Yejing Wang

In situ green synthesis and characterization of sericin-silver nanoparticle composite with effective antibacterial activity and good biocompatibility

Silver nanoparticle has been widely applied to a variety of fields for its outstanding antimicrobial activity. However, the stability of silver nanoparticle limits its application under certain conditions. Thus, improving the stability of silver nanoparticle via biosynthesis is a promising shortcut to expand its application. Sericin from silkworm cocoon has good hydrophilicity, reaction activity, biocompatibility and biodegradability. In this study, we developed a novel, simple, one-step biosynthesis method to prepare sericin-silver nanoparticle composite in situ in solution. Sericin served as the reductant of silver ion, the dispersant and stabilizer of the prepared sericin-silver nanoparticle composite. Natural light was the only power source used to catalyze the synthesis of silver nanoparticle in situ in solution. The novel sericin-silver nanoparticle composite was characterized by ultraviolet-visible and fluorescence spectroscopy, X-ray diffraction, transmission electron microscopy and fourier transform infrared spectroscopy. The results showed silver nanoparticle could be synthesized through the reduction of AgNO3 by the phenolic hydroxyl group of tyrosine residues of sericin under the catalysis of natural light. The synthesized silver nanoparticle had good crystalline, size distribution and long-term stability at room temperature. Light irradiation was essential for the preparation of sericin-silver nanoparticle composite. The antibacterial activity assay showed 25mg/L and 100mg/L were the minimum concentrations of sericin-silver nanoparticle composite required to inhibit the growth of Staphylococcus aureus and kill this bacterium, respectively. The cytotoxicity assay showed cell viability and cell growth were almost not affected by sericin-silver nanoparticle composite under the concentration of 25mg/L. Our study suggested the preparation of sericin-silver nanoparticle composite was environmentally friendly and energy conservation, and the prepared sericin-silver nanoparticle composite had long-term stability, effective antibacterial activity and good biocompatibility. This novel sericin-silver nanoparticle composite has shown great potentials for biomedical application such as antibacterial agent and wound care.

Preparation and characterization of silk sericin/PVA blend film with silver nanoparticles for potential antimicrobial application

Sericin has great potentials in biomedical applications for its good reactive activity, biocompatibility and biodegradability. However, the undesirable mechanical performance limits its application. Here, we developed a green, facile and economic approach to prepare sericin/polyvinyl alcohol (PVA) blend film. Further, silver nanoparticles (AgNPs) were synthesized in situ on the surface of sericin/PVA film via UV-assisted green synthesis method. Mechanical performance, swelling, mass losing and water retention tests showed the blend film had good mechanical performance, hygroscopicity, water retention capacity and low mass losing ratio. Scanning electron microscopy, fourier transfer infrared spectroscopy, X-ray diffractometry diffraction and X-ray photoelectron spectroscopy indicated the blending of PVA and sericin promoted the formation of hydrogen bond network between sericin and PVA, thus enhanced the mechanical performance and the stability of sericin, as well as the hygroscopicity and water retention capacity. UV irradiation and AgNPs modification did not affect the inner crystalline structure of sericin/PVA blend film. The inhibition zone and bacteria growth curve assay suggested AgNPs-sericin/PVA film had good antibacterial activities against E. coli and S. aureus. This novel AgNPs-sericin/PVA film shows great potentials in biomedical materials such as wound dressing and skin tissue engineering.

One-Step Synthesis of Silver Nanoparticles on Polydopamine-Coated Sericin/Polyvinyl Alcohol Composite Films for Potential Antimicrobial Applications

Silk sericin has great potential as a biomaterial for biomedical applications due to its good hydrophilicity, reactivity, and biodegradability. To develop multifunctional sericin materials for potential antibacterial application, a one-step synthesis method for preparing silver nanoparticles (AgNPs) modified on polydopamine-coated sericin/polyvinyl alcohol (PVA) composite films was developed. Polydopamine (PDA) acted as both metal ion chelating and reducing agent to synthesize AgNPs in situ on the sericin/PVA composite film. Scanning electron microscopy and energy dispersive spectroscopy analysis revealed that polydopamine could effectively facilitate the high-density growth of AgNPs as a 3-D matrix. X-ray diffractometry studies suggested the synthesized AgNPs formed good face-centered cubic crystalline structures. Contact angle measurement and mechanical test indicated AgNPs modified PDA-sericin/PVA composite film had good hydrophilicity and mechanical property. The bacterial growth curve and inhibition zone assays showed the AgNPs modified PDA-sericin/PVA composite film had long-term antibacterial activities. This work develops a new method for the preparation of AgNPs modified PDA-sericin/PVA film with good hydrophilicity, mechanical performance and antibacterial activities for the potential antimicrobial application in biomedicine.

Biosynthesis and Characterization of AgNPs–Silk/PVA Film for Potential Packaging Application

Bionanocomposite packaging materials have a bright future for a broad range of applications in the food and biomedical industries. Antimicrobial packaging is one of the bionanocomposite packaging materials. Silver nanoparticle (AgNP) is one of the most attractive antimicrobial agents for its broad spectrum of antimicrobial activity against microorganisms. However, the traditional method of preparing AgNPs-functionalized packaging material is cumbersome and not environmentally friendly. To develop an efficient and convenient biosynthesis method to prepare AgNPs-modified bionanocomposite material for packaging applications, we synthesized AgNPs in situ in a silk fibroin solution via the reduction of Ag+ by the tyrosine residue of fibroin, and then prepared AgNPs–silk/poly(vinyl alcohol) (PVA) composite film by blending with PVA. AgNPs were synthesized evenly on the surface or embedded in the interior of silk/PVA film. The prepared AgNPs–silk/PVA film exhibited excellent mechanical performance and stability, as well as good antibacterial activity against both Gram-negative and Gram-positive bacteria. AgNPs–silk/PVA film offers more choices to be potentially applied in the active packaging field.

In Situ Synthesis of Silver Nanoparticles on the Polyelectrolyte-Coated Sericin/PVA Film for Enhanced Antibacterial Application

To develop silk sericin (SS) as a potential antibacterial biomaterial, a novel composite of polyelectrolyte multilayers (PEMs) coated sericin/poly(vinyl alcohol) (SS/PVA) film modified with silver nanoparticles (AgNPs) has been developed using a layer-by-layer assembly technique and ultraviolet-assisted AgNPs synthesis method. Ag ions were enriched by PEMs via the electrostatic attraction between Ag ions and PEMs, and then reduced to AgNPs in situ with the assistance of ultraviolet irradiation. PEMs facilitated the high-density growth of AgNPs and protected the synthesized AgNPs due to the formation of a 3D matrix, and thus endowed SS/PVA film with highly effective and durable antibacterial activity. Scanning electron microscopy, energy dispersive spectroscopy, X-ray diffractometry, Fourier transfer infrared spectroscopy, water contact angle, mechanical property and thermogravimetric analysis were applied to characterize SS/PVA, PEMs-SS/PVA and AgNPs-PEMs-SS/PVA films, respectively. AgNPs-PEMs-SS/PVA film has exhibited good mechanical performance, hydrophilicity, water absorption capability as well as excellent and durable antibacterial activity against Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa and good stability and degradability. This study has developed a simple method to design and prepare AgNPs-PEMs-SS/PVA film for potential antibacterial application.

Fabrication of sericin/agrose gel loaded lysozyme and its potential in wound dressing application

Sericin is a biomaterial resource for its significant biodegradability, biocompatibility, hydrophilicity, and reactivity. Designing a material with superabsorbent, antiseptic, and non-cytotoxic wound dressing properties is advantageous to reduce wound infection and promote wound healing. Herein, we propose an environment-friendly strategy to obtain an interpenetrating polymer network gel through blending sericin and agarose and freeze-drying. The physicochemical characterizations of the sericin/agarose gel including morphology, porosity, swelling behavior, crystallinity, secondary structure, and thermal property were well characterized. Subsequently, the lysozyme loaded sericin/agarose composite gel was successfully prepared by the solution impregnation method. To evaluate the potential of the lysozyme loaded sericin/agarose gel in wound dressing application, we analyzed the lysozyme loading and release, antimicrobial activity, and cytocompatibility of the resulting gel. The results showed the lysozyme loaded composite gel had high porosity, excellent water absorption property, and good antimicrobial activities against Escherichia coli and Staphylococcus aureus. Also, the lysozyme loaded gel showed excellent cytocompatibility on NIH3T3 and HEK293 cells. So, the lysozyme loaded sericin/agarose gel is a potential alternative biomaterial for wound dressing.

A Novel AgNPs/Sericin/Agar Film with Enhanced Mechanical Property and Antibacterial Capability

Silk sericin is a protein from a silkworm’s cocoon. It has good biocompatibility, hydrophilicity, bioactivity, and biodegradability. However, sericin could not be used in biomedical materials directly because of its frangible characteristic. To develop multifunctional sericin-based materials for biomedical purposes, we prepared a sericin/agar (SS/agar) composite film through the blending of sericin and agar and repetitive freeze-thawing. Then, we synthesized silver nanoparticles (AgNPs) in situ on the surface of the composite film to endow it with antibacterial activity. Water contact angle, swelling and losing ratio, and mechanical properties analysis indicated that the composite film had excellent mechanical property, hydrophilicity, hygroscopicity, and stability. Scanning electron microscopy and X-ray photoelectron spectroscopy analysis confirmed the successful modification of AgNPs on the composite film. X-ray powder diffraction showed the face-centered cubic structures of the AgNPs. This AgNPs modified composite film exhibited an excellent antibacterial capability against Escherichia coli and Staphylococcus aureus. Our study develops a novel AgNPs/sericin/agar composite film with enhanced mechanical performance and an antimicrobial property for potential biomedical applications.

Preparation and Characterization of AgNPs In Situ Synthesis on Polyelectrolyte Membrane Coated Sericin/Agar Film for Antimicrobial Applications

Antibacterial materials are of great importance in preventing bacterial adhesion and reproduction in daily life. Silver nanoparticle (AgNP) is a broad-spectrum antibacterial nanomaterial that has attracted significant attentions for its ability to endow natural materials with antibacterial ability. Silk sericin (SS) has a great advantage for biomaterial application, as it is a natural protein with excellent hydrophilicity and biodegradability. In this study, we prepared AgNPs and polyelectrolyte membrane (PEM) modified SS/Agar films through the layer-by-layer adsorption technique and ultraviolet-assisted AgNPs synthesis method. The film was well characterized by scanning electron microscopy, energy dispersive spectroscopy, X-ray diffraction, fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy. Other properties such as water contact angle, wettability and tensile strength, the release of silver were also studied. The antimicrobial activity of AgNPs-PEM-SS/Agar film was investigated against Escherichia coli and Staphylococcus aureus as the model microorganisms by the inhibition zone and bacterial growth curve assays. The results suggested that the AgNPs-PEM-SS/Agar film had excellent mechanical performance, high hydrophilicity, prominent water absorption ability, as well as outstanding and durable antibacterial activity. Therefore, the prepared novel AgNPs-PEM-SS/Agar composite film is proposed as a potentially favorable antibacterial biomaterial for biomedical applications.

LBD1 of Vitellogenin Receptor Specifically Binds to the Female-Specific Storage Protein SP1 via LBR1 and LBR3

Storage proteins are the major protein synthesized in the fat body, released into hemolymph and re-sequestered into the fat body before pupation in most insect species. Storage proteins are important amino acid and nutrition resources during the non-feeding pupal period and play essential roles for the metamorphosis and oogenesis of insects. The sequestration of storage protein is a selective, specific receptor-mediated process. However, to date, the potential receptor mediating the sequestration of storage protein has not been determined in Bombyx mori. In this study, we expressed and purified the first ligand binding domain of Bombyx mori vitellogenin receptor (BmVgR), LBD1, and found LBD1 could bind with an unknown protein from the hemolymph of the ultimate silkworm larval instar via pull-down assay. This unknown protein was subsequently identified to be the female-specific storage protein SP1 by mass spectrometry. Furthermore, far western blotting assay, immunoprecipitation and isothermal titration calorimetry analysis demonstrated LBD1 specifically bound with the female-specific SP1, rather than another unisex storage protein SP2. The specific binding of LBD1 with SP1 was dependent on the presence of Ca2+ as it was essential for the proper conformation of LBD1. Deletion mutagenesis and ITC analysis revealed the first and third ligand binding repeats LBR1 and LBR3 were indispensable for the binding of LBD1 with SP1, and LBR2 and LBR4 also had a certain contribution to the specific binding. Our results implied BmVgR may mediate the sequestration of SP1 from hemolymph into the fat body during the larval-pupal transformation of Bombyx mori.

Biochemical characterization and functional analysis of the POU transcription factor POU-M2 of Bombyx mori

POU-M2 is a homeodomain transcription factor which plays important roles in the development and silk synthesis of Bombyx mori. In this study, we expressed, purified and characterized POU-M2 and studied its transcription regulation on fibroin heavy chain gene of Bombyx mori. Gel filtration showed POU-M2 existed as a dimer in solution. Far-UV circular dichroism spectra indicated POU-M2 had a well-defined α-helix structure and the α-helix content was about 26.4%. The thermal unfolding transition of POU-M2 was a cooperative process. Tm, ΔH and ΔS were 45.15 ± 0.2 °C, 138.4 ± 0.5 KJ/mol and 0.4349 ± 0.04 KJ/(mol·K), respectively. Western blotting analysis indicated the expression level of POU-M2 increased slightly from day 3 to day 7 of the fifth instar larvae in the posterior silk gland. POU-M2 was positioned in the nucleus of cells. The luciferase reporter assay demonstrated POU-M2 could stimulate the promoter activity of fibroin heavy chain gene, and the activation effect was dependent on the amount of POU-M2. Our study suggested POU-M2 may be involved in the transcriptional regulation of fibroin heavy chain gene. These findings expand toward a better understanding of the structure of POU-M2 and its function in silk synthesis of Bombyx mori.