk to weave a vest and scarf; this was the first application of spider silk fibers from transgenic silkworms. PMID:25162624

Kuwana, Yoshihiko; Sezutsu, Hideki; Nakajima, Ken-ichi; Tamada, Yasushi; Kojima, Katsura

2014-01-01

73

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

E-print Network

IDENTIFICATION, CHARACTERIZATION AND ANTI-FUNGAL ACTVITIES OF SILK PROTEINS IN ASPERGILLUS FLAVUS 2006 #12;IDENTIFICATION, CHARACTERIZATION AND ANTI-FUNGAL ACTVITIES OF SILK PROTEINS IN ASPERGILLUS: IDENTIFICATION, CHARACTERIZATION AND ANTIFUNGAL ACTIVITIES OF SILK PROTEINS IN ASPERGILLUS FLAVUS RESISTANT

Ray, David

74

Sericin supplementation improves semen freezability of buffalo bulls by minimizing oxidative stress during cryopreservation.  

PubMed

The variety of mammalian cells has been successfully cryopreserved by use of the silk protein sericin due to its strong free-radical-scavenging and potent antioxidant activity. The present study was conducted to examine the protective role of sericin on buffalo spermatozoa during cryopreservation. Semen of four breeding bulls was collected twice a week using artificial vagina technique. The ejaculates of four bulls were pooled, divided into five equal fractions, diluted with the extender supplemented with different concentrations of sericin (0, 0.25, 0.5, 1.5 and 2%) and then cryopreserved. Post-thawed motility was objectively assessed by computer assisted sperm analyzer. Sperm plasma membrane integrity was assessed by hypo-osmotic swelling test (HOST). Malondialdehyde (MDA) concentration, glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities were determined in frozen-thawed extended seminal plasma by spectrophotometry. The extender supplemented with 0.25, 0.5 and 1% sericin resulted in the higher sperm motility and GPx acivity. Furthermore, plasma membrane integrity and SOD activity were found to be higher (P<0.05) in group supplemented with 0.25 and 0.5% sericin (P<0.05). The MDA concentration was found to be significantly lower (P<0.05) in 0.25 and 0.5% sericin treated groups than control and other treated groups. In conclusion, the supplementation of 0.25-0.5% sericin in semen extender improves frozen-thawed semen quality through protecting sperm from oxidative stress. PMID:25497424

Kumar, Pradeep; Kumar, Dharmendra; Sikka, P; Singh, P

2015-01-01

75

Production And Characterization Of Synthetic Spider Silks Based On Nephila Clavipes Major Ampullate Silk Proteins  

NASA Astrophysics Data System (ADS)

The extraordinary mechanical properties of orb-weaving spider silks have served spiders for over 400 million years. However, only in the late 20th century did we start to understand the molecular nature of spider silk that contributes to its incredible properties as biomaterials. Among all seven types of spider silks, major ampullate silk from typical orb-weaving spiders is the toughest of all, it consists of primarily two proteins: MaSp1 and MaSp2. Variable ratios and conserved motifs of these two proteins in all the native spider silks demonstrate the significant role of MaSp1 and MaSp2 in controlling the mechanical properties of the fiber. The amino acid sequences of the orb weaving spider silk proteins have remained almost unchanged for more than 100 million years. Interestingly, MaSp1 and MaSp2 are the only two components in all studied dragline silk fibers from these spiders. The mechanical properties of native dragline silk vary slightly between species, which are believed to relate to the ratio of MaSp1 to MaSp2 in the silk. Both of these facts clearly indicate the importance of these two proteins to the mechanical properties of the fiber. Various types of synthetic spider silk fibers have been produced and studied in an effort to mass-produce man-made fibers with qualities comparable to native spider silk. To investigate the roles of MaSp1 and MaSp2 in silk fiber, synthetic MaSp1 (major abundant protein in Nephila clavipes major ampullate silks) only fibers, MaSp1/MaSp2 protein mixture fibers and chimeric protein fibers with both MaSp1 and MaSp2 sequence features have been produced and tested for mechanical properties. Solid-State Nuclear Magnetic Resonance was used to characterize the structure of silk fibers and reveal the relation between fiber spatial structure and mechanical properties.

An, Bo

76

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

77

Silks produced by insect labial glands  

PubMed Central

Insect silks are secreted from diverse gland types; this chapter deals with the silks produced by labial glands of Holometabola (insects with pupa in their life cycle). Labial silk glands are composed of a few tens or hundreds of large polyploid cells that secrete polymerizing proteins which are stored in the gland lumen as a semi-liquid gel. Polymerization is based on weak molecular interactions between repetitive amino acid motifs present in one or more silk proteins; cross-linking by disulfide bonds may be important in the silks spun under water. The mechanism of long-term storage of the silk dope inside the glands and its conversion into the silk fiber during spinning is not fully understood. The conversion occurs within seconds at ambient temperature and pressure, under minimal drawing force and in some cases under water. The silk filament is largely built of proteins called fibroins and in Lepidoptera and Trichoptera coated by glue-type proteins known as sericins. Silks often contain small amounts of additional proteins of poorly known function. The silk components controlling dope storage and filament formation seem to be conserved at the level of orders, while the nature of polymerizing motifs in the fibroins, which determine the physical properties of silk, differ at the level of family and even genus. Most silks are based on fibroin ?-sheets interrupted with other structures such as ?-helices but the silk proteins of certain sawflies have predominantly a collagen-like or polyglycine II arrangement and the silks of social Hymenoptera are formed from proteins in a coiled coil arrangement. PMID:19221523

Sutherland, Tara

2008-01-01

78

Bioengineered chimeric spider silk-uranium binding proteins.  

PubMed

Heavy metals constitute a source of environmental pollution. Here, novel functional hybrid biomaterials for specific interactions with heavy metals are designed by bioengineering consensus sequence repeats from spider silk of Nephila clavipes with repeats of a uranium peptide recognition motif from a mutated 33-residue of calmodulin protein from Paramecium tetraurelia. The self-assembly features of the silk to control nanoscale organic/inorganic material interfaces provides new biomaterials for uranium recovery. With subsequent enzymatic digestion of the silk to concentrate the sequestered metals, options can be envisaged to use these new chimeric protein systems in environmental engineering, including to remediate environments contaminated by uranium. PMID:23212989

Krishnaji, Sreevidhya Tarakkad; Kaplan, David L

2013-02-01

79

Bioengineered Chimeric Spider Silk-Uranium Binding Proteins  

PubMed Central

Heavy metals constitute a source of environmental pollution. Here, novel functional hybrid biomaterials for specific interactions with heavy metals are designed by bioengineering consensus sequence repeats from spider silk of Nephila clavipes with repeats of a uranium peptide recognition motif from a mutated 33-residue of calmodulin protein from Paramecium tetraurelia. The self-assembly features of the silk to control nanoscale organic/inorganic material interfaces provides new biomaterials for uranium recovery. With subsequent enzymatic digestion of the silk to concentrate the sequestered metals, options can be envisaged to use these new chimeric protein systems in environmental engineering, including to remediate environments contaminated by uranium. PMID:23212989

Krishnaji, Sreevidhya Tarakkad; Kaplan, David L.

2014-01-01

80

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

PubMed

Cocoon, a shelter for larva development to silk moth, contains the fibrous protein fibroin, which is coated by the globular protein sericin. Emergence of the silk moth requires the action of cocoonase, a protease secreted by the pupa. The full-length prococoonase cDNA, with 780 bp open reading frame encoding 260 amino acids, was cloned by reverse transcription from total RNA of the head of 6-day-old Thai-silk Bombyx mori pupa. Only the gene fragment lacking the propeptide encoding sequence was successfully expressed in Pichia pastoris, yielding an extracellularly active cocoonase. The recombinant cocoonase was purified to homogeneity by 80% ammonium-sulfate fractionation and CM-Sepharose chromatography, and its internal peptide sequences were analyzed by nano liquid chromatography-mass spectrometry/mass spectrometry. This monomeric protein has native molecular weight of 26 kDa by gel exclusion analysis and 25 kDa subunit size by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The enzyme hydrolyses sericin but does not hydrolyse fibroin, as shown by radial diffusion on thin-layer enzyme assay (RD-TEA). Scanning electron microscopy showed that purified recombinant cocoonase could remove sericin from natural silk completely in 24 h, without damaging fibroin, using only 1 immobilized sericin unit (ISU) of enzyme as determined by RD-TEA. Natural cocoonase isolated from B. mori pupa could also digest sericin effectively, but required more enzymes (2 ISU) and longer time (48 h). In comparison, a commercial enzyme, alcalase, with the same activity not only showed less complete digestion of sericin but also caused damage of fibroin. These results suggest that recombinant B. mori cocoonase is potentially useful for silk degumming. PMID:23169343

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

2012-12-01

81

Spider silks: recombinant synthesis, assembly, spinning, and engineering of synthetic proteins  

PubMed Central

Since thousands of years humans have utilized insect silks for their own benefit and comfort. The most famous example is the use of reeled silkworm silk from Bombyx mori to produce textiles. In contrast, despite the more promising properties of their silk, spiders have not been domesticated for large-scale or even industrial applications, since farming the spiders is not commercially viable due to their highly territorial and cannibalistic nature. Before spider silks can be copied or mimicked, not only the sequence of the underlying proteins but also their functions have to be resolved. Several attempts to recombinantly produce spider silks or spider silk mimics in various expression hosts have been reported previously. A new protein engineering approach, which combines synthetic repetitive silk sequences with authentic silk domains, reveals proteins that closely resemble silk proteins and that can be produced at high yields, which provides a basis for cost-efficient large scale production of spider silk-like proteins. PMID:15546497

Scheibel, Thomas

2004-01-01

82

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

83

Sericin accelerates the production of hyaluronan and decreases the incidence of polyspermy fertilization in bovine oocytes during in vitro maturation.  

PubMed

Fetal bovine serum (FBS) has been widely used as a supplement in the maturation medium of bovine oocytes in vitro. However, serum contains many undefined factors and is potentially infectious to humans and animals. As a serum replacement, we evaluated the feasibility of using the silk protein, sericin, derived from the cocoons of silkworm. To examine the rates of oocyte maturation and fertilization, cumulus-oocyte complexes were cultured in TCM-199 supplemented with 0.01%, 0.05%, 0.1% or 0.15% sericin or 5% FBS. The sizes of the perivitelline space that might relate to polyspermy, the expressions of Has2 and CD44 mRNA, the amount of hyaluronan (hyaluronic acid: HA) contained in the oocytes and the rates of blastocyst formation following insemination were then compared between the oocytes cultured with 0.05% sericin and 5% FBS, because the polyspermy rates in oocytes cultured with 0.05% sericin were significantly lower than in those cultured with 5% FBS. After in vitro maturation (IVM), the mean size of the perivitelline space was significantly greater in oocytes cultured with sericin than in those cultured with FBS, although the rates of nuclear maturation, fertilization and blastocyst formation of oocytes under both IVM conditions were not significantly different. The expression of HAS2 and CD44 mRNA and the amount of HA in the denuded oocytes cultured with 0.05% sericin were significantly greater than in those cultured with FBS. These results indicate the feasibility of sericin as an alternative protein supplement for IVM in bovine oocytes. PMID:24748396

Hosoe, Misa; Yoshida, Nao; Hashiyada, Yutaka; Teramoto, Hidetoshi; Takahashi, Toru; Niimura, Sueo

2014-01-01

84

Sericin Accelerates the Production of Hyaluronan and Decreases the Incidence of Polyspermy Fertilization in Bovine Oocytes During In Vitro Maturation  

PubMed Central

Fetal bovine serum (FBS) has been widely used as a supplement in the maturation medium of bovine oocytes in vitro. However, serum contains many undefined factors and is potentially infectious to humans and animals. As a serum replacement, we evaluated the feasibility of using the silk protein, sericin, derived from the cocoons of silkworm. To examine the rates of oocyte maturation and fertilization, cumulus-oocyte complexes were cultured in TCM-199 supplemented with 0.01%, 0.05%, 0.1% or 0.15% sericin or 5% FBS. The sizes of the perivitelline space that might relate to polyspermy, the expressions of Has2 and CD44 mRNA, the amount of hyaluronan (hyaluronic acid: HA) contained in the oocytes and the rates of blastocyst formation following insemination were then compared between the oocytes cultured with 0.05% sericin and 5% FBS, because the polyspermy rates in oocytes cultured with 0.05% sericin were significantly lower than in those cultured with 5% FBS. After in vitro maturation (IVM), the mean size of the perivitelline space was significantly greater in oocytes cultured with sericin than in those cultured with FBS, although the rates of nuclear maturation, fertilization and blastocyst formation of oocytes under both IVM conditions were not significantly different. The expression of HAS2 and CD44 mRNA and the amount of HA in the denuded oocytes cultured with 0.05% sericin were significantly greater than in those cultured with FBS. These results indicate the feasibility of sericin as an alternative protein supplement for IVM in bovine oocytes. PMID:24748396

HOSOE, Misa; YOSHIDA, Nao; HASHIYADA, Yutaka; TERAMOTO, Hidetoshi; TAKAHASHI, Toru; NIIMURA, Sueo

2014-01-01

85

Folding behavior of four silks of giant honey bee reflects the evolutionary conservation of aculeate silk proteins.  

PubMed

Multiple gene duplication events in the precursor of the Aculeata (bees, ants, hornets) gave rise to four silk genes. Whilst these homologs encode proteins with similar amino acid composition and coiled coil structure, the retention of all four homologs implies they each are important. In this study we identified, produced and characterized the four silk proteins from Apis dorsata, the giant Asian honeybee. The proteins were readily purified, allowing us to investigate the folding behavior of solutions of individual proteins in comparison to mixtures of all four proteins at concentrations where they assemble into their native coiled coil structure. In contrast to solutions of any one protein type, solutions of a mixture of the four proteins formed coiled coils that were stable against dilution and detergent denaturation. The results are consistent with the formation of a heteromeric coiled coil protein complex. The mechanism of silk protein coiled coil formation and evolution is discussed in light of these results. PMID:25712559

Maitip, Jakkrawut; Trueman, Holly E; Kaehler, Benjamin D; Huttley, Gavin A; Chantawannakul, Panuwan; Sutherland, Tara D

2015-04-01

86

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 for the first time to determine quantitatively the orientation of the -sheets in silk monofilaments from Bombyx, for systems with uniaxial symmetry such as silk, it is possible to determine the order parameters P2 and P4

Pezolet, Michel

87

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

88

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

NASA Astrophysics Data System (ADS)

Here we present a set of measurements using Differential Scanning Fluorimetry (DSF) as an inexpensive, high throughput screening method to investigate the folding of silk protein molecules as they abandon their first native melt conformation, dehydrate and denature into their final solid filament conformation. Our first data and analyses comparing silks from spiders, mulberry and wild silkworms as well as reconstituted `silk' fibroin show that DSF can provide valuable insights into details of silk denaturation processes that might be active during spinning. We conclude that this technique and technology offers a powerful and novel tool to analyse silk protein transitions in detail by allowing many changes to the silk solutions to be tested rapidly with microliter scale sample sizes. Such transition mechanisms will lead to important generic insights into the folding patterns not only of silks but also of other fibrous protein (bio)polymers.

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

2014-07-01

89

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

PubMed Central

Here we present a set of measurements using Differential Scanning Fluorimetry (DSF) as an inexpensive, high throughput screening method to investigate the folding of silk protein molecules as they abandon their first native melt conformation, dehydrate and denature into their final solid filament conformation. Our first data and analyses comparing silks from spiders, mulberry and wild silkworms as well as reconstituted ‘silk' fibroin show that DSF can provide valuable insights into details of silk denaturation processes that might be active during spinning. We conclude that this technique and technology offers a powerful and novel tool to analyse silk protein transitions in detail by allowing many changes to the silk solutions to be tested rapidly with microliter scale sample sizes. Such transition mechanisms will lead to important generic insights into the folding patterns not only of silks but also of other fibrous protein (bio)polymers. PMID:25004800

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

2014-01-01

90

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

91

Stability of Silk and Collagen Protein Materials in Space  

NASA Astrophysics Data System (ADS)

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.

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

2013-12-01

92

Mechanical performance of spider silk is robust to nutrient-mediated changes in protein composition.  

PubMed

Spider major ampullate (MA) silk is sought after as a biomimetic because of its high strength and extensibility. While the secondary structures of MA silk proteins (spidroins) influences silk mechanics, structural variations induced by spinning processes have additional effects. Silk properties may be induced by spiders feeding on diets that vary in certain nutrients, thus providing researchers an opportunity to assess the interplay between spidroin chemistry and spinning processes on the performance of MA silk. Here, we determined the relative influence of spidroin expression and spinning processes on MA silk mechanics when Nephila pilipes were fed solutions with or without protein. We found that spidroin expression differed across treatments but that its influence on mechanics was minimal. Mechanical tests of supercontracted fibers and X-ray diffraction analyses revealed that increased alignment in the amorphous region and to a lesser extent in the crystalline region led to increased fiber strength and extensibility in spiders on protein rich diets. PMID:25764227

Blamires, Sean J; Liao, Chen-Pan; Chang, Chung-Kai; Chuang, Yu-Chun; Wu, Chung-Lin; Blackledge, Todd A; Sheu, Hwo-Shuenn; Tso, I-Min

2015-04-13

93

Bombyx mori silk protein films microprocessing with a nanosecond ultraviolet laser and a femtosecond laser workstation: theory and experiments  

NASA Astrophysics Data System (ADS)

Laser microprocessing of several biopolymers from renewable resources is studied. Three proteinic materials were either extracted from the extracellular matrix like Silk Fibroin/Sericin and collagen, or coming from a commercial source like gelatin. All can find future applications in biomedical experimentation, in particular for cell scaffolding. Films of ˜hundred of microns thick were made by aqueous solution drying and laser irradiation. Attention is paid to the properties making them processable with two laser sources: the ultraviolet and nanosecond (ns) KrF (248 nm) excimer and the infrared and femtosecond (fs) Yb:KGW laser. The UV radiation is absorbed in a one-photon resonant process to yield ablation and the surface foaming characteristics of a laser-induced pressure wave. To the contrary, resonant absorption of the IR photons of the fs laser is not possible and does not take place. However, the high field of the intense I>˜1012 W/cm2 femtosecond laser pulse ionizes the film by the multiphoton absorption followed by the electron impact mechanism, yielding a dense plasma capable to further absorb the incident radiation of the end of the pulse. The theoretical model of this absorption is described in detail, and used to discuss the presented experimental effects (cutting, ablation and foaming) of the fs laser. The ultraviolet laser was used to perform simultaneous multiple spots experiments in which energetic foaming yields melt ejection and filament spinning. Airborne nanosize filaments "horizontally suspended by both ends" (0.25 ?m diameter and 10 ?m length) of silk biopolymer were observed upon irradiation with large fluences.

Lazare, S.; Sionkowska, A.; Zaborowicz, M.; Planecka, A.; Lopez, J.; Dijoux, M.; Louména, C.; Hernandez, M.-C.

2012-01-01

94

Electroresponsive Aqueous Silk Protein As “Smart” Mechanical Damping Fluid  

PubMed Central

Here we demonstrate the effectiveness of an electroresponsive aqueous silk protein polymer as a smart mechanical damping fluid. The aqueous polymer solution is liquid under ambient conditions, but is reversibly converted into a gel once subjected to an electric current, thereby increasing or decreasing in viscosity. This nontoxic, biodegradable, reversible, edible fluid also bonds to device surfaces and is demonstrated to reduce friction and provide striking wear protection. The friction and mechanical damping coefficients are shown to modulate with electric field exposure time and/or intensity. Damping coefficient can be modulated electrically, and then preserved without continued power for longer time scales than conventional “smart” fluid dampers. PMID:24750065

2015-01-01

95

Biological responses to spider silk-antibiotic fusion protein  

PubMed Central

The development of a new generation of multifunctional biomaterials is a continual goal for the field of materials science. The in vivo functional behaviour of a new fusion protein that combines the mechanical properties of spider silk with the antimicrobial properties of hepcidin was addressed in this study. This new chimeric protein, termed 6mer+hepcidin, fuses spider dragline consensus sequences (6mer) and the antimicrobial peptide hepcidin as we have recently described, with retention of bactericidal activity and low cytotoxicity. In the present study mice subcutaneous implants were studied to access the in vivo biological response to the 6mer+hepcidin, which were compared with controls of silk alone (6mer), poly-lactic-glycolic-acid (PLGA) films and empty defects. Along with visual observations, flow cytometry and histology analyses were used to determine the number and type of inflammatory cells at the implantation site. The results show a mild to low inflammatory reaction to the implanted materials and no apparent differences between the 6mer+hepcidin films and the other experimental controls, demonstrating that the new fusion protein has good in vivo biocompatibility, while maintaining antibiotic function. PMID:22514077

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

2011-01-01

96

Hierarchical structures made of proteins. The complex architecture of spider webs and their constituent silk proteins.  

PubMed

Biopolymers fulfil a variety of different functions in nature. They conduct various processes inside and outside cells and organisms, with a functionality ranging from storage of information to stabilization, protection, shaping, transport, cellular division, or movement of whole organisms. Within the plethora of biopolymers, the most sophisticated group is of proteinaceous origin: the cytoskeleton of a cell is made of protein filaments that aid in pivotal processes like intracellular transport, movement, and cell division; geckos use a distinct arrangement of keratin-like filaments on their toes which enable them to walk up smooth surfaces, such as walls, and even upside down across ceilings; and spiders spin silks that are extra-corporally used for protection of offspring and construction of complex prey traps. The following tutorial review describes the hierarchical organization of protein fibers, using spider dragline silk as an example. The properties of a dragline silk thread originate from the strictly controlled assembly of the underlying protein chains. The assembly procedure leads to protein fibers showing a complex hierarchical organization comprising three different structural phases. This structural organization is responsible for the outstanding mechanical properties of individual fibers, which out-compete even those of high-performance artificial fibers like Kevlar. Web-weaving spiders produce, in addition to dragline silk, other silks with distinct properties, based on slightly variant constituent proteins--a feature that allows construction of highly sophisticated spider webs with well designed architectures and with optimal mechanical properties for catching prey. PMID:20023846

Heim, Markus; Römer, Lin; Scheibel, Thomas

2010-01-01

97

Silk: A Potential Medium for Tissue Engineering  

PubMed Central

Objective: Human skin is a complex bilayered organ that serves as a protective barrier against the environment. The loss of integrity of skin by traumatic experiences such as burns and ulcers may result in considerable disability or ultimately death. Therefore, in skin injuries, adequate dermal substitutes are among primary care targets, aimed at replacing the structural and functional properties of native skin. To date, there are very few single application tissue-engineered dermal constructs fulfilling this criterion. Silk produced by the domestic silkworm, Bombyx mori, has a long history of use in medicine. It has recently been increasingly investigated as a promising biomaterial for dermal constructs. Silk contains 2 fibrous proteins, sericin and fibroin. Each one exhibits unique mechanical and biological properties. Methods: Comprehensive review of randomized-controlled trials investigating current dermal constructs and the structures and properties of silk-based constructs on wound healing. Results: This review revealed that silk-fibroin is regarded as the most promising biomaterial, providing options for the construction of tissue-engineered skin. Conclusion: The research available indicates that silk fibroin is a suitable biomaterial scaffold for the provision of adequate dermal constructs. PMID:18997857

Sobajo, Cassandra; Behzad, Farhad; Yuan, Xue-Feng; Bayat, Ardeshir

2008-01-01

98

Enhancing analysis of cells and proteins by fluorescence imaging on silk-based biomaterials: modulating the autofluorescence of silk.  

PubMed

Silk is a versatile and established biomaterial for various tissue engineering purposes. However, it also exhibits strong autofluorescence signals-thereby hindering fluorescence imaging analysis of cells and proteins on silk-derived biomaterials. Sudan Black B (SB) is a lysochrome dye commonly used to stain lipids in histology. It has also been reported to be able to quench autofluorescence of tissues in histology and has been tested on artificial biomedical polymers in recent years. It was hypothesized that SB would exert similar quenching effects on silk, modulating the autofluorescence signals, and thereby enabling improved imaging analysis of cells and molecules of interests. The quenching effect of SB on the intrinsic fluorescence properties of silk and on commercial fluorescent dyes were first investigated in this study. SB was then incorporated into typical fluorescence-based staining protocols to study its effectiveness in improving fluorescence-based imaging of the cells and proteins residing with the silk-based biomaterials. Silk processed into various forms of biomaterials (e.g., films, sponges, fibers, and electrospun mats) was seeded with cells and cultured in vitro. At sacrificial time points, specimens were harvested, fixed, and prepared for fluorescence staining. SB, available commercially as a powder, was dissolved in 70% ethanol (0.3% [w/v]) to form staining solutions. SB treatment was introduced at the last step of typical immunofluorescence staining protocols for 15-120?min. For actin staining protocols by phalloidin toxin, SB staining solutions were added before and after permeabilization with Triton-X for 15-30?min. Results showed that ideal SB treatment duration is about 15?min. Apart from being able to suppress the autofluorescence of silk, this treatment duration was also not too long to adversely affect the fluorescent labeling probes used. The relative improvement brought about by SB treatment was most evident in the blue and green emission wavelengths compared with the red emission wavelength. This study has showed that the use of SB is a cost and time effective approach to enhance fluorescence-based imaging analyses of cell-seeded silk biomaterials, which otherwise would have been hindered by the unmodulated autofluorescence signals. PMID:25050876

Neo, Puay Yong; Tan, Daryl Jian-An; Shi, Pujiang; Toh, Siew Lok; Goh, James Cho-Hong

2015-02-01

99

Sericin protects against diabetes-induced injuries in sciatic nerve and related nerve cells.  

PubMed

Sericin from discarded silkworm cocoons of silk reeling has been used in different fields, such as cosmetology, skin care, nutrition, and oncology. The present study established a rat model of type 2 diabetes by consecutive intraperitoneal injections of low-dose (25 mg/kg) streptozotocin. After intragastrical perfusion of sericin for 35 days, blood glucose levels significantly declined, and the expression of neurofilament protein in the sciatic nerve and nerve growth factor in L4-6 spinal ganglion and anterior horn cells significantly increased. However, the expression of neuropeptide Y in spinal ganglion and anterior horn cells significantly decreased in model rats. These findings indicate that sericin protected the sciatic nerve and related nerve cells against injury in a rat type 2 diabetic model by upregulating the expression of neurofilament protein in the sciatic nerve and nerve growth factor in spinal ganglion and anterior horn cells, and downregulating the expression of neuropeptide Y in spinal ganglion and anterior horn cells. PMID:25206693

Song, Chengjun; Yang, Zhenjun; Zhong, Meirong; Chen, Zhihong

2013-02-25

100

Sericin protects against diabetes-induced injuries in sciatic nerve and related nerve cells?  

PubMed Central

Sericin from discarded silkworm cocoons of silk reeling has been used in different fields, such as cosmetology, skin care, nutrition, and oncology. The present study established a rat model of type 2 diabetes by consecutive intraperitoneal injections of low-dose (25 mg/kg) streptozotocin. After intragastrical perfusion of sericin for 35 days, blood glucose levels significantly declined, and the expression of neurofilament protein in the sciatic nerve and nerve growth factor in L4–6 spinal ganglion and anterior horn cells significantly increased. However, the expression of neuropeptide Y in spinal ganglion and anterior horn cells significantly decreased in model rats. These findings indicate that sericin protected the sciatic nerve and related nerve cells against injury in a rat type 2 diabetic model by upregulating the expression of neurofilament protein in the sciatic nerve and nerve growth factor in spinal ganglion and anterior horn cells, and downregulating the expression of neuropeptide Y in spinal ganglion and anterior horn cells. PMID:25206693

Song, Chengjun; Yang, Zhenjun; Zhong, Meirong; Chen, Zhihong

2013-01-01

101

Processing and modification of films made from recombinant spider silk proteins  

NASA Astrophysics Data System (ADS)

Protein films represent an interesting class of materials with various possibilities for applications. We investigated films made of two different synthetic spider silk proteins derived from the garden spider’s (Araneus diadematus) two dragline silk proteins ADF-3 and ADF-4. Protein films cast from hexafluoroisopropanol solutions displayed a predominantly ?-helical secondary structure. Processing such films with potassium phosphate or methanol resulted in a transition to a ?-sheet rich structure. While as-cast films could be dissolved in water, processed ?-sheet rich films were water insoluble. The chemical stability of processed films depended on the amino acid sequence of the respective protein employed. As a proof of principle, fluorescent probes or enzymes were covalently attached to the film surface. The presented approach provides a basis for designing tailor-made protein films using silk proteins as scaffold, in which the film properties can be controlled by genetic engineering of the underlying silks.

Huemmerich, D.; Slotta, U.; Scheibel, T.

2006-02-01

102

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

103

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

104

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

PubMed

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

105

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

106

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

107

Review the role of terminal domains during storage and assembly of spider silk proteins.  

PubMed

Fibrous proteins in nature fulfill a wide variety of functions in different structures ranging from cellular scaffolds to very resilient structures like tendons and even extra-corporal fibers such as silks in spider webs or silkworm cocoons. Despite their different origins and sequence varieties many of these fibrous proteins share a common building principle: they consist of a large repetitive core domain flanked by relatively small non-repetitive terminal domains. Amongst protein fibers, spider dragline silk shows prominent mechanical properties that exceed those of man-made fibers like Kevlar. Spider silk fibers assemble in a spinning process allowing the transformation from an aqueous solution into a solid fiber within milliseconds. Here, we highlight the role of the non-repetitive terminal domains of spider dragline silk proteins during storage in the gland and initiation of the fiber assembly process. PMID:22057429

Eisoldt, Lukas; Thamm, Christopher; Scheibel, Thomas

2012-06-01

108

Optically Transparent Recombinant Silk-Elastinlike Protein Polymer Films  

PubMed Central

Recombinant protein polymers, evaluated extensively as biomaterials for applications in drug delivery and tissue engineering, are rarely reported as being optically transparent. Here we report the notable optical transparency of films composed of a genetically engineered silk-elastinlike protein polymer SELP-47K. SELP-47K films of 100 ?m in thickness display a transmittance of 93% in the wavelength range of 350–800 nm. While covalent cross-linking of SELP-47K via glutaraldehyde decreases its transmittance to 77% at the wavelength of 800 nm, noncovalent cross-linking using methanol slightly increases it to 95%. Non- and covalent cross-linking of SELP-47K films also influences their secondary structures and water contents. Cell viability and proliferation analyses further reveal the excellent cytocompatibility of both non- and covalently cross-linked SELP-47K films. The combination of high optical transparency and cytocompatibility of SELP-47K films, together with their previously reported outstanding mechanical properties, suggests that this protein polymer may be useful in unique, new biomedical applications. PMID:21288001

Teng, Weibing; Huang, Yiding; Cappello, Joseph; Wu, Xiaoyi

2011-01-01

109

Processing of recombinant spider silk proteins into tailor-made materials for biomaterials applications.  

PubMed

Spider silk has extraordinary mechanical properties, is biocompatible and biodegradable, and therefore an ideal material for biomedical applications. However, a drawback for any application is the inhomogeneity of spider silk, as seen for other natural materials, as well as the low availability due to the cannibalism of most spiders. Recently, developed recombinant spider silk proteins ensure constant material properties, as well as scalable production, and further the processing into morphologies other than fibres. Biotechnology enables genetic modification, broadening the range of applications, such as implant coatings, scaffolds for tissue engineering, wound dressing devices as well as drug delivery systems. PMID:24657706

Schacht, Kristin; Scheibel, Thomas

2014-10-01

110

Brown widow (Latrodectus geometricus) major ampullate silk protein and its material properties.  

PubMed

Major ampullate (dragline) silk is the main web component as well as the silk that spiders use for a lifeline when they fall. This silk has a breaking stress of 4.6 GPa, which is similar to that of Kevlar. The majority of the previous mechanical testing studies involved the major ampullate silk from orb-weaving spiders. To date, there have been no reports on dragline silk mechanical properties from a cob-weaver, brown widow Latrodectus geometricus. L. geometricus dragline was found to be composed of MaSp1, MaSp2, and MaSp-like proteins all of which have highly conserved amino acid motifs, especially the GGX, GA and poly A for MaSp1 and GPGGX and poly A for MaSp2. These sequences are the same as those found in the silks of orb-weaving spiders. To determine if protein sequences influence the material properties of the silk, mechanical testing was performed on single strands of silk fibers from adult female L. geometricus spiders. The 3 cm long silk fibers were tested for breaking stress and strain with a MTS Synergie 100 mechanical testing system using a 50 g load cell with the cross-head speed set at 10 mm/min. The breaking stress and strain were measured for 20 replicate samples and averaged. The values of 0.83 +/- 0.19 GPa for stress and 0.14 +/- 0.06 for strain shows that brown widow dragline is weaker than the orb-weaving spiders. PMID:15133936

Motriuk-Smith, Dagmara; Lewis, Randolph V

2004-01-01

111

An improvement in the attaching capability of cryopreserved human hepatocytes by a proteinaceous high molecule, sericin, in the serum-free solution.  

PubMed

The methodology of cryopreservation of human hepatocytes remains unsatisfactory. Even when the viability of thawed cells is tolerable, the cells often lose the attaching capability to a culture dish, resulting in the cells' inability to survive. Previously, we described the effectiveness of maltose on the attachment of hepatocytes. This article demonstrates that a silk-derived high molecular protein, sericin, improves the cell-attaching capability in the serum-free freezing medium. When human hepatocytes [initial viability: 60.9 ± 3.1% (mean ± SD, n = 3)] were frozen with serum-free Dulbecco's modified Eagle medium (DMEM) containing 10% dimethyl sulfoxide (DMSO), the viability was 29.4 ± 3.2% and the cell-attaching capability 20.4 ± 4.1%. On the other hand, DMEM containing 10% DMSO and 1% sericin increased the values to 45.0 ± 0.8% and 26.2 ± 3.2%. Moreover, the addition of 0.1 mol/L maltose to the sericin-containing medium improved to 42.2 ± 3.2% and 51.1 ± 1.0%, as we demonstrated in a previous report. The present results indicated that sericin combined with maltose is a novel additive in the serum-free freezing medium for human hepatocytes. PMID:20525438

Miyamoto, Yoshitaka; Teramoto, Naozumi; Hayashi, Shuji; Enosawa, Shin

2010-01-01

112

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

113

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

114

Bio-inspired Silicification of Silica-binding Peptide-Silk Protein Chimeras: Comparison of Chemically and Genetically Produced Proteins  

PubMed Central

Novel protein chimeras constituted of ‘silk’ and a silica-binding peptide (KSLSRHDHIHHH) were synthesized by genetic or chemical approaches and their influence on silica-silk based chimera composite formation evaluated. Genetic chimeras were constructed from 6 or 15 repeats of the 32 amino acid consensus sequence of Nephila clavipes spider silk ([SGRGGLGGQG AGAAAAAGGA GQGGYGGLGSQG]n) to which one silica binding peptide was fused at the N terminus. For the chemical chimera, 25 equivalents of the silica binding peptide were chemically coupled to natural Bombyx mori silk after modification of tyrosine groups by diazonium coupling and EDC/NHS activation of all acid groups. After silica formation under mild, biomaterial compatible conditions the effect of peptide addition on the properties of the silk and chimeric silk-silica composite materials was explored. The composite biomaterial properties could be related to the extent of silica condensation and to the higher number of silica binding sites in the chemical chimera as compared to the genetically derived variants. In all cases, the structure of the protein / chimera in solution dictated the type of composite structure that formed with the silica deposition process having little effect on the secondary structural composition of the silk based materials. Similarly to our study of genetic silk based chimeras containing the R5 peptide (SSKKSGSYSGSKGSKRRIL), the role of the chimeras (genetic and chemical) used in the present study resided more in aggregation and scaffolding than in the catalysis of condensation. The variables of peptide identity, silk construct (number of consensus repeats or silk source) and approach to synthesis (genetic or chemical) can be used to ‘tune’ the properties of the composite materials formed and is a general approach which can be used to prepare a range of materials for biomedical and sensor based applications. PMID:22229696

Canabady-Rochelle, Laetitia L.S.; Belton, David J.; Deschaume, Olivier; Currie, Heather A.; Kaplan, David L.; Perry, Carole C.

2012-01-01

115

Control of silicification by genetically engineered fusion proteins: Silk-silica binding peptides.  

PubMed

In the present study, an artificial spider silk gene, 6mer, derived from the consensus sequence of Nephila clavipes dragline silk gene, was fused with different silica-binding peptides (SiBPs), A1, A3 and R5, to study the impact of the fusion protein sequence chemistry on silica formation and the ability to generate a silk-silica composite in two different bioinspired silicification systems: solution-solution and solution-solid. Condensed silica nanoscale particles (600-800nm) were formed in the presence of the recombinant silk and chimeras, which were smaller than those formed by 15mer-SiBP chimeras [1], revealing that the molecular weight of the silk domain correlated to the sizes of the condensed silica particles in the solution system. In addition, the chimeras (6mer-A1/A3/R5) produced smaller condensed silica particles than the control (6mer), revealing that the silica particle size formed in the solution system is controlled by the size of protein assemblies in solution. In the solution-solid interface system, silicification reactions were performed on the surface of films fabricated from the recombinant silk proteins and chimeras and then treated to induce ?-sheet formation. A higher density of condensed silica formed on the films containing the lowest ?-sheet content while the films with the highest ?-sheet content precipitated the lowest density of silica, revealing an inverse correlation between the ?-sheet secondary structure and the silica content formed on the films. Intriguingly, the 6mer-A3 showed the highest rate of silica condensation but the lowest density of silica deposition on the films, compared with 6mer-A1 and -R5, revealing antagonistic crosstalk between the silk and the SiBP domains in terms of protein assembly. These findings offer a path forward in the tailoring of biopolymer-silica composites for biomaterial related needs. PMID:25462851

Zhou, Shun; Huang, Wenwen; Belton, David J; Simmons, Leo O; Perry, Carole C; Wang, Xiaoqin; Kaplan, David L

2015-03-01

116

Transgenic protein production in silkworm silk glands requires cathepsin and chitinase of Autographa californica multicapsid nucleopolyhedrovirus.  

PubMed

The silkworm Bombyx mori represents an established in vivo system for the production of recombinant proteins. Baculoviruses have been extensively investigated and optimised for the expression of high protein levels inside the haemolymph of larvae and pupae of this lepidopteran insect. Current technology includes deletion of genes responsible for the activity of virus-borne proteases, which in wild-type viruses, cause liquefaction of the host insect and enhance horizontal transmission of newly synthesised virus particles. Besides the haemolymph, the silk gland of B. mori provides an additional expression system for recombinant proteins. In this paper, we investigated how silk gland can be efficiently infected by a Autographa californica multicapsid nuclear polyhedrosis virus (AcMNPV). We demonstrated that the viral chitinase and the cysteine protease cathepsin are necessary to permit viral entry into the silk gland cells of intrahaemocoelically infected B. mori larvae. Moreover, for the first time, we showed AcMNPV crossing the basal lamina of silk glands in B. mori larvae, and we assessed a new path of infection of silk gland cells that can be exploited for protein production. PMID:24477386

Wöltje, Michael; Böbel, Melanie; Rheinnecker, Michael; Tettamanti, Gianluca; Franzetti, Eleonora; Saviane, Alessio; Cappellozza, Silvia

2014-05-01

117

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

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

2009-01-01

118

Rheology of reconstituted silk fibroin protein gels: the epitome of extreme mechanics.  

PubMed

In nature, silk fibroin proteins assemble into hierarchical structures with dramatic mechanical properties. With the hope of creating new classes of on demand silk-based biomaterials, Bombyx mori silk is reconstituted back into stable aqueous solutions that can be reassembled into functionalized materials; one strategy for reassembly is electrogelation. Electrogels (e-gels) are particularly versatile and can be produced using electrolysis with small DC electric fields. We characterize the linear and nonlinear rheological behavior of e-gels to provide fundamental insights into these distinct protein-based materials. We observe that e-gels form robust biopolymer networks that exhibit distinctive strain hardening and are recoverable from strains as large as ?=27, i.e. 2700%. We propose a simple microscopic model that is consistent with local restructuring of single proteins within the e-gel network. PMID:25489795

Tabatabai, A Pasha; Kaplan, David L; Blair, Daniel L

2015-01-28

119

Anti-fungal activity of maize silk proteins and role of chitinases in Aspergillus flavus resistance  

Technology Transfer Automated Retrieval System (TEKTRAN)

Proteins were extracted from silks of two Aspergillus flavus resistant maize (Zea mays L.) inbreds, two susceptible inbreds, and one intermediately-resistant inbred grown in the field. Two-dimensional gel electrophoresis was used to identify and compare expression patterns of the proteins in the m...

120

Charge-Tunable Silk-Tropoelastin Protein Alloys That Control Neuron Cell Responses  

PubMed Central

Tunable protein composites are important for constructing extracellular matrix mimics of human tissues with control of biochemical, structural, and mechanical properties. Molecular interaction mechanisms between silk fibroin protein and recombinant human tropoelastin, based on charge, are utilized to generate a new group of multifunctional protein alloys (mixtures of silk and tropoelastin) with different net charges. These new biomaterials are then utilized as a biomaterial platform to control neuron cell response. With a +38 net charge in water, tropoelastin molecules provide extraordinary elasticity and selective interactions with cell surface integrins. In contrast, negatively charged silk fibroin protein (net charge ?36) provides remarkable toughness and stiffness with morphologic stability in material formats via autoclaving-induced beta-sheet crystal physical crosslinks. The combination of these properties in alloy format extends the versatility of both structural proteins, providing a new biomaterial platform. The alloys with weak positive charges (silk/tropoelastin mass ratio 75/25, net charge around +16) significantly improved the formation of neuronal networks and maintained cell viability of rat cortical neurons after 10 days in vitro. The data point to these protein alloys as an alternative to commonly used poly-L-lysine (PLL) coatings or other charged synthetic polymers, particularly with regard to the versatility of material formats (e.g., gels, sponges, films, fibers). The results also provide a practical example of physically designed protein materials with control of net charge to direct biological outcomes, in this case for neuronal tissue engineering. PMID:25093018

Hu, Xiao; Tang-Schomer, Min D.; Huang, Wenwen; Xia, Xiao-Xia; Weiss, Anthony S.

2014-01-01

121

Properties and antityrosinase activity of sericin from various extraction methods.  

PubMed

The present study investigated the chemical properties and antityrosinase activities of SS (silk sericin) extracted from different Thai silk strains via various extraction methods. Different silk strains contain distinct SS with various amino acid compositions, which are significantly influenced by the extraction method used. Urea extraction of SS was the only method that provided clearly distinguishable bands and had the most significant impact on SS conformation as illustrated by FTIR (Fourier-transform infrared) spectra. The use of urea or either acidic or alkaline chemicals in the extraction process also influenced SS thermal behaviour. With regard to biological activity, SS extracted using urea exhibited the highest antityrosinase activity, whereas alkali-degraded SS showed no inhibition of mushroom tyrosinase. Pigments, primarily flavonoids and carotenoids from silk cocoons, were also found to enhance tyrosinase inhibition of SS. PMID:20055756

Aramwit, Pornanong; Damrongsakkul, Siriporn; Kanokpanont, Sorada; Srichana, Teerapol

2010-02-01

122

Sericin for resistance switching device with multilevel nonvolatile memory.  

PubMed

Resistance switching characteristics of natural sericin protein film is demonstrated for nonvolatile memory application for the first time. Excellent memory characteristics with a resistance OFF/ON ratio larger than 10(6) have been obtained and a multilevel memory based on sericin has been achieved. The environmentally friendly high performance biomaterial based memory devices may hold a place in the future of electronic device development. PMID:23893500

Wang, Hong; Meng, Fanben; Cai, Yurong; Zheng, Liyan; Li, Yuangang; Liu, Yuanjun; Jiang, Yueyue; Wang, Xiaotian; Chen, Xiaodong

2013-10-11

123

Snmp1, a Novel Membrane Protein of Olfactory Neurons of the Silk Moth Antheraea polyphemus with Homology to the CD36 Family of Membrane Proteins  

Microsoft Academic Search

While olfactory neurons of silk moths are well known for their exquisite sensitivity to sex pheromone odor- ants, 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 mem- branes of the wild silk moth Antheraea polyphemus .W e have purified and cloned a

Matthew E. Rogers; Ming Sun; Michael R. Lerner; Richard G. Vogt

1997-01-01

124

Protein unfolding versus ?-sheet separation in spider silk nanocrystals  

NASA Astrophysics Data System (ADS)

In this communication a mechanism for spider silk strain hardening is proposed. Shear failure of ?-sheet nanocrystals is the first failure mode that gives rise to the creation of smaller nanocrystals, which are of higher strength and stiffness. ?-sheet unfolding requires more energy than nanocrystal separation in a shear mode of failure. As a result, unfolding occurs after the nanocrystals separate in shear. ?-sheet unfolding yields a secondary strain hardening effect once the ?-sheet conformation is geometrically stable and acts like a unidirectional fibre in a fibre reinforced composite. The mechanism suggested herein is based on molecular dynamics calculations of residual inter-?-sheet separation strengths against residual intra-?-sheet unfolding strengths.

Alam, Parvez

2014-03-01

125

Freeze-gelled silk fibroin protein scaffolds for potential applications in soft tissue engineering.  

PubMed

Recently tissue engineering has escalated much interest in biomedical and biotechnological applications. In this regard, exploration of new and suitable biomaterials is needed. Silk fibroin protein is used as one of the most preferable biomaterials for fabrication of scaffolds and several new techniques are being adopted to fabricate silk scaffolds with greater ease, efficiency and perfection. In this study, a freeze gelation technique is used for fabrication of silk fibroin protein 3D scaffolds, which is both time and energy efficient as compared to the conventional freeze drying technique. The fabricated silk fibroin freeze-gelled scaffolds are evaluated micro structurally for morphology with scanning electron microscopy which reveals relatively homogeneous pore structure and good interconnectivity. The pore sizes and porosity of these scaffolds ranges between 60-110?m and 90-95%, respectively. Mechanical test shows that the compressive strength of the scaffolds is in the range of 20-40kPa. The applicability to cell culture of the freeze gelled scaffolds has been examined with human keratinocytes HaCat cells which show the good cell viability and proliferation of cells after 5 days of culture suggesting the cytocompatibility. The freeze-gelled 3D scaffolds show comparable results with the conventionally prepared freeze dried 3D scaffolds. Thus, this technique may be used as an alternative method for 3D scaffolds preparation and may also be utilized for tissue engineering applications. PMID:21557966

Bhardwaj, Nandana; Chakraborty, Sagar; Kundu, Subhas C

2011-10-01

126

Silk and collagen scaffolds for tendon reconstruction.  

PubMed

In this study, silk thread (Bombyx mori) was braided to a tube-like shape and sericin was removed from the silk tube. Thereafter, collagen/chondroitin-6-sulfate solution was poured into the silk tube, and the lyophilization process was performed. To assess the inflammatory response in vivo, raw silk and sericin-free silk tubes were implanted in the subcutaneous layer of mice. After 10 days of in vivo implantation, mild inflammatory responses were observed around the sericin-free silk tubes, and severe inflammation with the presence of neutrophils and macrophages was observed around the raw silk tubes. At 24 weeks post implantation, the regenerated tendon had a thick, cylindrical, grayish fibrous structure and a shiny white appearance, similar to that of the native tendon in the rabbit model of tendon defect. The average tensile strength of the native tendons was 220 ± 20 N, whereas the average tensile strength of the regenerated tendons was 167 ± 30 N and the diameter of the regenerated tendon (3 ± 0.2 mm) was similar to that of the native tendons (4 ± 0.3 mm). Histologically, the regenerated tendon resembled the native tendon, and all the regenerated tissues showed organized bundles of crimped fibers. Masson trichrome staining was performed for detecting collagen synthesis, and it showed that the artificial tendon was replaced by new collagen fibers and extracellular matrix. However, the regenerated tendon showed fibrosis to a certain degree. In conclusion, the artificial tendon, comprising a braided silk tube and lyophilized collagen sponge, was optimal for tendon reconstruction. Thus, this study showed an improved regeneration of neo-tendon tissues, which have the structure and tensile strength of the native tendon, with the use of the combination of collagen and silk scaffold. PMID:24705339

Kwon, Soon-Yong; Chung, Jin-Wha; Park, Hee-Jung; Jiang, Yuan-Yuan; Park, Jung-Keug; Seo, Young-Kwon

2014-04-01

127

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

128

Involvement of trypsin-digested silk peptides in the induction of RAW264.7 macrophage activation.  

PubMed

The activation of macrophages by trypsin-digested silk peptides was investigated by considering CD1 lb and CD40 expression in the RAW264.7 cell, a murine macrophage. Silk protein hydrolysates were digested with trypsin, following by centrifugal purification using the Centriprep 30k concentrator. Trypsin-digested total silk peptides and its centrifugal fractions were tested for macrophage activation in vitro. The functional peptide of fractionated silk peptides was examined by LC/MS/MS analysis. Trypsin-digested and fractionated silk peptides of more than 30 kDa induced an increase in the activation markers CD1 lb and CD40 in RAW264.7 cells. These results are supported by morphological changes reflecting an increase in the number of dendrites in activated cells. The fractionated silk peptides examined by LC/MS/MS contained partial peptides of Bombyx mori fibroin. These results suggest that the activation of RAW264.7 macrophages may be induced not by sericin-derived peptides but by fibroin-derived ones. PMID:24555292

Pyo, Kyoung-Ho; Kim, Min-Ki; Shin, Kwang-Soon; Chun, Hyang Sook; Shin, Eun-Hee

2013-12-01

129

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

130

Novel genes differentially expressed between posterior and median silk gland identified by SAGE-aided transcriptome analysis.  

PubMed

Serial analysis of gene expression (SAGE) profiles, from posterior and median cells of the silk gland of Bombyx mori, were analyzed and compared, so as to identify their respective distinguishing functions. The annotation of the SAGE libraries was performed with a B. mori reference tag collection, which was extracted from a novel set of Bombyx ESTs, sequenced from the 3' side. Most of the tags appeared at similar relative concentration within the two libraries, and corresponded with region-specific and highly abundant silk proteins. Strikingly, in addition to tags from silk protein mRNAs, 19 abundant tags were found (? 0.1%), in the median cell library, which were absent in the posterior cell tag collection. With the exception of tags from SP1 mRNA, no PSG specific tags were found in this subset class. The analysis of some of the MSG-specific transcripts, suggested that middle silk gland cells have diversified functions, in addition to their well characterized role in silk sericins synthesis and secretion. PMID:21078388

Royer, Corinne; Briolay, Jérôme; Garel, Annie; Brouilly, Patrick; Sasanuma, Shun-ichi; Sasanuma, Motoe; Shimomura, Michihiko; Keime, Céline; Gandrillon, Olivier; Huang, Yongping; Chavancy, Gérard; Mita, Kazuei; Couble, Pierre

2011-02-01

131

Comparative architecture of silks, fibrous proteins and their encoding genes in insects and spiders  

Microsoft Academic Search

The known silk fibroins and fibrous glues are thought to be encoded by members of the same gene family. All silk fibroins sequenced to date contain regions of long-range order (crystalline regions) and\\/or short-range order (non-crystalline regions). All of the sequenced fibroin silks (Flag or silk from flagelliform gland in spiders; Fhc or heavy chain fibroin silks produced by Lepidoptera

Catherine L. Craig; Christian Riekel

2002-01-01

132

Potential mode of protection of silkworm pupae from environmental stress by harboring the bacterial biofilm on the surfaces of silk cocoons.  

PubMed

The silkworm forms cocoon to protect its pupa that survives for months inside the cocoon without being affected by various environmental stresses. To understand the possible mode of pupal survival within the cocoon encasement, we investigate the cause that protects the cocoon. During the end of the spinning process, we have isolated different bacterial species from the cocoon surface. These are identified using molecular techniques and checked for their abilities to form biofilm in vitro. The bacteria are able to form biofilm either individually or in consortia. Of which, Bacillus and Erwinia species are prominent biofilm formers. Interestingly, these bacteria have the ability to form biofilm on the cocoon mimetic surface of the silk protein Sericin Hope that contains only sericin. The origin and the behavior of the bacteria lead us to hypothesize the possible role of biofilm layer on the cocoon surface, which provides protection from adverse environmental conditions. PMID:25292249

Halder, Pranab K; Naskar, Deboki; Kumar, Akash; Yao, Juming; Kundu, Subhas C; Ghosh, Anindya S

2015-02-01

133

Effect of sericin on diabetic hippocampal growth hormone/insulin-like growth factor 1 axis.  

PubMed

Previous studies have shown that sericin extracted from silk cocoon significantly reduces blood glucose levels and protects the nervous system against diabetes mellitus. In this study, a rat type 2 diabetes mellitus model was established by intraperitoneal injection of 25 mg/kg streptozotocin for 3 successive days, following which the rats were treated with sericin for 35 days. After treatment, the blood glucose levels of the diabetic rats decreased significantly, the growth hormone level in serum and its expression in the hippocampus decreased significantly, while the insulin-like growth factor-1 level in serum and insulin-like growth factor-1 and growth hormone receptor expression in the hippocampus increased significantly. The experimental findings indicate that sericin improves disorders of the growth hormone/insulin-like growth factor 1 axis to alleviate hippocampal damage in diabetic rats. PMID:25206472

Chen, Zhihong; Yang, Songhe; He, Yaqiang; Song, Chengjun; Liu, Yongping

2013-07-01

134

Effect of sericin on diabetic hippocampal growth hormone/insulin-like growth factor 1 axis  

PubMed Central

Previous studies have shown that sericin extracted from silk cocoon significantly reduces blood glucose levels and protects the nervous system against diabetes mellitus. In this study, a rat type 2 diabetes mellitus model was established by intraperitoneal injection of 25 mg/kg streptozotocin for 3 successive days, following which the rats were treated with sericin for 35 days. After treatment, the blood glucose levels of the diabetic rats decreased significantly, the growth hormone level in serum and its expression in the hippocampus decreased significantly, while the insulin-like growth factor-1 level in serum and insulin-like growth factor-1 and growth hormone receptor expression in the hippocampus increased significantly. The experimental findings indicate that sericin improves disorders of the growth hormone/insulin-like growth factor 1 axis to alleviate hippocampal damage in diabetic rats. PMID:25206472

Chen, Zhihong; Yang, Songhe; He, Yaqiang; Song, Chengjun; Liu, Yongping

2013-01-01

135

Characterization of Silk Fibroin Modified Surface: A Proteomic View of Cellular Response Proteins Induced by Biomaterials  

PubMed Central

The purpose of this study was to develop the pathway of silk fibroin (SF) biopolymer surface induced cell membrane protein activation. Fibroblasts were used as an experimental model to evaluate the responses of cellular proteins induced by biopolymer material using a mass spectrometry-based profiling system. The surface was covered by multiwalled carbon nanotubes (CNTs) and SF to increase the surface area, enhance the adhesion of biopolymer, and promote the rate of cell proliferation. The amount of adhered fibroblasts on CNTs/SF electrodes of quartz crystal microbalance (QCM) greatly exceeded those on other surfaces. Moreover, analyzing differential protein expressions of adhered fibroblasts on the biopolymer surface by proteomic approaches indicated that CD44 may be a key protein. Through this study, utilization of mass spectrometry-based proteomics in evaluation of cell adhesion on biopolymer was proposed. PMID:24818131

Yang, Ming-Hui; Yuan, Shyng-Shiou; Chung, Tze-Wen; Jong, Shiang-Bin; Lu, Chi-Yu; Tsai, Wan-Chi; Chen, Wen-Cheng; Lin, Po-Chiao; Chiang, Pei-Wen; Tyan, Yu-Chang

2014-01-01

136

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. PMID:24288266

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

2014-11-01

137

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

PubMed

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

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

2015-01-01

138

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

PubMed Central

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

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

2015-01-01

139

E-spun composite fibers of collagen and dragline silk protein: fiber mechanics, biocompatibility, and application in stem cell differentiation.  

PubMed

Biocomposite matrices with high mechanical strength, high stability, and the ability to direct matrix-specific stem cell differentiation are essential for the reconstruction of lesioned tissues in tissue engineering and cell therapeutics. Toward this end, we used the electrospinning technique to fabricate well-aligned composite fibers from collagen and spider dragline silk protein, obtained from the milk of transgenic goats, mimicking the native extracellular matrix (ECM) on a similar scale. Collagen and the dragline silk proteins were found to mix homogeneously at all ratios in the electrospun (E-spun) fibers. As a result, the ultimate tensile strength and elasticity of the fibers increased monotonically with silk percentage, whereas the stretchability was slightly reduced. Strikingly, we found that the incorporation of silk proteins to collagen dramatically increased the matrix stability against excessive fiber swelling and shape deformation in cell culture medium. When human decidua parietalis placental stem cells (hdpPSCs) were seeded on the collagen-silk matrices, the matrices were found to support cell proliferation at a similar rate as that of the pure collagen matrix, but they provided cell adhesion with reduced strengths and induced cell polarization at varied levels. Matrices containing 15 and 30 wt % silk in collagen (CS15, CS30) were found to induce a level of neural differentiation comparable to that of pure collagen. In particular, CS15 matrix induced the highest extent of cell polarization and promoted the development of extended 1D neural filaments strictly in-line with the aligned fibers. Taking the increased mechanical strength and fiber stability into consideration, CS15 and CS30 E-spun fibers offer better alternatives to pure collagen fibers as scaffolds that can be potentially utilized in neural tissue repair and the development of future nanobiodevices. PMID:25405355

Zhu, Bofan; Li, Wen; Lewis, Randolph V; Segre, Carlo U; Wang, Rong

2015-01-12

140

Use of Sericin in Feltproofing of Wool  

Microsoft Academic Search

A new process for feltproofing of wool fibers based on the biopolymer sericin is reported. The susceptibility of wool to sericin has been enhanced by creation of new active sites along the wool keratin molecules, such as cysteic acid and S-cystine sulphonate residues, by pretreatment with hydrogen peroxide and sodium sulphite. The sericin-combining capacity of wool has been even enhanced

O. G. Allam; H. El-Sayed; A. Kantouch; K. Haggag

2009-01-01

141

Self-assembly of silk-elastinlike protein polymers into three-dimensional scaffolds for biomedical applications  

NASA Astrophysics Data System (ADS)

Production of brand new protein-based materials with precise control over the amino acid sequences at single residue level has been made possible by genetic engineering, through which artificial genes can be developed that encode protein-based materials with desired features. As an example, silk-elastinlike protein polymers (SELPs), composed of tandem repeats of amino acid sequence motifs from Bombyx mori (silkworm) silk and mammalian elastin, have been produced in this approach. SELPs have been studied extensively in the past two decades, however, the fundamental mechanism governing the self-assembly process to date still remains largely unresolved. Further, regardless of the unprecedented success when exploited in areas including drug delivery, gene therapy, and tissue augmentation, SELPs scaffolds as a three-dimensional cell culture model system are complicated by the inability of SELPs to provide the embedded tissue cells with appropriate biochemical stimuli essential for cell survival and function. In this dissertation, it is reported that the self-assembly of silk-elastinlike protein polymers (SELPs) into nanofibers in aqueous solutions can be modulated by tuning the curing temperature, the size of the silk blocks, and the charge of the elastin blocks. A core-sheath model was 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. The assembled nanofibers further form nanofiber clusters on the microscale, and the nanofiber clusters then coalesce into nanofiber micro-assemblies, interconnection of which eventually leads to the formation of three-dimensional scaffolds with distinct nanoscale and microscale features. SELP-Collagen hybrid scaffolds were also fabricated to enable independent control over the scaffolds' biochemical input and matrix stiffness. It is reported herein that in the hybrid scaffolds, collagen provides essential biochemical cues needed to promote cell attachment and function while SELP imparts matrix stiffness tunability. To obtain tissue-specificity in matrix stiffness that spans over several orders of magnitude covering from soft brain to stiff cartilage, the hybrid SELP-Collagen scaffolds were crosslinked by transglutaminase at physiological conditions compatible for simultaneous cell encapsulation. The effect of the increase in matrix stiffness induced by such enzymatic crosslinking on cellular viability and proliferation was also evaluated using in vitro cell assays.

Zeng, Like

142

Hydrogen bonding-assisted thermal conduction in ?-sheet crystals of spider silk protein  

NASA Astrophysics Data System (ADS)

Using atomistic simulations, we demonstrate that ?-sheet, an essential component of spider silk protein, has a thermal conductivity 1-2 orders of magnitude higher than that of some other protein structures reported in the literature. In contrast to several other nanostructured materials of similar bundled/layered structures (e.g. few-layer graphene and bundled carbon nanotubes), the ?-sheet is found to uniquely feature enhanced thermal conductivity with an increased number of constituting units, i.e. ?-strands. Phonon analysis identifies inter-?-strand hydrogen bonding as the main contributor to the intriguing phenomenon, which prominently influences the state of phonons in both low- and high-frequency regimes. A thermal resistance model further verifies the critical role of hydrogen bonding in thermal conduction through ?-sheet structures.Using atomistic simulations, we demonstrate that ?-sheet, an essential component of spider silk protein, has a thermal conductivity 1-2 orders of magnitude higher than that of some other protein structures reported in the literature. In contrast to several other nanostructured materials of similar bundled/layered structures (e.g. few-layer graphene and bundled carbon nanotubes), the ?-sheet is found to uniquely feature enhanced thermal conductivity with an increased number of constituting units, i.e. ?-strands. Phonon analysis identifies inter-?-strand hydrogen bonding as the main contributor to the intriguing phenomenon, which prominently influences the state of phonons in both low- and high-frequency regimes. A thermal resistance model further verifies the critical role of hydrogen bonding in thermal conduction through ?-sheet structures. Electronic supplementary information (ESI) available: Structure of the ?-sheets, computational model, determination of area and temperature gradient, and additional phonon DOS results. See DOI: 10.1039/c4nr01195c

Zhang, Lin; Chen, Teli; Ban, Heng; Liu, Ling

2014-06-01

143

Ancient properties of spider silks revealed by the complete gene sequence of the prey-wrapping silk protein (AcSp1).  

PubMed

Spider silk fibers have impressive mechanical properties and are primarily composed of highly repetitive structural proteins (termed spidroins) encoded by a single gene family. Most characterized spidroin genes are incompletely known because of their extreme size (typically >9 kb) and repetitiveness, limiting understanding of the evolutionary processes that gave rise to their unusual gene architectures. The only complete spidroin genes characterized thus far form the dragline in the Western black widow, Latrodectus hesperus. Here, we describe the first complete gene sequence encoding the aciniform spidroin AcSp1, the primary component of spider prey-wrapping fibers. L. hesperus AcSp1 contains a single enormous (?19 kb) exon. The AcSp1 repeat sequence is exceptionally conserved between two widow species (?94% identity) and between widows and distantly related orb-weavers (?30% identity), consistent with a history of strong purifying selection on its amino acid sequence. Furthermore, the 16 repeats (each 371-375 amino acids long) found in black widow AcSp1 are, on average, >99% identical at the nucleotide level. A combination of stabilizing selection on amino acid sequence, selection on silent sites, and intragenic recombination likely explains the extreme homogenization of AcSp1 repeats. In addition, phylogenetic analyses of spidroin paralogs support a gene duplication event occurring concomitantly with specialization of the aciniform glands and the tubuliform glands, which synthesize egg-case silk. With repeats that are dramatically different in length and amino acid composition from dragline spidroins, our L. hesperus AcSp1 expands the knowledge base for developing silk-based biomimetic technologies. PMID:23155003

Ayoub, Nadia A; Garb, Jessica E; Kuelbs, Amanda; Hayashi, Cheryl Y

2013-03-01

144

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

145

New and highly efficient expression systems for expressing selectively foreign protein in the silk glands of transgenic silkworm  

Microsoft Academic Search

We constructed three different fibroin H-chain expression systems to estimate the efficacy of producing recombinant proteins\\u000a in the cocoon of transgenic silkworms. The results showed that the three different EGFP\\/H-chain fusion genes were all expressed\\u000a selectively in the posterior silk gland of the transgenic silkworm. The recombinant protein content of transgenic silkworm\\u000a cocoons is up to 15% (w\\/w) when using

Aichun Zhao; Tianfu Zhao; Yuansong Zhang; Qingyou Xia; Cheng Lu; Zeyang Zhou; Zhonghuai Xiang; M. Nakagaki

2010-01-01

146

Toward spinning artificial spider silk.  

PubMed

Spider silk is strong and extensible but still biodegradable and well tolerated when implanted, making it the ultimate biomaterial. Shortcomings that arise in replicating spider silk are due to the use of recombinant spider silk proteins (spidroins) that lack native domains, the use of denaturing conditions under purification and spinning and the fact that the understanding of how spiders control silk formation is incomplete. Recent progress has unraveled the molecular mechanisms of the spidroin N- and C-terminal nonrepetitive domains (NTs and CTs) and revealed the pH and ion gradients in spiders' silk glands, clarifying how spidroin solubility is maintained and how silk is formed in a fraction of a second. Protons and CO2, generated by carbonic anhydrase, affect the stability and structures of the NT and CT in different ways. These insights should allow the design of conditions and devices for the spinning of recombinant spidroins into native-like silk. PMID:25885958

Rising, Anna; Johansson, Jan

2015-05-01

147

Ubiquitous distribution of salts and proteins in spider glue enhances spider silk adhesion.  

PubMed

Modern orb-weaving spiders use micron-sized glue droplets on their viscid silk to retain prey in webs. A combination of low molecular weight salts and proteins makes the glue viscoelastic and humidity responsive in a way not easily achieved by synthetic adhesives. Optically, the glue droplet shows a heterogeneous structure, but the spatial arrangement of its chemical components is poorly understood. Here, we use optical and confocal Raman microscopy to show that salts and proteins are present ubiquitously throughout the droplet. The distribution of adhesive proteins in the peripheral region explains the superior prey capture performance of orb webs as it enables the entire surface area of the glue droplet to act as a site for prey capture. The presence of salts throughout the droplet explains the recent Solid-State NMR results that show salts directly facilitate protein mobility. Understanding the function of individual glue components and the role of the droplet's macro-structure can help in designing better synthetic adhesives for humid environments. PMID:25761668

Amarpuri, Gaurav; Chaurasia, Vishal; Jain, Dharamdeep; Blackledge, Todd A; Dhinojwala, Ali

2015-01-01

148

Ubiquitous distribution of salts and proteins in spider glue enhances spider silk adhesion  

NASA Astrophysics Data System (ADS)

Modern orb-weaving spiders use micron-sized glue droplets on their viscid silk to retain prey in webs. A combination of low molecular weight salts and proteins makes the glue viscoelastic and humidity responsive in a way not easily achieved by synthetic adhesives. Optically, the glue droplet shows a heterogeneous structure, but the spatial arrangement of its chemical components is poorly understood. Here, we use optical and confocal Raman microscopy to show that salts and proteins are present ubiquitously throughout the droplet. The distribution of adhesive proteins in the peripheral region explains the superior prey capture performance of orb webs as it enables the entire surface area of the glue droplet to act as a site for prey capture. The presence of salts throughout the droplet explains the recent Solid-State NMR results that show salts directly facilitate protein mobility. Understanding the function of individual glue components and the role of the droplet's macro-structure can help in designing better synthetic adhesives for humid environments.

Amarpuri, Gaurav; Chaurasia, Vishal; Jain, Dharamdeep; Blackledge, Todd A.; Dhinojwala, Ali

2015-03-01

149

Ubiquitous distribution of salts and proteins in spider glue enhances spider silk adhesion  

PubMed Central

Modern orb-weaving spiders use micron-sized glue droplets on their viscid silk to retain prey in webs. A combination of low molecular weight salts and proteins makes the glue viscoelastic and humidity responsive in a way not easily achieved by synthetic adhesives. Optically, the glue droplet shows a heterogeneous structure, but the spatial arrangement of its chemical components is poorly understood. Here, we use optical and confocal Raman microscopy to show that salts and proteins are present ubiquitously throughout the droplet. The distribution of adhesive proteins in the peripheral region explains the superior prey capture performance of orb webs as it enables the entire surface area of the glue droplet to act as a site for prey capture. The presence of salts throughout the droplet explains the recent Solid-State NMR results that show salts directly facilitate protein mobility. Understanding the function of individual glue components and the role of the droplet's macro-structure can help in designing better synthetic adhesives for humid environments. PMID:25761668

Amarpuri, Gaurav; Chaurasia, Vishal; Jain, Dharamdeep; Blackledge, Todd A.; Dhinojwala, Ali

2015-01-01

150

CHANGES IN FINE STRUCTURE DURING SILK PROTEIN PRODUCTION IN THE AMPULLATE GLAND OF THE SPIDER ARANEUS SERICATUS  

PubMed Central

The ampullate silk gland of the spider, Araneus sericatus, produces the silk fiber for the scaffolding of the web. The fine structure of the various parts of the gland is described. The distal portion of the duct consist of a tube of epithelial cells which appear to secrete a substance which forms the tunica intima of the duct wall. At the proximal end of the duct there is a region of secretory cells. The epithelium of the sac portion contains five morphologically distinct types of granules. The bulk of the synthesis of silk occurs in the tail of the gland, and in this region only a single type of secretory droplet is seen in the epithelium. Protein synthesis can be stimulated by the injection of 1 mg/kg acetylcholine into the body fluids. 10 min after injection, much of the protein stored in the cytoplasm of the epithelial cells has been secreted into the lumen. 20 min after stimulation, the ergastoplasmic sacs form large whorls in the cytoplasm. Protein, similar in electron-opacity to protein found in the lumen, begins to form in that portion of the cytoplasm which is enclosed by the whorls. The limiting membrane of these droplets is formed by ergastoplasmic membranes which lose their ribosomes. No Golgi material has been found in these cells. Protein appears to be manufactured in the cytoplasm of the tail cells in a form which is ready for secretion. PMID:5786985

Bell, Allen L.; Peakall, David B.

1969-01-01

151

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

152

More than just fibers: an aqueous method for the production of innovative recombinant spider silk protein materials.  

PubMed

Spider silk is a striking and robust natural material that has an unrivaled combination of strength and elasticity. There are two major problems in creating materials from recombinant spider silk proteins (rSSps): expressing sufficient quantities of the large, highly repetitive proteins and solvating the naturally self-assembling proteins once produced. To address the second problem, we have developed a method to rapidly dissolve rSSps in water in lieu of traditional organic solvents and accomplish nearly 100% solvation and recovery of the protein. Our method involves generating pressure and temperature in a sealed vial by using short, repetitive bursts from a conventional microwave. The method is scalable and has been successful with all rSSps used to date. From these easily generated aqueous solutions of rSSps, a wide variety of materials have been produced. Production of fibers, films, hydrogels, lyogels, sponges, and adhesives and studies of their mechanical and structural properties are reported. To our knowledge, ours is the only method that is cost-effective and scalable for mass production. This solvation method allows a choice of the physical form of product to take advantage of spider silks' mechanical properties without using costly and problematic organic solvents. PMID:25789668

Jones, Justin A; Harris, Thomas I; Tucker, Chauncey L; Berg, Kyle R; Christy, Stacia Y; Day, Breton A; Gaztambide, Danielle A; Needham, Nate J C; Ruben, Ashley L; Oliveira, Paula F; Decker, Richard E; Lewis, Randolph V

2015-04-13

153

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

PubMed

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

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

2015-01-01

154

Expression of a Truncated ATHB17 Protein in Maize Increases Ear Weight at Silking  

PubMed Central

ATHB17 (AT2G01430) is an Arabidopsis gene encoding a member of the ?-subclass of the homeodomain leucine zipper class II (HD-Zip II) family of transcription factors. The ATHB17 monomer contains four domains common to all class II HD-Zip proteins: a putative repression domain adjacent to a homeodomain, leucine zipper, and carboxy terminal domain. However, it also possesses a unique N-terminus not present in other members of the family. In this study we demonstrate that the unique 73 amino acid N-terminus is involved in regulation of cellular localization of ATHB17. The ATHB17 protein is shown to function as a transcriptional repressor and an EAR-like motif is identified within the putative repression domain of ATHB17. Transformation of maize with an ATHB17 expression construct leads to the expression of ATHB17?113, a truncated protein lacking the first 113 amino acids which encodes a significant portion of the repression domain. Because ATHB17?113 lacks the repression domain, the protein cannot directly affect the transcription of its target genes. ATHB17?113 can homodimerize, form heterodimers with maize endogenous HD-Zip II proteins, and bind to target DNA sequences; thus, ATHB17?113 may interfere with HD-Zip II mediated transcriptional activity via a dominant negative mechanism. We provide evidence that maize HD-Zip II proteins function as transcriptional repressors and that ATHB17?113 relieves this HD-Zip II mediated transcriptional repression activity. Expression of ATHB17?113 in maize leads to increased ear size at silking and, therefore, may enhance sink potential. We hypothesize that this phenotype could be a result of modulation of endogenous HD-Zip II pathways in maize. PMID:24736658

Creelman, Robert A.; Griffith, Cara; Ahrens, Jeffrey E.; Taylor, J. Philip; Murphy, Lesley R.; Manjunath, Siva; Thompson, Rebecca L.; Lingard, Matthew J.; Back, Stephanie L.; Larue, Huachun; Brayton, Bonnie R.; Burek, Amanda J.; Tiwari, Shiv; Adam, Luc; Morrell, James A.; Caldo, Rico A.; Huai, Qing; Kouadio, Jean-Louis K.; Kuehn, Rosemarie; Sant, Anagha M.; Wingbermuehle, William J.; Sala, Rodrigo; Foster, Matt; Kinser, Josh D.; Mohanty, Radha; Jiang, Dongming; Ziegler, Todd E.; Huang, Mingya G.; Kuriakose, Saritha V.; Skottke, Kyle; Repetti, Peter P.; Reuber, T. Lynne; Ruff, Thomas G.; Petracek, Marie E.; Loida, Paul J.

2014-01-01

155

Science Nation: Got Silk?  

NSDL National Science Digital Library

Spider silk fibers are stronger than almost any other man-made fiber and they're also elastic. There are a lot of potential applications for the fiber, but how do you come up with enough raw material? With help from the National Science Foundation (NSF), researchers have figured out a way to put the spider's silk-making genes into goats in a way that they only make the protein in their milk.

156

Cre-mediated targeted gene activation in the middle silk glands of transgenic silkworms (Bombyx mori).  

PubMed

Cre-mediated recombination is widely used to manipulate defined genes spatiotemporally in vivo. The present study evaluated the Cre/loxP system in Bombyx mori by establishing two transgenic lines. One line contained a Cre recombinase gene controlled by a sericin-1 gene (Ser1) promoter. The other line contained a loxP-Stop-loxP-DsRed cassette driven by the same Ser1 promoter. The precise deletion of the Stop fragment was found to be triggered by Cre-mediated site-specific excision, and led to the expression of DsRed fluorescence protein in the middle silk glands of all double-transgenic hybrids. This result was also confirmed by phenotypical analysis. Hence, the current study demonstrated the feasibility of Cre-mediated site-specific recombination in B. mori, and opened a new window for further refining genetic tools in silkworms. PMID:23264031

Duan, Jianping; Xu, Hanfu; Ma, Sanyuan; Guo, Huizhen; Wang, Feng; Zhao, Ping; Xia, Qingyou

2013-06-01

157

Nanostructure and molecular mechanics of spider dragline silk  

E-print Network

Nanostructure and molecular mechanics of spider dragline silk protein assemblies Sinan Keten1, Massachusetts Institute of Technology, 77 Massachusetts Ave, Cambridge, MA 02139, USA Spider silk is a self structure of silk proteins has a direct influence on the stiffness, toughness and failure strength of silk

Buehler, Markus J.

158

Effects of degumming conditions on electro-spinning rate of regenerated silk.  

PubMed

Electro-spun silk webs are potentially good candidates as tissue engineering scaffolds owing to their good bio- and cyto-compatibility. However, the low fabrication rate of electro-spun silk mats has been one of the obstacles in the mass production of such nanofibrous silk mats in applications to the biomedical field. In this study, the effects of degumming ratio and silk concentration on the electro-spinning process were investigated by using regenerated silk with different residual sericin contents and different silk concentrations in terms of the morphology and structure of the electro-spun silk web. The rate of production of electro-spun silk mats could be increased by approximately 5 fold at a degumming ratio of 19.5%. The electro-spinning rate of silk was affected by two main factors: (1) dope solution viscosity and (2) degumming ratio of silk. The conductivity of the silk dope solution, however, had little effects on the electro-spinning of regenerated silk. A constant spun fiber morphology was observed within the electro-spinning rate range (0.3-1.4 ml/h). Fourier transform infrared spectroscopy showed that partial ?-sheet crystallization occurred during electro-spinning. The molecular conformation was relatively unaffected by the electro-spinning rate of silk. PMID:23817099

Yoon, Kyunghwan; Lee, Ha Ni; Ki, Chang Seok; Fang, Dufei; Hsiao, Benjamin S; Chu, Benjamin; Um, In Chul

2013-10-01

159

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

160

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

161

Engineering of recombinant spider silk proteins allows defined uptake and release of substances.  

PubMed

Drug delivery carriers stabilize drugs and control their release, expanding the therapeutic window, and avoiding side effects of otherwise freely diffusing drugs in the human body. Materials used as carrier vehicles have to be biocompatible, biodegradable, nontoxic, and nonimmunogenic. Previously, particles made of the recombinant spider silk protein eADF4(C16) could be effectively loaded with positively and neutrally charged model substances. Here, a new positively charged variant thereof, named eADF4(?16), has been engineered. Its particle formation is indistinguishable to that of polyanionic eADF4(C16), but in contrast polycationic eADF4(?16) allows incorporation of negatively charged substances. Both high-molecular-weight substances, such as nucleic acids, and low-molecular-weight substances could be efficiently loaded onto eADF4(?16) particles, and release of nucleic acids was shown to be well controlled. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 104:988-994, 2015. PMID:25546241

Doblhofer, Elena; Scheibel, Thomas

2015-03-01

162

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

163

Free radical scavenging and tyrosinase inhibition activity of oils and sericin extracted from Thai native silkworms (Bombyx mori).  

PubMed

Oils and sericin were extracted from pupae and silk cocoons, respectively, of the five Thai native silkworms (Bombyx mori, Linnaeus (Bombycidae)), namely, Keawsakol, Nangnoi, Somrong, Nangleung, and Noneruesee, which are variations of the same species. The oils were extracted by a hot process using Soxhlet apparatus and a cold process using petroleum ether, while sericin was extracted by basic hydrolysis and autoclaving. Sericin from the five Thai native silkworms showed free radical scavenging activity lower than the standard antioxidants (vitamin C, vitamin E, and BHT) by about 20-100-fold, but all oils gave higher activity than that of the standard linoleic acid by 11-22-fold. Oil extracted from Noneruesee by the cold process gave the highest DPPH scavenging activity, compared with other oil samples. All sericin samples showed tyrosinase inhibition activity with IC(50) values in the range of 1.2-18.76 mg/mL, but only oils from Noneruesee extracted by the hot process, and Nangleung, Somrong, and Noneruesee extracted by the cold process, showed this activity. Oil extracted by the hot process and sericin by basic hydrolysis from Noneruesee gave the highest tyrosinase inhibition activity, but lower than that of the standards vitamin C and kojic acid by 20-49 and 3-8 times, respectively. This study has suggested that sericin and oil from Noneruesee extracted by basic hydrolysis and the cold process, which gave the highest tyrosinase inhibition and free radical scavenging activity, respectively, can be applied in antiaging and whitening cosmetic products. PMID:20673171

Manosroi, Aranya; Boonpisuttinant, Korawinwich; Winitchai, Supanida; Manosroi, Worapaka; Manosroi, Jiradej

2010-08-01

164

Long-range periodic sequence of the cement/silk protein of Stenopsyche marmorata: purification and biochemical characterisation.  

PubMed

The long-range periodic amino acid sequence of the bifunctional silk/cement protein from larvae of the caddisfly, Stenopsyche marmorata, is discussed in this study. The protein, named the S. marmorata silk protein (Smsp-1), was first purified to electrophoretic homogeneity. The results of Edman-based sequencing of Smsp-1 tryptic digests were consistent with the amino acid sequence deduced from a cDNA clone of the Smsp-1 gene. All undetected amino acids in the Edman-based sequencing were encoded as Ser, suggesting the presence of O-phospho-Ser. (31)P-NMR and an O-phospho-amino acid analysis successfully showed that the O-phospho-Ser residue occurred in a clustered manner, serving a cement function for Smsp-1. Two patterns of non-phosphorylated repeats, -SLGPYGDPRGDXLGPYGG- (X = V, G or D) and -GVGPYGDGLGPYGG-, were enriched in Smsp-1 compared with the O-phospho-Ser cluster, and have fibre-forming functions. PMID:23574115

Ohkawa, Kousaku; Miura, Yumi; Nomura, Takaomi; Arai, Ryoichi; Abe, Koji; Tsukada, Masuhiro; Hirabayashi, Kimio

2013-01-01

165

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

166

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

167

Biomimetic magnetic silk scaffolds.  

PubMed

Magnetic silk fibroin protein (SFP) scaffolds integrating magnetic materials and featuring magnetic gradients were prepared for potential utility in magnetic-field assisted tissue engineering. Magnetic nanoparticles (MNPs) were introduced into SFP scaffolds via dip-coating methods, resulting in magnetic SFP scaffolds with different strengths of magnetization. Magnetic SFP scaffolds showed excellent hyperthermia properties achieving temperature increases up to 8 °C in about 100 s. The scaffolds were not toxic to osteogenic cells and improved cell adhesion and proliferation. These findings suggest that tailored magnetized silk-based biomaterials can be engineered with interesting features for biomaterials and tissue-engineering applications. PMID:25734962

Samal, Sangram K; Dash, Mamoni; Shelyakova, Tatiana; Declercq, Heidi A; Uhlarz, Marc; Bañobre-López, Manuel; Dubruel, Peter; Cornelissen, Maria; Herrmannsdörfer, Thomas; Rivas, Jose; Padeletti, Giuseppina; De Smedt, Stefaan; Braeckmans, Kevin; Kaplan, David L; Dediu, V Alek

2015-03-25

168

Influence of sericin/TiO? nanocomposite on cotton fabric: part 1. Enhanced antibacterial effect.  

PubMed

Sericin as a biological material was extracted from raw silk by boiling in hot water and nano-TiO2 was dispersed in its solution. The prepared finishing agents with and without polycarboxylic acid cross-linking agents were treated on cotton fabric using pad-dry-cure. Presence of sericin, nano-TiO2, and cross-linking agents on cotton fabric was confirmed by at least one of the following experimental FTIR, SEM, EDX, and XRD. The antibacterial activity and the durability of modified cotton fabrics were investigated against one Gram-positive bacterium (Staphylococcus aureus) and one Gram-negative bacterium (Escherichia coli). The finishing treatment on the cotton fabric was more effective against S. aureus than E. coli. The fabrics treated with nano-TiO2 were possessed more activity against bacteria as compared to sericin and also considerably improved with given nanocomposite. The antibacterial activity of treated fabrics with cross-linking agents has not been considerably changed after 20 and 40 launderings. The fabrics treated with given nanocomposites did not dramatically affect the breaking strength. PMID:23544628

Doakhan, S; Montazer, M; Rashidi, A; Moniri, R; Moghadam, M B

2013-05-15

169

The molecular structure of spider dragline silk: Folding and orientation of the protein backbone  

PubMed Central

The design principles of spider dragline silk, nature's high-performance fiber, are still largely unknown, in particular for the noncrystalline glycine-rich domains, which form the bulk of the material. Here we apply two-dimensional solid-state NMR to determine the distribution of the backbone torsion angles (?,?) as well as the orientation of the polypeptide backbone toward the fiber at both the glycine and alanine residues. Instead of an “amorphous matrix,” suggested earlier for the glycine-rich domains, these new data indicate that all domains in dragline silk have a preferred secondary structure and are strongly oriented, with the chains predominantly parallel to the fiber. As proposed previously, the alanine residues are predominantly found in a ? sheet conformation. The glycine residues are partly incorporated into the ? sheets and otherwise form helical structures with an approximate 3-fold symmetry. PMID:12149440

van Beek, J. D.; Hess, S.; Vollrath, F.; Meier, B. H.

2002-01-01

170

From Micelles to Fibers: Balancing Self-Assembling and Random Coiling Domains in pH-Responsive Silk-Collagen-Like Protein-Based Polymers  

PubMed Central

We study the self-assembly of genetically engineered protein-based triblock copolymers consisting of a central pH-responsive silk-like middle block (SHn, where SH is a silk-like octapeptide, (GA)3GH and n is the number of repeats) flanked by hydrophilic random coil outer blocks (C2). Our previous work has already shown that triblocks with very long midblocks (n = 48) self-assemble into long, stiff protein filaments at pH values where the middle blocks are uncharged. Here we investigate the self-assembly behavior of the triblock copolymers for a range of midblock lengths, n = 8, 16, 24, 48. Upon charge neutralization of SHn by adjusting the pH, we find that C2SH8C2 and C2SH16C2 form spherical micelles, whereas both C2SH24C2 and C2SH48C2 form protein filaments with a characteristic beta-roll secondary structure of the silk midblocks. Hydrogels formed by C2SH48C2 are much stronger and form much faster than those formed by C2SH24C2. Enzymatic digestion of much of the hydrophilic outer blocks is used to show that with much of the hydrophilic outer blocks removed, all silk-midblocks are capable of self-assembling into stiff protein filaments. In that case, reduction of the steric repulsion by the hydrophilic outer blocks also leads to extensive fiber bundling. Our results highlight the opposing roles of the hydrophilic outer blocks and central silk-like midblocks in driving protein filament formation. They provide crucial information for future designs of triblock protein-based polymers that form stiff filaments with controlled bundling, that could mimick properties of collagen in the extracellular matrix. PMID:25133990

2014-01-01

171

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

172

TNF-? Blocker Effect of Naringenin-Loaded Sericin Microparticles that Are Potentially Useful in the Treatment of Psoriasis  

PubMed Central

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

173

Reproducing Natural Spider Silks’ Copolymer Behavior in Synthetic Silk Mimics  

PubMed Central

Dragline silk from orb-weaving spiders is a copolymer of two large proteins, major ampullate spidroin 1 (MaSp1) and 2 (MaSp2). The ratio of these proteins is known to have a large variation across different species of orb-weaving spiders. NMR results from gland material of two different species of spiders, N. clavipes and A. aurantia, indicates that MaSp1 proteins are more easily formed into ?-sheet nanostructures, while MaSp2 proteins form random coil and helical structures. To test if this behavior of natural silk proteins could be reproduced by recombinantly produced spider silk mimic protein, recombinant MaSp1/MaSp2 mixed fibers as well as chimeric silk fibers from MaSp1 and MaSp2 sequences in a single protein were produced based on the variable ratio and conserved motifs of MaSp1 and MaSp2 in native silk fiber. Mechanical properties, solid-state NMR, and XRD results of tested synthetic fibers indicate the differing roles of MaSp1 and MaSp2 in the fiber and verify the importance of postspin stretching treatment in helping the fiber to form the proper spatial structure. PMID:23110450

An, Bo; Jenkins, Janelle E.; Sampath, Sujatha; Holland, Gregory P.; Hinman, Mike; Yarger, Jeffery L.; Lewis, Randolph

2012-01-01

174

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

Pulse of the Planet

2005-11-23

175

Transmission Electron Microscopy of Bombyx Mori Silk Fibers  

NASA Astrophysics Data System (ADS)

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

Shen, Y.; Martin, D. C.

1997-03-01

176

Silk Gland Factor-2, Involved in Fibroin Gene Transcription, Consists of LIM Homeodomain, LIM-interacting, and Single-stranded DNA-binding Proteins*  

PubMed Central

SGF-2 binds to promoter elements governing posterior silk gland-specific expression of the fibroin gene in Bombyx mori. We purified SGF-2 and showed that SGF-2 contains at least four gene products: the silkworm orthologues of LIM homeodomain protein Awh, LIM domain-binding protein (Ldb), a sequence-specific single-stranded DNA-binding protein (Lcaf), and the silk protein P25/fibrohexamerin (fhx). Using co-expression of these factors in Sf9 cells, Awh, Ldb, and Lcaf proteins were co-purified as a ternary complex that bound to the enhancer sequence in vitro. Lcaf interacts with Ldb as well as Awh through the conserved regions to mediate transcriptional activation in yeast. Misexpression of Awh in transgenic silkworms induces ectopic expression of the fibroin gene in the middle silk glands, where Ldb and Lcaf are expressed. Taken together, this study demonstrates that SGF-2 is a multisubunit activator complex containing Awh. Moreover, our results suggest that the Ldb·Lcaf protein complex serves as a scaffold to facilitate communication between transcriptional control elements. PMID:24022586

Ohno, Kaoru; Sawada, Jun-ichi; Takiya, Shigeharu; Kimoto, Mai; Matsumoto, Akiko; Tsubota, Takuya; Uchino, Keiro; Hui, Chi-chung; Sezutsu, Hideki; Handa, Hiroshi; Suzuki, Yoshiaki

2013-01-01

177

The elaborate structure of spider silk  

PubMed Central

Biomaterials, having evolved over millions of years, often exceed man-made materials in their properties. Spider silk is one outstanding fibrous biomaterial which consists almost entirely of large proteins. Silk fibers have tensile strengths comparable to steel and some silks are nearly as elastic as rubber on a weight to weight basis. In combining these two properties, silks reveal a toughness that is two to three times that of synthetic fibers like Nylon or Kevlar. Spider silk is also antimicrobial, hypoallergenic and completely biodegradable. This article focuses on the structure-function relationship of the characterized highly repetitive spider silk spidroins and their conformational conversion from solution into fibers. Such knowedge is of crucial importance to understanding the intrinsic properties of spider silk and to get insight into the sophisticated assembly processes of silk proteins. This review further outlines recent progress in recombinant production of spider silk proteins and their assembly into distinct polymer materials as a basis for novel products. PMID:19221522

Römer, Lin

2008-01-01

178

Super Spider Silk  

E-print Network

to sox, spider silk can be used for sutures and fishing line. Well, now that they've made silk sox out of a spider's thread what's next? A silk purse out of a sow's ear? #ceas #hacker #japan #tsutsui...

Hacker, Randi; Tsutsui, William

2008-02-06

179

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

180

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

181

Pulsed laser deposition of silk protein: Effect of photosensitized-ablation on the secondary structure in thin deposited films  

SciTech Connect

Silk fibroin is a simple protein expected to have functional applications in medicine and bioelectronics. The primary structure of this protein is quite simple, and the main secondary structures are {beta}-sheet crystals and amorphous random coils. In the present study, we investigated pulsed laser deposition (PLD) of fibroin with the {beta}-sheet structures as targets. The primary and secondary structures in films deposited were analyzed using infrared spectroscopy. Normal laser deposition at 351 nm using neat fibroin targets produced thin films of fibroin with a random coiled structure. Ablation was triggered by two-photonic excitation of the peptide chains, which resulted in the destruction of {beta}-sheet structure in PLD. In order to avoid the two-photonic excitation, we adopted a PLD method utilizing anthracene (5{endash}0.1 wt%) in a photosensitized reaction involving doped fibroin targets. Laser light (351 or 355 nm) was absorbed only by anthracene, which plays an important role converting photon energy to thermal energy with great ablation efficiency. Thin fibroin films deposited by this method had both random coil and {beta}-sheet structures. As the dopant concentration and laser fluence decreased, the ratio of {beta}-sheet domain to random coil increased in thin deposited films. {copyright} 2001 American Institute of Physics.

Tsuboi, Yasuyuki; Goto, Masaharu; Itaya, Akira

2001-06-15

182

Sericin Enhances the Bioperformance of Collagen-Based Matrices Preseeded with Human-Adipose Derived Stem Cells (hADSCs).  

PubMed

Current clinical strategies for adipose tissue engineering (ATE), including autologous fat implants or the use of synthetic surrogates, not only are failing in the long term, but also can't face the latest requirements regarding the aesthetic restoration of the resulted imperfections. In this context, modern strategies in current ATE applications are based on the implantation of 3D cell-scaffold bioconstructs, designed for prospective achievement of in situ functional de novo tissue. Thus, in this paper, we reported for the first time the evaluation of a spongious 60% collagen and 40% sericin scaffold preseeded with human adipose-derived stem cells (hADSCs) in terms of biocompatibility and adipogenic potential in vitro. We showed that the addition of the sticky protein sericin in the composition of a classical collagen sponge enhanced the adhesion and also the proliferation rate of the seeded cells, thus improving the biocompatibility of the novel scaffold. In addition, sericin stimulated PPAR?2 overexpression, triggering a subsequent upregulated expression profile of FAS, aP2 and perilipin adipogenic markers. These features, together with the already known sericin stimulatory potential on cellular collagen production, promote collagen-sericin biomatrix as a good candidate for soft tissue reconstruction and wound healing applications. PMID:23325052

Dinescu, Sorina; Galateanu, Bianca; Albu, Madalina; Cimpean, Anisoara; Dinischiotu, Anca; Costache, Marieta

2013-01-01

183

Sericin Enhances the Bioperformance of Collagen-Based Matrices Preseeded with Human-Adipose Derived Stem Cells (hADSCs)  

PubMed Central

Current clinical strategies for adipose tissue engineering (ATE), including autologous fat implants or the use of synthetic surrogates, not only are failing in the long term, but also can’t face the latest requirements regarding the aesthetic restoration of the resulted imperfections. In this context, modern strategies in current ATE applications are based on the implantation of 3D cell-scaffold bioconstructs, designed for prospective achievement of in situ functional de novo tissue. Thus, in this paper, we reported for the first time the evaluation of a spongious 60% collagen and 40% sericin scaffold preseeded with human adipose-derived stem cells (hADSCs) in terms of biocompatibility and adipogenic potential in vitro. We showed that the addition of the sticky protein sericin in the composition of a classical collagen sponge enhanced the adhesion and also the proliferation rate of the seeded cells, thus improving the biocompatibility of the novel scaffold. In addition, sericin stimulated PPAR?2 overexpression, triggering a subsequent upregulated expression profile of FAS, aP2 and perilipin adipogenic markers. These features, together with the already known sericin stimulatory potential on cellular collagen production, promote collagen-sericin biomatrix as a good candidate for soft tissue reconstruction and wound healing applications. PMID:23325052

Dinescu, Sorina; Galateanu, Bianca; Albu, Madalina; Cimpean, Anisoara; Dinischiotu, Anca; Costache, Marieta

2013-01-01

184

Spider Webs and Silks.  

ERIC Educational Resources Information Center

Compares the attributes of the silk from spiders with those of the commercially harvested silk from silkworms. Discusses the evolution, design, and effectiveness of spider webs; the functional mechanics of the varieties of silk that can be produced by the same spider; and the composite, as well as molecular, structure of spider silk thread. (JJK)

Vollrath, Fritz

1992-01-01

185

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

186

Skin Equivalent Tissue-Engineered Construct: Co-Cultured Fibroblasts/ Keratinocytes on 3D Matrices of Sericin Hope Cocoons  

PubMed Central

The development of effective and alternative tissue-engineered skin replacements to autografts, allografts and xenografts has became a clinical requirement due to the problems related to source of donor tissue and the perceived risk of disease transmission. In the present study 3D tissue engineered construct of sericin is developed using co-culture of keratinocytes on the upper surface of the fabricated matrices and with fibroblasts on lower surface. Sericin is obtained from “Sericin Hope” silkworm of Bombyx mori mutant and is extracted from cocoons by autoclave. Porous sericin matrices are prepared by freeze dried method using genipin as crosslinker. The matrices are characterized biochemically and biophysically. The cell proliferation and viability of co-cultured fibroblasts and keratinocytes on matrices for at least 28 days are observed by live/dead assay, Alamar blue assay, and by dual fluorescent staining. The growth of the fibroblasts and keratinocytes in co-culture is correlated with the expression level of TGF-?, b-FGF and IL-8 in the cultured supernatants by enzyme-linked immunosorbent assay. The histological analysis further demonstrates a multi-layered stratified epidermal layer of uninhibited keratinocytes in co-cultured constructs. Presence of involucrin, collagen IV and the fibroblast surface protein in immuno-histochemical stained sections of co-cultured matrices indicates the significance of paracrine signaling between keratinocytes and fibroblasts in the expression of extracellular matrix protein for dermal repair. No significant amount of pro inflammatory cytokines (TNF-?, IL-1? and nitric oxide) production are evidenced when macrophages grown on the sericin matrices. The results all together depict the potentiality of sericin 3D matrices as skin equivalent tissue engineered construct in wound repair. PMID:24058626

Nayak, Sunita; Dey, Sancharika; Kundu, Subhas C.

2013-01-01

187

Skin equivalent tissue-engineered construct: co-cultured fibroblasts/ keratinocytes on 3D matrices of sericin hope cocoons.  

PubMed

The development of effective and alternative tissue-engineered skin replacements to autografts, allografts and xenografts has became a clinical requirement due to the problems related to source of donor tissue and the perceived risk of disease transmission. In the present study 3D tissue engineered construct of sericin is developed using co-culture of keratinocytes on the upper surface of the fabricated matrices and with fibroblasts on lower surface. Sericin is obtained from "Sericin Hope" silkworm of Bombyx mori mutant and is extracted from cocoons by autoclave. Porous sericin matrices are prepared by freeze dried method using genipin as crosslinker. The matrices are characterized biochemically and biophysically. The cell proliferation and viability of co-cultured fibroblasts and keratinocytes on matrices for at least 28 days are observed by live/dead assay, Alamar blue assay, and by dual fluorescent staining. The growth of the fibroblasts and keratinocytes in co-culture is correlated with the expression level of TGF-?, b-FGF and IL-8 in the cultured supernatants by enzyme-linked immunosorbent assay. The histological analysis further demonstrates a multi-layered stratified epidermal layer of uninhibited keratinocytes in co-cultured constructs. Presence of involucrin, collagen IV and the fibroblast surface protein in immuno-histochemical stained sections of co-cultured matrices indicates the significance of paracrine signaling between keratinocytes and fibroblasts in the expression of extracellular matrix protein for dermal repair. No significant amount of pro inflammatory cytokines (TNF-?, IL-1? and nitric oxide) production are evidenced when macrophages grown on the sericin matrices. The results all together depict the potentiality of sericin 3D matrices as skin equivalent tissue engineered construct in wound repair. PMID:24058626

Nayak, Sunita; Dey, Sancharika; Kundu, Subhas C

2013-01-01

188

An innovative bi-layered wound dressing made of silk and gelatin for accelerated wound healing.  

PubMed

In this study, the novel silk fibroin-based bi-layered wound dressing was developed. Wax-coated silk fibroin woven fabric was introduced as a non-adhesive layer while the sponge made of sericin and glutaraldehyde-crosslinked silk fibroin/gelatin was fabricated as a bioactive layer. Wax-coated silk fibroin fabrics showed improved mechanical properties compared with the non-coated fabrics, but less adhesive than the commercial wound dressing mesh. This confirmed by results of peel test on both the partial- and full-thickness wounds. The sericin-silk fibroin/gelatin spongy bioactive layers showed homogeneous porous structure and controllable biodegradation depending on the degree of crosslinking. The bi-layered wound dressings supported the attachment and proliferation of L929 mouse fibroblasts, particularly for the silk fibroin/gelatin ratio of 20/80 and 0.02% GA crosslinked. Furthermore, we proved that the bi-layered wound dressings promoted wound healing in full-thickness wounds, comparing with the clinically used wound dressing. The wounds treated with the bi-layered wound dressings showed the greater extent of wound size reduction, epithelialization, and collagen formation. The superior properties of the silk fibroin-based bi-layered wound dressings compared with those of the clinically used wound dressings were less adhesive and had improved biological functions to promote cell activities and wound healing. This novel bi-layered wound dressing should be a good candidate for the healing of full-thickness wounds. PMID:22771972

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

2012-10-15

189

Structural characterization of nanofiber silk produced by embiopterans (webspinners)†  

PubMed Central

Embiopterans produce silken galleries and sheets using exceptionally fine silk fibers in which they live and breed. In this study, we use electron microscopy (EM), Fourier-transform infrared (FT-IR) spectroscopy, wide angle X-ray diffraction (WAXD) and solid-state nuclear magnetic resonance (ssNMR) techniques to elucidate the molecular level protein structure of webspinner (embiid) silks. Silks from two species Antipaluria urichi and Aposthonia ceylonica are studied in this work. Electron microscopy images show that the fibers are about 90–100 nm in diameter, making webspinner silks among the finest of all known animal silks. Structural studies reveal that the silk protein core is dominated by ?-sheet structures, and that the protein core is coated with a hydrophobic alkane-rich surface coating. FTIR spectra of native embiid silk shows characteristic alkane CH2 stretchings near 2800–2900 cm?1, which decrease approximately 50% after washing the silk with 2 : 1 CHCl3 : MeOH. Furthermore, 13C ssNMR data shows a significant CH2 resonance that is strongly affected by the presence of water, supporting the idea that the silk fibers are coated with a hydrocarbon-rich layer. Such a layer is likely used to protect the colonies from rain. FTIR data also suggests that embiid silks are dominated by ?-sheet secondary structures similar to spider and silkworm silk fibers. NMR data confirms the presence of ?-sheet nanostructures dominated by serine-rich repetitive regions. A deconvolution of the serine C? NMR resonance reveals that approximately 70% of all seryl residues exist in a ?-sheet structure. This is consistent with WAXD results that suggest webspinner silks are 70% crystalline, which is the highest crystalline fraction reported for any animal silks. The work presented here provides a molecular level structural picture of silk fibers produced by webspinners. PMID:25383190

Addison, J. Bennett; Popp, Thomas M. Osborn; Weber, Warner S.; Edgerly, Janice S.; Holland, Gregory P.; Yarger, Jeffery L.

2014-01-01

190

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

191

Silkmapin of Hyriopsis cumingii, a novel silk-like shell matrix protein involved in nacre formation.  

PubMed

Understanding the role of matrix proteins in nacre formation and biomineralization in mollusks is important for the pearl industry. In this study, the gene encoding the novel Hyriopsis cumingii shell matrix protein silkmapin was characterized. The gene encodes a protein of 30.89kDa in which Gly accounts for 34.41% of the amino acid content, and the C-terminal region binds Ca(2+). Secondary structure prediction indicated a predominantly ?-fold and a structure typical of filamentous proteins. Real-time quantitative PCR and in situ hybridization showed that silkmapin was expressed in epithelial cells at the edge and pallial of mantle tissue, indicated that silkmapin play roles in the shell nacreous and prismatic layer formation. Further real-time PCR results indicated an involvement in pearl formation via nucleation of calcium carbonate prior to formation of the nacre. PMID:25447895

Liu, Xiaojun; Dong, Shaojian; Jin, Can; Bai, Zhiyi; Wang, Guiling; Li, Jiale

2015-01-25

192

Silk fibroin rods for sustained delivery of breast cancer therapeutics.  

PubMed

A silk-protein based reservoir rod was developed for zero-order and long-term sustained drug delivery applications. Silk reservoir rod formulations were processed in three steps. First, a regenerated silk fibroin solution, rich in random-coil content was transformed into a tubular silk film with controllable dimensions, uniform film morphology and a structure rich in silk II, ?-sheet content via "film-spinning." Second, the drug powder was loaded into swollen silk tubes followed by tube end clamping. Last, clamped silk tube ends were sealed completely via dip coating. Anastrozole, an FDA approved active ingredient for the treatment of breast cancer, was used as a model drug to investigate viability of the silk reservoir rod technology for sustained drug delivery. The in vitro and in vivo pharmacokinetic data (in a female Sprague-Dawley rat model) analyzed via liquid chromatography-tandem mass spectroscopy indicated zero-order release for 91 days. Both in vitro and in vivo anastrozole release rates could be controlled simply by varying silk rod dimensions. The swelling behavior of silk films and zero-order anastrozole release kinetics indicated practically immediate film hydration and formation of a linear anastrozole concentration gradient along the silk film thickness. The dependence of anastrozole release rate on the overall silk rod dimensions was in good agreement with an essentially diffusion-controlled sustained release from a reservoir cylindrical geometry. In vivo results highlighted a strong in vitro-in vivo pharmacokinetic correlation and a desirable biocompatibility profile of silk reservoir rods. During a 6-month implantation in rats, the apparent silk molecular weight values decreased gradually, while rod dry mass and ?-sheet crystal content values remained essentially constant, providing a suitable timeframe for controlled, long-term sustained delivery applications. Overall, the silk reservoir rod may be a viable candidate for sustained delivery of breast cancer therapeutics. PMID:25009069

Yucel, Tuna; Lovett, Michael L; Giangregorio, Raffaella; Coonahan, Erin; Kaplan, David L

2014-10-01

193

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

194

History of Silk  

NSDL National Science Digital Library

Nicely illustrated history of silk and explanation of silk production. A fine discussion of the moths, life history, and feeding are included, as well as the extensive cultural history of this outstanding insect product.

0000-00-00

195

Stylized Silk Painting.  

ERIC Educational Resources Information Center

Presents an art activity inspired by a workshop "Surrounded by Silk" given by Susan Skvoe in which the students create silk paintings. Explains that the students first sketch their floral design on paper, trace the design on the silk's surface, and apply liquid dye for color. Provides an easier activity for younger students. (CMK)

Skophammer, Karen

1998-01-01

196

Proteomic profiling of the photo-oxidation of silk fibroin: implications for historic tin-weighted silk.  

PubMed

The stability of silk proteins to ultraviolet light is an issue of significant concern in both the appearance retention of silk-derived products and the preservation of historic silk textiles. Until now, evaluation of silk degradation has only been performed at the holistic, rather than molecular level. This article describes the first proteomic profiling of silk photo-oxidation, characterizing protein primary level modification leading to coloration changes, and evaluating the effects of tin weighting on photodegradation. Heavy-chain fibroin, the main proteinaceous component of the silk thread, is a repetitive, highly crystalline protein with a content rich in tyrosine. Photoproducts of tyrosine were characterized and the levels of oxidative modification at the protein primary structural level correlated with changes in coloration and tensile strength. The effect of tin as a weighting agent used on historical fabrics was examined. Tin-weighted fabrics were evaluated following two treatments (pink and dynamite) and proteomic analysis revealed a significant increase in oxidatively modified amino acid residues within the pink-treated silk. These findings offer new insight into the molecular-level oxidation of silk proteins under UV exposure, and the effects of silk treatments in either exacerbating or ameliorating this degradation. PMID:22554154

Solazzo, Caroline; Dyer, Jolon M; Deb-Choudhury, Santanu; Clerens, Stefan; Wyeth, Paul

2012-01-01

197

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

PubMed

To prepare a bioactive bone substitute, which integrates biphasic calcium phosphate (BCP) and rhBMP-2/silk fibroin (SF) microsphere, and to evaluate its characteristics. Hydroxyapatite and ?-tricalcium phosphate were integrated with a ratio of 60–40%. RhBMP-2/SF (0.5 ?g/1 mg) microsphere was prepared, and its rhBMP-2-release kinetics was assed. After joining pore-forming agent (Sodium chloride, NaCl), porous BCP/rhBMP-2/SF were manufactured, and its characteristics and bioactivity in vitro were evaluated. Mean diameter of rhBMP-2/SF microsphere was 398.7 ± 99.86 nm, with a loading rate of 4.53 ± 0.08%. RhBMP-2 was released in a dual-phase pattern, of which fast-release (nearly half of protein released) focused on the initial 3 days, and slow-release sustained more than 28 days. With the increase in concentration of NaCl, greater was porosity and pore size, but smaller mechanical strength of BCP/rhBMP-2/SF. Material with 150% (w/v) NaCl had an optimal performance, with a porosity of 78.83%, pore size of 293.25 ± 42.77?m and mechanical strength of 31.03 MPa. Proliferation of human placenta-derived mesenchymal stem cells (hPMSCs) on leaching extract medium was similar to the normal medium (P = 0.89), which was better than that on control group (P = 0.03). Activity of alkaline phosphatase on BCP/rhBMP-2/SF surface was higher than on pure BCP at each time point except at 1 day (P < 0.05). RhBMP-2 has a burst release on early times and a sustaining release on later times. BCP/rhBMP-2/SF with 150% (w/v) pore-forming agent has excellent porosity, pore size and mechanical strength. The biomaterial induces proliferation and differentiation hPMSCs effectively. PMID:24659100

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

2014-07-01

198

Materials Fabrication from Bombyx mori Silk Fibroin  

PubMed Central

Silk fibroin, derived from Bombyx mori cocoons, is a widely used and studied protein polymer for biomaterial applications. Silk fibroin has remarkable mechanical properties when formed into different materials, demonstrates biocompatibility, has controllable degradation rates from hours to years, and it can be chemically modified to alter surface properties or to immobilize growth factors. A variety of aqueous or organic solvent processing methods can be used to generate silk biomaterials for a range of applications. In this protocol we include methods to extract silk from B. mori cocoons in order to fabricate hydrogels, tubes, sponges, composites, fibers, microspheres and thin films. These materials can be used directly as biomaterials for implants, as scaffolding in tissue engineering and in vitro disease models, and for drug delivery. PMID:21959241

Rockwood, Danielle N.; Preda, Rucsanda C.; Yücel, Tuna; Wang, Xiaoqin; Lovett, Michael L.; Kaplan, David L.

2013-01-01

199

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

200

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

201

Structure-Function-Property-Design Interplay in Biopolymers: Spider Silk  

PubMed Central

Spider silks have been a focus of research for almost two decades due to their outstanding mechanical and biophysical properties. Recent advances in genetic engineering have led to the synthesis of recombinant spider silks, thus helping to unravel a fundamental understanding of structure-function-property relationships. The relationships between molecular composition, secondary structures, and mechanical properties found in different types of spider silks are described, along with a discussion of artificial spinning of these proteins and their bioapplications, including the role of silks in biomineralization and fabrication of biomaterials with controlled properties. PMID:23962644

Tokareva, Olena; Jacobsen, Matthew; Buehler, Markus; Wong, Joyce; Kaplan, David L.

2013-01-01

202

9422 Stratospheric ice catalyzes chlorine reactions 9428 Fusing silk and silica  

E-print Network

9422 Stratospheric ice catalyzes chlorine reactions 9428 Fusing silk and silica 9482 Identifying, and Mario J. Molina (see pages 9422­9427) CHEMISTRY Fusing silk and silica An engineered protein combining the flexibility and tensile strength of spider silk with the resilience of silica has been developed. The result

McFadden, Geoff

203

Construction of Silk Fiber Core in Lepidoptera Frantisek Sehnal* and Michal Zurovec  

E-print Network

Construction of Silk Fiber Core in Lepidoptera Frantisek Sehnal* and Michal Zurovec Entomological silk fibers depend on amino acid repeats in the principal protein, heavy chain fibroin (H. Introduction Silk fibers are produced from various types of ectodermal glands in the mites, spiders

Â?urovec, Michal

204

Silk Fibroin\\/Sodium Carboxymethylcellulose Blended Films for Biotechnological Applications  

Microsoft Academic Search

The potential of silk protein is increased because of its importance as natural biopolymer for biotechnological and biomedical applications. The main disadvantage of silk fibroin films is their high brittleness. Thus, we studied blends of fibroin with other polymers to improve the film properties. Considering the possible applications of films in biomedical applications, we used a natural and biodegradable polymer

Joydip Kundu; Riti Mohapatra; S. C. Kundu

2011-01-01

205

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

NASA Astrophysics Data System (ADS)

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.

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

2012-10-01

206

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

2012-01-01

207

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

208

Bone Tissue Engineering with Premineralized Silk Scaffolds  

PubMed Central

Silks fibroin biomaterials are being explored as novel protein-based systems for cell and tissue culture. In the present study, biomimetic growth of calcium phosphate on porous silk fibroin polymeric scaffolds was explored to generate organic/inorganic composites as scaffolds for bone tissue engineering. Aqueous-derived silk fibroin scaffolds were prepared with the addition of polyaspartic acid during processing, followed by the controlled deposition of calcium phosphate by exposure to CaCl2 and Na2HPO4. These mineralized protein-composite scaffolds were subsequently seeded with human bone marrow stem cells (hMSC) and cultured in vitro for 6 weeks under osteogenic conditions with or without BMP-2. The extent of osteoconductivity was assessed by cell numbers, alkaline phosphatase and calcium deposition, along with immunohistochemistry for bone related outcomes. The results suggest increased osteoconductive outcomes with an increase in initial content of apatite and BMP-2 in the silk fibroin porous scaffolds. The premineralization of these highly porous silk fibroin protein scaffolds provided enhanced outcomes for the bone tissue engineering. PMID:18387349

Kim, Hyeon Joo; Kim, Ung-Jin; Kim, Hyun Suk; Li, Chunmei; Wada, Masahisa; Leisk, Gary G.; Kaplan, David L.

2009-01-01

209

Bone tissue engineering with premineralized silk scaffolds.  

PubMed

Silk fibroin biomaterials are being explored as novel protein-based systems for cell and tissue culture. In the present study, biomimetic growth of calcium phosphate on porous silk fibroin polymeric scaffolds was explored to generate organic/inorganic composites as scaffolds for bone tissue engineering. Aqueous-derived silk fibroin scaffolds were prepared with the addition of polyaspartic acid during processing, followed by the controlled deposition of calcium phosphate by exposure to CaCl(2) and Na(2)HPO(4). These mineralized protein-composite scaffolds were subsequently seeded with human bone marrow stem cells (hMSC) and cultured in vitro for 6 weeks under osteogenic conditions with or without BMP-2. The extent of osteoconductivity was assessed by cell numbers, alkaline phosphatase and calcium deposition, along with immunohistochemistry for bone-related outcomes. The results suggest increased osteoconductive outcomes with an increase in initial content of apatite and BMP-2 in the silk fibroin porous scaffolds. The premineralization of these highly porous silk fibroin protein scaffolds provided enhanced outcomes for the bone tissue engineering. PMID:18387349

Kim, Hyeon Joo; Kim, Ung-Jin; Kim, Hyun Suk; Li, Chunmei; Wada, Masahisa; Leisk, Gary G; Kaplan, David L

2008-06-01

210

Production of Bombyx mori silk fibroin incorporated with unnatural amino acids.  

PubMed

Silk fibroin incorporated with unnatural amino acids was produced by in vivo feeding of p-chloro-, p-bromo-, and p-azido-substituted analogues of L-phenylalanine (Phe) to transgenic silkworms (Bombyx mori) that expressed a mutant of phenylalanyl-tRNA synthetase with expanded substrate recognition capabilities in silk glands. Cutting down the content of Phe in the diet was effective for increasing the incorporation of Phe analogues but simultaneously caused a decrease of fibroin production. The azide groups incorporated in fibroin were active as chemical handles for click chemistry in both the solubilized and the solid (fibrous) states. The azides survived degumming in the boiling alkaline solution that is required for complete removal of the sericin layer, demonstrating that AzPhe-incorporated silk fibroin could be a versatile platform to produce "clickable" silk materials in various forms. This study indicates the huge potential of UAA mutagenesis as a novel methodology to alter the characteristics of B. mori silk. PMID:24884258

Teramoto, Hidetoshi; Kojima, Katsura

2014-07-14

211

Art on Silk Hoops  

ERIC Educational Resources Information Center

Painting on silk has a magic all its own. Versions of painting on silk can be found throughout the world from Japan and Europe to the United States. Themes for the paintings can be most any type of design or imagery. Applying the liquid dyes is exciting, as the vivid liquid colors flow and blend into the fabric. The process captures students'…

Padrick, Deborah

2012-01-01

212

Silk Road Seattle  

NSDL National Science Digital Library

Silk Road Seattle is a "collaborative public education project using the 'Silk Road' theme to explore cultural interaction across Eurasia from the beginning of the Common Era (A.D.) to the Sixteenth Century." The Silk Road is a term understood to have been the overland trade route from China to the Mediterranean, opened first in the 2nd century and coming to an end between the 15th and 17th centuries. Traditional discussions of the Silk Road, however, recognize that there were branches that went into South Asia, or extended from Central Asia north of the Caspian Sea to the Black Sea. Sponsored primarily by the Simpson Center for the Humanities at the University of Washington and directed by Professors Daniel C. Waugh, Joel Walker, and Cynthea Bogel, this Web site contains a vast range of Silk Road materials. These materials include a list of texts that may be used in teaching and learning about the Silk Road, a section on cities and architecture along the Silk Road, information on traditional culture in Central Asia, a page of teaching and learning guides that list and annotate Silk Road materials, and a virtual art exhibit. This site is still in a constructive stage; therefore, every link is not yet accessible. In short, this site may be of value for a range of audiences, including school children, teachers, college and graduate students, and independent adult learners.

213

A silk fibroin based hepatocarcinoma model and the assessment of the drug response in hyaluronan-binding protein 1 overexpressed HepG2 cells.  

PubMed

Microenvironment around tumor cells plays an important role in its malignancy or invasiveness. Hyaluronan (HA), a major component of extracellular matrix is found to be elevated in most of cancerous niche/microenvironment and performs regulatory role in the progression of tumors and metastasis. Overexpression of the hyaladherin, hyaluronan-binding protein 1 (HABP1) in the hepatocarcinoma cells (HepG2) termed as HepR21 leads to enhanced cell proliferation with increased HA 'pool' associated with HA 'cables' indicating elevated tumorous potential under 2D culture conditions. For in vitro experimentation, scaffold based three dimensional niche modeling may have greater acceptance than conventional 2D culture condition. Thus, we have examined the influence of intrinsic properties of non-mulberry tropical tasar silk fibroin on the HepR21 cells in order to develop a 3D hepatocarcinoma construction to act as model. The scaffold of tasar silk fibroin of Antheraea mylitta when efficiently loaded with transformed hepatocarcinoma cells, HepR21; exhibits enhanced adhesiveness, viability, metabolic activity, proliferation and enlarged cellular morphology in 3D compared to its parent cell line HepG2, supporting the earlier observation made in 2D system. In addition, formation of multicellular aggregates, the indicator of tumor progression is also revealed in silk based 3D culture conditions. Further, the use of 4-MU (a hyaluronan synthase inhibitor) on HepR21 cells reduces the HA level and downregulates the expression of growth promoting factors like pAKT and PKC; while upregulating the expression of the tumor suppressor p53. Thus, 4-MU efficiently reduces the tumor potency associated with increased HA pool as well as HA cables and the effect of 4-MU doubling up as an anticancer agent in 2D and 3D are also comparable. The in vitro 3D multicellular model demonstrates the insight of hepatocarcinoma progression and offers the predictability of cellular response to transfection efficacy, drug treatment and therapeutic intervention. PMID:24016853

Kundu, Banani; Saha, Paramita; Datta, Kasturi; Kundu, Subhas C

2013-12-01

214

Silk Batik using Cochineal Dye  

Technology Transfer Automated Retrieval System (TEKTRAN)

The history of silk, including sericulture (the production of raw silk, which requires the raising of silkworms on their natural diet, mulberry leaves) and silk manufacturing, is rich and extensive. It encompasses several famous “silk roads” (trade routes), various cultures and technologies, ideas,...

215

Production of scFv-conjugated affinity silk film and its application to a novel enzyme-linked immunosorbent assay  

PubMed Central

Bombyx mori (silkworm) silk proteins have been utilized as unique biomaterials for various medical applications. To develop a novel affinity silk material, we generated a transgenic silkworm that spins silk protein containing the fibroin L-chain linked with the single-chain variable fragment (scFv) as a fusion protein. Previously, the scFv-conjugated “affinity” silk powder specifically immunoprecipitated its target protein, Wiskott-Aldrich syndrome protein. To expand the applicability of affinity silk materials, we processed the scFv-conjugated silk protein into a thin film by dissolving it in lithium bromide, then drying it in the wells of 96-well plates. Enzyme-linked immunosorbent assay demonstrated specific detection of Wiskott-Aldrich syndrome protein, both as a recombinant protein and in its native form extracted from mouse macrophages. These findings suggest that this scFv-conjugated silk film serves as the basis for an alternative immunodetection system. PMID:24518284

Sato, Mitsuru; Kojima, Katsura; Sakuma, Chisato; Murakami, Maria; Tamada, Yasushi; Kitani, Hiroshi

2014-01-01

216

Biofabrication of Cell-Loaded 3D Spider Silk Constructs.  

PubMed

Biofabrication is an emerging and rapidly expanding field of research in which additive manufacturing techniques in combination with cell printing are exploited to generate hierarchical tissue-like structures. Materials that combine printability with cytocompatibility, so called bioinks, are currently the biggest bottleneck. Since recombinant spider silk proteins are non-immunogenic, cytocompatible, and exhibit physical crosslinking, their potential as a new bioink system was evaluated. Cell-loaded spider silk constructs can be printed by robotic dispensing without the need for crosslinking additives or thickeners for mechanical stabilization. Cells are able to adhere and proliferate with good viability over at least one week in such spider silk scaffolds. Introduction of a cell-binding motif to the spider silk protein further enables fine-tuned control over cell-material interactions. Spider silk hydrogels are thus a highly attractive novel bioink for biofabrication. PMID:25640578

Schacht, Kristin; Jüngst, Tomasz; Schweinlin, Matthias; Ewald, Andrea; Groll, Jürgen; Scheibel, Thomas

2015-02-23

217

Safety evaluation of silk protein film (a novel wound healing agent) in terms of acute dermal toxicity, acute dermal irritation and skin sensitization.  

PubMed

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

218

Controlling silk fibroin particle features for drug delivery  

PubMed Central

Silk proteins are a promising material for drug delivery due to their aqueous processability, biocompatibility, and biodegradability. A simple aqueous preparation method for silk fibroin particles with controllable size, secondary structure and zeta potential is reported. The particles were produced by salting out a silk fibroin solution with potassium phosphate. The effect of ionic strength and pH of potassium phosphate solution on the yield and morphology of the particles was determined. Secondary structure and zeta potential of the silk particles could be controlled by pH. Particles produced by salting out with 1.25 M potassium phosphate pH 6 showed a dominating silk II (crystalline) structure whereas particles produced at pH 9 were mainly composed of silk I (less crystalline). The results show that silk I rich particles possess chemical and physical stability and secondary structure which remained unchanged during post treatments even upon exposure to 100% ethanol or methanol. A model is presented to explain the process of particle formation based on intra- and intermolecular interactions of the silk domains, influenced by pH and kosmotrope salts. The reported silk fibroin particles can be loaded with small molecule model drugs, such as alcian blue, rhodamine B, and crystal violet, by simple absorption based on electrostatic interactions. In vitro release of these compounds from the silk particles depends on charge – charge interactions between the compounds and the silk. With crystal violet we demonstrated that the release kinetics are dependent on the secondary structure of the particles. PMID:20219241

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

2010-01-01

219

An Unlikely Silk: The Composite Material of Green Lacewing Cocoons  

SciTech Connect

Spiders routinely produce multiple types of silk; however, common wisdom has held that insect species produce one type of silk each. This work reports that the green lacewing (Mallada signata, Neuroptera) produces two distinct classes of silk. We identified and sequenced the gene that encodes the major protein component of the larval lacewing cocoon silk and demonstrated that it is unrelated to the adult lacewing egg-stalk silk. The cocoon silk protein is 49 kDa in size and is alanine rich (>40%), and it contains an {alpha}-helical secondary structure. The final instar lacewing larvae spin protein fibers of {approx}2 {mu}m diameter to construct a loosely woven cocoon. In a second stage of cocoon construction, the insects lay down an inner wall of lipids that uses the fibers as a scaffold. We propose that the silk protein fibers provide the mechanical strength of the composite lacewing cocoon whereas the lipid layer provides a barrier to water loss during pupation.

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

2009-01-15

220

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

PubMed

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

221

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

222

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

223

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

224

Lyophilized Silk Fibroin Hydrogels for the Sustained Local Delivery of Therapeutic Monoclonal Antibodies  

PubMed Central

The development of sustained delivery systems compatible with protein therapeutics continues to be a significant unmet need. A lyophilized silk fibroin hydrogel matrix (lyogel) for the sustained release of pharmaceutically relevant monoclonal antibodies is described. Sonication of silk fibroin prior to antibody incorporation avoids exposing the antibody to the sol-gel transition inducing shear stress. Fourier Transform Infrared (FTIR) analysis showed no change in silk structural composition between hydrogel and lyogel or with increasing silk fibroin concentration. Antibody release from hydrogels occurred rapidly over 10 days regardless of silk concentration. Upon lyophilization, sustained antibody release was observed over 38 days from lyogels containing 6.2% (w/w) silk fibroin and above. In 3.2% (w/w) silk lyogels, antibody release was comparable to hydrogels. Swelling properties of lyogels followed a similar threshold behavior. Lyogels at 3.2% (w/w) silk recovered approximately 90% of their fluid mass upon rehydration, while approximately 50% fluid recovery was observed at 6.2% (w/w) silk and above. Antibody release was primarily governed by hydrophobic/hydrophilic silk-antibody interactions and secondarily altered by the hydration resistance of the lyogel. Hydration resistance was controlled by altering ?-sheet (crystalline) density of the matrix. The antibody released from lyogels maintained biological activity. Silk lyogels offer an advantage as a delivery matrix over other hydrogel materials for the slow release of the loaded protein, making lyogels suitable for long-term sustained release applications. PMID:21216004

Guziewicz, Nicholas; Best, Annie; Perez-Ramirez, Bernardo; Kaplan, David L.

2011-01-01

225

Molecular and Mechanical Characterization of Aciniform Silk: Uniformity of Iterated Sequence Modules in a Novel Member of the  

E-print Network

Molecular and Mechanical Characterization of Aciniform Silk: Uniformity of Iterated Sequence Modules in a Novel Member of the Spider Silk Fibroin Gene Family Cheryl Y. Hayashi,* Todd A. Blackledge to seven different protein-based silks/glues that have diverse physical properties. The fibroin sequences

Blackledge, Todd

226

Evaluation of sericin as a fetal bovine serum-replacing cryoprotectant during freezing of human mesenchymal stromal cells and human osteoblast-like cells.  

PubMed

A reliable, cryoprotective, xeno-free medium suitable for different cell types is highly desirable in regenerative medicine. There is danger of infection or allergic reaction with the use of fetal bovine serum (FBS), making it problematic for medical applications. The aim of the present study was to develop an FBS-free cryoprotective medium for human mesenchymal stromal cells (hMSCs; primary cells) and immortalized human osteoblasts (SAOS-2 cell line). Furthermore, we endeavored to eliminate or reduce the presence of dimethyl sulfoxide (DMSO) in the medium. Sericin, a sticky protein derived from the silkworm cocoon, was investigated as a substitute for FBS and DMSO in the freezing medium. Cell viability (24 hours after thawing, both hMSC and SAOS-2) and colony-forming ability (2 weeks after thawing, only for hMSCs) were both determined. The FBS-free medium with 1% sericin in 10% DMSO was found to be a suitable freezing medium for primary hMSCs, in contrast to immortalized human osteoblasts. Surprisingly, the storage of hMSCs in a cultivation medium with only 10% DMSO also provided satisfactory results. Any drop in DMSO concentration led to significantly worse survival of cells, with little improvement in hMSC survival in the presence of sericin. Thus, sericin may substitute for FBS in the freezing medium for primary hMSCs, but cannot substitute for DMSO. PMID:24749876

Verdanova, Martina; Pytlik, Robert; Kalbacova, Marie Hubalek

2014-04-01

227

Mandibular repair in rats with premineralized silk scaffolds and BMP2-modified bMSCs  

Microsoft Academic Search

Premineralized silk fibroin protein scaffolds (mSS) were prepared to combine the osteoconductive properties of biological apatite with aqueous-derived silk scaffold (SS) as a composite scaffold for bone regeneration. The aim of present study was to evaluate the effect of premineralized silk scaffolds combined with bone morphogenetic protein-2 (BMP-2) modified bone marrow stromal cells (bMSCs) to repair mandibular bony defects in

Xinquan Jiang; Jun Zhao; Shaoyi Wang; Xiaojuan Sun; Xiuli Zhang; Jake Chen; David L. Kaplan; Zhiyuan Zhang

2009-01-01

228

Effect of sequence features on assembly of spider silk block copolymers.  

PubMed

Bioengineered spider silk block copolymers were studied to understand the effect of protein chain length and sequence chemistry on the formation of secondary structure and materials assembly. Using a combination of in vitro protein design and assembly studies, we demonstrate that silk block copolymers possessing multiple repetitive units self-assemble into lamellar microstructures. Additionally, the study provides insights into the assembly behavior of spider silk block copolymers in concentrated salt solutions. PMID:24613991

Tokareva, Olena S; Lin, Shangchao; Jacobsen, Matthew M; Huang, Wenwen; Rizzo, Daniel; Li, David; Simon, Marc; Staii, Cristian; Cebe, Peggy; Wong, Joyce Y; Buehler, Markus J; Kaplan, David L

2014-06-01

229

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

230

Rapid communication: Computational simulation and analysis of a candidate for the design of a novel silk-based biopolymer.  

PubMed

This work theoretically investigates the mechanical properties of a novel silk-derived biopolymer as polymerized in silico from sericin and elastin-like monomers. Molecular Dynamics simulations and Steered Molecular Dynamics were the principal computational methods used, the latter of which applies an external force onto the system and thereby enables an observation of its response to stress. The models explored herein are single-molecule approximations, and primarily serve as tools in a rational design process for the preliminary assessment of properties in a new material candidate. © 2014 Wiley Periodicals, Inc. Biopolymers 101: 915-923, 2014. PMID:24723330

Golas, Ewa I; Czaplewski, Cezary

2014-09-01

231

A three-dimensional multiporous fibrous scaffold fabricated with regenerated spider silk protein/poly(l-lactic acid) for tissue engineering.  

PubMed

An axially aligned three-dimensional (3-D) fibrous scaffold was fabricated with regenerated spider silk protein (RSSP)/poly (l-lactic acid) (PLLA) through electrospinning and post treatment. The morphology, mechanical and degradation properties of the scaffold were controlled through the weight ratio of RSSP to PLLA, the thickness of the scaffold and the treatment time. The scaffold with a weight ratio of 2:3 (RSSP:PLLA) had a nanoleaves-on-nanofibers hierarchical nanostructure; the length and thickness of the nanoleaves were about 400 and 30 nm, respectively. The holes of the scaffolds ranged from hundreds of nanometers to several microns. The scaffold showed an ideal mechanical property that it was stiff when dry, but became soft once hydrated in the culture medium. Its degradation rate was very slow in the first 2 months, and then accelerated in the following 2 months. The pH values of the degradation mediums of all the samples remained in the range of 7.40-7.12 during degradation for 6 months. It had good biocompatibility with PC 12 cells. The aligned hierarchical nanostructure could guide the directions of the axon extension. This scaffold has a potential application in Tissue Engineering and controlled release. This study provides a method to produce synthetic or natural biodegradable polymer scaffold with tailored morphology, mechanical, and degradation properties. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 721-729, 2015. PMID:24825592

Yu, Qiaozhen; Sun, Chengjun

2015-02-01

232

An Australian webspinner species makes the finest known insect silk fibers  

SciTech Connect

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

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

2009-01-15

233

Sericins exhibit ROS-scavenging, anti-tyrosinase, anti-elastase, and in vitro immunomodulatory activities.  

PubMed

Some biological properties of Bombyx mori sericins from twenty strains were investigated, fourteen fed with artificial diet, two with fresh mulberry leaves and four with both diets. Sericin exhibited ROS-scavenging, anti-tyrosinase and anti-elastase properties, the strain significantly influenced these properties, while diet only influenced the anti-tyrosinase activity. Sericins were clustered into 5 groups and one sericin from each group was further studied: sericins showed anti-proliferative activity on in vitro stimulated peripheral blood mononuclear cells; some strains decreased in vitro secretion of IFN?, while no effects were observed on TNF? and IL10 release. Therefore, a mixture of sericins extracted from the most promising strains may be useful for dermatological and cosmetic use. PMID:23541552

Chlapanidas, Theodora; Faragò, Silvio; Lucconi, Giulia; Perteghella, Sara; Galuzzi, Marta; Mantelli, Melissa; Avanzini, Maria Antonietta; Tosca, Marta Cecilia; Marazzi, Mario; Vigo, Daniele; Torre, Maria Luisa; Faustini, Massimo

2013-07-01

234

Inhibitory effect of corn silk on skin pigmentation.  

PubMed

In this study, the inhibitory effect of corn silk on melanin production was evaluated. This study was performed to investigate the inhibitory effect of corn silk on melanin production in Melan-A cells by measuring melanin production and protein expression. The corn silk extract applied on Melan-A cells at a concentration of 100 ppm decreased melanin production by 37.2% without cytotoxicity. This was a better result than arbutin, a positive whitening agent, which exhibited a 26.8% melanin production inhibitory effect at the same concentration. The corn silk extract did not suppress tyrosinase activity but greatly reduced the expression of tyrosinase in Melan-A cells. In addition, corn silk extract was applied to the human face with hyperpigmentation, and skin color was measured to examine the degree of skin pigment reduction. The application of corn silk extract on faces with hyperpigmentation significantly reduced skin pigmentation without abnormal reactions. Based on the results above, corn silk has good prospects for use as a material for suppressing skin pigmentation. PMID:24595276

Choi, Sang Yoon; Lee, Yeonmi; Kim, Sung Soo; Ju, Hyun Min; Baek, Ji Hwoon; Park, Chul-Soo; Lee, Dong-Hyuk

2014-01-01

235

Spider genomes provide insight into composition and evolution of venom and silk  

PubMed Central

Spiders are ecologically important predators with complex venom and extraordinarily tough silk that enables capture of large prey. Here we present the assembled genome of the social velvet spider and a draft assembly of the tarantula genome that represent two major taxonomic groups of spiders. The spider genomes are large with short exons and long introns, reminiscent of mammalian genomes. Phylogenetic analyses place spiders and ticks as sister groups supporting polyphyly of the Acari. Complex sets of venom and silk genes/proteins are identified. We find that venom genes evolved by sequential duplication, and that the toxic effect of venom is most likely activated by proteases present in the venom. The set of silk genes reveals a highly dynamic gene evolution, new types of silk genes and proteins, and a novel use of aciniform silk. These insights create new opportunities for pharmacological applications of venom and biomaterial applications of silk. PMID:24801114

Sanggaard, Kristian W.; Bechsgaard, Jesper S.; Fang, Xiaodong; Duan, Jinjie; Dyrlund, Thomas F.; Gupta, Vikas; Jiang, Xuanting; Cheng, Ling; Fan, Dingding; Feng, Yue; Han, Lijuan; Huang, Zhiyong; Wu, Zongze; Liao, Li; Settepani, Virginia; Thøgersen, Ida B.; Vanthournout, Bram; Wang, Tobias; Zhu, Yabing; Funch, Peter; Enghild, Jan J.; Schauser, Leif; Andersen, Stig U.; Villesen, Palle; Schierup, Mikkel H; Bilde, Trine; Wang, Jun

2014-01-01

236

Structural Properties of Silk Electro-Gels  

NASA Astrophysics Data System (ADS)

The interest in Bombyx Mori silk emerges from its biocompatibility and its structural superiority to synthetic polymers. Our particular interest lies in understanding the capabilities of silk electro-gels because of their reversibility and tunable adhesion. We create an electro-gel by applying a DC electric potential across a reconstituted silk fibroin solution derived directly from Bombyx Mori cocoons. This process leads to the intermolecular self-assembly of fibroin proteins into a weak gel. In this talk we will present our results on the effects of applied shear on electro-gels. We quantify the structural properties while dynamically imaging shear induced fiber formation; known as fibrillogenesis. It is observed that the mechanical properties and microstructure of these materials are highly dependent on shear history. We will also discuss the role of surface modification, through micro-patterning, on the observed gel structure. Our results provide an understanding of both the viscoelastiticity and microstucture of reconstituted silks that are being utilized as tissue scaffolds.

Tabatabai, A. P.; Urbach, J. S.; Blair, D. L.; Kaplan, D. L.

2013-03-01

237

The critical role of water in spider silk and its consequence for protein Cameron P. Brown,*abcd  

E-print Network

to the presence of proline, which twists away from simple, ordered configurations.3 This combination of proteins, Canada. E-mail: rosei@emt.inrs. ca d Italy-Quebec Joint Laboratory in Nanostructured Materials for Energy

238

Optically switchable natural silk  

NASA Astrophysics Data System (ADS)

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

Krasnov, Igor; Krekiehn, Nicolai R.; Krywka, Christina; Jung, Ulrich; Zillohu, Ahnaf U.; Strunskus, Thomas; Elbahri, Mady; Magnussen, Olaf M.; Müller, Martin

2015-03-01

239

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

240

The elaborate structure of spider silk: structure and function of a natural high performance fiber.  

PubMed

Biomaterials, having evolved over millions of years, often exceed man-made materials in their properties. Spider silk is one outstanding fibrous biomaterial which consists almost entirely of large proteins. Silk fibers have tensile strengths comparable to steel and some silks are nearly as elastic as rubber on a weight to weight basis. In combining these two properties, silks reveal a toughness that is two to three times that of synthetic fibers like Nylon or Kevlar. Spider silk is also antimicrobial, hypoallergenic and completely biodegradable. This article focuses on the structure-function relationship of the characterized highly repetitive spider silk spidroins and their conformational conversion from solution into fibers. Such knowedge is of crucial importance to understanding the intrinsic properties of spider silk and to get insight into the sophisticated assembly processes of silk proteins. This review further outlines recent progress in recombinant production of spider silk proteins and their assembly into distinct polymer materials as a basis for novel products. PMID:19221522

Römer, Lin; Scheibel, Thomas

2008-01-01

241

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

2005-01-24

242

Stabilization of Enzymes in Silk Films  

PubMed Central

Material systems are needed that promote stabilization of entrained molecules, such as enzymes or therapeutic proteins, without destroying their activity. We demonstrate that the unique structure of silk fibroin protein, when assembled into the solid state, establishes an environment that is conducive to the stabilization of entrained proteins. Enzymes (glucose oxidase, lipase and horseradish peroxidase) entrapped in these films over ten months retained significant activity, even when stored at 37°C, and in the case of glucose oxidase did not lose any activity. Further, the mode of processing of the silk protein into the films could be correlated to the stability of the enzymes. The relationship between processing and stability offers a large suite of conditions within which to optimize such stabilization processes. Overall, the techniques reported here result in materials that stabilize enzymes to a remarkable extent, without the need for cryoprotectants, emulsifiers, covalent immobilization or other treatments. Further, these systems are amenable to optical characterization, environmental distribution without refrigeration, are ingestible, and offer potential use in vivo, since silk materials are biocompatible and FDA approved, degradable with proteases and currently used in biomedical devices. PMID:19323497

Lu, Shenzhou; Wang, Xiaoqin; Lv, Qiang; Hu, Xiao; Uppal, Neha; Omenetto, Fiorenzo

2009-01-01

243

Hypotensive and vasorelaxant effects of sericin-derived oligopeptides in rats.  

PubMed

Sericin-derived oligopeptides obtained from silk cocoons were investigated for the in vivo hypotensive effect and investigated for the underlying mechanism involved in vasodilation in isolated rat thoracic aorta. In normotensive anesthetized rats, oligopeptides induced an immediate and transient hypotensive activity. In rat aortic rings, oligopeptides induced a concentration-dependent vasorelaxation in vessels precontracted with both KCl and phenylephrine (PE) with endothelium-intact or endothelium-denuded rings. In endothelium-intact rings, pretreatment with N ? -Nitro-L-arginine methyl ester hydrochloride (L-NAME, 100?µM), an inhibitor of the NO synthase (NOS) or 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ, 1?µM), a selective inhibitor of the guanylyl cyclase enzyme, significantly reduced the relaxant effect of oligopeptides. However, indomethacin, an inhibitor of the cyclooxygenase, had no effect on oligopeptides-induced relaxation. In addition, pretreatment with tetraethylammonium (TEA, 5?mM) reduced the maximal relaxant effect induced by oligopeptides. By contrast, relaxation was not affected by 4-aminopyridine (4-AP, 1?mM), glibenclamide (10?µM), or barium chloride (BaCl2, 1?mM). In depolarization Ca(2+)-free solution, oligopeptides inhibited calcium chloride- (CaCl2-) induced contraction in endothelium-denuded rings in a concentration-dependent manner. Nevertheless, oligopeptides attenuated transient contractions in Ca(2+)-free medium containing EGTA (1?mM) induced by 1?µM PE, but they were not affected by 20?mM caffeine. It is obvious that potent vasodilation effect of oligopeptides is mediated through both the endothelium and the vascular smooth muscle. PMID:24312733

Onsa-Ard, Amnart; Shimbhu, Dawan; Tocharus, Jiraporn; Sutheerawattananonda, Manote; Pantan, Rungusa; Tocharus, Chainarong

2013-01-01

244

Hypotensive and Vasorelaxant Effects of Sericin-Derived Oligopeptides in Rats  

PubMed Central

Sericin-derived oligopeptides obtained from silk cocoons were investigated for the in vivo hypotensive effect and investigated for the underlying mechanism involved in vasodilation in isolated rat thoracic aorta. In normotensive anesthetized rats, oligopeptides induced an immediate and transient hypotensive activity. In rat aortic rings, oligopeptides induced a concentration-dependent vasorelaxation in vessels precontracted with both KCl and phenylephrine (PE) with endothelium-intact or endothelium-denuded rings. In endothelium-intact rings, pretreatment with N?-Nitro-L-arginine methyl ester hydrochloride (L-NAME, 100?µM), an inhibitor of the NO synthase (NOS) or 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ, 1?µM), a selective inhibitor of the guanylyl cyclase enzyme, significantly reduced the relaxant effect of oligopeptides. However, indomethacin, an inhibitor of the cyclooxygenase, had no effect on oligopeptides-induced relaxation. In addition, pretreatment with tetraethylammonium (TEA, 5?mM) reduced the maximal relaxant effect induced by oligopeptides. By contrast, relaxation was not affected by 4-aminopyridine (4-AP, 1?mM), glibenclamide (10?µM), or barium chloride (BaCl2, 1?mM). In depolarization Ca2+-free solution, oligopeptides inhibited calcium chloride- (CaCl2-) induced contraction in endothelium-denuded rings in a concentration-dependent manner. Nevertheless, oligopeptides attenuated transient contractions in Ca2+-free medium containing EGTA (1?mM) induced by 1?µM PE, but they were not affected by 20?mM caffeine. It is obvious that potent vasodilation effect of oligopeptides is mediated through both the endothelium and the vascular smooth muscle. PMID:24312733

Pantan, Rungusa; Tocharus, Chainarong

2013-01-01

245

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

246

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

247

Electrospun Silk Biomaterial Scaffolds for Regenerative Medicine  

PubMed Central

Electrospinning is a versatile technique that enables the development of nanofiber-based biomaterial scaffolds. Scaffolds can be generated that are useful for tissue engineering and regenerative medicine since they mimic the nanoscale properties of certain fibrous components of the native extracellular matrix in tissues. Silk is a natural protein with excellent biocompatibility, remarkable mechanical properties as well as tailorable degradability. Integrating these protein polymer advantages with electrospinning results in scaffolds with combined biochemical, topographical and mechanical cues with versatility for a range of biomaterial, cell and tissue studies and applications. This review covers research related to electrospinning of silk, including process parameters, post treatment of the spun fibers, functionalization of nanofibers, and the potential applications for these material systems in regenerative medicine. Research challenges and future trends are also discussed. PMID:19643154

Zhang, Xiaohui; Reagan, Michaela R; Kaplan, David L.

2009-01-01

248

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

249

Spider silk reduces insect herbivory  

PubMed Central

The role of predators in food webs extends beyond their ability to kill and consume prey. Such trait-mediated effects occur when signals of the predator influence the behaviour of other animals. Because all spiders are silk-producing carnivores, we hypothesized that silk alone would signal other arthropods and enhance non-lethal effects of spiders. We quantified the herbivory inflicted by two beetle species on green bean plants (Phaseolus vulgaris) in the presence of silkworm silk and spider silk along with no silk controls. Single leaflets were treated and enclosed with herbivores in the laboratory and field. Another set of leaflets were treated and left to experience natural herbivory in the field. Entire plants in the field were treated with silk and enclosed with herbivores or left exposed to herbivory. In all cases, the lowest levels of herbivory occurred with spider silk treatments and, in general, silkworm silk produced intermediate levels of leaf damage. These results suggest that silk may be a mechanism for the trait-mediated impacts of spiders and that it might contribute to integrated pest management programmes. PMID:23193048

Rypstra, Ann L.; Buddle, Christopher M.

2013-01-01

250

Spider silk reduces insect herbivory.  

PubMed

The role of predators in food webs extends beyond their ability to kill and consume prey. Such trait-mediated effects occur when signals of the predator influence the behaviour of other animals. Because all spiders are silk-producing carnivores, we hypothesized that silk alone would signal other arthropods and enhance non-lethal effects of spiders. We quantified the herbivory inflicted by two beetle species on green bean plants (Phaseolus vulgaris) in the presence of silkworm silk and spider silk along with no silk controls. Single leaflets were treated and enclosed with herbivores in the laboratory and field. Another set of leaflets were treated and left to experience natural herbivory in the field. Entire plants in the field were treated with silk and enclosed with herbivores or left exposed to herbivory. In all cases, the lowest levels of herbivory occurred with spider silk treatments and, in general, silkworm silk produced intermediate levels of leaf damage. These results suggest that silk may be a mechanism for the trait-mediated impacts of spiders and that it might contribute to integrated pest management programmes. PMID:23193048

Rypstra, Ann L; Buddle, Christopher M

2013-02-23

251

Ingrowth of Human Mesenchymal Stem Cells into Porous Silk Particle Reinforced Silk Composite Scaffolds: An In Vitro Study  

PubMed Central

Silk fibroin protein is biodegradable and biocompatible, exhibiting excellent mechanical properties for various biomedical applications. However, porous 3D silk fibroin scaffolds, or silk sponges, usually fall short in matching the initial mechanical requirements for bone tissue engineering. In the present study, silk sponge matrices were reinforced with silk microparticles to generate protein-protein composite scaffolds with desirable mechanical properties for in vitro osteogenic tissue formation. It was found that increasing the silk microparticle loading led to a substantial increase in the scaffold compressive modulus from 0.3 MPa (nonreinforced) to 1.9 MPa for 1:2 (matrix:particle) reinforcement loading by dry mass. Biochemical, gene expression, and histological assays were employed to study the possible effects of increasing composite scaffold stiffness, due to microparticle reinforcement, on in vitro osteogenic differentiation of human mesenchymal stem cells (hMSCs). Increasing silk microparticle loading increased the osteogenic capability of hMSCs in the presence of bone morphogenic protein-2 (BMP-2) and other osteogenic factors in static culture for up to six weeks. The calcium adsorption increased dramatically with increasing loading, as observed from biochemical assays, histological staining, and microCT (?CT) analysis. Specifically, calcium content in the scaffolds increased by 0.57, 0.71, and 1.27 mg (per ?g of DNA) from 3 to 6 weeks for matrix to particle dry mass loading ratios of 1:0, 1:1 and 1:2, respectively. In addition, ?CT imaging revealed that at 6 weeks, bone volume fraction increased from 0.78% for nonreinforced to 7.1% and 6.7% for 1:1 and 1:2 loading, respectively. Our results support the hypothesis that scaffold stiffness may strongly influence the 3D in vitro differentiation capabilities of hMSCs, providing a means to improve osteogenic outcomes. PMID:20656075

Rockwood, Danielle N.; Gil, Eun Seok; Park, Sang-Hyug; Kluge, Jonathan A.; Grayson, Warren; Bhumiratana, Sarindr; Rajkhowa, Rangam; Wang, Xungai; Kim, Sung Jun; Vunjak-Novakovic, Gordana; Kaplan, David L

2010-01-01

252

Relationships between supercontraction and mechanical properties of spider silk  

NASA Astrophysics Data System (ADS)

Typical spider dragline silk tends to outperform other natural fibres and most man-made filaments. However, even small changes in spinning conditions can have large effects on the mechanical properties of a silk fibre as well as on its water uptake. Absorbed water leads to significant shrinkage in an unrestrained dragline fibre and reversibly converts the material into a rubber. This process is known as supercontraction and may be a functional adaptation for the silk's role in the spider's web. Supercontraction is thought to be controlled by specific motifs in the silk proteins and to be induced by the entropy-driven recoiling of molecular chains. In analogy, in man-made fibres thermal shrinkage induces changes in mechanical properties attributable to the entropy-driven disorientation of `unfrozen' molecular chains (as in polyethylene terephthalate) or the `broken' intermolecular hydrogen bonds (as in nylons). Here we show for Nephila major-ampullate silk how in a biological fibre the spinning conditions affect the interplay between shrinkage and mechanical characteristics. This interaction reveals design principles linking the exceptional properties of silk to its molecular orientation.

Liu, Yi; Shao, Zhengzhong; Vollrath, Fritz

2005-12-01

253

Transdermal Delivery Devices: Fabrication, Mechanics and Drug Release from Silk**  

PubMed Central

Microneedles are a relatively simple, minimally invasive and painless approach to deliver drugs across the skin. However, there remain limitations with this approach because of the materials most commonly utilized for such systems. Silk protein, with tunable and biocompatibility properties, is a useful biomaterial to overcome the current limitations with microneedles. Silk devices preserve drug activity, offer superior mechanical properties and biocompatibility, can be tuned for biodegradability, and can be processed under aqueous, benign conditions. In the present work, we report the fabrication of dense microneedle arrays from silk with different drug release kinetics. The mechanical properties of the microneedle patches are tuned by post-fabrication treatments or by loading the needles with silk microparticles to increase capacity and mechanical strength. Drug release is further enhanced by the encapsulation of the drugs in the silk matrix and coating with a thin dissolvable drug layer. The microneedles are used on human cadaver skin and drugs were delivered successfully. The various attributes demonstrated suggest that silk-based microneedle devices can provide significant benefit as a platform material for transdermal drug delivery. PMID:23653252

Raja, Waseem K.; MacCorkle, Scott; Diwan, Izzuddin M.; Abdurrob, Abdurrahman; Lu, Jessica; Omenetto, Fiorenzo G.; Kaplan, David L.

2013-01-01

254

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

255

21 CFR 184.1262 - Corn silk and corn silk extract.  

Code of Federal Regulations, 2012 CFR

...Drugs 3 2012-04-01 2012-04-01 false Corn silk and corn silk extract. 184.1262 Section 184.1262 Food...Specific Substances Affirmed as GRAS § 184.1262 Corn silk and corn silk extract. (a) Corn silk is...

2012-04-01

256

21 CFR 184.1262 - Corn silk and corn silk extract.  

Code of Federal Regulations, 2010 CFR

...Drugs 3 2010-04-01 2009-04-01 true Corn silk and corn silk extract. 184.1262 Section 184.1262 Food...Specific Substances Affirmed as GRAS § 184.1262 Corn silk and corn silk extract. (a) Corn silk is...

2010-04-01

257

21 CFR 184.1262 - Corn silk and corn silk extract.  

Code of Federal Regulations, 2011 CFR

...Drugs 3 2011-04-01 2011-04-01 false Corn silk and corn silk extract. 184.1262 Section 184.1262 Food...Specific Substances Affirmed as GRAS § 184.1262 Corn silk and corn silk extract. (a) Corn silk is...

2011-04-01

258

21 CFR 184.1262 - Corn silk and corn silk extract.  

Code of Federal Regulations, 2014 CFR

...Drugs 3 2014-04-01 2014-04-01 false Corn silk and corn silk extract. 184.1262 Section 184.1262 Food...Specific Substances Affirmed as GRAS § 184.1262 Corn silk and corn silk extract. (a) Corn silk is...

2014-04-01

259

21 CFR 184.1262 - Corn silk and corn silk extract.  

Code of Federal Regulations, 2013 CFR

...Drugs 3 2013-04-01 2013-04-01 false Corn silk and corn silk extract. 184.1262 Section 184.1262 Food...Specific Substances Affirmed as GRAS § 184.1262 Corn silk and corn silk extract. (a) Corn silk is...

2013-04-01

260

Tuning chemical and physical cross-links in silk electrogels for morphological analysis and mechanical reinforcement.  

PubMed

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 cross-linking as well as thermal, mechanical, and morphological stabilities. In the present work, the tuning of covalent and physical ?-sheet cross-links 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-cross-linking 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 cross-links. The selective incorporation of either chemical or physical or both cross-links 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

2013-08-12

261

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

262

Preparation of hexagonal GeO? 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

2014-11-28

263

Synthetic Spider Silk Production on a Laboratory Scale  

PubMed Central

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

264

Molecular dynamics of spider dragline silk fiber investigated by (2)h MAS NMR.  

PubMed

The molecular dynamics of the proteins that comprise spider dragline silk were investigated with solid-state (2)H magic angle spinning (MAS) NMR line shape and spin-lattice relaxation time (T1) analysis. The experiments were performed on (2)H/(13)C/(15)N-enriched N. clavipes dragline silk fibers. The silk protein side-chain and backbone dynamics were probed for Ala-rich regions (?-sheet and 31-helical domains) in both native (dry) and supercontracted (wet) spider silk. In native (dry) silk fibers, the side chains in all Ala containing regions undergo similar fast methyl rotations (>10(9) s(-1)), while the backbone remains essentially static (<10(2) s(-1)). When the silk is wet and supercontracted, the presence of water initiates fast side-chain and backbone motions for a fraction of the ?-sheet region and 31-helicies. ?-Sheet subregion 1 ascribed to the poly(Ala) core exhibits slower dynamics, while ?-sheet subregion 2 present in the interfacial, primarily poly(Gly-Ala) region that links the ?-sheets to disordered 31-helical motifs, exhibits faster motions when the silk is supercontracted. Particularly notable is the observation of microsecond backbone motions for ?-sheet subregion 2 and 31-helicies. It is proposed that these microsecond backbone motions lead to hydrogen-bond disruption in ?-sheet subregion 2 and helps to explain the decrease in silk stiffness when the silk is wet and supercontracted. In addition, water mobilizes and softens 31-helical motifs, contributing to the increased extensibility observed when the silk is in a supercontracted state. The present study provides critical insight into the supercontraction mechanism and corresponding changes in mechanical properties observed for spider dragline silks. PMID:25619304

Shi, Xiangyan; Holland, Gregory P; Yarger, Jeffery L

2015-03-01

265

Effect of sericin supplementation during in vitro maturation on the maturation, fertilization and development of porcine oocytes.  

PubMed

This study aimed to examine the effects of sericin supplementation during in vitro oocyte maturation on the nuclear maturation, fertilization and development of porcine oocytes. Cumulus-oocyte complexes (COCs) were cultured in maturation medium supplemented with 0 (control), 0.1, 0.5, 1.0, 2.5 or 5.0% sericin and were then subjected to in vitro fertilization and embryo culture. More COCs matured with 1.0% sericin underwent germinal vesicle breakdown and reached metaphase II compared with the control COCs matured without sericin (p < 0.01). The proportions of oocytes with DNA-fragmented nuclei did not differ between the groups, regardless of the sericin level. The total fertilization rate of oocytes matured with 1.0% sericin was higher (p < 0.05) than that of oocytes matured with 0.1%, 2.5% and 5.0% sericin. Supplementation with more than 1.0% sericin decreased the DNA fragmentation index of the blastocysts compared with the control group (p < 0.05). However, the supplementation of the maturation medium with sericin had no beneficial effects on the cleavage, development to the blastocyst stage and the total cell number of the embryos. Our findings indicate that supplementation with 1.0% sericin during maturation culture may improve the nuclear maturation and the quality of the embryos but does not affect blastocyst formation. PMID:24467637

Do, L T K; Namula, Z; Luu, V V; Sato, Y; Taniguchi, M; Isobe, T; Kikuchi, K; Otoi, T

2014-04-01

266

Silk electrogel coatings for titanium dental implants.  

PubMed

The aim of this study was to develop biocompatible, biodegradable dental implant coatings capable of withstanding the mechanical stresses imparted during implant placement. Two techniques were developed to deposit uniform silk fibroin protein coatings onto dental implants. Two novel coating techniques were implemented to coat titanium shims, studs, and implants. One technique involved electrodeposition of the silk directly onto the titanium substrates. The second technique consisted of melting electrogels and dispensing the melted gels onto the titanium to form the coatings. Both techniques were tested for coating reproducibility using a stylus profilometer and a dial thickness gauge. The mechanical strength of adhered titanium studs was assessed using a universal mechanical testing machine. Uniform, controllable coatings were obtained from both the electrodeposition and melted electrogel coating techniques, tunable from 35 to 1654?µm thick under the conditions studied, and able to withstand delamination during implantation into implant socket mimics. Mechanical testing revealed that the adhesive strength of electrogel coatings, 0.369?±?0.09?MPa, rivaled other biologically derived coating systems such as collagen, hydroxyapatite, and chitosan (0.07-4.83?MPa). These novel silk-based techniques offer a unique approach to the deposition of safe, simple, mechanically robust, biocompatible, and degradable implant coatings. PMID:25425563

Elia, Roberto; Michelson, Courtney D; Perera, Austin L; Harsono, Masly; Leisk, Gray G; Kugel, Gerard; Kaplan, David L

2015-04-01

267

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

268

Immunolocalization of pheromone-binding protein and general odorant-binding protein in olfactory sensilla of the silk moths Antheraea and Bombyx  

Microsoft Academic Search

The distribution of odorant-binding proteins among olfactory sensilla of three moth species was studied by immuno-electron microscopy. Two polyclonal antisera were used in a post-embedding labelling protocol on sections of cryo-substituted antennae. The first was directed against the pheromone-binding protein (PBP) of Antheraea polyphemus, the second against the general odorant-binding protein (GOBP) of the same species. Immunoblots showed that these

R. A. Steinbrecht; M. Laue; G. Ziegelberger

1995-01-01

269

Nutrient Deprivation Induces Property Variations in Spider Gluey Silk  

PubMed Central

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

270

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

271

Increasing silk fibre strength through heterogeneity of bundled fibrils  

PubMed Central

Can naturally arising disorder in biological materials be beneficial? Materials scientists are continuously attempting to replicate the exemplary performance of materials such as spider silk, with detailed techniques and assembly procedures. At the same time, a spider does not precisely machine silk—imaging indicates that its fibrils are heterogeneous and irregular in cross section. While past investigations either focused on the building material (e.g. the molecular scale protein sequence and behaviour) or on the ultimate structural component (e.g. silk threads and spider webs), the bundled structure of fibrils that compose spider threads has been frequently overlooked. Herein, I exploit a molecular dynamics-based coarse-grain model to construct a fully three-dimensional fibril bundle, with a length on the order of micrometres. I probe the mechanical behaviour of bundled silk fibrils with variable density of heterogenic protrusions or globules, ranging from ideally homogeneous to a saturated distribution. Subject to stretching, the model indicates that cooperativity is enhanced by contact through low-force deformation and shear ‘locking’ between globules, increasing shear stress transfer by up to 200 per cent. In effect, introduction of a random and disordered structure can serve to improve mechanical performance. Moreover, addition of globules allows a tuning of free volume, and thus the wettability of silk (with implications for supercontraction). These findings support the ability of silk to maintain near-molecular-level strength at the scale of silk threads, and the mechanism could be easily adopted as a strategy for synthetic fibres. PMID:23486175

Cranford, Steven W.

2013-01-01

272

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

273

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

274

Neural Responses to Electrical Stimulation on Patterned Silk Films  

PubMed Central

Peripheral nerve injury is a critical issue for trauma patients. Following injury, incomplete axon regeneration or misguided axon innervation into tissue will result in loss of sensory and motor functions. The objective of this study was to examine axon outgrowth and axon alignment in response to surface patterning and electrical stimulation. To accomplish our objective, metal electrodes with dimensions of 1.5 mm × 4 cm, were sputter coated onto micropatterned silk protein films, with surface grooves 3.5 ?m wide × 500 nm deep. P19 neurons were seeded on the patterned electronic silk films and stimulated at 120 mV, 1 kHz, for 45 minutes each day for 7 days. Responses were compared to neurons on flat electronic silk films, patterned silk films without stimulation, and flat silk films without stimulation. Significant alignment was found on the patterned film groups compared to the flat film groups. Axon outgrowth was greater (p < 0.05) on electronic films on day 5 and day 7 compared to the unstimulated groups. In conclusion, electrical stimulation, at 120 mV, 1 kHz, for 45 minutes daily, in addition to surface patterning, of 3.5 ?m wide × 500 nm deep grooves, offered control of nerve axon outgrowth and alignment. PMID:23401351

Hronik-Tupaj, Marie; Raja, Waseem Khan; Tang-Schomer, Min; Omenetto, Fiorenzo G.; Kaplan, David L.

2013-01-01

275

Neural responses to electrical stimulation on patterned silk films.  

PubMed

Peripheral nerve injury is a critical issue for patients with trauma. Following injury, incomplete axon regeneration or misguided axon innervation into tissue will result in loss of sensory and motor functions. The objective of this study was to examine axon outgrowth and axon alignment in response to surface patterning and electrical stimulation. To accomplish our objective, metal electrodes with dimensions of 1.5 mm × 4 cm, were sputter coated onto micropatterned silk protein films, with surface grooves 3.5 ?m wide × 500 nm deep. P19 neurons were seeded on the patterned electronic silk films and stimulated at 120 mV, 1 kHz, for 45 min each day for 7 days. Responses were compared with neurons on flat electronic silk films, patterned silk films without stimulation, and flat silk films without stimulation. Significant alignment was found on the patterned film groups compared with the flat film groups. Axon outgrowth was greater (p < 0.05) on electronic films on days 5 and 7 compared with the unstimulated groups. In conclusion, electrical stimulation, at 120 mV, 1 kHz, for 45 min daily, in addition to surface patterning, of 3.5 ?m wide × 500 nm deep grooves, offered control of nerve axon outgrowth and alignment. PMID:23401351

Hronik-Tupaj, Marie; Raja, Waseem Khan; Tang-Schomer, Min; Omenetto, Fiorenzo G; Kaplan, David L

2013-09-01

276

INTRODUCTION Most spiders spin a variety of different silks that are utilized for  

E-print Network

qualities that make it a focus of biomimetic research (Hakimi et al., 2007; Vollrath and Porter, 2006b of evolutionary history, the amino acid sequences of dragline silk proteins, called major ampullate spidroins (Ma

Agnarsson, Ingi

277

All-water-based electron-beam lithography using silk as a resist  

NASA Astrophysics Data System (ADS)

Traditional nanofabrication techniques often require complex lithographic steps and the use of toxic chemicals. To move from the laboratory scale to large scales, nanofabrication should be carried out using alternative procedures that are simple, inexpensive and use non-toxic solvents. Recent efforts have focused on nanoimprinting and the use of organic resists (such as quantum dot-polymer hybrids, DNA and poly(ethylene glycol)), which still require, for the most part, noxious chemicals for processing. Significant advances have been achieved using `green' resists that can be developed with water, but so far these approaches have suffered from low electron sensitivity, line edge roughness and scalability constraints. Here, we present the use of silk as a natural and biofunctional resist for electron-beam lithography. The process is entirely water-based, starting with the silk aqueous solution and ending with simple development of the exposed silk film in water. Because of its polymorphic crystalline structure, silk can be used either as a positive or negative resist through interactions with an electron beam. Moreover, silk can be easily modified, thereby enabling a variety of `functional resists', including biologically active versions. As a proof of principle of the viability of all-water-based silk electron-beam lithography (EBL), we fabricate nanoscale photonic lattices using both neat silk and silk doped with quantum dots, green fluorescent proteins (GFPs) or horseradish peroxidase (HRP).

Kim, Sunghwan; Marelli, Benedetto; Brenckle, Mark A.; Mitropoulos, Alexander N.; Gil, Eun-Seok; Tsioris, Konstantinos; Tao, Hu; Kaplan, David L.; Omenetto, Fiorenzo G.

2014-04-01

278

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

279

Biomaterials 24 (2003) 401416 Silk-based biomaterials  

E-print Network

Biomaterials 24 (2003) 401­416 Silk-based biomaterials Gregory H. Altman, Frank Diaz, Caroline with other commonly used biomaterials such as polylactic acid and collagen. Furthermore, the unique rationale for the exploration of this family of fibrous proteins for biomaterial applications. For example

Lu, Helen H.

280

Silk matrix for tissue engineered anterior cruciate ligaments  

Microsoft Academic Search

A silk-fiber matrix was studied as a suitable material for tissue engineering anterior cruciate ligaments (ACL). The matrix was successfully designed to match the complex and demanding mechanical requirements of a native human ACL, including adequate fatigue performance. This protein matrix supported the attachment, expansion and differentiation of adult human progenitor bone marrow stromal cells based on scanning electron microscopy,

Gregory H Altman; Rebecca L Horan; Helen H Lu; Jodie Moreau; Ivan Martin; John C Richmond; David L Kaplan

2002-01-01

281

Carbon nanotubes on a spider silk scaffold  

NASA Astrophysics Data System (ADS)

Understanding the compatibility between spider silk and conducting materials is essential to advance the use of spider silk in electronic applications. Spider silk is tough, but becomes soft when exposed to water. Here we report a strong affinity of amine-functionalised multi-walled carbon nanotubes for spider silk, with coating assisted by a water and mechanical shear method. The nanotubes adhere uniformly and bond to the silk fibre surface to produce tough, custom-shaped, flexible and electrically conducting fibres after drying and contraction. The conductivity of coated silk fibres is reversibly sensitive to strain and humidity, leading to proof-of-concept sensor and actuator demonstrations.

Steven, Eden; Saleh, Wasan R.; Lebedev, Victor; Acquah, Steve F. A.; Laukhin, Vladimir; Alamo, Rufina G.; Brooks, James S.

2013-09-01

282

Carbon nanotubes on a spider silk scaffold.  

PubMed

Understanding the compatibility between spider silk and conducting materials is essential to advance the use of spider silk in electronic applications. Spider silk is tough, but becomes soft when exposed to water. Here we report a strong affinity of amine-functionalised multi-walled carbon nanotubes for spider silk, with coating assisted by a water and mechanical shear method. The nanotubes adhere uniformly and bond to the silk fibre surface to produce tough, custom-shaped, flexible and electrically conducting fibres after drying and contraction. The conductivity of coated silk fibres is reversibly sensitive to strain and humidity, leading to proof-of-concept sensor and actuator demonstrations. PMID:24022336

Steven, Eden; Saleh, Wasan R; Lebedev, Victor; Acquah, Steve F A; Laukhin, Vladimir; Alamo, Rufina G; Brooks, James S

2013-01-01

283

Carbon nanotubes on a spider silk scaffold  

PubMed Central

Understanding the compatibility between spider silk and conducting materials is essential to advance the use of spider silk in electronic applications. Spider silk is tough, but becomes soft when exposed to water. Here we report a strong affinity of amine-functionalised multi-walled carbon nanotubes for spider silk, with coating assisted by a water and mechanical shear method. The nanotubes adhere uniformly and bond to the silk fibre surface to produce tough, custom-shaped, flexible and electrically conducting fibres after drying and contraction. The conductivity of coated silk fibres is reversibly sensitive to strain and humidity, leading to proof-of-concept sensor and actuator demonstrations. PMID:24022336

Steven, Eden; Saleh, Wasan R.; Lebedev, Victor; Acquah, Steve F. A.; Laukhin, Vladimir; Alamo, Rufina G.; Brooks, James S.

2013-01-01

284

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.

285

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

E-print Network

Unraveled mechanism in silk engineering: Fast reeling induced silk toughening Xiang Wu,1,2 Xiang response of silkworm and spider silks against stretching and the relationship with the underlying and textile applications. © 2009 American Institute of Physics. DOI: 10.1063/1.3216804 Spider silk is superior

Li, Baowen

286

In vitro study on silk fibroin textile structure for anterior cruciate ligament regeneration.  

PubMed

A novel hierarchical textile structure made of silk fibroin from Bombyx mori capable of matching the mechanical performance requirements of anterior cruciate ligament (ACL) and in vitro cell ingrowth is described. This sericin-free, Silk Fibroin Knitted Sheath with Braided Core (SF-KSBC) structure was fabricated using available textile technologies. Micro-CT analysis confirmed that the core was highly porous and had a higher degree of interconnectivity than that observed for the sheath. The in vivo cell colonization of the scaffolds is thus expected to penetrate even the internal parts of the structure. Tensile mechanical tests demonstrated a maximum load of 1212.4±56.4 N (under hydrated conditions), confirming the scaffold's suitability for ACL reconstruction. The absence of cytotoxic substances in the extracts of the SF-KSBC structure in culture medium was verified by in vitro tests with L929 fibroblasts. In terms of extracellular matrix production, Human Periodontal Ligament Fibroblasts (HPdLFs) cultured in direct contact with SF-KSBC, compared to control samples, demonstrated an increased secretion of aggrecan (PG) and fibronectin (FBN) at 3 and 7 days of culture, and no change in IL-6 and TNF-? secretion. Altogether, the outcomes of this investigation confirm the significant utility of this novel scaffold for ACL tissue regeneration. PMID:23910255

Farè, Silvia; Torricelli, Paola; Giavaresi, Gianluca; Bertoldi, Serena; Alessandrino, Antonio; Villa, Tomaso; Fini, Milena; Tanzi, Maria Cristina; Freddi, Giuliano

2013-10-01

287

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

PubMed Central

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.

2013-01-01

288

Optically probing torsional superelasticity in spider silks  

SciTech Connect

We investigate torsion mechanics of various spider silks using a sensitive optical technique. We find that spider silks are torsionally superelastic in that they can reversibly withstand great torsion strains of over 10{sup 2?3} rotations per cm before failure. Among various silks from a spider, we find the failure twist-strain is greatest in the sticky capture silk followed by dragline and egg-case silk. Our in situ laser-diffraction measurements reveal that torsional strains on the silks induce a nano-scale transverse compression in its diameter that is linear and reversible. These unique torsional properties of the silks could find applications in silk-based materials and devices.

Kumar, Bhupesh; Thakur, Ashish; Panda, Biswajit; Singh, Kamal P. [Department of Physical Sciences, IISER Mohali, Sector 81, Manauli, Mohali 140306 (India)] [Department of Physical Sciences, IISER Mohali, Sector 81, Manauli, Mohali 140306 (India)

2013-11-11

289

Bioconjugation of silk fibroin nanoparticles with enzyme and Peptide and their characterization.  

PubMed

Bombyx mori silk fibroin is a type of protein-based polymer with unique characteristics that is widely used in the research and development of medical biomaterials. The degummed filament of silk fibroin can be dissolved in a highly concentrated salt solution. After desalination, the regenerated liquid silk fibroin (LSF) solution could be made into various forms of silk biomaterials, such as powder, fiber, film, porous matrix, 3D scaffold, and hydrogel, depending on its application. In this study, we mixed the liquid silk solution with enzymes, including oxidase and hydrolase, and rapidly injected the mixture into an excess of acetone. The enzyme retained most of its enzymatic activity and was also captured in silk fibroin nanoparticles (SFNs), which instantly formed via a configuration transition of the regenerated silk protein from a random coil and ?-helix to a ?-sheet. The resulting enzyme-captured SFNs displayed a fine crystal structure with a high activity recovery and good thermal stability. Moreover, the affinities of these modified enzymes to their substrate did not evidently suffer from the capture. When only the liquid silk solution was rapidly injected into acetone, the resulting globular SFNs with the same crystallinity were also a good carrier that was covalently conjugated to enzymes and insulin. Thus, silk protein nanoparticles are of potential value as an enzyme or peptide delivery system for the research and development of medical biomaterials. In this report, the bioconjugation of SFNs with glucose oxidase, superoxidase, ?-glucosidase, l-asparaginase, neutral protease, and insulin and their characterization are described in detail. PMID:25819282

Wang, Fei; Zhang, Yu-Qing

2015-01-01

290

Fifty Years Later: The Sequence, Structure and Function of Lacewing Cross-beta Silk  

SciTech Connect

Classic studies of protein structure in the 1950s and 1960s demonstrated that green lacewing egg stalk silk possesses a rare native cross-beta sheet conformation. We have identified and sequenced the silk genes expressed by adult females of a green lacewing species. The two encoded silk proteins are 109 and 67 kDa in size and rich in serine, glycine and alanine. Over 70% of each protein sequence consists of highly repetitive regions with 16-residue periodicity. The repetitive sequences can be fitted to an elegant cross-beta sheet structural model with protein chains folded into regular 8-residue long beta strands. This model is supported by wide-angle X-ray scattering data and tensile testing from both our work and the original papers. We suggest that the silk proteins assemble into stacked beta sheet crystallites bound together by a network of cystine cross-links. This hierarchical structure gives the lacewing silk high lateral stiffness nearly threefold that of silkworm silk, enabling the egg stalks to effectively suspend eggs and protect them from predators.

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

2009-12-01

291

Enhanced Cellular Adhesion on Titanium by Silk Functionalized with titanium binding and RGD peptides  

PubMed Central

Soft tissue adhesion on titanium represents a challenge for implantable materials. In order to improve adhesion at the cell/material interface we used a new approach based on the molecular recognition of titanium by specific peptides. Silk fibroin protein was chemically grafted with titanium binding peptide (TiBP) to increase adsorption of these chimeric proteins to the metal surface. Quartz Crystal Microbalance was used to quantify the specific adsorption of TiBP-functionalized silk and an increase in protein deposition by more than 35% was demonstrated due to the presence of the binding peptide. A silk protein grafted with TiBP and fibronectin-derived RGD peptide was then prepared. The adherence of fibroblasts on the titanium surface modified with the multifunctional silk coating demonstrated an increase in the number of adhering cells by 60%. The improved adhesion was demonstrated by Scanning Electron Microscopy and immunocytochemical staining of focal contact points. Chick embryo organotypic culture also revealed strong adhesion of endothelial cells expanding on the multifunctional silk-peptide coating. These results demonstrated that silk functionalized with TiBP and RGD represents a promising approach to modify cell-biomaterial interfaces, opening new perspectives for implantable medical devices, especially when reendothelialization is required. PMID:22975628

Vidal, Guillaume; Blanchi, Thomas; Mieszawska, Aneta J.; Calabrese, Rossella; Rossi, Claire; Vigneron, Pascale; Duval, Jean-Luc; Kaplan, David L.; Egles, Christophe

2012-01-01

292

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

293

Bacterial cellulose nanocrystals-embedded silk nanofibers.  

PubMed

Nanofibrous Bacterial cellulose nanocrystals (BCNs)-embedded silk fibroin were successfully fabricated using electrospinning. The morphology, structure and mechanical properties of the silk fibroin nanofibers were investigated at various BCNs concentrations from 0 to 7 wt%. SEM, TEM and XRD analyses were conducted to confirm the incorporation of the BCNs in the electrospun silk fibroin nanofibers. The average diameter of the silk fibroin/BCNs nanofibers increased from 230 to 430 nm according to the increasing of the BCNs ratio due to the rising solute content. The FT-IR spectra confirmed the conformational transition of the silk fibroin, from a random coil to a beta-sheet structure, which shows the enhanced mechanical properties of silk fibroin based nanofibers even with small amounts of the BCNs. Moreover, it was observed that the Young's modulus of the silk fibroin/BCNs nanofibers unexpectedly increased with the formation of BCNs with a percolation structure at a concentration between 3 and 5 wt%. PMID:22966722

Park, Doo Jin; Choi, Youngeun; Heo, Semi; Cho, Se Youn; Jin, Hyung-Joon

2012-07-01

294

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

295

Lumbar interbody fusion with porous biphasic calcium phosphate enhanced by recombinant bone morphogenetic protein-2/silk fibroin sustained-released microsphere: an experimental study on sheep model.  

PubMed

Biphasic calcium phosphate (BCP) has been investigated extensively as a bone substitute nowadays. However, the bone formation capacity of BCP is limited owing to lack of osteoinduction. Silk fibroin (SF) has a structure similar to type I collagen, and could be developed to a microsphere for the sustained-release of rhBMP-2. In our previous report, bioactivity of BCP could be enhanced by rhBMP-2/SF microsphere (containing 0.5 µg rhBMP-2) in vitro. However, the bone regeneration performance of the composite in vivo was not investigated. Thus, the purpose of this study was to evaluate the efficacy of BCP/rhBMP-2/SF in a sheep lumbar fusion model. A BCP and rhBMP-2/SF microsphere was developed, and then was integrated into a BCP/rhBMP-2/SF composite. BCP, BCP/rhBMP-2 and BCP/rhBMP-2/SF were implanted randomly into the disc spaces of 30 sheep at the levels of L1/2, L3/4 and L5/6. After sacrificed, the fusion segments were evaluated by manual palpation, CT scan, biomechanical testing and histology at 3 and 6 months, respectively. The composite demonstrated a burst-release of rhBMP-2 (39.1 ± 2.8 %) on the initial 4 days and a sustained-release (accumulative 81.3 ± 4.9 %) for more than 28 days. The fusion rates, semi-quantitative CT scores, fusion stiffness in bending in all directions and histologic scores of BCP/rhBMP-2/SF were significantly greater than BCP and BCP/rhBMP-2 at each time point, respectively (P < 0.05). These findings indicate that the SF microspheres containing a very low dose of rhBMP-2 improve fusion in sheep using BCP constructs. PMID:25690620

Chen, Liang; Liu, Hai-Long; Gu, Yong; Feng, Yu; Yang, Hui-Lin

2015-03-01

296

Decrease in corneal damage due to benzalkonium chloride by the addition of sericin into timolol maleate eye drops.  

PubMed

We investigated the protective effects of sericin on corneal damage due to benzalkonium chloride (BAC) used as a preservative in commercially available timolol maleate eye drops using rat debrided corneal epithelium and a human cornea epithelial cell line (HCE-T). Corneal wounds were monitored using a fundus camera TRC-50X equipped with a digital camera; eye drops were instilled into the rat eyes five times a day after corneal epithelial abrasion. The viability of HCE-T cells was calculated by TetraColor One; and Escherichia coli (ATCC 8739) were used to measure antimicrobial activity. The reducing effects on transcorneal penetration and intraocular pressure (IOP) of the eye drops were determined using rabbits. The corneal wound healing rate and rate constants (kH) as well as cell viability were higher following treatment with 0.005% BAC solution containing 0.1% sericin than in the case of treatment with BAC solution alone; the antimicrobial activity was approximately the same for BAC solutions with and without sericin. In addition, the kH for rat eyes instilled with commercially available timolol maleate eye drops containing 0.1% sericin was significantly higher than that of eyes instilled with timolol maleate eye drops without sericin, and the addition of sericin did not affect the corneal penetration or IOP reducing effect of commercially available timolol maleate eye drops. A preservative system comprising BAC and sericin may provide effective therapy for glaucoma patients requiring long-term anti-glaucoma agents. PMID:23470443

Nagai, Noriaki; Ito, Yoshimasa; Okamoto, Norio; Shimomura, Yoshikazu

2013-01-01

297

Thromboelastometric and platelet responses to silk biomaterials  

PubMed Central

Silkworm's silk is natural biopolymer with unique properties including mechanical robustness, all aqueous base processing and ease in fabrication into different multifunctional templates. Additionally, the nonmulberry silks have cell adhesion promoting tri-peptide (RGD) sequences, which make it an immensely potential platform for regenerative medicine. The compatibility of nonmulberry silk with human blood is still elusive; thereby, restricts its further application as implants. The present study, therefore, evaluate the haematocompatibility of silk biomaterials in terms of platelet interaction after exposure to nonmulberry silk of Antheraea mylitta using thromboelastometry (ROTEM). The mulberry silk of Bombyx mori and clinically used Uni-Graft W biomaterial serve as references. Shortened clotting time, clot formation times as well as enhanced clot strength indicate the platelet mediated activation of blood coagulation cascade by tested biomaterials; which is comparable to controls. PMID:24824624

Kundu, Banani; Schlimp, Christoph J.; Nürnberger, Sylvia; Redl, Heinz; Kundu, S. C.

2014-01-01

298

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

E-print Network

Polarized light microscopy, variability in spider silk diameters, and the mechanical characterization of spider silk Todd A. Blackledge,a Richard A. Cardullo, and Cheryl Y. Hayashi Department of Biology, University of California, Riverside, California 92521, USA Abstract. Spider silks possess

Blackledge, Todd

299

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

300

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

301

Extraction conditions of Antheraea mylitta sericin with high yields and minimum molecular weight degradation.  

PubMed

Although the technique for extracting the Bombyx mori sericin has been extensively known, the extraction of sericin from wild-silkworm cocoons is not yet standardized. The aim of this study was to find the optimal conditions for the extraction of sericin from Antheraea mylitta cocoons, with high yields and minimum degradation. We attempted to apply various protocols for the extraction of the A. mylitta sericin (AmS). Among these, we found that the extraction of AmS with a sodium carbonate solution exhibited the highest yield except the conventional soap-alkali extraction. To find the optimal conditions for the AmS extraction with the sodium carbonate, we changed the concentration of sodium carbonate and the treatment time. With an increase in the sodium carbonate concentration and the extraction time, the yield of AmS increased, but the molecular weight (MW) of AmS decreased. Considering the yield, molecular weight distribution (MWD) and amino acid composition of AmS, we suggest that the optimal conditions for the AmS extraction require treatment with 0.02 M sodium carbonate and boiling for 60 min. PMID:23026092

Yun, Haesung; Oh, Hanjin; Kim, Moo Kon; Kwak, Hyo Won; Lee, Jeong Yun; Um, In Chul; Vootla, Shyam Kumar; Lee, Ki Hoon

2013-01-01

302

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

303

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

PubMed Central

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

2013-01-01

304

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.

305

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

306

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; Köster, Sarah; Hu, Xiao; Evans, Heather M.; Pfohl, Thomas; Kaplan, David L.; Wong, Joyce

2011-01-01

307

Construction of transgenic silkworm spinning antibacterial silk with fluorescence.  

PubMed

A targeting vector consisting of a fusion gene of the green fluorescent protein (GFP) gene gfp and the antimicrobial peptide cecropin gene cec flanked by pieces of the 5' and 3' sequences of the fibroin L chain gene fib-L of the silkworm (Bombyx mori) and a negative selection DsRed marker gene driven by the baculovirus immediate early gene 1 (i.e.-1) promoter, was used to target the silkworm genome in order to explore the possibility of improving the performance of silk. A transgenic silkworm with a green fluorescent cocoon was obtained and PCR analysis of its genome confirmed that the target genes had been integrated into the silkworm genome correctly. Furthermore, in the posterior silk glands of the G6 generation transformation silkworm, a band representing the fusion protein Fib-L-GFP-Cec with a molecular mass of 68.7 kDa was detected by western blotting with an antibody against GFP. An investigation of the number of bacteria attached to a cocoon showed the transgenic silkworm cocoon possessed antibacterial properties. These results suggested the performance of silk can be improved by modifying the fibroin gene. PMID:25223857

Li, Zhen; Jiang, Yue; Cao, Guangli; Li, Jingzhi; Xue, Renyu; Gong, Chengliang

2015-01-01

308

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

309

Adult spiders use tougher silk: ontogenetic changes in web architecture and silk biomechanics in the orb-weaver spider  

E-print Network

Adult spiders use tougher silk: ontogenetic changes in web architecture and silk biomechanics provide an interesting case, where a relatively homogeneous foraging strategy, aerial silk webs are also likely to increase as spiders develop. Here, we examine how relative silk investment, web

Agnarsson, Ingi

310

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

311

Gel spinning of silk tubes for tissue engineering  

PubMed Central

Tubular vessels for tissue engineering are typically fabricated using a molding, dipping, or electrospinning technique. While these techniques provide some control over inner and outer diameters of the tube, they lack the ability to align the polymers or fibers of interest throughout the tube. This is an important aspect of biomaterial composite structure and function for mechanical and biological impact of tissue outcomes. We present a novel aqueous process system to spin tubes from biopolymers and proteins such as silk fibroin. Using silk as an example, this method of winding an aqueous solution around a reciprocating rotating mandrel offers substantial improvement in the control of the tube properties, specifically with regard to winding pattern, tube porosity, and composite features. Silk tube properties are further controlled via different post-spinning processing mechanisms such as methanol-treatment, air-drying, and lyophilization. This approach to tubular scaffold manufacture offers numerous tissue engineering applications such as complex composite biomaterial matrices, blood vessel grafts and nerve guides, among others. PMID:18801570

Lovett, Michael; Cannizzaro, Christopher; Vunjak-Novakovic, Gordana; Kaplan, David L.

2011-01-01

312

Structural study of Bombyx mori silk fibroin during processing for regeneration  

NASA Astrophysics Data System (ADS)

Bombyx mori silk fibroin has excellent mechanical properties combined with flexibility, tissue compatibility, and high oxygen permeability in the wet condition. This important material should be dissolved and regenerated to be utilized as useful forms such as gel, film, fiber, powder, or non-woven. However, it has long been a problem that the regenerated fibroin materials show poor mechanical properties and brittleness. These problems were technically solved by improving a fiber processing method reported here. The regenerated fibroin fibers showed much better mechanical properties compared to the original silk fibers. This improved technique for the fiber processing of Bombyx mori silk fibroin may be used as a model system for other semi-crystalline fiber forming proteins, becoming available through biotechnology. The physical and chemical properties of the regenerated fibers were characterized by SinTechRTM tensile testing, X-ray diffraction, solid state 13C NMR spectroscopy, and SEM. Unlike synthetic polymers, the molecular weight distribution of Bombyx mori silk fibroin is mono-disperse because silk fibroin is synthesized from DNA template. Genetic studies have revealed the entire amino acid sequence of Bombyx mori silk fibroin. It is known that the crystalline silk II structure is composed of hexa-amino acid sequences, GAGAGS. However, in the amino acid sequence of Bombyx mori silk fibroin heavy chain, there are present 11 chemically irregular but evolutionarily conserved sequences with about 31 amino acid residues (irregular GT˜GT sequences). The structure and role of these irregular sequences have remained unknown. One of the most frequently appearing irregular sequences was synthesized by a peptide synthesizer. The three-dimensional structure of this irregular silk peptide was studied by the high resolution two-dimensional NMR technique. The three-dimensional structure of this peptide shows that it makes a turn or loop structure (distorted O shape), which means the proceeding backbone direction is changed 180° by this sequence. This may facilitate the beta-sheet formation of the crystal forming building blocks, GAGAGS/GY˜GY sequences, in fibroin heavy chain. It may also facilitate the solubilization of the fibroin heavy chain within the silk gland.

Ha, Sung-Won

313

Spider silk gut: Development and characterization of a novel strong spider silk fiber  

PubMed Central

Spider silk fibers were produced through an alternative processing route that differs widely from natural spinning. The process follows a procedure traditionally used to obtain fibers directly from the glands of silkworms and requires exposure to an acid environment and subsequent stretching. The microstructure and mechanical behavior of the so-called spider silk gut fibers can be tailored to concur with those observed in naturally spun spider silk, except for effects related with the much larger cross-sectional area of the former. In particular spider silk gut has a proper ground state to which the material can revert independently from its previous loading history by supercontraction. A larger cross-sectional area implies that spider silk gut outperforms the natural material in terms of the loads that the fiber can sustain. This property suggests that it could substitute conventional spider silk fibers in some intended uses, such as sutures and scaffolds in tissue engineering. PMID:25475975

Jiang, Ping; Marí-Buyé, Núria; Madurga, Rodrigo; Arroyo-Hernández, María; Solanas, Concepción; Gañán, Alfonso; Daza, Rafael; Plaza, Gustavo R.; Guinea, Gustavo V.; Elices, Manuel; Cenis, José Luis; Pérez-Rigueiro, José

2014-01-01

314

Evaluation of gel spun silk-based biomaterials in a murine model of bladder augmentation.  

PubMed

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 poly-glycolic 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 and voided volume 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; Di Vizio, Dolores; Gomez, Pablo; Kaplan, David L; Adam, Rosalyn M; Estrada, Carlos R

2011-01-01

315

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), 22 kV voltage, 10 cm spinning distance at 0.25 mL/h feed rate to prepare nanofibers with small diameter and narrow size distribution without beads. The mean diameter of nanofibers was about 180 nm while introducing AgNO3 led to smaller nanofibers diameter about 95 nm. 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

316

Self-assembly of nucleic acids, silk and hybrid materials thereof.  

PubMed

Top-down approaches based on etching techniques have almost reached their limits in terms of dimension. Therefore, novel assembly strategies and types of nanomaterials are required to allow technological advances. Self-assembly processes independent of external energy sources and unlimited in dimensional scaling have become a very promising approach. Here,we highlight recent developments in self-assembled DNA-polymer, silk-polymer and silk-DNA hybrids as promising materials with biotic and abiotic moieties for constructing complex hierarchical materials in ‘bottom-up’ approaches. DNA block copolymers assemble into nanostructures typically exposing a DNA corona which allows functionalization, labeling and higher levels of organization due to its specific addressable recognition properties. In contrast, self-assembly of natural silk proteins as well as their recombinant variants yields mechanically stable ?-sheet rich nanostructures. The combination of silk with abiotic polymers gains hybrid materials with new functionalities. Together, the precision of DNA hybridization and robustness of silk fibrillar structures combine in novel conjugates enable processing of higher-order structures with nanoscale architecture and programmable functions. PMID:25419786

Humenik, Martin; Scheibel, Thomas

2014-12-17

317

Self-assembly of nucleic acids, silk and hybrid materials thereof  

NASA Astrophysics Data System (ADS)

Top-down approaches based on etching techniques have almost reached their limits in terms of dimension. Therefore, novel assembly strategies and types of nanomaterials are required to allow technological advances. Self-assembly processes independent of external energy sources and unlimited in dimensional scaling have become a very promising approach. Here, we highlight recent developments in self-assembled DNA-polymer, silk-polymer and silk-DNA hybrids as promising materials with biotic and abiotic moieties for constructing complex hierarchical materials in ‘bottom-up’ approaches. DNA block copolymers assemble into nanostructures typically exposing a DNA corona which allows functionalization, labeling and higher levels of organization due to its specific addressable recognition properties. In contrast, self-assembly of natural silk proteins as well as their recombinant variants yields mechanically stable ?-sheet rich nanostructures. The combination of silk with abiotic polymers gains hybrid materials with new functionalities. Together, the precision of DNA hybridization and robustness of silk fibrillar structures combine in novel conjugates enable processing of higher-order structures with nanoscale architecture and programmable functions.

Humenik, Martin; Scheibel, Thomas

2014-12-01

318

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

319

2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim 1wileyonlinelibrary.com Silk Foam Terahertz Waveguides  

E-print Network

for biomedical applications. Silk material from worm cocoon can be processed into different forms-protein films with a two-dimensional square lattice of air holes were proposed and demonstrated to achieve

Skorobogatiy, Maksim

320

Beating the Heat - Fast Scanning Melts Silk Beta Sheet Crystals  

NASA Astrophysics Data System (ADS)

Beta-pleated-sheet crystals are among the most stable of protein secondary structures, and are responsible for the remarkable physical properties of many fibrous proteins, such as silk, or proteins forming plaques as in Alzheimer's disease. Previous thinking, and the accepted paradigm, was that beta-pleated-sheet crystals in the dry solid state were so stable they would not melt upon input of heat energy alone. Here we overturn that assumption and demonstrate that beta-pleated-sheet crystals melt directly from the solid state to become random coils, helices, and turns. We use fast scanning chip calorimetry at 2,000 K/s and report the first reversible thermal melting of protein beta-pleated-sheet crystals, exemplified by silk fibroin. The similarity between thermal melting behavior of lamellar crystals of synthetic polymers and beta-pleated-sheet crystals is confirmed. Significance for controlling beta-pleated-sheet content during thermal processing of biomaterials, as well as towards disease therapies, is envisioned based on these new findings.

Cebe, Peggy; Hu, Xiao; Kaplan, David L.; Zhuravlev, Evgeny; Wurm, Andreas; Arbeiter, Daniela; Schick, Christoph

2013-01-01

321

Beating the Heat - Fast Scanning Melts Silk Beta Sheet Crystals  

PubMed Central

Beta-pleated-sheet crystals are among the most stable of protein secondary structures, and are responsible for the remarkable physical properties of many fibrous proteins, such as silk, or proteins forming plaques as in Alzheimer's disease. Previous thinking, and the accepted paradigm, was that beta-pleated-sheet crystals in the dry solid state were so stable they would not melt upon input of heat energy alone. Here we overturn that assumption and demonstrate that beta-pleated-sheet crystals melt directly from the solid state to become random coils, helices, and turns. We use fast scanning chip calorimetry at 2,000?K/s and report the first reversible thermal melting of protein beta-pleated-sheet crystals, exemplified by silk fibroin. The similarity between thermal melting behavior of lamellar crystals of synthetic polymers and beta-pleated-sheet crystals is confirmed. Significance for controlling beta-pleated-sheet content during thermal processing of biomaterials, as well as towards disease therapies, is envisioned based on these new findings. PMID:23350037

Cebe, Peggy; Hu, Xiao; Kaplan, David L.; Zhuravlev, Evgeny; Wurm, Andreas; Arbeiter, Daniela; Schick, Christoph

2013-01-01

322

Silk Optics Mark Cronin-Golomb  

E-print Network

stem cell culture shows biocompatibility #12;1/27/14 4 Silk film biomaterials for cornea tissue) Bioreactors in incubator Commercialization · David Kaplan student Greg Altman with torn ACL, decides to do

Barthelat, Francois

323

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

324

Injectable silk-polyethylene glycol hydrogels.  

PubMed

Silk hydrogels for tissue repair are usually pre-formed via chemical or physical treatments from silk solutions. For many medical applications, it is desirable to utilize injectable silk hydrogels at high concentrations (>8%) to avoid surgical implantation and to achieve slow in vivo degradation of the gel. In the present study, injectable silk solutions that formed hydrogels in situ were generated by mixing silk with low-molecular-weight polyethylene glycol (PEG), especially PEG300 and 400 (molecular weight 300 and 400g mol(-1)). Gelation time was dependent on the concentration and molecular weight of PEG. When the concentration of PEG in the gel reached 40-45%, gelation time was less than 30min, as revealed by measurements of optical density and rheological studies, with kinetics of PEG400 faster than PEG300. Gelation was accompanied by structural changes in silk, leading to the conversion from random coil in solution to crystalline ?-sheets in the gels, based on circular dichroism, attenuated total reflection Fourier transform infrared spectroscopy and X-ray diffraction. The modulus (127.5kPa) and yield strength (11.5kPa) determined were comparable to those of sonication-induced hydrogels at the same concentrations of silk. The time-dependent injectability of 15% PEG-silk hydrogel through 27G needles showed a gradual increase of compression forces from ?10 to 50N within 60min. The growth of human mesenchymal stem cells on the PEG-silk hydrogels was hindered, likely due to the presence of PEG, which grew after a 5 day delay, presumably while the PEG solubilized away from the gel. When 5% PEG-silk hydrogel was subcutaneously injected in rats, significant degradation and tissue in-growth took place after 20 days, as revealed by ultrasound imaging and histological analysis. No significant inflammation around the gel was observed. The features of injectability, slow degradation and low initial cell attachment suggests that these PEG-silk hydrogels are of interest for many biomedical applications, such as anti-fouling and anti-adhesion. PMID:25449912

Wang, Xiaoqin; Partlow, Benjamin; Liu, Jian; Zheng, Zhaozhu; Su, Bo; Wang, Yansong; Kaplan, David L

2015-01-01

325

Mechanical testing of spider silk at cryogenic temperatures.  

PubMed

A method and results for mechanical testing of spider silk in extreme environments is presented. In particular, silk from the spider Steatoda triangulosa is harvested, and samples are subjected to cryogenic temperatures by means of liquid nitrogen submersion. Samples are destructively tested while immersed in liquid nitrogen, and the stress-strain characteristics are compared to those of silk at room temperature. The strength, elasticity, and toughness of the cryogenically submersed silk are determined. It is found that on average, silk is 64% stronger while immersed in liquid nitrogen (i.e., at -196°C). The testing method could also be used for testing of silk in chemically hostile environments. PMID:20933539

Pogozelski, E M; Becker, W L; See, B D; Kieffer, C M

2011-01-01

326

Supramolecular organization of regenerated silkworm silk fibers  

Microsoft Academic Search

The microstructures of N-methylmorpholine-N-oxide (NMMO) regenerated silk fibers have been characterized by atomic force microscopy from the micrometer to the nanometer scale and compared with those previously found from natural silks. Regenerated fibers show poor tensile properties and a brittle behavior, but their mechanical properties improve if subjected to post-spinning drawing. Consequently, it was hypothesized that post-spinning drawing would lead

J. Pérez-Rigueiro; L. Biancotto; P. Corsini; E. Marsano; M. Elices; G. R. Plaza; G. V. Guinea

2009-01-01

327

Studies on Silk Proteins. I. The Properties and Constitution of Fibroin. The Conversion of Fibroin into a Water-Soluble Form and Its Bearing on the Phenomenon of Denaturation  

Microsoft Academic Search

Some of the constituent amino-acids of fibroin (degummed silk) are determined. Special attention is directed to histidine, owing to its use in the calculation of the molecular weight of fibroin. A value of 0\\\\cdot 45% has been found by methods in which the histidine is isolated as nitranilate or di-(3:4-dichlorobenzenesulphonate). Other values obtained are serine 12\\\\cdot 6%, threonine 1\\\\cdot 5%,

D. Coleman; F. O. Howitt

1947-01-01

328

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

329

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. PMID:25396043

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

2014-01-01

330

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

331

pH-Dependent anticancer drug release from silk nanoparticles  

PubMed Central

Silk has traditionally been used as a suture material because of its excellent mechanical properties and biocompatibility. These properties have led to the development of different silk-based material formats for tissue engineering and regenerative medicine. Although there have been a small number of studies about the use of silk particles for drug delivery, none of these studies have assessed the potential of silk to act as a stimulus-responsive anticancer nanomedicine. This report demonstrates that an acetone precipitation of silk allowed the formation of uniform silk nanoparticles (98 nm diameter, polydispersity index 0.109), with an overall negative surface charge (-33.6 ±5.8 mV), in a single step. Silk nanoparticles were readily loaded with doxorubicin (40 ng doxorubicin/?g silk) and showed pH-dependent release (pH 4.5>> 6.0 > 7.4). In vitro studies with human breast cancer cell lines demonstrated that the silk nanoparticles were not cytotoxic (IC50 >120/?/ml) and that doxorubicin-loaded silk nanoparticles were able to overcome drug resistance mechanisms. Live cell fluorescence microscopy studies showed endocytic uptake and lysosomal accumulation of silk nanoparticles. In summary, the pH-dependent drug release and lysosomal accumulation of silk nanoparticles demonstrated the ability of drug-loaded silk nanoparticles to serve as a lysosomotropic anticancer nanomedicine. PMID:23625825

Seib, F. Philipp; Jones, Gregory T.; Rnjak-Kovacina, Jelena; Lin, Yinan; Kaplan, David L.

2013-01-01

332

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

333

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

334

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

335

The performance of silk scaffolds in a rat model of augmentation cystoplasty.  

PubMed

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, 5-50 ?m) lamellar-like sheets buttressed by a dense outer layer. Bi-layer FF scaffolds were comprised of porous foams (pore size, ~400 ?m) 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 3-4 mm. In contrast, rats augmented with FF scaffolds displayed substantially lower rates (20%) and stone size (2 mm), similar to the levels observed in controls (13%, 2 mm). 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

2013-07-01

336

Shotgun proteomic analysis of the Bombyx mori anterior silk gland: An insight into the biosynthetic fiber spinning process.  

PubMed

The Bombyx mori anterior silk gland (ASG) is a natural fiber manipulator for the material provided by the middle and posterior silk glands. In view of the significant role of the ASG in the liquid-crystal spinning process, a shotgun proteomics approach was taken to study the relationship between the function of proteins in the silkworm ASG and the spinning mechanism. A total of 1132 proteins with 7647 unique peptides were identified in the ASG dataset including some involved in the cuticle, ion transportation, energy metabolism, and apoptosis. Two putative cuticle-specific proteins were highly and specifically expressed in the ASG; therefore, the ASG dataset could provide clues for comprehensive understanding of the natural silk spinning mechanism in the silkworm. All MS data have been deposited in the ProteomeXchange with identifier PXD000090. PMID:23828816

Yi, Qiying; Zhao, Ping; Wang, Xin; Zou, Yong; Zhong, Xiaowu; Wang, Chen; Xiang, Zhonghuai; Xia, Qing-You

2013-09-01

337

Sericin can reduce hippocampal neuronal apoptosis by activating the Akt signal transduction pathway in a rat model of diabetes mellitus?  

PubMed Central

In the present study, a rat model of type 2 diabetes mellitus was established by continuous peritoneal injection of streptozotocin. Following intragastric perfusion of sericin for 35 days, blood glucose levels significantly reduced, neuronal apoptosis in the hippocampal CA1 region decreased, hippocampal phosphorylated Akt and nuclear factor kappa B expression were enhanced, but Bcl-xL/Bcl-2 associated death promoter expression decreased. Results demonstrated that sericin can reduce hippocampal neuronal apoptosis in a rat model of diabetes mellitus by regulating abnormal changes in the Akt signal transduction pathway.

Chen, Zhihong; He, Yaqiang; Song, Chengjun; Dong, Zhijun; Su, Zhejun; Xue, Jingfeng

2012-01-01

338

Sericin can reduce hippocampal neuronal apoptosis by activating the Akt signal transduction pathway in a rat model of diabetes mellitus.  

PubMed

In the present study, a rat model of type 2 diabetes mellitus was established by continuous peritoneal injection of streptozotocin. Following intragastric perfusion of sericin for 35 days, blood glucose levels significantly reduced, neuronal apoptosis in the hippocampal CA1 region decreased, hippocampal phosphorylated Akt and nuclear factor kappa B expression were enhanced, but Bcl-xL/Bcl-2 associated death promoter expression decreased. Results demonstrated that sericin can reduce hippocampal neuronal apoptosis in a rat model of diabetes mellitus by regulating abnormal changes in the Akt signal transduction pathway. PMID:25767499

Chen, Zhihong; He, Yaqiang; Song, Chengjun; Dong, Zhijun; Su, Zhejun; Xue, Jingfeng

2012-01-25

339

Version of July 8, 2009 Hyper Logic Programs in SILK  

E-print Network

Version of July 8, 2009 Hyper Logic Programs in SILK For Business and Science: An Overview Benjamin Inc., benjaming@vulcan.com http://silk.semwebcentral.org http://www.mit.edu/~bgrosof/ ** Workshop and Vulcan Inc. All rights reserved. #12;Outline of Talk · Intro to the SILK effort, and its parent Project

Polz, Martin

340

Silk Fiber Mechanics from Multiscale Force Distribution Analysis Murat Cetinkaya,  

E-print Network

Silk Fiber Mechanics from Multiscale Force Distribution Analysis Murat Cetinkaya, Senbo Xiao, Bernd the molecular determinants for the extreme toughness of spider silk fibers. Our bottom-up computational approach of the internal strain distribution and load-carrying motifs in silk fibers on scales of both molecular

Gräter, Frauke

341

Rapid Nanoimprinting of Silk Fibroin Films for Biophotonic Applications  

E-print Network

Rapid Nanoimprinting of Silk Fibroin Films for Biophotonic Applications By Jason J. Amsden, Peter-based material substrate would provide entirely new options for such devices. Silk fibroin is an appealing,10] and biocompatibility.[11] Biologically functionalized silk fibroin films can be patterned on the micro and nanoscale

White, Robert D.

342

Hyper Logic Programs in SILK: Redefining the KR Playing Field  

E-print Network

1 Hyper Logic Programs in SILK: Redefining the KR Playing Field for Business and VLKB Benjamin@vulcan.com http://silk.projects.semwebcentral.org http://www.mit.edu/~bgrosof/ ** The International Rule;2 Outline of Talk · Introduction · Project Halo · SILK · Overall Approach, Vision, Goals, and Plan · Hyper

Polz, Martin

343

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). Silk also provides ecologists with links between the behaviors of spiders as they spin webs). Molecular biologists are increasing the known diversity of genes that code for a variety of spider silk

Blackledge, Todd

344

Signature change events: A challenge for quantum gravity? Silke Weinfurtner  

E-print Network

Signature change events: A challenge for quantum gravity? By Silke Weinfurtner in collaboration with Angela White Matt Visser #12;Signature change events: A challenge for quantum gravity? By Silke for quantum gravity? By Silke Weinfurtner in collaboration with Angela White Matt Visser #12;Outlook Trans

Visser, Matt

345

SILK: Higher Level Rules with Defaults and Semantic Scalability  

E-print Network

SILK: Higher Level Rules with Defaults and Semantic Scalability Benjamin Grosof* October 26 2009://www.mit.edu/~bgrosof/ http://silk.semwebcentral.org ** 3rd International Conference on Web Reasoning and Rules, Chantilly, VA and Vulcan Inc. All rights reserved. #12;Outline of Talk · Overview of SILK effort · Vision, Origins

Polz, Martin

346

Ex vivo rheology of spider silk , C. Clasen3  

E-print Network

1 Ex vivo rheology of spider silk N. Koji1,2 , J. Bico1 , C. Clasen3 & G.H. McKinley1 1 Hatsopoulos a considerable source of questions1-3 . In order to probe the rheological properties of minute amounts or genetically-engineered silks to match those of the spider. During the past decade spider dragline silk has

347

Influence factors analysis on the formation of silk I structure.  

PubMed

Regenerated silk fibroin aqueous solution was used to study the crystalline structure of Bombyx mori silk fibroin in vitro. By controlling environmental conditions and concentration of silk fibroin solution, it provided a means for the direct preparing silk I structure and understanding the details of silk fibroin molecules interactions in formation process. In this study, silk fibroin molecules were assembled to form random coil at low concentration of solution and then, as the concentration increases, were converted to silk I at 55% relative humidity (RH). At the same time, the structure of silk fibroin forming below 45°C was mostly in silk I. A partial ternary phase diagram of temperature-humidity-concentration was constructed based on the results. The results showed silk I structure could be controlled by adjusting the external environmental conditions. The enhanced control over silk I structure, as embodied in phase diagram, could potentially be utilized to understand the molecular chain conformation of silk I in further research work. PMID:25677178

Ming, Jinfa; Pan, Fukui; Zuo, Baoqi

2015-04-01

348

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

349

Combining flagelliform and dragline spider silk motifs to produce tunable synthetic biopolymer fibers.  

PubMed

The two Flag/MaSp 2 silk proteins produced recombinantly were based on the basic consensus repeat of the dragline silk spidroin 2 protein (MaSp 2) from the Nephila clavipes orb weaving spider. However, the proline-containing pentapeptides juxtaposed to the polyalanine segments resembled those found in the flagelliform silk protein (Flag) composing the web spiral: (GPGGX(1) GPGGX(2))(2) with X(1) /X(2) = A/A or Y/S. Fibers were formed from protein films in aqueous solutions or extruded from resolubilized protein dopes in organic conditions when the Flag motif was (GPGGX(1) GPGGX(2))(2) with X(1) /X(2) = Y/S or A/A, respectively. Post-fiber processing involved similar drawing ratios (2-2.5×) before or after water-treatment. Structural (ssNMR and XRD) and morphological (SEM) changes in the fibers were compared to the mechanical properties of the fibers at each step. Nuclear magnetic resonance indicated that the fraction of ?-sheet nanocrystals in the polyalanine regions formed upon extrusion, increased during stretching, and was maximized after water-treatment. X-ray diffraction showed that nanocrystallite orientation parallel to the fiber axis increased the ultimate strength and initial stiffness of the fibers. Water furthered nanocrystal orientation and three-dimensional growth while plasticizing the amorphous regions, thus producing tougher fibers due to increased extensibility. These fibers were highly hygroscopic and had similar internal network organization, thus similar range of mechanical properties that depended on their diameters. The overall structure of the consensus repeat of the silk-like protein dictated the mechanical properties of the fibers while protein molecular weight limited these same properties. Subtle structural motif re-design impacted protein self-assembly mechanisms and requirements for fiber formation. PMID:22012252

Teulé, Florence; Addison, Bennett; Cooper, Alyssa R; Ayon, Joel; Henning, Robert W; Benmore, Chris J; Holland, Gregory P; Yarger, Jeffery L; Lewis, Randolph V

2012-06-01

350

Regeneration of Bombyx mori silk nanofibers and nanocomposite fibrils by the electrospinning process  

NASA Astrophysics Data System (ADS)

In recent years, there has been significant interest in the utilization of natural materials for novel nanoproducts such as tissue engineered scaffolds. Silkworm silk fibers represent one of the strongest natural fibers known. Silkworm silk, a protein-based natural biopolymer, has received renewed interest in recent years due to its unique properties (strength, toughness) and potential applications such as smart textiles, protective clothing and tissue engineering. The traditional 10--20 mum diameter, triangular-shaped Bombyx mori fibers have remained unchanged over the years. However, in our study, we examine the scientific implication and potential applications of reducing the diameter to the nanoscale, changing the triangular shape of the fiber and adding nanofillers in the form of single wall carbon nanotubes (SWNT) by the electrospinning process. The electrospinning process preserves the natural conformation of the silk (random and beta-sheet). The feasibility of changing the properties of the electrospun nanofibers by post processing treatments (annealing and chemical treatment) was investigated. B. mori silk fibroin solution (formic acid) was successfully electrospun to produce uniform nanofibers (as small as 12 nm). Response Surface Methodology (RSM) was applied for the first time to experimental results of electrospinning, to develop a processing window that can reproduce regenerated silk nanofibers of a predictable size (d < 100nm). SWNT-silk multifunctional nanocomposite fibers were fabricated for the first time with anticipated properties (mechanical, thermal and electrically conductive) that may have scientific applications (nerve regeneration, stimulation of cell-scaffold interaction). In order to realize these applications, the following areas need to be addressed: a systematic investigation of the dispersion of the nanotubes in the silk matrix, a determination of new methodologies for characterizing the nanofiber properties and establishing the nature of the silk-SWNT interactions. A new visualization system was developed to characterize the transport properties of the nanofibrous assemblies. The morphological, chemical, structural and mechanical properties of the nanofibers were determined by field emission environmental scanning microscopy, Fourier transform infrared and Raman spectroscopy, wide angle x-ray diffraction and microtensile tester respectively.

Ayutsede, Jonathan Eyitouyo

351

Synthesis and characterization of biocompatible nanodiamond-silk hybrid material  

PubMed Central

A new hybrid material consisting of nanodiamonds (NDs) and silk has been synthesized and investigated. NDs can contain bright fluorescence centers, important for bioprobes to image biological structures at the nanoscale and silk provides a transparent, robust matrix for these nanoparticles in-vivo or in-vitro. The ND-silk hybrid films were determined to be highly transparent in the visible to near infrared wavelength range. The NDs embedded in silk exhibited significant enhancement of emission relative to air, correlating with theoretical predictions. Furthermore, animal toxicity tests confirmed ND-silk films to be non-toxic in an in-vivo mice model. PMID:24575352

Khalid, Asma; Lodin, Rebecca; Domachuk, Peter; Tao, Hu; Moreau, Jodie E.; Kaplan, David L.; Omenetto, Fiorenzo G.; Gibson, Brant C.; Tomljenovic-Hanic, Snjezana

2014-01-01

352

The use of injectable sonication-induced silk hydrogel for VEGF 165 and BMP2 delivery for elevation of the maxillary sinus floor  

Microsoft Academic Search

Sonication-induced silk hydrogels were previously prepared as an injectable bone replacement biomaterial, with a need to improve osteogenic features. Vascular endothelial growth factor (VEGF165) and bone morphogenic protein-2 (BMP-2) are key regulators of angiogenesis and osteogenesis, respectively, during bone regeneration. Therefore, the present study aimed at evaluating in situ forming silk hydrogels as a vehicle to encapsulate dual factors for

Wenjie Zhang; Xiuli Wang; Shaoyi Wang; Jun Zhao; Lianyi Xu; Chao Zhu; Deliang Zeng; Jake Chen; Zhiyuan Zhang; David L. Kaplan; Xinquan Jiang

2011-01-01

353

Isolation and characterisation of sericin antifreeze peptides and molecular dynamics modelling of their ice-binding interaction.  

PubMed

This study aimed to isolate and characterise a novel sericin antifreeze peptide and investigate its ice-binding molecular mechanism. The thermal hysteresis activity of ice-binding sericin peptides (I-SP) was measured and their activity reached as high as 0.94 °C. A P4 fraction, with high hypothermia protective activity and inhibition activity of ice recrystallisation, was obtained from I-SP, and a purified sericin peptide, named SM-AFP, with the sequence of TTSPTNVSTT and a molecular weight of 1009.50 Da was then isolated from the P4 fraction. Treatment of Lactobacillus delbrueckii Subsp. bulgaricus LB340 LYO with 100 ?g/ml synthetic SM-AFP led to 1.4-fold increased survival (p < 0.05). Finally, an SM-AFP/ice binding model was constructed and results of molecular dynamics simulation suggested that the binding of SM-AFP with ice and prevention of ice crystal growth could be attributed to hydrogen bond formation, hydrophobic interaction and non-bond interactions. Sericin peptides could be developed into beneficial cryoprotectants and used in frozen food processing. PMID:25529728

Wu, Jinhong; Rong, Yuzhi; Wang, Zhengwu; Zhou, Yanfu; Wang, Shaoyun; Zhao, Bo

2015-05-01

354

Osteogenic and adipogenic differentiation of rat bone marrow cells on non-mulberry and mulberry silk gland fibroin 3D scaffolds.  

PubMed

This study investigates the potential of 3D silk scaffolds fabricated using tropical tasar non-mulberry, Antheraea mylitta and mulberry, Bombyx mori silk gland fibroin proteins as substrate for osteogenic and adipogenic differentiation of rat bone marrow cells (BMCs). The scaffolds are mechanically robust and show homogenous pore distribution with high porosity and interconnected pore walls. Low immunogenicity of fabricated silk scaffolds as estimated through TNF alpha release indicates its potential as future biopolymeric graft material. Rat bone marrow cells cultured on scaffolds for 28 days under static conditions in osteogenic and adipogenic media respectively led to induction of differentiation. Proliferation and spreading of fibroblasts and bone marrow cells on silk scaffolds were observed to be dependent on scaffold porosity as revealed through confocal microscopic observations. Histological analysis shows osteogenic differentiation within silk scaffolds resulting in extensive mineralization in the form of deposited nodules as observed through intense Alizarin Red S staining. Similarly, adipogenesis was marked by the presence of lipid droplets within scaffolds on staining with Oil Red O. Real-time PCR studies reveal higher transcript levels for osteopontin (Spp1), osteocalcin (Bglap2) and osteonectin (Sparc) genes under osteogenic conditions. Similarly, upregulated adipogenic gene expression was observed within A. mylitta and B. mori scaffolds under adipogenic conditions for Peroxisome proliferator activated receptor gamma (PPARgamma2), lipoprotein lipase (LPL) and adipocyte binding protein (aP2) genes. The results suggest suitability of silk fibroin protein 3D scaffolds as natural biopolymer for potential bone and adipose tissue engineering applications. PMID:19577292

Mandal, Biman B; Kundu, Subhas C

2009-10-01

355

Stimuli-responsive capsules prepared from regenerated silk fibroin microspheres.  

PubMed

Microcapsules are synthesized via the self-assembly of silk-fibroin microspheres with polycaprolactone in a colliodosome preparation procedure. The microsphere building blocks with controlled diameter are prepared from the naturally occurring biopolymer, silk fibroin, and then act as stabilizers for oil-in-water (O/W) emulsion and organized themselves on the surface of chloroform droplets to form the capsules. The concentration of the protein-based microspheres and the binding polycacrolactone is used to tailor the size, as well as the permeability of the resultant capsules. Furthermore, microencapsulation of fluorescently labeled macromolecules (20-2000 KDa) is demonstrated. The permeability of the capsules is observed to be stimuli responsive, making it possible to incorporate a pH-triggered payload release from the capsules. The relatively simple preparation of capsules with controlled dimensions and tunable permeability, alongside the biocompatibility and biodegradability of both polymer components makes these promising materials for potential use in smart drug-delivery systems. PMID:24532252

Cheng, Cheng; Teasdale, Ian; Brüggemann, Oliver

2014-06-01

356

Comprehensive characterization of well-defined silk fibroin surfaces: Toward multitechnique studies of surface modification effects.  

PubMed

The study aims at a comprehensive surface characterization of untreated and oxygen plasma-treated silk fibroin with a particular focus on phenomena relevant to biointeraction and cell adhesion. For that purpose, a range of advanced surface diagnostic techniques is employed to thoroughly investigate well-defined and especially clean silk fibroin samples in a comparable setting. This includes surface chemistry and surface charges as factors, which control protein adsorption, but also hydration and swelling of the material as important parameters, which govern the mechanical stiffness at the interface with aqueous media. Oxygen plasma exposure of silk fibroin surfaces reveals that material ablation strongly predominates over the introduction of functional groups even for mild plasma conditions. A substantial increase in mechanical stiffness is identified as the most prominent effect upon this kind of plasma treatment. Regarding the experimental approach and the choice of techniques, the work goes beyond previous studies in this field and paves the way for well-founded investigations of other surface-selective modification procedures that enhance the applicability of silk fibroin in biomedical applications. PMID:25899685

Amornsudthiwat, Phakdee; Nitschke, Mirko; Zimmermann, Ralf; Friedrichs, Jens; Grundke, Karina; Pöschel, Kathrin; Damrongsakkul, Siriporn; Werner, Carsten

2015-01-01

357

Mechanical and physical properties of recombinant spider silk films using organic and aqueous solvents.  

PubMed

Spider silk has exceptional mechanical and biocompatibility properties. The goal of this study was optimization of the mechanical properties of synthetic spider silk thin films made from synthetic forms of MaSp1 and MaSp2, which compose the dragline silk of Nephila clavipes. We increased the mechanical stress of MaSp1 and 2 films solubilized in both HFIP and water by adding glutaraldehyde and then stretching them in an alcohol based stretch bath. This resulted in stresses as high as 206 MPa and elongations up to 35%, which is 4× higher than the as-poured controls. Films were analyzed using NMR, XRD, and Raman, which showed that the secondary structure after solubilization and film formation in as-poured films is mainly a helical conformation. After the post-pour stretch in a methanol/water bath, the MaSp proteins in both the HFIP and water-based films formed aligned ?-sheets similar to those in spider silk fibers. PMID:25030809

Tucker, Chauncey L; Jones, Justin A; Bringhurst, Heidi N; Copeland, Cameron G; Addison, J Bennett; Weber, Warner S; Mou, Qiushi; Yarger, Jeffery L; Lewis, Randolph V

2014-08-11

358

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

359

Porous silk scaffolds for delivery of growth factors and stem cells to enhance bone regeneration.  

PubMed

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

Zhang, Wenjie; Zhu, Chao; Ye, Dongxia; Xu, Ling; Zhang, Xiaochen; Wu, Qianju; Zhang, Xiuli; Kaplan, David L; Jiang, Xinquan

2014-01-01

360

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

361

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

362

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

363

The Ancient Art of Silk Painting  

ERIC Educational Resources Information Center

In this article, the author describes a silk-painting project with a sea-creature theme for eighth-grade students. Other themes can be used such as geometric quilt designs, tropical rain forest, large flowers, Art Nouveau motifs, portraits and more. (Contains 2 resources.)

Yonker, Kim

2010-01-01

364

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

365

A green salt-leaching technique to produce sericin/PVA/glycerin scaffolds with distinguished characteristics for wound-dressing applications.  

PubMed

Sericin/PVA/glycerin scaffolds could be fabricated using the freeze-drying technique; they showed good physical and biological properties and can be applied as wound dressings. However, freeze-drying is an energy- and time-consuming process with a high associated cost. In this study, an alternative, solvent-free, energy- and time-saving, low-cost salt-leaching technique is introduced as a green technology to produce sericin/PVA/glycerin scaffolds. We found that sericin/PVA/glycerin scaffolds were successfully fabricated without any crosslinking using a salt-leaching technique. The salt-leached sericin/PVA/glycerin scaffolds had a porous structure with pore interconnectivity. The sericin in the salt-leached scaffolds had a crystallinity that was as high as that of the freeze-dried scaffolds. Compared to the freeze-dried scaffolds with the same composition, the salt-leached sericin/PVA/glycerin scaffolds has larger pores, a lower Young's modulus, and faster rates of biodegradation and sericin release. When cultured with L929 mouse fibroblast cells, a higher number of cells were found in the salt-leached scaffolds. Furthermore, the salt-leached scaffolds were less adhesive to the wound, which would reduce pain upon removal. Therefore, salt-leached sericin/PVA/glycerin scaffolds with distinguished characteristics were introduced as another choice of wound dressing, and their production process was simpler, more energy efficient, and saved time and money compared to the freeze-dried scaffolds. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2014. PMID:25175958

Aramwit, Pornanong; Ratanavaraporn, Juthamas; Ekgasit, Sanong; Tongsakul, Duangta; Bang, Nipaporn

2014-08-30

366

Antibiotic-Releasing Silk Biomaterials for Infection Prevention and Treatment.  

PubMed

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

2013-02-18

367

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

368

Thermal Analysis, Structural Studies and Morphology of Spider Silk-like Block Copolymers  

NASA Astrophysics Data System (ADS)

Spider silk is a remarkable natural block copolymer, which offers a unique combination of low density, excellent mechanical properties, and thermal stability over a wide range of temperature, along with biocompatibility and biodegrability. The dragline silk of Nephila clavipes, is one of the most well understood and the best characterized spider silk, in which alanine-rich hydrophobic blocks and glycine-rich hydrophilic blocks are linked together generating a functional block copolymer with potential uses in biomedical applications such as guided tissue repair and drug delivery. To provide further insight into the relationships among peptide amino acid sequence, block length, and physical properties, in this thesis, we studied synthetic proteins inspired by the genetic sequences found in spider dragline silks, and used these bioengineered spider silk block copolymers to study thermal, structural and morphological features. To obtain a fuller understanding of the thermal dynamic properties of these novel materials, we use a model to calculate the heat capacity of spider silk block copolymer in the solid or liquid state, below or above the glass transition temperature, respectively. We characterize the thermal phase transitions by temperature modulated differential scanning calorimetry (TMDSC) and thermogravimetric analysis (TGA). We also determined the crystallinity by TMDSC and compared the result with Fourier transform infrared spectroscopy (FTIR) and wide angle X-ray diffraction (WAXD). To understand the protein-water interactions with respect to the protein amino acid sequence, we also modeled the specific reversing heat capacity of the protein-water system, Cp(T), based on the vibrational, rotational and translational motions of protein amino acid residues and water molecules. Advanced thermal analysis methods using TMDSC and TGA show two glass transitions were observed in all samples during heating. The low temperature glass transition, Tg(1), is related to both the bound water removal induced conformational change and the hydrophobicity of the protein sequences, while the high temperature glass transition, Tg( 2), above 130 °C is the now dry protein glass transition. Real-time Fourier transform infrared spectroscopy (FTIR) confirmed that conformational change occurred during the two glass transition, with a random coils to beta turns transition during Tg(1) and alpha helices to beta turns transition during Tg( 2). Due to the hydrophobic and hydrophilic nature of the blocks, the spider silk block copolymers tend to self-assemble into various microstructures. To study the morphological features, the spider silk-like block copolymers were treated with hexafluoroisopropanol or methanol, or subjected to thermal treatment. Using scanning electron microscopies, micelles were observed in thermally treated films. Fibrillar networks and hollow vesicles were observed in methanol-cast samples, while no micro-structures were formed in HFIP-cast films, indicating that morphology and crystallinity can be tuned by thermal treatments. Results indicate when we increase the number of repeating unit of A-block in the protein, sample films crystallize more easily and are more thermally stable. Moreover, when samples crystallize, the secondary structure of A-block and B-block become different, thus it will be easier to form bilayer structures which could fold into vesicles or tube structures during drying.

Huang, Wenwen

369

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

370

CO2-assisted synthesis of silk fibroin hydrogels and aerogels.  

PubMed

Biocompatible and biodegradable porous materials based on silk fibroin (SF), a natural protein derived from the Bombyx mori silkworm, are being extensively investigated for use in biomedical applications including mammalian cell bioprocessing, tissue engineering and drug delivery applications. In this work, low-pressure, gaseous CO2 is used as an acidifying agent to fabricate SF hydrogels. This low-pressure CO2 acidification method is compared to an acidification method using high-pressure CO2 to demonstrate the effect of CO2 mass transfer and pressure on SF sol-gel kinetics. The effect of SF molecular weight on the sol-gel kinetics is determined using the low-pressure CO2 method. The results from these studies demonstrate that low-pressure CO2 processing proves to be a facile method for synthesizing 3-D SF hydrogels. PMID:24954908

Mallepally, Rajendar R; Marin, Michael A; McHugh, Mark A

2014-10-01

371

Prof. Dr. Klaus-Peter Wild Dr. Silke Schworm  

E-print Network

Prof. Dr. Klaus-Peter Wild Dr. Silke Schworm Stephanie Hiltmann, M. A. Dipl.-Päd. Magdalena Würfl Schritten Prof. Dr. K.-P. Wild #12;Sekretariat des Lehrstuhls Prof. Wild Prof. Dr. K.-P. Wild #12;silke.schworm@paedagogik.uni-r.de Dr. Silke Schworm klaus-peter.wild@paedagogik.uni-r.de Prof. Dr. Klaus-Peter Wild stephanie

Schubart, Christoph

372

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

373

Silk-based biomaterials for sustained drug delivery.  

PubMed

Silk presents a rare combination of desirable properties for sustained drug delivery, including aqueous-based purification and processing options without chemical cross-linkers, compatibility with common sterilization methods, controllable and surface-mediated biodegradation into non-inflammatory by-products, biocompatibility, utility in drug stabilization, and robust mechanical properties. A versatile silk-based toolkit is currently available for sustained drug delivery formulations of small molecule through macromolecular drugs, with a promise to mitigate several drawbacks associated with other degradable sustained delivery technologies in the market. Silk-based formulations utilize silk's well-defined nano- through microscale structural hierarchy, stimuli-responsive self-assembly pathways and crystal polymorphism, as well as sequence and genetic modification options towards targeted pharmaceutical outcomes. Furthermore, by manipulating the interactions between silk and drug molecules, near-zero order sustained release may be achieved through diffusion- and degradation-based release mechanisms. Because of these desirable properties, there has been increasing industrial interest in silk-based drug delivery systems currently at various stages of the developmental pipeline from pre-clinical to FDA-approved products. Here, we discuss the unique aspects of silk technology as a sustained drug delivery platform and highlight the current state of the art in silk-based drug delivery. We also offer a potential early development pathway for silk-based sustained delivery products. PMID:24910193

Yucel, Tuna; Lovett, Michael L; Kaplan, David L

2014-09-28

374

Spider Silk as Guiding Biomaterial for Human Model Neurons  

PubMed Central

Over the last years, a number of therapeutic strategies have emerged to promote axonal regeneration. An attractive strategy is the implantation of biodegradable and nonimmunogenic artificial scaffolds into injured peripheral nerves. In previous studies, transplantation of decellularized veins filled with spider silk for bridging critical size nerve defects resulted in axonal regeneration and remyelination by invading endogenous Schwann cells. Detailed interaction of elongating neurons and the spider silk as guidance material is unknown. To visualize direct cellular interactions between spider silk and neurons in vitro, we developed an in vitro crossed silk fiber array. Here, we describe in detail for the first time that human (NT2) model neurons attach to silk scaffolds. Extending neurites can bridge gaps between single silk fibers and elongate afterwards on the neighboring fiber. Culturing human neurons on the silk arrays led to an increasing migration and adhesion of neuronal cell bodies to the spider silk fibers. Within three to four weeks, clustered somata and extending neurites formed ganglion-like cell structures. Microscopic imaging of human neurons on the crossed fiber arrays in vitro will allow for a more efficient development of methods to maximize cell adhesion and neurite growth on spider silk prior to transplantation studies. PMID:24949480

Roloff, Frank; Strauß, Sarah; Vogt, Peter M.; Bicker, Gerd; Radtke, Christine

2014-01-01

375

Spider silk as guiding biomaterial for human model neurons.  

PubMed

Over the last years, a number of therapeutic strategies have emerged to promote axonal regeneration. An attractive strategy is the implantation of biodegradable and nonimmunogenic artificial scaffolds into injured peripheral nerves. In previous studies, transplantation of decellularized veins filled with spider silk for bridging critical size nerve defects resulted in axonal regeneration and remyelination by invading endogenous Schwann cells. Detailed interaction of elongating neurons and the spider silk as guidance material is unknown. To visualize direct cellular interactions between spider silk and neurons in vitro, we developed an in vitro crossed silk fiber array. Here, we describe in detail for the first time that human (NT2) model neurons attach to silk scaffolds. Extending neurites can bridge gaps between single silk fibers and elongate afterwards on the neighboring fiber. Culturing human neurons on the silk arrays led to an increasing migration and adhesion of neuronal cell bodies to the spider silk fibers. Within three to four weeks, clustered somata and extending neurites formed ganglion-like cell structures. Microscopic imaging of human neurons on the crossed fiber arrays in vitro will allow for a more efficient development of methods to maximize cell adhesion and neurite growth on spider silk prior to transplantation studies. PMID:24949480

Roloff, Frank; Strauß, Sarah; Vogt, Peter M; Bicker, Gerd; Radtke, Christine

2014-01-01

376

Implantable silk composite microneedles for programmable vaccine release kinetics and enhanced immunogenicity in transcutaneous immunization  

PubMed Central

Microneedle vaccines mimic several aspects of cutaneous pathogen invasion by targeting antigen to skin-resident dendritic cells and triggering local inflammatory responses in the skin, which are correlated with enhanced immune responses. Here we tested whether control over vaccine delivery kinetics can enhance immunity through further mimicry of kinetic profiles present during natural acute infections. We report an approach for the fabrication of silk/poly(acrylic acid) (PAA) composite microneedles composed of a silk tip supported on a PAA base. On brief application of microneedle patches to skin, the PAA bases rapidly dissolved to deliver a protein subunit vaccine bolus, while also implanting persistent silk hydrogel depots into the skin for a low-level sustained cutaneous vaccine release over 1-2 weeks. Use of this platform to deliver a model whole-protein vaccine with optimized release kinetics resulted in >10-fold increases in antigen-specific T-cell and humoral immune responses relative to traditional parenteral needle-based immunization. PMID:23847143

Irvie, Darrell J.; Hammond, Paula T.

2014-01-01

377

Silk-regulated hierarchical hollow magnetite/carbon nanocomposite spheroids for lithium-ion battery anodes  

NASA Astrophysics Data System (ADS)

Hierarchical olive-like structured carbon-Fe3O4 nanocomposite particles composed of a hollow interior and a carbon coated surface are prepared by a facile, silk protein-assisted hydrothermal method. Silk nanofibers as templates and carbon precursors first regulate the formation of hollow Fe2O3 microspheres and then they are converted into carbon by a reduction process into Fe3O4. This process significantly simplifies the fabrication and carbon coating processes to form complex hollow structures. When tested as anode materials for lithium-ion batteries, these hollow carbon-coated particles exhibit high capacity (900 mAh g?1), excellent cycle stability (180 cycles) and rate performance due to their unique hierarchical hollow structure and carbon coating.

Sheng, Weiqin; Zhu, Guobin; Kaplan, David L.; Cao, Chuanbao; Zhu, Hesun; Lu, Qiang

2015-03-01

378

Silk-regulated hierarchical hollow magnetite/carbon nanocomposite spheroids for lithium-ion battery anodes.  

PubMed

Hierarchical olive-like structured carbon-Fe3O4 nanocomposite particles composed of a hollow interior and a carbon coated surface are prepared by a facile, silk protein-assisted hydrothermal method. Silk nanofibers as templates and carbon precursors first regulate the formation of hollow Fe2O3 microspheres and then they are converted into carbon by a reduction process into Fe3O4. This process significantly simplifies the fabrication and carbon coating processes to form complex hollow structures. When tested as anode materials for lithium-ion batteries, these hollow carbon-coated particles exhibit high capacity (900 mAh g(-1)), excellent cycle stability (180 cycles) and rate performance due to their unique hierarchical hollow structure and carbon coating. PMID:25706314

Sheng, Weiqin; Zhu, Guobin; Kaplan, David L; Cao, Chuanbao; Zhu, Hesun; Lu, Qiang

2015-03-20

379

Physical and biological regulation of neuron regenerative growth and network formation on recombinant dragline silks.  

PubMed

Recombinant spider silks produced in transgenic goat milk were studied as cell culture matrices for neuronal growth. Major ampullate spidroin 1 (MaSp1) supported neuronal growth, axon extension and network connectivity, with cell morphology comparable to the gold standard poly-lysine. In addition, neurons growing on MaSp1 films had increased neural cell adhesion molecule (NCAM) expression at both mRNA and protein levels. The results indicate that MaSp1 films present useful surface charge and substrate stiffness to support the growth of primary rat cortical neurons. Moreover, a putative neuron-specific surface binding sequence GRGGL within MaSp1 may contribute to the biological regulation of neuron growth. These findings indicate that MaSp1 could regulate neuron growth through its physical and biological features. This dual regulation mode of MaSp1 could provide an alternative strategy for generating functional silk materials for neural tissue engineering. PMID:25701039

An, Bo; Tang-Schomer, Min D; Huang, Wenwen; He, Jiuyang; Jones, Justin A; Lewis, Randolph V; Kaplan, David L

2015-04-01

380

P25 gene regulation in Bombyx mori silk gland: two promoter-binding factors have distinct tissue and developmental specificities.  

PubMed Central

The gene encoding the silk protein P25 is expressed in the posterior silk gland of Bombyx mori with strict territorial and developmental specificities. The cis-acting regulatory elements previously located within the 441-bp 5' proximal sequence of the gene were examined for protein-binding capacities. We identified two factors, BMFA and SGFB, that lead to prominent band shifts and the target sites for which are included in a region homologous to the fibroin gene enhancer sequence. Analysis of the tissue-specific incidence of both factors showed that BMFA is ubiquitous, whereas SGFB is restricted to the silk gland cells. However, SGFB was found in both posterior and middle silk gland cells and therefore likely directs organ-specific, but not territory-specific, expression. Developmental studies throughout the fourth larval molt, at which the P25 gene status changes from derepressed to repressed, revealed that BMFA is reversibly modified at the transition from intermolt to molt. Indeed, the preexisting BMFA is replaced by a structurally related factor, BMFA', during the 2 h following head capsule apolysis. The exact temporal coincidence of this conversion with the onset of gene repression suggests that BMFA' is involved in transcription inactivation and likely results from a transduction process initiated by the hormonal change at molting. Images PMID:1448104

Durand, B; Drevet, J; Couble, P

1992-01-01

381

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)

2013-01-01

382

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

383

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

384

Transgenic phenolic production in corn silks moderately enhances insect resistance  

Technology Transfer Automated Retrieval System (TEKTRAN)

Some phenolic compounds produced in corn silks, such as maysin, can promote resistance to caterpillar pests. We evaluated transgenic maize engineered to express a maize cDNA controlled by a putative silk specific promoter for secondary metabolite production and corn earworm resistance. Transgene e...

385

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

386

Innovative and Multidirectional Applications of Natural Fibre, Silk - A Review  

Microsoft Academic Search

Realization of current trends and innovative uses of insect fibre, silk is required to exploit its compatibility, eco friendly and value addition potential. The nutritive value as human diet for cardiac and diabetic patients, component for cosmetic preparations indicate silk application rate. The bio-compatibility made it a base material for tissue wall, membrane, muscle ligament, blood vessel, nerve gadget, cartilage

R. Manohar Reddy

387

First investigation of spider silk as a braided microsurgical suture.  

PubMed

Inhibition of axonal outgrowth accompanied by neuroma formation appears in microsurgical nerve repair as reaction to common microsuture materials like silk, nylon, or polyglycolic acid. In contrast, recent findings revealed advantages of spider silk fibers in guiding Schwann cells in nerve regeneration. Here, we asked if we could braid microsutures from native spider silk fibers. Microsutures braided of native spider dragline silk were manufactured, containing either 2 × 15 or 3 × 10 single fibres strands. Morphologic appearance was studied and tensile strength and stress-strain ratio (SSR) were calculated. The constructed spider silk sutures showed a median thickness of 25 ?m, matching the USP definition of 10-0. Maximum load and tensile strength for both spider silk microsutures were significantly more than 2-fold higher than for nylon suture; SSR was 1.5-fold higher. All values except elasticity were higher in 3 × 10 strand sutures compared to 2 × 15 strand sutures, but not significantly. In this pilot study, we demonstrate the successful manufacture of microsutures from spider silk. With regards to the mechanical properties, these sutures were superior to nylon sutures. As spider silk displays high biocompatibility in nerve regeneration, its usage in microsurgical nerve repair should be considered. PMID:21432995

Kuhbier, Joern W; Reimers, Kerstin; Kasper, Cornelia; Allmeling, Christina; Hillmer, Anja; Menger, Björn; Vogt, Peter M; Radtke, Christine

2011-05-01

388

Mechanical Response of Silk Crystalline Units from Force-Distribution Analysis  

E-print Network

Mechanical 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-strain relationships of four different models, from spider and Bombyx mori silk peptides, in antiparallel and parallel

Gräter, Frauke

389

Nephila clavipes Spider Dragline Silk Microstructure Studied by Scanning Transmission X-ray Microscopy  

E-print Network

Nephila clavipes Spider Dragline Silk Microstructure Studied by Scanning Transmission X-mail: michel.pezolet@chm.ulaval.ca Abstract: Nephila clavipes dragline silk microstructure has been parameter P2 have been derived for spider silk for the first time. Dragline silk presents a very fine

Hitchcock, Adam P.

390

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

E-print Network

1990 INTRODUCTION Most of the world's 40,000 species of spiders produce dragline silk from major ampullate silk glands to spin lifelines and frames of webs (Fig.1A,B). Dragline silk's impressive toughness spider silk an important model for biomimetic research (Gosline et al., 1986; Vadlamudi, 1995; Vollrath

Agnarsson, Ingi

391

Effects of crystalline subunit size on silk fiber mechanics Murat Cetinkaya,*a  

E-print Network

Effects of crystalline subunit size on silk fiber mechanics Murat Cetinkaya,*a Senbo Xiaob subunits as they occur in most spider silks on silk fiber mechanics. We vary the crystal size in terms with Finite Element simulations of a two phase silk fiber model in order to determine the stress

Gräter, Frauke

392

In vivo study of anterior cruciate ligament regeneration using mesenchymal stem cells and silk scaffold  

Microsoft Academic Search

Although most in vitro studies indicate that silk is a suitable biomaterial for ligament tissue engineering, in vivo studies of implanted silk scaffolds for ligament reconstruction are still lacking. The objective of this study is to investigate anterior cruciate ligament (ACL) regeneration using mesenchymal stem cells (MSCs) and silk scaffold. The scaffold was fabricated by incorporating microporous silk sponges into

Hongbin Fan; Haifeng Liu; Eugene J. W. Wong; Siew L. Toh; James C. H. Goh

2008-01-01

393

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.

394

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

Microsoft Academic Search

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

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

2005-01-01

395

Structural studies of Bombyx mori silk fibroin during regeneration from solutions and wet fiber spinning.  

PubMed

Regenerated silk fibroin materials show properties dependent on the methods used to process them. The molecular structures of B. mori silk fibroin both in solution and in solid states were studied and compared using X-ray diffraction, FTIR, and (13)C NMR spectroscopy. Some portion of fibroin protein molecules dissolved in formic acid already have a beta-sheet structure, whereas those dissolved in TFA have some helical conformation. Moreover, fibroin molecules were spontaneously assembled into an ordered structure as the acidic solvents were removed from the fibroin-acidic solvent systems. This may be responsible for the improved physical properties of regenerated fibroin materials from acidic solvents. Regenerated fibroin materials have shown poor mechanical properties and brittleness compared to their original form. These problems were technically solved by improving the fiber forming process according to a method reported here. The regenerated fibroin fibers showed much better mechanical properties compared to the native silk fiber and their physical and chemical properties were characterized by X-ray diffraction, solid state (13)C NMR spectroscopy, SinTech tensile testing, and SEM. PMID:15877399

Ha, Sung-Won; Tonelli, Alan E; Hudson, Samuel M

2005-01-01

396

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

397

?-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

398

Electricity from the Silk Cocoon Membrane  

NASA Astrophysics Data System (ADS)

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.

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

2014-06-01

399

Local Structure and Dynamics of Serine in the Heterogeneous Structure of the Crystalline Domain of Bombyx mori Silk Fibroin in  

E-print Network

of Bombyx mori Silk Fibroin in Silk II Form Studied by 2D 13 C-13 C Homonuclear Correlation NMR Information ABSTRACT: The crystalline fraction (Cp fraction) of silk fibroin in silk II form from the silkworm packing geometries that form separate but small domains. INTRODUCTION Silks continue to attract

Williamson, Mike P.

400

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. PMID:24616219

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

2015-01-01

401

The effect of steam sterilization on recombinant spider silk particles.  

PubMed

In this work, the recombinant spider silk protein eADF4(C16) was used to fabricate particles in the submicron range using a micromixing method. Furthermore, particles in the micrometer range were produced using an ultrasonic atomizer system. Both particle species were manufactured by an all-aqueous process. The submicroparticles were 332nm in average diameter, whereas 6.70?m was the median size of the microparticles. Both particle groups showed a spherical shape and exhibited high ?-sheet content in secondary structure. Submicro- and microparticles were subsequently steam sterilized and investigated with respect