Decoding silkworm spinning programmed by pH and metal ions.

Silk is one of the toughest fibrous materials known despite spun at ambient temperature and pressure with water as a solvent. It is a great challenge to reproduce high-performance artificial fibers comparable to natural silk by bionic for the incomplete understanding of silkworm spinning in vivo. Here, we found that amphipol and digitonin stabilized the structure of natural silk fibroin (NSF) by a large-scale screening in vitro, and then studied the close-to-native ultrastructure and hierarchical assembly of NSF in the silk gland lumen. Our study showed that NSF formed reversible flexible nanofibrils mainly composed of random coils with a sedimentation coefficient of 5.8 S and a diameter of about 4 nm, rather than a micellar or rod-like structure assembled by the aggregation of globular NSF molecules. Metal ions were required for NSF nanofibril formation. The successive pH decrease from posterior silk gland (PSG) to anterior silk gland (ASG) resulted in a gradual increase in NSF hydrophobicity, thus inducing the sol-gelation transition of NSF nanofibrils. NSF nanofibrils were randomly dispersed from PSG to ASG-1, and self-assembled into anisotropic herringbone patterns at ASG-2 near the spinneret ready for silkworm spinning. Our findings reveal the controlled self-assembly mechanism of the multi-scale hierarchical architecture of NSF from nanofibrils to herringbone patterns programmed by metal ions and pH gradient, which provides novel insights into the spinning mechanism of silk-secreting animals and bioinspired design of high-performance fibers.

Mn-Anti-CTLA4-CREKA-Sericin Nanotheragnostics for Enhanced Magnetic Resonance Imaging and Tumor Immunotherapy.

The integration of magnetic resonance imaging (MRI), cGAS-STING, and anti-CTLA-4 (aCTLA-4) based immunotherapy offers new opportunities for tumor precision therapy. However, the precise delivery of aCTLA-4 and manganese (Mn), an activator of cGAS, to tumors remains a major challenge for enhanced MRI and active immunotherapy. Herein, a theragnostic nanosphere Mn-CREKA-aCTLA-4-SS (MCCS) is prepared by covalently assembling Mn2+, silk sericin (SS), pentapeptide CREKA, and aCTLA-4. MCCS are stable with an average size of 160 nm and is almost negatively charged or neutral at pH 5.5/7.4. T1-weighted images showed MCCS actively targeted tumors to improve the relaxation rate r1 and contrast time of MRI. This studies demonstrated MCCS raises reactive oxygen species levels, activates the cGAS-STING pathway, stimulates effectors CD8+ and CD80+ T cells, reduces regulatory T cell numbers, and increases IFN-γ and granzyme secretion, thereby inducing tumor cells autophagy and apoptosis in vitro and in vivo. Also, MCCS are biocompatible and biosafe. These studies show the great potential of Mn-/SS-based integrative material MCCS for precision and personalized tumor nanotheragnostics.

POUM2 homeostasis regulates intimal remodeling and cells fate in the anterior silk gland of the silkworm.

Tissue/organ remodeling and cells fate determination play key roles in the life cycle of animals. However, they are still poorly understood in insects, especially in the silkworm. The anterior silk gland (ASG) of the silkworm is essential for the formation and performance of silk fibers, but the regulatory mechanism of ASG remodeling and cells fate determination is less known. Here we found that silencing of POUM2 caused shorter ASG length, intimal structural defects, silkworm spinning failure, and the resultant naked pupae death, but cells number was not affected. Cells staining showed that DNA endoreduplication was not affected in the ASG. Transmission electron microscopy and chitin staining showed cuticle proteins and chitin were greatly reduced in the ASG during the molting period. Transcriptional analysis showed the expression profiles of cuticle proteins and chitin synthase were similar to that of POUM2 during the molting period, and POUM2 down-regulation reduced the expression of cuticle proteins, chitin synthase, autophagy and apoptosis-related genes. While the phenotype resulting from POUM2 over-expression was similar to that of POUM2 down-regulation. Cells staining revealed marked cells apoptosis with cells number reduction and inhibition of DNA endoreduplication in the ASG. Transcriptional analysis showed the expression of autophagy and apoptosis-related genes, and some cuticle proteins and chitin synthase were significantly up-regulated. The results suggest that POUM2 homeostasis regulates ASG intimal remodeling and cells fate, thus affecting ASG development, silkworm spinning and metamorphosis. Our studies not only offer potential molecular targets for genetic improvement of silk performance and molecular breeding of the silkworm, but also provide new insights into POU factor-mediated tissue remodeling and cells fate determination in insects.

POU-M2 promotes juvenile hormone biosynthesis by directly activating the transcription of juvenile hormone synthetic enzyme genes in Bombyx mori.

Juvenile hormone (JH) plays a key role in preventing larval precocious metamorphosis, maintaining larval state, controlling adult sexual development and promoting insect egg maturation. Genetic studies have shown that POU factor ventral veins lacking regulates JH synthesis to control the timing of insect metamorphosis. However, how POU factor regulates JH synthesis is largely unknown. Here, we found POU-M2 was highly expressed in corpora allata (CA) and specifically localized in the nucleus of CA. The overexpression of POU-M2 promoted the expression of JH synthase genes and kr-h1 and enhanced the activity of JH synthase genes promoter. Further, POU-M2 promoted the transcription of JH acid O-methyltransferase (JHAMT) by directly binding to the key cis-regulatory elements -207, -249 and -453 within the proximal regions of JHAMT promoter. Both the POU domain and homeodomain were vital for the activation of POU-M2 on JHAMT transcription. Our study reveals the mechanism by which POU-M2 regulates JHAMT transcription.

Fabrication of antibacterial sericin based hydrogel as an injectable and mouldable wound dressing.

Hydrogel-based wound dressings are ideal materials as they could provide a moist environment to promote wound healing. Silk sericin is a natural macromolecular protein produced by silkworms and has positive effects on fibroblasts and keratinocytes. Sodium alginate is a natural polysaccharide known as a mouldable matrix. To develop novel hydrogel wound dressings, sodium alginate was functionalized with silk sericin to yield an injectable and mouldable semi-interpenetrating network hydrogel (SA/Se) using calcium ions as the crosslinking agents. Meanwhile, silver nanoparticles (AgNPs) were synthesized in situ by the phenolic hydroxyl group of tyrosine in sericin to enhance the antibacterial activity of the hybrid hydrogel. UV-visible spectroscopy, fluorescence spectroscopy, transmission electron microscope, X-ray photoelectron spectroscopy and energy dispersive spectroscopy demonstrated the successful synthesis of AgNPs with average size of 15.7 nm in SA/Se-Ag0.2. Field emission scanning electron microscopy showed the macroporous structure of the hydrogel. The hybrid hydrogel had a swelling ratio of 32 and more than 5% of retention ratio after 50 h. It exhibited effective antibacterial activity through irreversibly disrupting bacterial membrane structure to result in bacterial lysis and death. Cell counting kit-8 assay suggested the hybrid hydrogel was highly cytocompatible. The animal experiment showed the wound contraction ratio in SA/Se-Ag0.2 group was 99% after 12 days, indicating it can effectively promote wound healing. This novel hydrogel is promising for wound healing and tissue engineering.

Rational Design of Ag/ZnO Hybrid Nanoparticles on Sericin/Agarose Composite Film for Enhanced Antimicrobial Applications.

Silver-based hybrid nanomaterials are receiving increasing attention as potential alternatives for traditional antimicrobial agents. Here, we proposed a simple and eco-friendly strategy to efficiently assemble zinc oxide nanoparticles (ZnO) and silver nanoparticles (AgNPs) on sericin-agarose composite film to impart superior antimicrobial activity. Based on a layer-by-layer self-assembly strategy, AgNPs and ZnO were immobilized on sericin-agarose films using the adhesion property of polydopamine. Scanning electron microscopy, energy-dispersive X-ray spectroscopy, and X-ray powder diffraction spectroscopy were used to show the morphology of AgNPs and ZnO on the surface of the composite film and analyze the composition and structure of AgNPs and ZnO, respectively. Water contact angle, swelling ratio, and mechanical property were determined to characterize the hydrophilicity, water absorption ability, and mechanical properties of the composite films. In addition, the antibacterial activity of the composite film was evaluated against Gram-positive and Gram-negative bacteria. The results showed that the composite film not only has desirable hydrophilicity, high water absorption ability, and favorable mechanical properties but also exhibits excellent antimicrobial activity against both Gram-positive and Gram-negative bacteria. It has shown great potential as a novel antimicrobial biomaterial for wound dressing, artificial skin, and tissue engineering.

Impact of a group-based intervention program on physical activity and health-related outcomes in worksite settings.

BACKGROUND: The benefits of physical activity (PA) have been well documented, and the worksite is a promising setting for PA promotion. The aims of this study were as follows: 1. To evaluate the effect of a group-based worksite intervention on PA and health-related outcomes by using pedometers. 2. To examine the associations between the change in vigorous physical activity (VPA)/moderate physical activity (MPA)/walking and health related outcomes. METHODS: A total of 398 participants (221 in the intervention group (IG) and 177 in the control group (CG)) from 17 worksites were recruited for a prospective self-controlled trial of a worksite physical activity intervention program in China. In the IG, a pedometer was utilized to self-monitor the PA, together with group competition, goal setting, and other incentives. No intervention was applied to the CG. Physical activity, sedentary behavior, and health-related outcomes were measured at baseline and immediately after the 100-day period intervention. RESULTS: A total of 262 participants completed the program (68.3% adherence). Adherence in the intervention group was 67.9% (n = 150/221). Improvements between baseline and follow-up among intervention participants were observed in the following parameters: VPA (+ 109.7 METs/week; p < 0.05), walking (+ 209.2 METs/week; p < 0.01), systolic blood pressure (SBP; - 2.1 mmHg; p < 0.01), waist circumference (WC; - 2.3 cm; p < 0.01), body fat percentage (BF); - 1.0%; p < 0.01), and body mass index (BMI; - 0.5 kg/m2; p < 0.01). VPA was related to changes in body fat percentage (p < 0.05) and body mass index (p < 0.05). CONCLUSION: This integrated group-based intervention program contributed to comprehensive improvement in health-related outcomes. The study was useful for establishing associations between change in VPA/MPA/walking and health-related outcomes in a natural setting. Long-term evaluation is required to examine the potential of such an integrated intervention to promote PA. REGISTRATION: This study was prospectively registered in the Chinese Clinical Trial Registry. TRIAL REGISTRATION NUMBER: ChiCTR-1,800,015,529. Date of registration: April 5, 2018.

Effects of SDS on the activity and conformation of protein tyrosine phosphatase from thermus thermophilus HB27.

Deciphering the activity-conformation relationship of PTPase is of great interest to understand how PTPase activity is determined by its conformation. Here we studied the activity and conformational transitions of PTPase from thermus thermophilus HB27 in the presence of sodium dodecyl sulfate (SDS). Activity assays showed the inactivation of PTPase induced by SDS was in a concentration-dependent manner. Fluorescence and circular dichroism spectra suggested SDS induced significant conformational transitions of PTPase, which resulted in the inactivation of PTPase, and the changes of α-helical structure and tertiary structure of PTPase. Structural analysis revealed a number of hydrophobic and charged residues around the active sites of PTPase may be involved in the hydrophobic and ionic bonds interactions of PTPase and SDS, which are suggested to be the major driving force to result in PTPase inactivation and conformational transitions induced by SDS. Our results suggested the hydrophobic and charged residues around the active sites were essential for the activity and conformation of PTPase. Our study promotes a better understanding of the activity and conformation of PTPase.

Factors Associated With Colonoscopy Compliance Based on Health Belief Model in a Community-Based Colorectal Cancer Screening Program Shanghai, China.

Screening can help early detection of colorectal cancer (CRC) in the general population. However, colonoscopy compliance in screening program is low in China. The study aimed to identify factors associated with colonoscopy compliance based on Health Belief Model (HBM). An investigation was conducted in Huangpu District, Shanghai in 2015. High-risk individuals of CRC received an in-person interview with physicians to fill out a questionnaire. The questionnaires assessing predictors of colonoscopy compliance were collected, and status of colonoscopy participation was determined. Univariate and multiple logistic regression analyses were conducted. Among 2,568 high-risk population (20.68%), 531 subjects underwent colonoscopy. Participants with both risk assessment and fecal immunochemical test positive were most likely to undergo colonoscopy. Based on HBM, colonoscopy compliance was positively associated with higher perceived severity (odds ratio [OR] = 1.05, 95% confidence interval [CI] = [1.00, 1.10]). Higher perceived barriers (OR = 0.97, 95% CI = [0.94, 0.99]), without prior colonoscopy (OR = 0.35, 95% CI = [0.26, 0.47]), not knowing anyone who underwent colonoscopy before (OR = 0.74, 95% CI = [0.58, 0.96]), without health-care provider recommendation on colonoscopy (OR = 0.58, 95% CI = [0.44, 0.77]), and without psychosocial support from someone for colonoscopy (OR = 0.27, 95% CI = [0.21, 0.35]) were shown to be associated with colonoscopy noncompliance. The colonoscopy compliance was low in this CRC screening program in Shanghai, China. The influencing factors were positive results in primary screening, perceived severity, perceived barriers, personal or others' experiences in colonoscopy, health-care provider recommendation, and psychosocial supports. Effective education campaign and facilitated communication between health-care providers and high-risk population were suggested in the future interventions.

Rational Design and Fabrication of ZnONPs Functionalized Sericin/PVA Antimicrobial Sponge.

The interests of developing antimicrobial biomaterials based on silk sericin from Bombyx mori cocoon, have been shooting up in the last decades. Sericin is a valuable natural protein owing to its hydrophilicity, biodegradability, and biocompatibility. Here, we fabricated a sponge with antibacterial capacities for potential wound dressing application. By co-blending of sericin, polyvinyl alcohol (PVA) and zinc oxide nanoparticles (ZnONPs), the ZnONPs-sericin/PVA composite sponge (ZnONPs-SP) was successfully prepared after freeze-drying. Scanning electron microscopy showed the porous structure of ZnONPs-SP. Energy dispersive spectroscopy indicated the existence of Zn in the sponge. X-ray diffractometry revealed the hexagonal wurtzite structure of ZnONPs. Fourier transform infrared spectroscopy showed the biologic coupling of ZnONPs and sericin resulted in a decrease of α-helix and random coil contents, and an increase of ß-sheet structure in the sponge. The swelling experiment suggested ZnONPs-SP has high porosity, good hydrophilicity, and water absorption capability. The plate bacterial colony counting coupled with growth curve assays demonstrated that the composite sponge has an efficiently bacteriostatic effect against Staphylococcus aureus and Escherichia coli, respectively. Furthermore, the cell compatibility analysis suggested the composite sponge has excellent cytocompatibility on NIH3T3 cells. In all, ZnONPs-SP composite sponge has significant potentials in biomaterials such as wound dressing and tissue engineering.

A Novel Adenosine Kinase from Bombyx mori: Enzymatic Activity, Structure, and Biological Function.

Adenosine kinase (ADK) is the first enzyme in the adenosine remediation pathway that catalyzes adenosine phosphorylation into adenosine monophosphate, thus regulating adenosine homeostasis in cells. To obtain new insights into ADK from Bombyx mori (BmADK), we obtained recombinant BmADK, and analyzed its activity, structure, and function. Gel-filtration showed BmADK was a monomer with molecular weight of approximately 38 kDa. Circular dichroism spectra indicated BmADK had 36.8% α-helix and 29.9% ß-strand structures, respectively. The structure of BmADK was stable in pH 5.0-11.0, and not affected under 30 °C. The melting temperature and the enthalpy and entropy changes in the thermal transition of BmADK were 46.51 ± 0.50 °C, 253.43 ± 0.20 KJ/mol, and 0.79 ± 0.01 KJ/(mol·K), respectively. Site-directed mutagenesis demonstrated G68, S201, E229, and D303 were key amino acids for BmADK structure and activity. In particular, S201A mutation significantly increased the α-helix content of BmADK and its activity. BmADK was located in the cytoplasm and highly expressed in the silk gland during the pre-pupal stage. RNA interference revealed the downregulation of BmADK decreased ATG-8, Caspase-9, Ec-R, E74A, and Br-C expression, indicating it was likely involved in 20E signaling, apoptosis, and autophagy to regulate silk gland degeneration and silkworm metamorphosis. Our study greatly expanded the knowledge on the activity, structure, and role of ADK.

Design and performance of sericin/poly(vinyl alcohol) hydrogel as a drug delivery carrier for potential wound dressing application.

Developing a non-toxic, super-absorbent and antibacterial hydrogel as a skin wound dressing is of significant importance. Sericin hydrogel is an ideal dressing material as it has excellent biocompatibility, biodegradability, moisture retention, high affinity to biomolecules, self-healing and promoting cell proliferation activity. Here, we blended silk sericin (SS) with poly(vinyl alcohol) (PVA) to prepare a SS/PVA hydrogel through repetitive freeze-thawing. The photoluminescence of SS/PVA hydrogel indicated PVA was well blended with sericin. SS/PVA hydrogel showed excellent hydrophilicity and swelling behavior for its porous structure. PVA blending effectively improved the thermostability of sericin and enhanced the mechanical property of sericin, but did not affect the crystallinity of sericin and PVA. SS/PVA hydrogel exhibited the ability to load and release small molecule drugs and silver nanoparticles. Cytotoxicity and immuno-toxicity assays suggested the gentamicin loaded SS/PVA hydrogel had excellent cytocompatibility on mammalian cells. Bacterial inhibition assay and wound infection model demonstrated the gentamicin loaded SS/PVA hydrogel could effectively inhibit bacterial growth, thus maintain cells viability. This novel hydrogel with antimicrobial activity has shown great potential in wound dressing.

Cross-linking of dialdehyde carboxymethyl cellulose with silk sericin to reinforce sericin film for potential biomedical application.

Developing biomaterials based on the natural biomacromolecule silk sericin from Bombyx mori cocoon is of great interest for biomedical application. Dialdehyde carboxymethyl cellulose (DCMC) is derived from periodate oxidation of carboxy- methyl cellulose. Here, we developed a novel strategy of cross-linking of sericin with DCMC via the Schiff's base reaction. Fourier transform infrared spectroscopy and scanning electron microscopy indicated the formation of Schiff's base via the blending of sericin and DCMC. The mechanical properties tests suggested the covalent cross-linking effectively enhanced the tensile strength of sericin. The swelling test and water contact angle indicated the DCMC/SS film had excellent hydrophilicity, swellability. Additionally, we demonstrated the DCMC/SS film had excellent blood compatibility, cytocompatibility and promoting cell proliferation activity by the hemolysis ratio analysis, cell adhesion, cells viability and proliferation assays. The prepared DCMC/SS film has shown great promise in biomedical applications such as wound dressing, artificial skin and tissue engineering.

A Convenient, Rapid, Sensitive, and Reliable Spectrophotometric Assay for Adenylate Kinase Activity.

Enzymatic activity assays are essential and critical for the study of enzyme kinetics. Adenylate kinase (Adk) plays a fundamental role in cellular energy and nucleotide homeostasis. To date, assays based on different principles have been used for the determination of Adk activity. Here, we show a spectrophotometric analysis technique to determine Adk activity with bromothymol blue as a pH indicator. We analyzed the effects of substrates and the pH indicator on the assay using orthogonal design and then established the most optimal assay for Adk activity. Subsequently, we evaluated the thermostability of Adk and the inhibitory effect of KCl on Adk activity with this assay. Our results show that this assay is simple, rapid, and precise. It shows great potential as an alternative to the conventional Adk activity assay. Our results also suggest that orthogonal design is an effective approach, which is very suitable for the optimization of complex enzyme reaction conditions.

Polydopamine-Based Surface Modification of ZnO Nanoparticles on Sericin/Polyvinyl Alcohol Composite Film for Antibacterial Application.

Silk sericin (SS) is a type of natural macromolecular protein with excellent hydrophilicity, biocompatibility and biodegradability, but also has very poor mechanical properties. To develop sericin-based wound dressings, we utilized polyvinyl alcohol (PVA) to reinforce the mechanical property of sericin by blending PVA and sericin, then modified zinc oxide nanoparticles (ZnO NPs) on SS/PVA film with the assistance of polydopamine (PDA) to endow SS/PVA film with antibacterial activity. Scanning electron microscopy, energy dispersive spectroscopy and X-ray powder diffraction demonstrated ZnO NPs were well grafted on PDA-SS/PVA film. Fourier transform infrared spectra suggested PDA coating and ZnONPs modification did not alter the structure of sericin and PVA. Water contact angle and swelling tests indicated the excellent hydrophilicity and swellability of ZnO NPs-PDA-SS/PVA composite film. Mass loss analysis showed ZnO NPs-PDA-SS/PVA film had excellent stability. The mechanical performance test suggested the improved tensile strength and elongation at break could meet the requirement of ZnO NPs-PDA-SS/PVA film in biomaterial applications. The antibacterial assay suggested the prepared ZnO NPs-PDA-SS/PVA composite film had a degree of antimicrobial activity against Escherichia coli and Staphylococcus aureus. The excellent hydrophilicity, swellability, stability, mechanical property and antibacterial activity greatly promote the possibility of ZnO NPs-PDA-SS/PVA composite film in antibacterial biomaterials application.

Cross-talk between juvenile hormone and ecdysone regulates transcription of fibroin modulator binding protein-1 in Bombyx mori.

Juvenile hormone (JH) and 20-hydroxyecdysone (20E) are the most important hormones in silkworm and play vital roles in silkworm development, metamorphosis, and silk protein synthesis. Fibroin modulator binding protein-1 (FMBP-1) is a novel transcription factor regulating fibroin heavy chain (fib-H) transcription in Bombyx mori. The roles of JH and 20E on FMBP-1 transcription are less known. Here, we show FMBP-1 transcription is repressed by juvenile hormone analog (JHA) and activated by 20E. We identify two Krüppel homolog 1 (Kr-h1) binding sites (KBS1 and KBS2) and an E74A binding site (EBS) in the promoter of FMBP-1. We demonstrate Kr-h1 directly binds to KBS1 and KBS2 to repress FMBP-1 transcription, and 20E promotes FMBP-1 transcription through E74A. In the presence of JH and 20E, E74A abolishes the repression of Kr-h1 and activates FMBP-1 transcription through direct binding to EBS between KBS1 and KBS2 in FMBP-1 promoter. Further, JHA and 20E treatment and RNA interference confirm the effects of JH and 20E on FMBP-1 transcription in vivo, thus affecting fib-H transcription. Our results reveal the molecular mechanism of FMBP-1 transcription regulated by the cross-talk between JH and 20E in Bombyx mori, and provide novel insights into FMBP-1 transcriptional regulation and silk protein synthesis.

Insights into the repression of fibroin modulator binding protein-1 on the transcription of fibroin H-chain during molting in Bombyx mori.

Fibroin modulator binding protein-1 (FMBP-1) is a novel DNA-binding protein containing a conserved score and three amino acid peptide repeat (STPR) domain. The roles of factors containing STPR domain are less known. Although multiple transcription factors are involved in the transcriptional regulation of silk protein genes during the development of silkworm, the mechanism of transcriptional repression of silk protein genes during molting remains unclear. Here, we found that FMBP-1 expression was contrary to that of fibroin heavy chain (fib-H) during the fourth molting period of Bombyx mori. FMBP-1 repressed fib-H promoter activity by directly binding to the -130 element in the fib-H promoter region. We also identified two proteins, Bmsage and Bmdimm, that interacted with FMBP-1 in the posterior silk gland of silkworm larvae, and further verified these interactions by far western blotting and microscale thermophoresis in vitro, as well as co-immunoprecipitation and bimolecular fluorescence complementation at the cellular level. The luciferase reporter assay showed that the interaction between FMBP-1 and Bmdimm antagonized the activation of Bmdimm on fib-H transcription, but did not affect FMBP-1-mediated transcriptional repression on fib-H gene. Therefore, we proposed the following mechanism of fib-H transcriptional repression by FMBP-1 during the molting of silkworm larvae: 1) FMBP-1 directly binds to the -130 element in the fib-H promoter to repress fib-H transcription; 2) FMBP-1 interacts with Bmdimm to antagonize the activation of Bmdimm on fib-H transcription. Our findings promote a better understanding of fib-H transcriptional regulation and provide novel insights into the transcriptional repression of fib-H by FMBP-1 and basic helix-loop-helix factors Bmdimm during the molting of silkworm larvae. Our study also provides valuable information regarding the biological function of factors containing STPR domain.

Polydopamine-Assisted Silver Nanoparticle Self-Assembly on Sericin/Agar Film for Potential Wound Dressing Application.

Silver nanoparticles (AgNPs) are extensively applied for their broad-spectrum and excellent antibacterial ability in recent years. Polydopamine (PDA) has great advantages for synthesizing large amounts of AgNPs, as it has multiple sites for silver ion binding and phenolic hydroxyl structure to reduce silver ions to AgNPs. Here, we mixed sericin and agar solution and dried at 65 °C to prepare a sericin (SS)/Agar composite film, and then coated polydopamine (PDA) on the surface of SS/Agar film by soaking SS/Agar film into polydopamine solution, subsequently synthesizing high-density AgNPs with the assistance of PDA to yield antibacterial AgNPs-PDA- SS/Agar film. Scanning electron microscope (SEM), Fourier transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD) spectra indicated the successful synthesis of high-density AgNPs on the surface of PDA-SS/Agar film. PDA coating and AgNPs modification did not affect the structure of sericin and agar. Furthermore, water contact angle, water absorption and mechanical property analysis showed that AgNPs-PDA-SS/Agar film had excellent hydrophilicity and proper mechanical properties. Inhibition zone and growth curve assays suggested the prepared film had excellent and long-lasting antibacterial ability. In addition, it had excellent cytocompatibility on the fibroblast NIH/3T3 cells. The film shows great potential as a novel kind of wound dressing.

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.

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.