Piezoelectric fibers based on silk fibroin with excellent output performance

The self-powered tissue engineering scaffold with good biocompatibility is of great significance for stimulating nerve cell growth.In this study,silk fibroin(SF)-based fibers with regulatable structure and piezoelectric performance are fabricated by dry-spinning and post-treatment.The concentration of SF and calcium ion in spinning dope and the post-treatment affect the conformation transition and crystallinity of SF.As a result,the SF fibers exhibit high piezoelectric coefficient d_(33)(3.24 pm/V)and output voltage(~27 V).Furthermore,these piezoelectric fibers promote the growth of PC-12 cells,demonstrating the promising potential for nerve repair and other energy harvester.

Engineering biomimetic silk fibroin hydrogel scaffolds with“organic-inorganic assembly”strategy for rapid bone regeneration

Although natural polymers have been widely used in constructing bone scaffolds,it still remains challenging to fabricate natural polymer-derived bone scaffolds with biomimetic mechanical properties as well as outstanding osteogenic properties for large-size and weight-bearing bone defects regeneration.Herein,an“organic-inorganic assembly”strategy is developed to construct silk fibroin(SF)-based bone scaffolds with the aforementioned merits.After secondary structure reshuffling,the 3.3-fold increment ofβ-sheet structures in SF hydrogel resulted in a 100-fold improvement of mineral-assembly efficacy via influencing the ion adsorption process and providing templates for mineral growth.Notably,abundant minerals were deposited within the hydrogel and also on the surface,which indicated entire mineral-assembly,which ensured the biomimetic mechanical properties of the digital light processing 3D printed SF hydrogel scaffolds with haversian-mimicking structure.In vitro experiments proved that the assembly between the mineral and SF results in rapid adhesion and enhanced osteogenic differentiation of human bone marrow-derived mesenchymal stem cells.In vivo experiments further proved that the mineral-assembled SF hydrogel scaffold could significantly enhance integration and bone regeneration at the weight-bearing site within one month.This SF-based“organic-inorganic assembly”strategy sheds light on constructing cell-free,growth factor-free and natural polymer-derived bone scaffolds with biomimetic 3D structure,mechanical properties and excellent osteogenic properties.

Research on the Development of Fibroin and Nano-Fiber from Silk Cocoons for Regenerated Tissue Engineering Applications by Electro-Spinning

In this paper, the main goal is to prepare silk fibroin nano-fiber, which is used for regenerated tissue applications. Silk scaffold nano-fibers made by electro-spinning technology can be used in regenerated tissue applications. The purpose of the research is to prepare a silk-fibroin nano-fiber solution for potential applications in tissue engineering. Using a degumming process, pure silk fibroin protein is extracted from silk cocoons. The protein solution for fibroin is purified, and the protein content is determined. The precise chemical composition, exact temperature, time, voltage, distance, ratio, and humidity all have a huge impact on degumming, solubility, and electro-spinning nano-fibers. The SEM investigates the morphology of silk fibroin nano-fibres at different magnifications. It also reveals the surface condition, fiber orientation, and fiber thickness of the silk fibroin nano-fiber. The results show that regenerated silk fibroin and nano-fiber can be used in silk fibroin scaffolds for various tissue engineering applications.

Preparation of Regenerated Silk Fibroin Hybrid Fibers with Hydrogen Peroxide Sensing Properties by Wet Spinning

Silk is widely used in the production of high-quality textiles.At the same time,the amount of silk textiles no longer in use and discarded is increasing,resulting in significant waste and pollution.This issue is of great concern in many countries where silk is used.Hydrogen peroxide as a naturally occurring compound is an important indicator of detection in both biology and the environment.This study aims to develop a composite fiber with hydrogen peroxide-sensing properties using discarded silk materials.To achieve this goal,firstly,polydopamine(PDA)was used to encapsulate the ZnFe_(2)O_(4) NPs to achieve the improvement of dispersion,and then regenerated silk fibroin(RSF)and PDA@ZnFe_(2)O_(4)/RSF hybrid fibers are prepared by wet spinning.Research has shown that PDA@ZnFe_(2)O_(4)/RSF demonstrates exceptional sensitivity,selectivity,and stability in detecting hydrogen peroxide,while maintaining high mechanical strength.Furthermore,the complete hybridization of PDA@ZnFe_(2)O_(4) with silk fibroin not only results in the combination of the durability of silk fibroin and PDA@ZnFe_(2)O_(4)’s rigidity,ensuring a reliable service life,but also makes PDA@ZnFe_(2)O_(4)/RSF exhibit excellent catalytic activity and biocompatibility.Therefore,the composite fiber exhibits exceptional mechanical properties and reliable hydrogen peroxide sensing capabilities,making it a promising material for biological and medical applications.

A Review on Silk Fibroin as a Biomaterial in Tissue Engineering

Regenerative medicine progress is based on the development of cell and tissue bioengineering. One of the aims of tissue engineering is the development of scaffolds, which should substitute the functions of the replaced organ after their implantation into the body. The tissue engineering material must meet a range of requirements, including biocompatibility, mechanical strength, and elasticity. Furthermore, the materials have to be attractive for cell growth: stimulate cell adhesion, migration, proliferation and differentiation. One of the natural biomaterials is silk and its component (silk fibroin). An increasing number of scientists in the world are studying silk and silk fibroin. The purpose of this review article is to provide information about the properties of natural silk (silk fibroin), as well as its manufacture and clinical application of each configuration of silk fibroin in medicine. Materials and research methods. Actual publications of foreign authors on resources PubMed, Medline, E-library have been analyzed. The selection criteria were materials containing information about the structure and components of silk, methods of its production in nature. This article placed strong emphasis on silk fibroin, the ways of artificial modification of it for use in various sphere of medicine.

Preparation and release of curcumin/silk fibroin/sodium alginate film

The aim of this study was to prepare silk fibroin/sodium alginate composite film containing curcumin by casting method.Orthogonal test was used to optimize the formulation according to the values of tensile strength and elongation at break.The release of curcumin in the optimal film was studied in order to explore its application as wound dressing.The results showed that the optimum composition of curcumin/silk fibroin/sodium alginate composite film was as follows:Silk fibroin(70 mg/mL)2.7 g,sodium alginate(24 mg/mL)0.84 g,span 40(5.0 mg/mL)0.4 g,glycerol(3.75%,V/V)3 mL,curcumin(0.2 mg/mL)0.016 g.The optimum film showed the tensile strength and the elongation at break was(0.628±0.032)MPa and(0.794±0.046)%,respectively.

IrO_(2)@SF纳米粒子的合成及其抗肿瘤应用

缺氧肿瘤微环境通常会促进肿瘤增殖和导致肿瘤细胞对活性氧(ROS)治疗的抵抗.选择丝素蛋白(SF)为模板,通过模拟生物矿化作用在SF表面矿化一层氧化铱(IrO_(2)),得到丝素蛋白和氧化铱的纳米复合物IrO_(2)@SF(IOS).天然SF的加载赋予了IOS多重抗肿瘤活性.实验结果表明:IOS具有较高的光热转换效率、过氧化氢酶和过氧化物酶活性以及较好的声动力效果.IOS能够通过分解肿瘤微环境过表达的过氧化氢,从而逆转肿瘤微环境的缺氧状态.体外细胞实验表明:IOS实现了光热联合纳米酶催化动力学抗肿瘤与声动力联合纳米酶催化动力学抗肿瘤治疗,2种多模式结合的治疗方式均展现了优异的抗肿瘤效果.同时,由于SF这种天然蛋白的加载,IOS还具有较高的生物相容性.

Structure Changes of Silk Fibroin(SF) by Blending with Poly(ε-caprolactone)(PCL):Characterization of SF and PCL Blended Electrospinning Films

The mechanical properties and water solubility of electrospinning SF films limit their use as biomaterials. In order to develop a tissue engineering biomaterial with both satisfying biological properties and sufficient biomechanical properties,blended films composed of silk fibroin( SF) and poly( ε-caprolactone)( PCL) were fabricated by electrospinning in this study. Scanning electron microscope( SEM), X-ray diffraction( XRD),thermal analysis,Fourier transform-infrared( FT-IR),Raman spectra,mechanical testing,and water solubility were used to characterize the morphological, structural and mechanical properties of the blended electrospinning films. Results showed that the diameter of the blended fiber was distributed between 600 and1000 nm,and the fiber diameter increased as the PCL content increased. There is no obvious phase separation due to the similarity and intermiscibility,as well as the interactions( mainly hydrogen bonds), between the two polymers. Meanwhile, the secondary structures of SF changed from random coils and Silk I to Silk II because of the interactions between SF and PCL. For this reason,the tensile strength and elongation at break of the electrospinning films improved significantly,and the water solubility decreased. In conclusion,the blended electrospinning films fabricated in this study showed satisfying mechanical properties and water insolubilities,and they may be promising biomaterials for applications in tissue engineering for blood vessels,nerve conduits,tendons,ligaments and other tissues.

Effects of Human Insulin Gene Transfection on the Adipogenic Differentiation of Human Umbilical Cord Mesenchymal Stem Cells in Silk Fibroin Scaffolds <i>in Vitro</i>

The resorption of the transplanted fat over time limited the use of autologous fat for the reconstruction of soft tissue defect. Tissue engineering (TE) adipose with silk fibroin scaffold could be a promising substitute for soft tissue filling. In this study, we try to develop a tissue engineering adipose in vitro by seeding silk fibroin scaffold with human umbilical cord mesenchymal stem cells (hUCMSCs) after transfected with recombinant human insulin gene lentivirus. Our aim was to observe the effects of the insulin gene transfection on the adipogenesis of hUCMSCs when cultured with silk fibroin scaffolds. The hUCMSCs infected with recombinant lentiviral pLenti6.3-insulin-IRES-EGFP were seeded on silk fibroin scaffolds and cultured in adipogenic differentiation medium for 5 - 7 days. The expression of adipogenic gene PPARγ-2 was tested by RT-PCR after 7 days culture of adipogenic induction. The accumulation of cytoplasmic droplets of neutral lipids was assessed by Oil Red O staining. The RNA and protein expression of transfected insulin gene in hUCMSCs were detected by QPCR and western blot. The effect of recombinant lentivirus transfection on the growth and proliferation of hUCMSCs was observed by MTT test. We observed that the 2-ΔΔCt value of insulin gene expression of hUCMSCs in the transfected group was 300.25 times higher than that in the untransfected group. The western blot showed that a positive band was discerned at the site of a relative molecular mass of 8 × 103 Dalton in transfected group. After adipogenic culture for 7 days, under the fluorescent inverted phase-contrast microscope, after Oil Red O staining, a lot of adipocytes appeared in silk fibroin scaffold;round adipose droplets showed intracellularly;the size of the adipocytes was not homogenous, and the density of adipocytes in transfected group was significantly higher than that in untransfected group (P = 0.007, P < 0.01). RT-PCR results showed that the expression of adipogenic gene PPARγ-2 in transfected group was much stronger than that in untransfected group. MTT test showed that there was no significant difference in optical density (A) at each time point between transfected group and nontransfected group (P = 0.056, P > 0.05). And there was also no significant difference in optical density (A) between cell group and cell-scalffold group (P = 0.066, P > 0.05). We concluded that insulin gene could obviously promote the adipogenic differentiation of hUCMSCs, and a tissue engineering adipose could be constructed by the silk fibroin scaffolds seeded with human insulin gene-modified hUCMSCs effectively in vitro.

Three-dimensional bioprinting collagen/silk fibroin scaffold combined with neural stem cells promotes nerve regeneration after spinal cord injury

Many studies have shown that bio-scaffolds have important value for promoting axonal regeneration of injured spinal cord.Indeed,cell transplantation and bio-scaffold implantation are considered to be effective methods for neural regeneration.This study was designed to fabricate a type of three-dimensional collagen/silk fibroin scaffold (3D-CF) with cavities that simulate the anatomy of normal spinal cord.This scaffold allows cell growth in vitro and in vivo.To observe the effects of combined transplantation of neural stem cells (NSCs) and 3D-CF on the repair of spinal cord injury.Forty Sprague-Dawley rats were divided into four groups: sham (only laminectomy was performed),spinal cord injury (transection injury of T10 spinal cord without any transplantation),3D-CF (3D scaffold was transplanted into the local injured cavity),and 3D-CF + NSCs (3D scaffold co-cultured with NSCs was transplanted into the local injured cavity.Neuroelectrophysiology,imaging,hematoxylin-eosin staining,argentaffin staining,immunofluorescence staining,and western blot assay were performed.Apart from the sham group,neurological scores were significantly higher in the 3D-CF + NSCs group compared with other groups.Moreover,latency of the 3D-CF + NSCs group was significantly reduced,while the amplitude was significantly increased in motor evoked potential tests.The results of magnetic resonance imaging and diffusion tensor imaging showed that both spinal cord continuity and the filling of injury cavity were the best in the 3D-CF + NSCs group.Moreover,regenerative axons were abundant and glial scarring was reduced in the 3D-CF + NSCs group compared with other groups.These results confirm that implantation of 3D-CF combined with NSCs can promote the repair of injured spinal cord.This study was approved by the Institutional Animal Care and Use Committee of People’s Armed Police Force Medical Center in 2017 (approval No.2017-0007.2).

Human amniotic epithelial cells combined with silk fibroin scaffold in the repair of spinal cord injury

Treatment and functional reconstruction after central nervous system injury is a major medical and social challenge. An increasing number of researchers are attempting to use neural stem cells combined with artificial scaffold materials, such as fibroin, for nerve repair. However, such approaches are challenged by ethical and practical issues. Amniotic tissue, a clinical waste product, is abundant, and amniotic epithe- lial cells are pluripotent, have low immunogenicity, and are not the subject of ethical debate. We hypothesized that amniotic epithelial cells combined with silk fibroin scaffolds would be conducive to the repair of spinal cord injury. To test this, we isolated and cultured amniotic epithelial cells, and constructed complexes of these cells and silk fibroin scaffolds. Implantation of the cell-scaffold complex into a rat model of spinal cord injury resulted in a smaller glial scar in the damaged cord tissue than in model rats that received a blank scaffold, or amniotic epithelial cells alone. In addition to a milder local immunological reaction, the rats showed less inflammatory cell infiltration at the trans- plant site, milder host-versus-graft reaction, and a marked improvement in motor function. These findings confirm that the transplantation of amniotic epithelial ceils combined with silk fibroin scaffold can promote the repair of spinal cord injury. Silk fibroin scaffold can provide a good nerve regeneration microenvironment for amniotic epithelial cells.

Novel conductive polypyrrole/silk fibroin scaffold for neural tissue repair

Three dimensional（3D） bioprinting, which involves depositing bioinks（mixed biomaterials） layer by layer to form computer-aided designs, is an ideal method for fabricating complex 3D biological structures. However, it remains challenging to prepare biomaterials with micro-nanostructures that accurately mimic the nanostructural features of natural tissues. A novel nanotechnological tool, electrospinning, permits the processing and modification of proper nanoscale biomaterials to enhance neural cell adhesion, migration, proliferation, differentiation, and subsequent nerve regeneration. The composite scaffold was prepared by combining 3D bioprinting with subsequent electrochemical deposition of polypyrrole and electrospinning of silk fibroin to form a composite polypyrrole/silk fibroin scaffold. Fourier transform infrared spectroscopy was used to analyze scaffold composition. The surface morphology of the scaffold was observed by light microscopy and scanning electron microscopy. A digital multimeter was used to measure the resistivity of prepared scaffolds. Light microscopy was applied to observe the surface morphology of scaffolds immersed in water or Dulbecco＇s Modified Eagle＇s Medium at 37℃ for 30 days to assess stability. Results showed characteristic peaks of polypyrrole and silk fibroin in the synthesized conductive polypyrrole/silk fibroin scaffold, as well as the structure of the electrospun nanofiber layer on the surface. The electrical conductivity was 1 × 10^-5–1 × 10^-3 S/cm, while stability was 66.67%. A 3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide assay was employed to measure scaffold cytotoxicity in vitro. Fluorescence microscopy was used to observe Ed U-labeled Schwann cells to quantify cell proliferation. Immunohistochemistry was utilized to detect S100β immunoreactivity, while scanning electron microscopy was applied to observe the morphology of adherent Schwann cells. Results demonstrated that the polypyrrole/silk fibroin scaffold was not cytotoxic and did not affect Schwann cell proliferation. Moreover, filopodia formed on the scaffold and Schwann cells were regularly arranged. Our findings verified that the composite polypyrrole/silk fibroin scaffold has good biocompatibility and may be a suitable material for neural tissue engineering.

Preparation of Nano Silver/Silk Fibroin Composite Films

In this study, the outstanding biocompatibility of silk fibroin （SF） and the highly efficient anti-bacterial effect of nano silver （NS） were utilized to prepare SF/NS composite film with anti- bacterial property. The structure and property of the film were characterized. The results showed that the structure of SF in the film was mainly silk I. SF in the film was almost insoluble in water. The tensile strength of film with NS was significantly lower than that of films without NS. When the addition of NS was within the range of 0%-0.6%, the elongation at break had no significant difference. The antibacterial rate of the film on staphylococcus aurens and escherichia coil increased with the amount of NS. The minimum amount of NS in the fdm was O. 1% and the maximum amount was 0.5%.

Preparation and Characterization of PEGDE Crosslinked Silk Fibroin Film

To obtain water-insoluble silk fibroin （SF） materials, polyethylene glycol diglycidyl ether （PEG-DE） was selected as a crosslinking agent to prepare SF films （blends）. The reaction conditions were optimized for the crosslinking of the SF molecules. The hot water stability of the blends was measured using BCA protein assay and gravimetric analysis. The molecular conformation and crystalline structure of the blends were analyzed by FTIR and XRD, respectively. When the mass ratio of SF：PEG-DE was 1.0：0.8, the hot water loss rate of the SF blends was minimized. PEG-DE could induce SF molecules to form fl-sheets during the gel reaction process, resulting in improved crystallinity and hot water dissolved resistance of the blend films. In order to demonstrate the eytotoxicity of the chemical reagents used to crosslink SF, L929 cells were seeded on the blend film （SF：PEG-DE = 1：1） and cultured for 3 days. Cells of L929 readily adhered and spread in the fusiform on the blend film resulting in high cell viability. The extracted liquid from the SF porous film did not inhibit cell proliferation, as estimated by the MTT assay.

SURFACE MODIFICATION OF BLEND FILMS COMPOSED OF SILK FIBROIN AND POLY(ETHYLENE GLYCOL) MACROMER AND THEIR IN VITRO ANTITHROMBOGENICITY

In order to improve the blood compatibility of silk fibroin (SF), poly(ethylene glycol) macromer (PEGM) in different amounts was added to the SF film to incorporate C=C group into the surface of blend films which were then modified by SO2 gas plasma treatment. ATR-FITR and XPS were used to analyze the chemical change which had occurred on the film's surface. When the content of sulfur on the surface of blend films surpasses 1.59%, the antithrombogenicity of plasma treated films increases remarkably due to surface sulfonation. This result implies that SF with blend of PEGM after SO2 plasma treatment have potential use for making blood-contacting biomaterials.

Electrospun silk fibroin nanofibers promote Schwann cell adhesion, growth and proliferation

In this study, Schwann cells, at a density of 1 x 105 cells/well, were cultured on regenerated silk fibroin nanofibers （305 ＋ 84 nm） prepared using the electrospinning method. Schwann cells cultured on the silk fibroin nanofibers appeared more ordered, their processes extended further, and they formed more extensive and complex interconnections. In addition, the silk fibroin nanofibers had no impact on the proliferation of Schwann cells or on the secretion of ciliary neurotrophic factor, brain-derived neurotrophic factor or nerve growth factor. These findings indicate that regenerated electrospun silk fibroin nanofibers can promote Schwann cell adhesion, growth and proliferation, and have excellent biocompatibility.

The degradation behavior of silk fibroin derived from different ionic liquid solvents

Establishing an appropriate degradation rate is critical for tissue engineering scaffolds. In this study, the degradation rate of silk fibroin three-dimensional scaffolds was regulated by changing the molecular weight (MW) of the silk fibroin. The solubility of silk fibroin depends primarily on the ionic ability of the slovent to dissolve silk fibroin, therefore, we regulated the MW of the silk fibroin using LiBr, Ca(NO3)2 and CaCl2 to dissolve the silk fibers. SDS-PAGE analysis showed that the MW of the CaCl2-derived silk fibroin was lower than the MW produced using LiBr and Ca(NO3)2. In vitro and in vivo degradation results showed that the scaffolds prepared by low-MW silk fibroin were more rapidly degraded. Furthermore, FTIR and amino acid analysis suggested that the amorphous regions were preferentially degraded by Collagenase IA, while the SDS-PAGE and amino acid analysis indicated that the scaffolds were degraded into polypeptides (mainly at 10-30 kDa) and amino acids. Because the CaCl2-derived scaffolds contained abundant low MW polypeptides, inter-intramolecular entanglement and traversing of molecular chains in the crystallites reduced, which resulted in rapid degradation. The in vivo degradation results suggested that the degradation rate of the CaCl2-derived scaffolds was better matched to dermis regeneration, indicating that the degradation rate of silk fibroin can be effectively regulated by changing the MW to achieve a suitable dermal tissue regeneration rate.

Mesoporous Bioglass/Silk Fibroin Scaffolds as a Drug Delivery System: Fabrication, Drug Loading and Release in vitro and Repair Calvarial Defects in vivo

The potential of combining bioactive glass（MBG） and silk fibroin（SF） together as a new drug delivery system was evaluated. The three-dimensional porous scaffolds were selected as the form of SF, and sol-gel method was adopted to fabricate MBG in this study. The characteristic of the synthesized material was measured by transmission electron microscopy and scanning electron microscopy. In vitro evaluation of drug delivery was carried out in terms of drug loading and drug release. And aspirin was chosen as the drug for scaffolds to carry out in vitro tests and repair BALB/C mice calvarial defects. Bone formation was examined by microcomputed tomography. The experimental results show that MBG/silk scaffolds have better physiochemical properties compared with silk scaffolds. In comparison to pure silk scaffolds, MBG/silk scaffolds enhance the drug loading efficiency, release rate in vitro and promote bone regeneration in vivo. Thus we conclude that MBG/silk scaffold is a more efficient drug delivery system than pure silk scaffolds.

Designing and Cloning of the Gene Sequence Encoding Silk Fibroin Amorphous Domain

To provide materials used in investigating the relationship between amino acid compositions of silk-like protein, structure, and functions, especially the biological functions, the motif genes encoding the silk fibroin amorphous domain, SGFGPVANGGSGEASSESDFGSSGFGPVANASSGEASSESDFAG(F) were designed and extended using a "head-to-tail" construction strategy. The designed genes were cloned into PSLFA1180FA and multimerized to form structures containing a two-timer, a four-timer, an eight-timer, and a twelve-timer. All the resulting plasmids were digested using the restriction enzyme BamHI and the double-enzymes BglII/HindIII. Restriction enzyme analysis and DNA sequencing revealed the motif was successfully cloned into PSLFA1180FA and multimerized to form a twelve-timer without gene deletion or mutation.

Silk Fibroin/Poly-L-lactide Lactone Bi-layered Membranes for Guided Bone Regeneration

After removal of the caries or diseased teeth,the alveolar ridge will undergo absorption and atrophy.When the amount of alveolar bone is insufficient,it will cause an inability to perform effective dental implant restoration.In order to control the absorption and promote the repair and regeneration of alveolar ridge,a method of implanting guided bone regeneration(GBR)membranes at the extraction site is often used.In this study,silk fibroin(SF)and poly-L-lactide lactone(PLCL)were used to prepare bilayered guided bone regeneration membranes,and its morphology,hydrophilicity,surface roughness and mechanical properties were studied.At the same time,the drug release behaviors and cell compatibility of the bilayered membranes were studied.The results showed that SF/PLCL bi-layered membranes had good mechanical properties and surface hydrophilicity,and the drug-loaded bi-layered membranes had good cell compatibility.The bilayered membranes fabricated in this study are of potential for applying in the oral health field to promote bone regeneration.