Silk fibroins modify the atmospheric low temperature plasma-treated poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) film for the application of cardiovascular tissue engineering

Tissue engineered scaffold is one of the hopeful therapies for the patients with organ or tissue damages. The key element for a tissue engineered scaffold material is high biocompatibility. Herein the poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) film was irradiated by the low temperature atmospheric plasma and then coated by the silk fibroins (SF). After plasma treatment, the surface of PHBHHx film became rougher and more hydrophilic than that of original film. The experiment of PHBHHx flushed by phosphate buffer solution (PBS) proves that the coated SF shows stronger immobilization on the plasma-treated film than that on the untreated film. The cell viability assay demonstrates that SF-coated PHBHHx films treated by the plasma significantly supports the proliferation and growth of the human smooth muscle cells (HSMCs). Furthermore, the scanning electron microscopy and hemotoylin and eosin (HE) staining show that HSMCs formed a cell sub-monolayer and secreted a large amount of extracellular matrix (ECM) on the films after one week's culture. The silk fibroins modify the plasma-treated PHBHHx film, providing a material potentially applicable in the cardiovascular tissue engi-neering.

蚕丝基生物材料精准和功能性组装的3D打印策略

In recent years,significant progress has been made in both three-dimensional(3D)printing technologies and the exploration of silk as an ink to produce biocompatible constructs.Combined with the unlimited design potential of 3D printing,silk can be processed into a broad range of functional materials and devices for various biomedical applications.The ability of silk to be processed into various materials,including solutions,hydrogels,particles,microspheres,and fibers,makes it an excellent candidate for adaptation to different 3D printing techniques.This review presents a didactic overview of the 3D printing of silk-based materials,major categories of printing techniques,and their prototyping mechanisms and structural features.In addition,we provide a roadmap for researchers aiming to incorporate silk printing into their own work by summarizing promising strategies from both technical and material aspects,to relate state-of-the-art silk-based material processing with fast-developing 3D printing technologies.Thus,our focus is on elucidating the techniques and strategies that advance the development of precise assembly strategies for silk-based materials.Precise printing(including high printing resolution,complex structure realization,and printing fidelity)is a prerequisite for the digital design capability of 3D printing technology and would definitely broaden the application era of silk,such as complex biomimetic tissue structures,vasculatures,and transdermal microneedles.

Injectable Nanorobot-Hydrogel Superstructure for Hemostasis and Anticancer Therapy of Spinal Metastasis

Surgery remains the standard treatment for spinal metastasis.However,uncontrolled intraoperative bleeding poses a significant challenge for adequate surgical resection and compromises surgical outcomes.In this study,we develop a thrombin(Thr)-loaded nanorobothydrogel hybrid superstructure by incorporating nanorobots into regenerated silk fibroin nanofibril hydrogels.This superstructure with superior thixotropic properties is injected percutaneously and dispersed into the spinal metastasis of hepatocellular carcinoma(HCC)with easy bleeding characteristics,before spinal surgery in a mouse model.Under near-infrared irradiation,the self-motile nanorobots penetrate into the deep spinal tumor,releasing Thr in a controlled manner.Thr-induced thrombosis effectively blocks the tumor vasculature and reduces bleeding,inhibiting tumor growth and postoperative recurrence with Au nanorod-mediated photothermal therapy.Our minimally invasive treatment platform provides a novel preoperative therapeutic strategy for HCC spinal metastasis effectively controlling intraoperative bleeding and tumor growth,with potentially reduced surgical complications and enhanced operative outcomes.

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.

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.

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.

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.

Humanoid Intelligent Display Platform for Audiovisual Interaction and Sound Identification

This study proposes a rational strategy for the design,fabrication and system integration of the humanoid intelligent display platform(HIDP)to meet the requirements of highly humanized mechanical properties and intelligence for human-machine interfaces.The platform’s sandwich structure comprises a middle lightemitting layer and surface electrodes,which consists of silicon elastomer embedded with phosphor and silk fibroin ionoelastomer,respectively.Both materials are highly stretchable and resilient,endowing the HIDP with skin-like mechanical properties and applicability in various extreme environments and complex mechanical stimulations.Furthermore,by establishing the numerical correlation between the amplitude change of animal sounds and the brightness variation,the HIDP realizes audiovisual interaction and successful identification of animal species with the aid of Internet of Things(IoT)and machine learning techniques.The accuracy of species identification reaches about 100%for 200 rounds of random testing.Additionally,the HIDP can recognize animal species and their corresponding frequencies by analyzing sound characteristics,displaying real-time results with an accuracy of approximately 99%and 93%,respectively.In sum,this study offers a rational route to designing intelligent display devices for audiovisual interaction,which can expedite the application of smart display devices in human-machine interaction,soft robotics,wearable sound-vision system and medical devices for hearing-impaired patients.

A multifunctional osteogenic system of ultrasonically spray deposited bone-active coatings on plasma-activated magnesium

Biomimetic bone-active coatings composed of inorganic nano-hydroxyapatite(i.e.,nHA)and organic silk fibroin(i.e.,SF)are layer-by-layer deposited on Mg-Zn-Ca alloy by a controllable ultrasonic spray method.Meanwhile,plasma activation is developed as a promising strategy to pretreat magnesium surfaces,which facilitates the direct adhesion of coatings with enhanced bonding interfaces.In this work,we engineer the nHA/SF composite coatings with excellent mechanical properties and adhesion force.The optimized parameters of ultrasonic spray bring significant influence on the surface morphologies of coatings.Assisted by hybrid plasma of oxygen and nitrogen(i.e.,O_(2)/N_(2) plasma),the activated Mg-Zn-Ca surfaces are uniformly covered by a robust and compact nHA/SF composite coating,establishing a multifunctional system with superior corrosion resistance and biological performance.Interestingly,secondary oxygen plasma treatment of nHA/SF coatings(A-nHA/SF)promotes the hydrophilicity,leading to a rapid self-repair effect from surface damage.The improvement of anti-corrosion and self-repair provides a dependable platform for better cell adhesion,proliferation,spreading and differentiation.These favorable factors contribute to the preferable in vivo biocompatibility and the promotion of newly formed bones for the A-nHA/SF-coated Mg implants.This study lays important foundations for coating strategy on biomedical magnesium alloy as multifunctional osteogenic system in bioactive implantable applications.

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 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.

IMPROVEMENT OF WET-CREASE RECOVERY PROPERTIES OF SILK FABRICS TREATED WITH EPOXIDES

Silk fabrics is finished with epoxide TDEA or TDEB. By the determination of washing fastness, DSC, IR spectrum and amino acid contents tests, TDEA or TDEB is confirmed to react with silk fibroin. By means of tests of silk fibroin swelling and solubility in some solvents, crosslinking reactions between TDEA or TDEB and silk fibroin arc further proved, which is obviously responsible to the increase of wet-crease recovery of silk fabrics finished with epoxide TDEA or TDEB.

Exploration of the enhanced performances for silk fibroin/sodium alginate composite coatings on biodegradable Mg-Zn-Ca alloy

To expand the future clinic applications of biodegradable magnesium alloy,polymer coatings with excellent biocompatibility are the keys to solve the local alkalinity and rapid hydrogen release.Natural-organic silk fibroin provides an approach to fabricate a protective coating on biomedical Mg-Zn-Ca alloy,however,the adhesion force and mechanical properties of the coating on substrates are ought to be further improved without any chemical conversion/intermediate layer.Hereby,based on VUV/O;surface activation,a hybrid of silk fibroin and sodium alginate is proposed to enhance the adhesion force and mechanical properties of the composite coatings on hydrophilic Mg-Zn-Ca alloy surfaces.Various mass ratios of sodium alginate addition were investigated to achieve the optimum coating strategy.The nanoscratch test and nanoindentation test confirmed that the adhesion force was tripled and mechanical properties index was significantly improved when the mass ratio of silk fibroin/sodium alginate was 70/30 compared to pure silk fibroin or sodium alginate coatings.Meanwhile,the corrosion rate of the coated Mg-Zn-Ca alloy was significantly delayed with the addition of sodium alginate,resulting in a reaction layer during corrosion process.Furthermore,the mechanisms for both adhesion and corrosion processes were discussed in detail.Our findings offer more possibilities for the controllable surface performance of degradable metals.

Pulling out a peptide chain from β-sheet crystallite: Propagation of instability of H-bonds under shear force

Anti-parallel β-sheet crystallite as the main component of silk fibroin has attracted much attention due to its superior mechanical properties. In this study, we examine the processes of pulling a peptide chain from β-sheet crystallite using steered molecular dynamics simulations to investigate the rupture behavior of the crystallite. We show that the failure of β-sheet crystallite was accompanied by a propagation of instability of hydrogen-bonds （H-bonds） in the crystallite. In addition, we find that there is an optimum size of the crystallite at which the H-bonds can work cooperatively to achieve the highest shear strength. In addition, we find that the stiffness of loading device and the loading rates have significant effects on the rupture behavior of β-sheet crystallite. The stiff loading device facilitates the rebinding of the Hbond network in the stick-slip motion between the chains, while the soft one suppresses it. Moreover, the rupture force of β-sheet crystallites decreases with loading rate. Particularly, when the loading rate decreases to a critical value, the rupture force of the β-sheet crystallite becomes independent of the loading rates. This study provides atomistic details of rupture behaviors of β-sheet crystallite, and, therefore, sheds valuable light on the underlying mechanism of the superior mechanical properties of silk fibroin.

Fabricating a reactive surface on the fibroin film by a room-temperature plasma jet array for biomolecule immobilization

A simple dielectric barrier discharge（DBD） jet array was designed with a liquid electrode and helium gas.The characteristics of the jet array discharge and the preliminary polymerization with acrylic acid（AA） monomer were presented.The plasma reactor can produce a cold jet array with a gas temperature lower than 315 K,using an applied discharge power between 6 W and 30 W（V dis × I dis）.A silk fibroin film（SFF） was modified using the jet array and AA monomer,and the treated SFF samples were characterized by atomic force microscopy（AFM）,scanning electron microscopy（SEM）,Fourier transform infrared spectroscopy（FTIR）,and contact angle（CA）.The deposition rate of the poly acrylic acid（PAA） was able to reach 300 nm/min,and the surface roughness and energy increased with the AA flow rate.The FTIR results indicate that the modified SFF had more carboxyl groups（-COOH） than the original SFF.This latter characteristic allowed the modified SFF to immobilize more quantities of antimicrobial peptide（AP,LL-37） which inhibited the Escherichia coli（E.Coli） effectively.