Antheraea Pernyi Silk Fibroin Research Articles

Preparation of Water-Insoluble Antheraea Pernyi Silk Fibroin Films

Antheraea pernyi silk fibroin (ASF) solution was prepared by dissolving Antheraea pernyi silk fiber in lithium thiocyanate solution. The ASF/glycol blend films were prepared by casting aqueous solution of ASF mixed with glycol. The structure of blend film was investigated by the X-ray diffraction method and infrared spectroscopy. The result showed that the structure of regenerated ASF film was α-helix and random coil conformation. After mixing with glycol, it resulted in significant increase in β-sheet structure with the improvement of water resistance of the films. This effect was strongly dependent on glycol content in the blend film. When the glycol content was more than 45 wt%, the structure of ASF changed to β-sheet and the film became water-insoluble. The breaking strength and elongation of ASF/glycol blend film were 30 Mpa and 50 %, respectively. In summary, the ASF/glycol blend film provided a great potential as a biological material.

Regenerated <i>Antheraea pernyi </i>Silk Fibroin Nanofiber Film

Regenerated Antheraea Pernyi silk fibroin (ASF) solution was prepared by dissolving Antheraea Pernyi (A. pernyi) silk fiber in lithium thiocyanate solution. Atomic force microscopy (AFM) examination showed that there were many short nanofibers in regenerated ASF solution, which were made up of 10-20 nm nanospheres. In this paper, we discussed the formation mechanism of these nanofibers. The results of the surface tension showed that ASF had surface active and can significantly decline the surface tension of water from 73 mN/m to 51 mN/m. The hydrophobic side chains of ASF molecular would try to break away from water environment and thereby undergo self-assembly into nanospheres. And then, these nanospheres arrange into a short nanofiber. Then, the ASF nanofiber film was prepared by casting regenerated ASF solution blended with glycol. The structure of ASF was β-sheet and the film was water-insoluble. This blend film not only had excellent mechanical properties, but also can be biodegradated by enzyme in vitro. All of this indicated that this new regenerated ASF nanofiber film not only provided excellent biocompatibility, but also had large surface as extracellular matrix (ECM) to cell adhesion. The film made of nanofiber was similar to ECM on the nanoscale, which promoted cell migration and proliferation. In summary, it provided a great potential as a biological material.

Preparation of <i>Antheraea pernyi</i> Silk Fibroin/Chitosan Blend Scaffolds

In this work, Antheraea pernyi silk fibroin (ASF) /chitosan (CS) blend scaffolds were prepared by freeze-drying. The relationship between preparation conditions and morphological structures of blend scaffolds was also studied. The results indicated that the porosity and pore diameter of the ASF /CS scaffolds decreased with increasing of the proportion of chitosan and decreasing of the freezing temperature. By changing the process conditions, the average pore diameter and porosity of ASF/CS scaffolds could be adjusted in the range of 134 - 527 μm and 71 - 91% respectively. Compared with the pure ASF or CS materials, the ASF / CS (60/40) blend materials were more favor of cell adhesion.

Study on Antheraea Pernyi Silk Fibroin Porous Materials

Antheraea pernyi silk fibroin (ASF) solution was prepared by dissolving Antheraea pernyi silk fiber in lithium thiocyanate solution. The ASF/1,4-butanediol (ASF/BDO) blend porous materials were prepared with freeze-drying method. The structure of ASF was investigated by the X-ray diffraction method. The result showed that the structure of regenerated ASF scaffold was α-helix and random coil conformation. After ASF mixed with 5 wt% BDO, the structure of ASF was changed to β-sheet and the ASF scaffolds became water-insoluble. There were some small pores and fibrous structure in the big pores and the surface of pores was rough with a great many raised particles. The scaffolds with the average pore size of 300-1000μm and the porosity of 82-92% can efficiently be produced in this paper. Due to avoid the use of organic solvents or harsh chemicals, these new ASF based porous materials provide much more excellent biocompatibility and is expected to be applied to tissue regeneration scaffolds.

Enhancement of Lipid Metabolism and Antioxidant Defense Status in Mice Fed with High Fat Diet Supplemented with Antheraea pernyi Silk Fibroin Powder

The effect of diet supplementation of Antheraea pernyi (A. pernyi) silk fibroin on the lipid metabolism and antioxidant defense status in high fat-fed mice was investigated. The animals were given normal control diet (NC group), high fat diet (HF group), or high fat diet supplemented with A. pernyi silk fibroin powder (HFS group) for 7 weeks. After the experimental period, the HF group showed significant increase in body weight, plasma and hepatic total cholesterol levels, and hepatic triglyceride concentration, and decreased activities of hepatic antioxidant enzymes relative to NC group. However, the HFS group exhibited marked reduction in body weight, plasma cholesterol and hepatic triglyceride levels, hepatic lipogenic enzyme activities, and lipid peroxidation rate, and higher high-density lipoprotein (HDL)-cholesterol level, fecal triglyceride content, and antioxidant enzymes activities compared with that of HF group. These findings demonstrate that dietary feeding of A. pernyi silk fibroin could improve the lipid metabolism and antioxidant defense system via regulation of hepatic antioxidant and lipogenic enzymes activities. Hence, this silk fibroin may be beneficial as a functional biomaterial for the development of therapeutic agent against high fat diet-induced hyperlipidemia and its related diseases.

Enzymatic degradation of Antheraea pernyi silk fibroin 3D scaffolds and fibers

In this study, the in vitro enzymatic degradation behavior of the regenerated Antheraea pernyi silk fibroin ( Ap-SF) three-dimensional (3D) scaffolds and the natural Ap-SF fibers exposed to enzyme solutions of α-chymotrypsin, collagenase IA and protease XIV were investigated. The results indicated that all three proteases could degrade the Ap-SF 3D scaffolds, and the degradation ability was in the order protease XIV > collagenase IA > α-chymotrypsin. The regenerated Ap-SF 3D scaffold could be degraded completely in 18 days when exposed to 1.0 U/ml protease XIV at 37 °C, whereas under the same condition, the natural Ap-SF fiber only lost 5.6% of its weight, revealing its long-term degradation characteristics. There were abundant peptides and some free amino acids in the Ap-SF degradation products, but no free alanine. We suggested that the polyalanine block in the regenerated Ap-SF 3D scaffolds had strong resistance to enzyme attack. The proteolytic attack occurred in the non-polyalanine block of Ap-SF. The degradation rate of Ap-SF materials depended on the molecular conformation of Ap-SF, which could be controlled in the manufacturing process.

Characterization of tyrosine-rich Antheraea pernyi silk fibroin hydrolysate

Antheraea pernyi silk fibroin (SF) hydrolysate were characterized using UV–VIS spectrometer, amino acid composition and heavy metal contents to explore its potential sources for food or cosmetic additives. The hydrolyzed A. pernyi SF was separated into two parts: (a) SFA, alanine-rich fraction and (b) SFB, tyrosine-rich fraction. SFB exhibited strong absorption peaks at 210 and 280 nm due to the presence of the tyrosine. Heavy metal analysis showed that arsenic and mercury did not detect. Other heavy metals, which includes lead, cadmium, etc., were recorded only a trace amount. Therefore, A. pernyi SF hydrolysate could be safely used as sources of food, cosmetic and pharmaceuticals.

Gelation behavior of Antheraea pernyi silk fibroin

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

Antheraea pernyi silk fibroin maintains the immunosupressive properties of human bone marrow mesenchymal stem cells

We reported previously that regenerated Antheraea pernyi silk fibroin (A. pernyi SF) could support the attachment and growth of human bone marrow mesenchymal stem cells (hBMSCs). In this work, the immunosupressive effects of hBMSCs cultured on the A. pernyi SF films on T-cells were investigated in vitro. The production of IL-6, CD80, CD86 and HLA-DR by the hBMSCs was also observed. The study showed that hBMSCs cultured on the regenerated A. pernyi SF films still kept their immunosupression on T-cell proliferation and IL-2 secretion. Moreover, regenerated A. pernyi SF like regenerated Bombyx mori SF and collagen did not elicit T-cell proliferation but it could support the expression of IL-6 and surface antigen of hBMSCs. Regenerated A. pernyi SF can maintain the function of hBMSCs in immunomodulation and cytokines production, which has the potential utility of hBMSCs combined with A. pernyi SF in tissue replacement and repair.

Condensed Structure of Regenerated Antheraea Pernyi Silk Fibroin Porous Materials Prepared by Freeze-drying

Condensed Structure of Regenerated Antheraea Pernyi Silk Fibroin Porous Materials Prepared by Freeze-drying

Structure and properties of regenerated Antheraea pernyi silk fibroin filaments

AbstractDegummed Antheraea pernyi silk fibroin (APSF) fibers were dissolved in a lithium thiocyanate solution that was dialyzed against distilled water for 3 days. Solution‐cast APSF films were dissolved in hexafluoroisopropanol to prepare a spinning solution. Wet spinning was employed to prepare APSF filaments. Scanning electron microscopy, X‐ray diffraction, Fourier transform infrared spectroscopy, Raman spectroscopy, solid‐state 13C cross‐polarization/magic angle spinning nuclear magnetic resonance, and differential scanning calorimetry were employed to study the regenerated APSF filaments. The results showed a uniform distribution of molecular weights, and the largest one may have been greater than 200 kDa. The regenerated filaments contained β‐sheet, α‐helix, and random‐coil conformations, and their breaking strength was 52.20% of the breaking strength of the native fibers. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009

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

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

Porous 3‐D scaffolds from regenerated Antheraea pernyi silk fibroin

AbstractIn this study, porous three‐dimensional (3‐D) materials were prepared with the regenerated Antheraea pernyi (A. pernyi) silk fibroin by freeze‐drying from a lithium thiocyanate solution of its fibers. The relationship between preparation conditions and morphological structures of 3‐D materials was also studied. We concluded that with the decrease in A. pernyi silk fibroin solution concentration and the increase in the freezing temperature, the porosity and the average pore diameter of the 3‐D materials were increased while the pore density was decreased. By adjusting the freezing temperature and the silk fibroin solution concentration, the 3‐D materials having the average pore diameter of 75–260 µm and the porosity of 70–90% can efficiently be produced. As a kind of new material with excellent biocompatibility and bioactivity, the material is expected to be applied to tissue regeneration scaffolds. Copyright © 2007 John Wiley & Sons, Ltd.

Preparation and Characterization of Antheraea pernyi Silk Fibroin Based Nanohydroxyapatite Composites

In the present study, Antheraea pernyi silk fibroin (Ap-SF) is employed to regulate the mineralization of hydroxyapatite (HAp) nanocrystals. Calcium phosphate crystals precipitate in the aqueous solution of regenerated Ap-SF at pH 7.4 and room temperature. The samples are charaterized with X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), and scanning electron microscopy (SEM). Based on the XRD results, the inorganic phase in the mineralized Ap-SF composites is HAp as small particles with low crystallinity. The FTIR analysis reveals that the HAp crystals are carbonate-substituted HAp and compounded with Ap-SF. The TEM images show mineralized nanofibrils in the composites as rod-like shapes 4—5 nm in diameter. Electron diffraction (SAED) patterns of selected areas of the composites exhibit polycrystalline rings that are ascribed to HAp. SEM images show mineralized Ap-SF composites with several mineralized rods 49—74 nm in diameter.

Structure and properties of regenerated Antheraea pernyi silk fibroin in aqueous solution

Antheraea pernyi silk fibroin fibers were dissolved by aqueous lithium thiocyanate to obtain regenerated A. pernyi silk fibroin solution. By means of circular dichroism, 13C NMR and Raman spectroscopy, the molecular conformation of regenerated A. pernyi silk fibroin in aqueous solution was investigated. The relationship of environmental factors and sol–gel transformation behavior of regenerated A. pernyi silk fibroin was also studied. The molecular conformations of regenerated A. pernyi silk fibroin mainly were α-helix and random coil in solution. There also existed a little β-sheet conformation. It was obviously different with Bombyx mori silk fibroin, whose molecular conformation in solution was only random coil but no α-helix existence. With the increase of temperature and solution concentration and with the decrease of solution pH value, the gelation velocity of regenerated A. pernyi silk fibroin solution increased. Especially, it showed that A. pernyi silk fibroin was more sensitive to temperature than B. mori silk fibroin during the sol–gel transformation. The velocity increased obviously when the temperature was above 30 °C. During the sol–gel transformation, the molecular conformation of regenerated A. pernyi silk fibroin changed from random coil to β-sheet structure. The results of these studies provided important insight into the preparation of new biomaterials by silk fibroin protein.

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

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

In Vitro Study of the Proteolytic Degradation of Antheraea pernyi Silk Fibroin

In this study, Antheraea pernyi silk fibroin (Ap-SF) films were incubated with Protease Type XXI from Streptomyces griseus, at 37 degrees C, to investigate the degradation behavior in an in vitro model system. The enzyme-resistant fractions of Ap-SF films and the soluble peptides formed by proteolytic degradation were collected at specified times, from 1 to 17 days, and analyzed by high performance liquid chromatography, differential scanning calorimetry, FT-Raman, and FT-IR spectroscopy. Proteolysis resulted in extensive weight loss and progressive fragmentation of films, especially at long degradation times. A range of soluble peptides was formed by proteolysis. By high performance-size exclusion chromatography it was found that their average molecular weight changed with the time of incubation. The chemical analysis of the enzyme-resistant fraction of Ap-SF films at different times of degradation indicated that the proteolytic attack preferentially occurred in the less ordered Gly rich sequences and that the contribution of the Ala rich crystalline regions to the composition of biodegraded films became progressively larger. Accordingly, DSC and spectroscopic results showed an enhancement of the crystalline character of the biodegraded films. From the behavior of the most important thermal transitions, it was deduced that the alpha-helix domains probably represent the most enzyme-resistant fraction. The in vitro approach used in the present study seems to be a valid tool for studying the rate and mechanism of degradation of Ap-SF films and of other biopolymers of potential biomedical utility.

Supercritical CO2‐assisted synthesis of poly(acrylic acid)/Antheraea pernyi SF blend

AbstractMonomer acrylic acid (AA) and initiator azo‐bis(isobutyronitrile)were carried into Antheraea pernyi silk fibroin (SF) fibers using supercritical CO2 as a solvent and carrier, followed by free radical polymerization at a suitable temperature, resulting in PAA/SF blends. The binary system of CO2/AA and the ternary system of CO2/AA/AIBN systems were studied. Different impregnation conditions, such as time, pressure, and concentration of AA in the fluid phase on mass uptake, were studied. Fourier transform infrared spectroscopy and X‐ray diffraction results confirmed that PAA was indeed present in the silk and that there were intermolecular hydrogen bonds between PAA and SF. According to thermogravimetry and DTG, blending with PAA could enhance the thermal stability of SF slightly. The water retention values indicated that the hydrophilic nature of the fibers was improved. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 98: 864–868, 2005

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

Raman spectroscopy was used to investigate various enzyme-catalyzed reactions onto silk fibroin, i.e. the biodegradation of Tussah ( Antheraea pernyi) silk fibroin films by a proteolytic enzyme, the oxidation of domestic ( Bombyx mori) silk fibroin by mushroom tyrosinase and the subsequent grafting of chitosan onto oxidized silk. The spectra of Tussah silk fibroin films exposed to a bacterial protease for different times demonstrated that the cleavage of sensitive peptide bonds in the amorphous glycine-rich domains resulted in the loss of various amino acid residues (Tyr, Trp, Asp, etc.). The bands attributed to the crystalline alanine-rich sequences increased in intensity, and the β-sheet molecular conformation was not affected by biodegradation. Following oxidation with mushroom tyrosinase, the tyrosine bands of Bombyx mori fibroin decreased in intensity but did not disappear. The increase of the I 853/ I 829 intensity ratio indicated that the Tyr residues not accessible to the enzyme were located in a strongly hydrophobic environment. Raman spectroscopy provided evidence that chitosan was effectively grafted onto oxidized silk, probably via the Schiff-base mechanism, as shown by the behavior of the imine band at about 1646 cm −1. Grafting chitosan onto silk fibroin resulted in a β-sheet→random coil conformational transition of the protein component in the bioconjugated product.

Controlling molecular conformation of regenerated wild silk fibroin by aqueous ethanol treatment

AbstractRegenerated Antheraea pernyi silk fibroin film was prepared by dissolution of native silk fiber in aqueous lithium thiocyanate. The influence of aqueous ethanol treatment of the dried regenerated film on molecular conformation was studied by X‐ray diffraction, infrared spectroscopy, differential scanning calorimetry, and thermogravimetric analysis. While the initial regenerated film consisted of α‐helix and random coil components, aqueous ethanol treatment of the film resulted in significant increase in β‐sheet component and improvement of water resistance of the film. This effect was strongly dependent on ethanol concentration, and 40–60% (w/w) ethanol was most effective due to balance of hydrophilic/hydrophobic action of the solvent. Copyright © 2003 John Wiley & Sons, Ltd.