Preparation and characterization of Bombyx mori silk fibroin and wool keratin

Silk and wool are well‐known protein‐based fibers. Their environmental stability, biocompatibility, and unique mechanical properties provide an important basis for using these natural proteins in biomedical applications. To use them as biomaterials in the form of fibers, films, or membranes, it is necessary to characterize these proteins in their solution and solid states because structural characteristics and morphological features have a great influence on the physical and mechanical properties of these new regenerated protein forms. Therefore, in the present study, silk fibroin and wool keratin were dissolved and their solution behaviors and secondary structures are analyzed and compared, using particle size distribution, molecular weight distribution (SDS‐PAGE), Fourier transform infrared, and X‐ray diffraction techniques. It was shown that keratin is more stable in solution and more amorphous in the solid state. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 100:4260–4264, 2006     

Chemical modification of Bombyx mori silk with calcium‐salt treatment and subsequent glycerin triglycidyl ether crosslinking

In this article, we propose a new modification method for obtaining porous silk fibers with excellent wet elastic resilience and flexibility. Bombyx mori silks were modified by calcium‐salt treatment and subsequent epoxy crosslinking with glycerin triglycidyl ether. The effects of temperature, time, and catalyst (sodium carbonate) on the crosslinking reaction of the silk fibers were investigated, and the best conditions of reaction were determined as a temperature of 120°C, a crosslinking agent concentration of 7%, and immersion for 1 h with 2% Na₂CO₃ solution before the crosslinking reaction. The change in the structure and the physical properties of the silk fibers after calcium‐salt treatment and epoxy crosslinking was studied. Separating behavior of the microfibers occurred on the surface of the silk fiber after calcium‐salt treatment, and a porous structure formed in the interior of the silk. This porous structure of the silk was enlarged by subsequent epoxy crosslinking, and accordingly, the moisture conduction of the silk fibers improved remarkably. The breaking strength, breaking elongation, and wet elastic resilience of the silk fibers increased evidently after modification, and the modified silks exhibited a better flexibility. The conformation of silk fibroin fibers changed from β sheet to random coil after calcium‐salt treatment, whereas the β‐sheet content in the silk fibers increased after subsequent epoxy crosslinking. The significant reductions in the crystallinity and crystalline sizes in the silk fibers after the crosslinking reaction indicated that the crosslinking reaction occurred within the crystalline region because the calcium‐salt treatment increased the reaction accessibility. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010     

Chemical modification of tussah silk with acid anhydrides

Tussah silk fibroin was chemically modified by acylation with aliphatic, aromatic, and hydrophobic acid anhydrides. The tussah silk fibers were pretreated by immersing them in a lithium thiocyanate (LiSCN) solution and then acylated in dimethylformamide (DMF) at elevated temperatures. Using this method, acylated tussah silk fibers with weight gains of 8–22% could be obtained. The pretreatment with LiSCN was necessary to promote the acylation. Without it, the reaction did not proceed. The optimum temperature and reaction time of the pretreatment was 55°C and 60 min, respectively. When examining the physical properties and the thermal behavior of both pretreated and acylated tussah silk, it was found that the mechanical properties and the position of the major DSC endothermic peak remained unchanged, regardless of pretreatment and acylation. The moisture regain of the pretreated tussah silk increased slightly while the moisture regain of the acylated silk decreased linearly with increasing weight gain. The chemical modification allows for a wide control of the tussah silk fiber's properties, making it possible to use tussah silk for the development and production of novel textile and biomaterials. © 2000 John Wiley & Sons, Inc. J Appl Polym Sci 78: 382–391, 2000     

Effect of moisture absorption on the thermal properties of Bombyx mori silk fibroin films

Films of regenerated Bombyx mori silk are strongly affected by absorbed moisture, a phenomenon studied here by differential scanning calorimetry (DSC). Exposure of previously dried films to environments of controlled relative humidity produces test samples of well-defined equilibrium moisture content. Ultimate moisture uptake is as high as 20–23% (by weight) at 75% relative humidity. The glass transition temperature, T_g, drops by 40°C at moisture uptakes as low as 2%, and T_g depressions as large as 140°C are observed at higher relative humidity. The moisture-induced decrease of T_g is completely reversible, as a film remoistened and then redried possesses an unchanged T_g. Trends in T_g with water uptake correspond reasonably well to predictions of a classical thermodynamic theory, indicating that the plasticization effect of moisture on the combined silk-water system can be satisfactorily explained from macroscopic properties of the constituents without any reference to specific interactions. © 1997 John Wiley & Sons, Inc. J Appl Polym Sci 63: 401–410, 1997     

聚乳酸/丝素共混膜的制备及其性能

用分子量为10万的聚乳酸（PLLA）对丝素膜进行改性,研究不同的聚乳酸加入量对丝素膜性能的影响,对聚乳酸/丝素共混膜进行了一系列表征。万能电子试验机的测试结果表明,经聚乳酸改性后,丝素膜的断裂强度,断裂伸长率有了较大的改善,当加入聚乳酸占丝素质量为5%时,丝素膜的强度可达到27.1 MPa,伸长率达4.4%; 改性后的丝素膜的亲水性有一定程度降低,溶失率则明显减小,透汽透湿性也有所提高;红外光谱测试表明,改性后的丝素膜含有较多的β构象成分。     

Curcumin loaded electrospun Bombyx mori silk nanofibers for drug delivery

Silk fibroin from Bombyx mori silk cocoons was electrospun into silk nanofibers (SNFs). SEM images show that 9% w/v of SNFs was smooth and beadless having an average diameter in the range 30–150 nm. Curcumin (0.5–1.5 wt%) was incorporated into the silk fibroin solution and electrospun to obtain curcumin incorporated silk nanofibers (CSNFs) with diameters between 50 and 200 nm. The dispersion of curcumin in the SNFs was confirmed by TEM. The amorphous nature of curcumin upon incorporation into SNFs was confirmed by XRD. The functional groups of SNF and CSNF were confirmed by Fourier transform infrared spectroscopy. The SNFs and CSNFs were thermally stable up to ca 350 °C as evidenced by TGA. The glass transition temperature (T_g) of SNFs (168 °C) increased to 184 °C in the case of CSNFs as confirmed by DSC. The storage modulus, loss modulus and tan δ were determined by dynamic mechanical analysis. The percentages of porosity and water uptake of SNFs were 85% and 150%, respectively. The percentage in vitro cumulative release of curcumin at the end of the tenth day for 0.5, 1 and 1.5 wt% formulations was 82%, 84% and 80%, respectively. © 2013 Society of Chemical Industry     

Production of silk sericin/silk fibroin blend nanofibers

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.     

丝素蛋白在离子液体中的溶解特性研究

文章以离子液体为溶剂,研究了丝素蛋白在离子液体中的溶解特性。首先研究了丝素蛋白在不同结构的离子液体1-烯丙基-3-甲基咪唑氯化物（[AMIM]C1）和1-丁基-3_-甲基咪唑氯化物（[BMIM]c1）中的溶解速率和溶解度。结果发现,相同条件下,丝素蛋白在[AMIM]cl中具有较快的溶解速率和较大的溶解度;随温度的升高,溶解速度加快,合适的温度为95～105℃;随着丝素蛋白浓度的增大,相同条件下丝素蛋白在离子液体中的溶解速度减慢。XRD结果表明,丝素蛋白的非晶相部分首先被溶解,晶相部分结构被破坏后与溶剂作用逐渐被溶解。采用粘度法测定了丝素蛋白分子质量随溶解时间和温度的变化,结果表明：随加热时间增加和温度升高,蛋白质分子质量减小。为保证丝素蛋白的稳定性和溶解速率,温度一般控制在100℃以下,溶解时间小于5h。     

从丝素水溶液中再生的丝纤维的结构与性能

通过使用表面皿直接拉伸、毛细管重力纺丝和人工拉伸3种不同的成丝方法,从高浓度再生丝素水溶液中制得了丝纤维。用偏光显微镜观察了丝纤维的取向,用拉曼光谱仪和Instron拉力仪表征了丝纤维的结构和力学性能。结果发现,经毛细管剪切流动后再拉伸有利于再生丝性能的提高,所得的丝有较好的取向和较多的β折叠结构,力学性能也相对较好。剪切在丝纤维的成形过程中起重要的作用。     

Comparison of three-dimensional printing for fabricating silk fibroin-blended scaffolds

In this study, two types of scaffolds were fabricated by 3D printing. Morphology, physical properties, biochemical were evaluated. Cell morphology and distribution were observed. It was found that the silk fibroin/collagen (SF/C) scaffold-based material had significantly higher values than the silk fibroin/chitosan (SF/CS) scaffold-based material. Hematoxylin and eosin staining of the scaffolds revealed that the number of cells in the SF/C scaffold was higher. Cells grew well inside the SF/C scaffold as measured by scanning electron microscope. Reverse Transcriotion-Polymerase Chain Reaction (RT-PCR) and Western blot showed that type II collagen and Sox9 can be found in SF/C scaffold. Therefore, the SF/C scaffold exhibited better overall performance compared with the SF/CS scaffold.     

丝素/羧甲基壳聚糖复合材料的制备及其在丝绸文物加固中的应用

为了对脆弱丝绸文物进行保护修复，延长其寿命，本研究以丝素蛋白（SF）、羧甲基壳聚糖（CMCS）为原料，谷氨酰胺转氨酶为交联剂，制备丝素/羧甲基壳聚糖复合材料（SF/CMCS），并将其应用于老化丝绸加固保护。选用傅里叶变换红外光谱（FT-IR）、X-射线衍射仪（XRD）、测色配色分光光度计、扫描电镜（SEM）及伺服高低温控制拉伸机对老化丝绸加固前后效果进行表征。结果表明，与老化丝绸相比，SF/CMCS加固丝绸的颜色无明显变化，物理机械性能有明显提升，抗张强度和断裂伸长率分别提升了379.12%、14.12%%，且有一定的抗菌性。     

离子液体对丝素蛋白溶解性能的影响

合成了一系列碱性离子液体,其中离子液体1-丁基-3-甲基咪唑乙酸盐([Bmim]AC)对丝素蛋白具有良好的溶解性能。考察了丝素蛋白在该离子液体中的溶解特性及在离子液体中的溶解温度和溶解时间,结果表明,在75℃下,溶解840 min,该离子液体对丝素蛋白的溶解能力达到15%(质量分数)。XRD和FTIR分析表明,再生丝素蛋白的构象以α-螺旋为主,丝素蛋白在溶解过程中没有发生衍生化。因此,[Bmim]AC离子液体是丝素蛋白的优良溶剂。     

Grafting vinyl monomers onto silk (Bombyx mori) using different initiators: Properties of grafted silk

The graft–copolymerization of silk with methacrylamide (MAA), 2‐hydroxyethyl methacrylate (HEMA), and methyl methacrylate (MMA) was studied using three different free‐radical initiators: an inorganic peroxide [ammonium persulfate (APS)] and two azo compounds [2,2′‐azobisisobutyronitrile (AIBN) and 2,2′‐azobis(2‐methylpropionamidine) dihydrochloride (ADC)]. The rate and yield of grafting followed the order: APS > AIBN > ADC. The performance of AIBN was close to that of APS in terms of weight gain attained. The degree of yellowing of grafted silk varied as a function of the initiator–monomer system used. APS induced the highest degree of yellowing, regardless of which monomer was used, whereas silk grafted with the MAA/AIBN system displayed the lowest level of yellowing. Moisture regain of grafted silk changed as a function of the hydrophilic/hydrophobic character of the grafted polymer, regardless of the kind of initiator used. Accordingly, tensile properties showed a tendency to decrease with increasing weight gain of grafted silk, the extent of which was independent of the kind of monomer and initiator used. The different initiators did not induce any appreciable change in the fine structure of silk, as demonstrated by optical measurements. Uneven surface deposition of homopolymer was detected to variable extent with MMA and HEMA grafting, whereas the surface of MAA‐grafted silk was completely free of any foreign deposit, independently of the initiator used. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 81: 1401–1409, 2001     

Preparation of nanosize cadmium sulfide particles with a silk fibroin membrane and their photocatalytic activity

We present a novel method for the synthesis of nanosize CdS particles with a silk fibroin membrane, which were characterized with UV spectrometry, X‐ray diffraction (XRD), and atomic force microscopy. The UV spectra showed a clear blueshift in the absorption edge for the CdS particles prepared at different concentrations. XRD showed the prevailing crystal were the cube ZnS crystals formed. The particle sizes were calculated from the band gaps with the effective mass approximate model, suggesting that they were dependent on the concentration of the precursors. Meanwhile, nanosize CdS had good photocatalytic activity throughout the photocatalytic degradation experiments. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 101: 2162–2166, 2006     

Compatibilization of acrylic polymer–silk fibroin blend fibers. I. Graft copolymerzation of acrylonitrile onto silk fibroin

To synthesize effective compatibilizers for the compatibilization of acrylic polymer–silk fibroin blend fibers, the homogeneous graft copolymerization of acrylonitrile onto silk fibroin, initiated by potassium persulfate–sodium bisulfite redox system in 60 wt % ZnCl₂ aqueous solution was investigated. The percentage graft, percentage efficiency, and molecular weight of the grafted polyacrylonitrile increase with the increase of the concentration of initiator, the concentration of acrylonitrile, and the temperature to optimum values, respectively, and then decrease. With the increase of reaction time, the percentage graft and percentage efficiency increase, whereas the molecular weight of the grafted polyacrylonitrile decreases. The influence of the reactionconditions on the molecular architecture of the graft copolymers is also discussed. © 1998 John Wiley & Sons, Inc. J Appl Polym Sci 69: 1089–1097, 1998     

Characterization of a Silk‐Resinified Compact Fabricated Using a Pulse‐Energizing Sintering Device

Pulse electric current sintering is used to prepare a compact from resinificated hydrous silk powder. Compacts with no remnant silk powders are formed with 20 wt% added water, 20–40 MPa molding pressure, and >353 K molding temperature. The latter two are much lower than those used for conventional hot pressing. No dependence on molding pressure and temperature are found in XRD or FT‐IR analysis, except for a compact molded at 473 K, for which silk fibroin decomposition is confirmed by DSC, EGA‐MS, and molecular weight measurements. The compact's three‐point bending strength depends on the molding temperature, except for the temperature at which silk fibroin decomposes. The maximum three‐point bending strength resembles that of general‐purpose epoxy resin and is much higher than that of PLA.

     

Biodegradation of Bombyx mori silk fibroin fibers and films

The in vitro biodegradation of Bombyx mori silk fibroin was studied by incubating fibers and films with proteolytic enzymes (collagenase type F, α‐chymotrypsin type I‐S, protease type XXI), for times ranging from 1 to 17 days. The changes in sample weight and degree of polymerization of silk fibers exposed to proteolytic attack were negligible. However, tensile properties were significantly affected, as shown by the drop of strength and elongation as a function of the degradation time. Upon incubation with proteolytic enzymes, silk films exhibited a noticeable decrease of sample weight and degree of polymerization, the extent of which depended on the type of enzyme, on the enzyme‐to‐substrate ratio, and on the degradation time. Protease was more aggressive than α‐chymotrypsin or collagenase. Film fragments resistant to enzymatic degradation were enriched in glycine and alanine. FT‐IR measurements showed that the degree of crystallinity of biodegraded films increased. Soluble degradation products of silk films consisted of a range of peptides widely differing in size, deriving from the amorphous sequences of the silk fibroin chains. Biodegraded fibers showed an increase of surface roughness, while films displayed surface cracks and cavities with internal voids separated by fiber‐like elements. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 91: 2383–2390, 2004     

Silk I structure formation through silk fibroin self-assembly

Regenerated silk fibroin films are normally produced by increasing the Silk II structure (β-sheet content). In the present study, silk fibroin films were prepared by controlling the environmental temperature and humidity, resulting in the formation of silk films with a predominant Silk I structure instead of Silk II structure. Wide angle X-ray diffraction indicated that when the relative humidity was 55%, the silk films prepared were mainly composed of Silk I structure, whereas silk films formed on other relative humidity had a higher Silk II structure. Fourier transform infrared analysis (FTIR) results also conformed that the secondary structure of silk fibroin can be controlled by changing the humidity of the films formed process. Thermal analysis results revealed Silk I structure was a stable crystal, and the degradation peak increased to 320°C, indicating a greater thermal stability of these films formed under the 55% relative humidity conditions. Atomic force microscopy (AFM) results depicted silk fibroin in the fresh solution had many nanospheres existing with 20–50 nm diameters and mainly maintained a random coil structure without specific nanostructures. At the same time, it also illustrated the self-assembly process of silk fibroin in the aqueous solution without any human intervention. In addition, this present study also provided additional support for self-assembly mechanism of silk fibroin films formation. © 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012     

再生丝素/丝胶蛋白水溶液的微流体静电纺丝研究

以再生丝素蛋白(SF)水溶液和丝胶蛋白(SS)水溶液为纺丝液,微流体多通道芯片为纺丝器,成功制备了SF/SS和SF/SS/SF纤维毡。制备SF/SS纤维毡时,上述两流体在微通道中以层流方式流动,到达芯片出口处既不共混也不分层,而是各自独立成丝,通过扫描电镜可以观察到粗细差异较大的两种纤维共存,形貌较好。以SS水溶液为微通道的中心流体,SF水溶液为两侧流体,通过微流体静电纺制得SF/SS/SF纤维毡,SF部分与SS部分可能形成了三明治螺旋特殊结构。纺丝液流速会对纤维毡形态结构产生影响,SF溶液流速保持不变,增加SS溶液的流速,纤维粘连严重且容易出现断裂;保持SS流速不变,增大SF溶液流速,纤维粘连性得到改善,且形貌变好。