Silver nanoparticles incorporated electrospun silk fibers

We present a simple and mass-producible method of incorporating silver nanoparticles on the surface of electrospun silk non-woven membranes for the fabrication of antimicrobial wound dressings. Nanofibrous silk membranes with fiber diameters of 460 +/- 40 nm were electrospun from an aqueous Bombyx mori fibroin solution. The electrospun membranes incorporating silver nanoparticles were prepared by dipping the membranes in aqueous silver nitrate (AgNO3) solution (0.5 or 1.0 wt%) followed by photoreduction. Field emission scanning and transmission electron microscopy showed that silver nanoparticles were generated on the electrospun silk fibroin nanofibers as well as inside them. The interaction between the silver nanoparticles and amide groups in the silk fibroin molecules was characterized using X-ray photoelectron spectroscopy.     

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

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

Fabrication and characterization of fibroin solution and nanoparticle from silk fibers of Bombyx mori

The present investigation aims to study the effect of degumming time on the structural property of silk fiber obtained by silk cocoons of Bombyx mori, followed by preparation of the regenerated silk fibroin (RSF) solution which can be subsequently molded into silk nanoparticles. Silk fibers degummed with different media at different time intervals were investigated for the degumming loss and were characterized using Ffourier transform infrared (FTIR), differential scanning calorimetry (DSC), x-ray diffraction (XRD), and scanning electron microscopy (SEM). Maximum degumming was observed when the fibers were treated with sodium carbonate for 60 min. SEM and atomic force microscopy (AFM) images of RSF solution showed aggregation of silk globules resulting in formation of solvated macrochains and giving it an appearance of island-like morphology. Blank silk nanoparticles prepared from the RSF solution showed a smooth and spherical surface devoid of any adhesion using SEM, AFM, and transmission electron microscopy (TEM). The prepared silk nanoparticles may further be explored for loading drug entities and targeting.     

The preparation of insoluble fibroin films induced by degummed fibroin or fibroin microspheres

Blending degummed fibroin (DF) or insoluble fibroin microspheres with concentrated fibroin solution, the insoluble films were obtained through drying the solution at 40–45 ^∘C. The conformation of silk fibroin films was analyzed by infrared spectrum and X-ray diffractometry. The results demonstrated that β-sheet conformation increased rapidly when the degummed silk or insoluble microspheres blended with fibroin, while the pure SF membrane was mainly composed of α/random coil conformation when the other conditions kept same. This suggested that fibroin microspheres and degummed fibroin could induce the formation of β-sheet crystal and the insoluble films, without methanol after-treatment, could be obtained at approximately room temperature. Although the fibroin films blending with DF had many protuberances, the films containing fibroin microspheres had the smooth surface and could be used effectively in biotechnological materials and biomedical application.     

A novel electrospun silk fibroin/hydroxyapatite hybrid nanofibers

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

Fabrication of novel biofibers by coating silk fibroin with chitosan impregnated with silver nanoparticles

Nanoparticle based agents often applied as coatings on biomaterials have shown promise in delivering the improved sterility against variety of microbes. In the present study, silk fibers (SF) were coated with chitosan impregnated with silver nanoparticles (Ag–C–SF). These Ag–C–SF fibers were characterized using scanning electron microscopy (SEM) with energy dispersive X-ray analysis, atomic force microscopy (AFM), Infra Red spectroscopy, Thermogravimetric Analysis and Microbiological assay techniques. AFM studies have confirmed the nano sized silver particles in chitosan solution; SEM pictures have exhibited the coating of chitosan along with silver nanoparticles on the silk fibroin. The modified fibers have also shown anti-microbial activity and improved thermal stability. The Ag–C–SF fibers may be explored as wound dressing and tendon reconstruction material in future.     

渗析分离丝素蛋白盐溶液的研究

采用浓差渗析器进行丝素蛋白盐溶液的脱盐试验，研究了操作参数对脱盐试验的影响，确定了原料液丝素蛋白浓度10％、料液和自来水流量40L／h为其较合理的工艺参数．采用此设备脱盐，脱盐效率较高，脱盐率达到99.36％，丝素蛋白回收率达到89．7％．     

纳米TiO_2改性丝素膜制备工艺优化及其性能研究

设计正交试验法并采用溶胶一凝胶法制备了性能良好纳米TiO2溶胶丝素复合膜,并利用SEM、FTIR及XRD等分析手段对丝素膜进行表征,结果表明:纳米TiO2均匀分散在丝素膜中,纳米TiO2与丝素分子相互作用,导致丝素分子的重排,并诱导丝素构象从Silk Ⅰ向SilkⅡ转变.     

Conductive Au nanowires regulated by silk fibroin nanofibers

Conductive Au-biopolymer composites have promising applications in tissue engineering such as nerve tissue regeneration. In this study, silk fibroin nanofibers were formed in aqueous solution by regulating silk self-assembly process and then used as template for Au nanowire fabrication. We performed the synthesis of Au seeds by repeating the seeding cycles for several times in order to increase the density of Au seeds on the nanofibers. After electroless plating, densely decorated Au seeds grew into irregularly shaped particles following silk nanofiber to fill the gaps between particles and finally form uniform continuous nanowires. The conductive property of the Au-silk fibroin nanowires was studied with current-voltage （I-V） measurement. A typical ohmic behavior was observed, which highlighted their potential applications in nerve tissue regeneration.     

Characterization and deposition behavior of silk hydrogels soaked in simulated body fluid

This paper reports the mineralization ability of semi-interpenetrating networks composed of regenerated silk fibroin and polyacrylamide hydrogels soaked in simulated body fluid (SBF1x). Hydrogels were prepared by polymerization of acrylamide and N,N′-methylenebisacrylamide in silk fibroin solution with a redox pair as initiator. The incorporation of the fibroin within the polyacrylamide matrix was proved by FTIR–ATR spectroscopy. Swelling measurements in saline solution were performed to evaluate the behavior of these hydrogels having various compositions. Mineralization assays in SBF1x solution revealed the presence of apatite-like crystals onto the surface of the silk fibroin/polyacrylamide hydrogels. Cytotoxicity test by extract method revealed that these hydrogels with various contents in silk fibroin have not developed cytotoxic effects on human fibroblast cultures which increases the possibility of their use in biomedical applications. Mechanical compressive tests revealed good strengths for silk fibroin/polyacrylamide hydrogels. In this way, organic–inorganic hybrids analogous to bone structure can be produced under biomimetic conditions and could be further used in biomedical applications.     

Bovine pericardium coated with biopolymeric films as an alternative to prevent calcification: In vitro calcification and cytotoxicity results

Bovine pericardium, for cardiac valve fabrication, was coated with either chitosan or silk fibroin film. In vitro calcification tests of coated and non coated bovine pericardium were performed in simulated body fluid solution in order to investigate potential alternatives to minimize calcification on implanted heart valves. Complementary, morphology was assessed by scanning electron microscopy — SEM; X-ray diffraction (XRD) and infrared spectroscopy (FTIR-ATR) were performed for structural characterization of coatings and biocompatibility of chitosan. Silk fibroin films were assayed by in vitro cytotoxicity and endothelial cell growth tests. Bovine pericardium coated with silk fibroin or chitosan did not present calcification during in vitro calcification tests, indicating that these biopolymeric coatings do not induce bovine pericardium calcification. Chitosan and silk fibroin films were characterized as non cytotoxic and silk fibroin films presented high affinity to endothelial cells. The results indicate that bovine pericardium coated with silk fibroin is a potential candidate for cardiac valve fabrication, since the affinity of silk fibroin to endothelial cells can be explored to induce the tissue endothelization and therefore, increase valve durability by increasing their mechanical resistance and protecting them against calcification.     

丝素在离子液体中的溶解及再生丝素纤维的结构

以离子液体1-丁基-3-甲基咪唑氯盐（[BMIM]Cl）为溶剂,研究了丝素蛋白在[BMIM]Cl中的溶解情况,结果表明,温度对丝素在[BMIM]Cl中的溶解有较大影响,当温度〈90℃时,即使延长溶解时间,丝素也难以溶解,而当温度≥90℃时,随着温度升高,溶解速率明显加快。考虑到高温会使丝素降解,因此,选择100℃为丝素...     

A novel growth process of calcium carbonate crystals in silk fibroin hydrogel system

We report an interesting finding of calcium carbonate (CaCO₃) crystal growth in the silk fibroin (SF) hydrogel with different concentrations by a simple ion diffusion method. The experimental results indicate that the CaCO₃ crystals obtained from silk fibroin gels with low and high concentrations are all calcites with unusual morphologies. Time-dependent growth study was carried out to investigate the crystallization process. It is believed that silk fibroin hydrogel plays an important role in the process of crystallization. The possible formation mechanism of CaCO₃ crystals is proposed. This study provides a better explanation of the influence of silk fibroin concentration and its structure on CaCO₃ crystals growth.     

Green synthesis of gold nanoparticles: Its effect on cocoon and silk traits of mulberry silkworm (Bombyx mori L.)

In the present century the small particles are unique phenomenon which can be developed by bottom-up and top-down processes. These small particles may be considered as nanoparticles which help to build up a technology called nanotechnology. Nanomaterials are those materials which possess the length scales below 100 nm and quite often they make a comparison with a human hair, which is about 80,000 nm wide. We have introduced this technology, specially the green synthesis of gold (Au) nanoparticles in silkworm (Bombyx mori L.). The gold nanoparticles clearly indicate that they have a tremendous effect on enhancement of silk proteins and thus the enhancement of the cocoon weight in silkworms. Gold nanoparticles were prepared from onion, Allium cepa L. The extracted green gold nanomaterials from A. cepa were confirmed by UV-Vis spectrophotometer, XRD, FTIR, SEM, TEM and AFM. The function of green gold nanomaterials extracted from A. cepa was tested on silkworm physiology. We have used UV for judgment of the nature of particles and spectrum peak wavelength showed an absorption peak at 535 nm and indicated the wavelength of the surface plasmon resonance in gold nanoparticles (Au NPs). In blank solutions no such absorption peak was observed at 535 nm. Moreover, the gold (Au) XRD spectrum is supposed to and does demonstrate (111), (200), (220), and (311) peaks in the assortment of superimpose on the background. The process includes the (002) trace graphite peaks, where the (111) peak appears to be exceptionally sharp and strong which helps to propose that it is gold in nature. The FTIR shows that the examined particles are gold in nature. In SEM where electrons interact with atoms in the sample, producing various signals that can be detected and that hold information about the sample's surface topography and composition. The electron beam in SEM is generally scanned in a raster scan pattern, and therefore the beam's position is combined and detects the signal to produce an image. SEM can attain a resolution better than 1 nanometer size. The transmission electron microscope helps to accelerate the electrons as a source of elucidation. The AFM measurement is made in three dimensions process and thus it may be measured as horizontal to X-Y plane. Therefore, decree (magnification) measured at Z–direction, which is normally higher than X-Y. The said repulsive force is major one in AFM. Thus the tip and sample may considered to be the specific force in AFM which may measured at Z–direction. The effect of green gold nanoparticles on mulberry silkworm (Bombyx mori L) can exaggerated the silkworm physiological function. Larvae at 50, 100, 200, and 300 ppm doses were studied right from 1st stage to 5th instar stage. Gold nano treatment resulted in significant alterations in the percentage of fibroin and sericin proteins in the 5th instar as compared to that of control. At a 300 ppm dose of green nano gold the percentage of fibroin was 78.07, while sericin decreased from 39.46 (control) to 21.92. It was observed that the green gold nanomaterials have the ability to not only alter the fibroin protein but also enhance the cocoon and silk traits. The aim of this study was to investigate the effect of extra foliation of mulberry leaves with G-GNPs extracted from A. cepa on larval duration, mature silk gland weight, pupal weight, cocoon weight, cocoon shell weight, fibroin and sericins contents, etc. Moreover, the enhanced production of fibroin will explore a new venture in bioengineering and also in biomedical field.     

原子力显微镜下蚕丝及蜘蛛丝的微观结构

利用原子力显微镜研究了蚕丝丝素、蜘蛛牵引丝及其内外层包卵丝的微观结构.研究表明,丝素和包卵丝纤维的纵向表面都有成丝过程中液态丝蛋白流动而形成的清晰的构槽和条纹,在低速下自然分泌的牵引丝的表面皮层相对比较细腻,而垂直下落蜘蛛在高速下分泌的牵引丝具有和丝素纤维比较相似的微观结构特征.这些丝纤维的断面内都分布有大量微细的原纤,形状基本为圆形,其中三种蜘蛛丝的微纤维直径相似,而丝素纤维内的微纤维要粗得多.     

SILK.IT project: Silk Italian Technology for industrial biomanufacturing

Silk fibroin is a natural protein that constitutes the core fibre of the silkworm cocoon. Recently, a great deal of attention is being paid to the use of the regenerated silk fibroin (RSF) solution that can be obtained by water-processing of the cocoon fibre. Indeed, thanks to its chemical and physical properties it has been shown that substrates obtained by RSF can be used as biomaterials for several biomedical and technological applications. In this context, silk fibroin could be exploited as raw material that can become a platform for eco-sustainable manufacturing.The project SILK.IT, coordinated by the CNR-ISOF, within the framework of the Flagship Project “Factory of the Future”, a research programme approved by the Interministerial Committee for Economic Planning (CIPE), aims to support this assumption by developing methods and protocols to use this silk fibroin as new material for advanced biotechnologies and sustainable manufacturing.The main goal of the project is to establish and control the whole-chain underpinning the silk fibroin-based technology, and to promote its up-scaling from the laboratory to industrial scale, targeting bio-photonics for biomedical application.The recent results obtained by the project are herein reported as key building block towards proposed achievement to establish a sustainable and process-controlled silk-based bio-manufacturing of the future.     

Production and properties of electrosprayed sericin nanopowder

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.     

Small-diameter vascular grafts of Bombyx mori silk fibroin prepared by a combination of electrospinning and sponge coating

The present study aimed to produce small-diameter grafts made of a silk fibroin by electrospinning. In order to reinforce the electrospun silk fibroin graft (ES), the graft was coated with a silk sponge (ESSC). Physical properties such as a diameter of the electrospun silk fibers were influenced by a concentration of fibroin solution. Ultimate tensile strength (UTS) of the ESSC graft was improved compared to the ES graft. However, the ESSC graft was less compliant than the ES graft. Importantly, water permeability of the ESSC graft was within the range of which endothelialization was promoted in previous studies.     

Production and properties of electrosprayed sericin nanopowder

Abstract

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.     

EDC/NHS交联改性再生桑蚕丝素纳米纤维

用静电纺丝技术制备了再生桑蚕丝素纳米纤维,并用1-(3-二甲基氨基丙基)-3-乙基碳化二亚胺(EDC)和N-羟基丁二酰亚胺(NHS)进行交联改性,考察了交联改性前后,桑蚕丝素纳米纤维微观形貌及聚集态结构等的变化,采用扫描电镜(SEM)、X射线衍射(XRD)及红外光谱(FT-IR)等测试方法对纳米纤维进行表征。研究结果表明,经EDC/NHS交联改性后,纤维直径由250 nm增加到320 nm,纤维表面变粗糙,且纤维发生弯曲变形;丝素的结构以Silk II为主,并含有部分无规卷曲或Silk I构象;桑蚕丝纳米纤维的力学性能和亲水性有所提高,且交联改性后的纳米纤维具有良好的细胞相容性。