Mesocrystals with the symmetry defying morphologies and highly ordered superstructures composed of primary units are of particular interest, but the fabrication has proved extremely challenging. A novel strategy based on biomineralization approach for the synthesis of hematite mesocrystals is developed by using silk fibroin as a biotemplate. The resultant hematite mesocrystals are uniform, highly crystalline, and porous nanostructures with tunable size and morphologies by simply varying the concentration of the silk fibroin and iron(III) chloride in this biomineralization system. In particular, we demonstrate a complex mesoscale biomineralization process induced by the silk fibroin for the formation of hematite mesocrystals. This biomimetic strategy features precisely tunable, high efficiency, and low-cost and opens up an avenue to access new novel functional mesocrystals with hierarchical structures in various practical applications.
A strategy to prepare doxorubicin-loaded magnetic silk fibroin nanoparticles is presented. The nanoparticles serve as a nanometer-scale drug-delivery system in the chemotherapy of multidrug-resistant cancer under the guidance of a magnetic field. The magnetic tumor-targeting ability broadens the range of biomedical applications of silk fibroin, and the nanoparticle-assisted preparation strategy is useful for the advancement of other biomacromolecule-based materials.
Variability is a common feature of natural silk fibres, caused by a range of natural processing conditions. Better understanding of variability will not only be favourable for explaining the enviable mechanical properties of animal silks but will provide valuable information for the design of advanced artificial and biomimetic silk-like materials. In this work, we have investigated the origin of variability in forcibly reeled Antheraea pernyi silks from different individuals using dynamic mechanical thermal analysis (DMTA) combined with the effect of polar solvent penetration. Quasi-static tensile curves in different media have been tested to show the considerable variability of tensile properties between samples from different silkworms. The DMTA profiles (as a function of temperature or humidity) through the glass transition region of different silks as well as dynamic mechanical properties after high temperature and water annealing are analysed in detail to identify the origin of silk variability in terms of molecular structures and interactions, which indicate that different hydrogen bonded structures exist in the amorphous regions and they are notably different for silks from different individuals. Solubility parameter effects of solvents are quantitatively correlated with the different glass transitions values. Furthermore, the overall ordered fraction is shown to be a key parameter to quantify the variability in the different silk fibres, which is consistent with DMTA and FTIR observations.
Fourier transform infrared (FTIR) and scanning transmission X-ray microscopy (STXM) spectroscopic imaging techniques are introduced to determine the structure of protein-based polymer blends, using the silk fibroin/polyethylene oxide (SF/PEO) blend as a model material. We demonstrate that FTIR and STXM imaging techniques provide complementary chemical sensitivities, resolution ranges and sample thickness requirements that can enable a greater understanding of SF/PEO blend films. From the FTIR images, we find that SF shows random coil and/or helical conformation in the SF-rich domains, and ?-sheet conformation in the PEO-rich matrix. In the meantime, the SF content in SF-rich domains is 74 ± 4%, and 38 ± 6% in the PEO-rich matrix from the STXM images. These findings support and give further evidence to the conclusions of the previous studies on SF/PEO blends in the literature. Our results strongly suggest that FTIR and STXM imaging techniques are two promising complementary approaches for the study of phase behaviour and molecular conformation in protein-based polymer blend materials.
Amyloid fibrils and silk fibroin (SF) fibrils are proteinaceous aggregates occurring either naturally or as artificially reconstituted fibrous systems, in which the constituent ?-strands are aligned either orthogonally or parallel to the fibril main axis, conferring complementary physical properties. Here, it is shown how the combination of these two classes of protein fibrils with orthogonally oriented ?-strands results in composite materials with controllable physical properties at the molecular, mesoscopic, and continuum length scales.
Application of artificial ligament in anterior cruciate ligament reconstruction is one of the research focuses of sports medicine but the biological tendon-bone healing still remains a problem. The preliminary study of hydroxyapatite (HAP) coating on the polyethylene terephthalate (PET) surface could effectively induce the osteoblast differentiation, but the tendon-bone healing was still not stable. As a green synthesis process, the biomimetic mineralization can simulate the natural bone growth in vitro and in vivo.
Polycations as gene carriers have attracted considerable attention over the past decade. Generally, polyplexes between polycations and deoxyribonucleic acid (DNA) are formed at low N/P ratios (the ratios of the numbers of nitrogen atoms in a polycation to the numbers of phosphorus atoms in DNA), but high transfection efficiency can only be obtained at much higher N/P ratios. Thus, many polycationic chains are still free in solution. In this study, we investigated the detailed functions of free polyethylenimine chains (PEI-F) and free N,N,N-trimethyl chitosan chains (TMC-F) using the same polyplex, ie, TMC polyplex (TMC-P), which has high stability in Dulbecco's Modified Eagle's Medium (DMEM). Meanwhile, PEI polyplex (PEI-P)/PEI-F was also evaluated rather than PEI-P/TMC-F because the stability of PEI-P is low in DMEM and, in the latter case, the TMC-F may replace the bound PEI chain in PEI-P to form TMC-P. The transfection results show that both TMC-F and PEI-F can significantly increase the transfection efficiency of TMC-P; however, PEI-F can upregulate the gene expression of TMC-P more efficiently than TMC-F. Further investigations on the endocytosis and intracellular trafficking show that PEI-P/PEI-F, TMC-P/PEI-F, and TMC-P/TMC-F exhibit similar cellular uptake efficiency. However, by shutting down the clathrin-mediated endocytosis or vacuolar proton pump, the transfection efficiency decreases in the order of PEI-P/PEI-F > TMC-P/PEI-F > TMC-P/TMC-F. These findings indicate that PEI-F and TMC-F may promote the transfection efficiency of the polyplex by affecting its cellular uptake pathway and intracellular trafficking.
Natural polymer Bombyx mori silk fibroin is used as a biotemplate to produce silver nanoparticles in situ under light (both incandescent light and sunlight) at room temperature. Silk fibroin provides multiple functions in the whole reaction system, serving as the reducing agent of silver, and the dispersing and stabilizing agent of the resulted silver nanoparticles. As the reaction needs not any other chemicals and only uses light as power source, the synthetic route of silver nanoparticles reported here is rather environment-friendly and energy-saving. The silk fibroin-silver nanoparticle composite prepared by this method can be stably stored in a usual environment (room temperature, exposure to light, and so forth) for at least one month. Such a silk fibroin-silver nanoparticle composite shows an effective antibacterial activity against the methicillin-resistant Staphylcoccus aureus (S. aureus) and subsequently inhibits the biofilm formation caused by the same bacterium. Moreover, a maturely formed biofilm created by methicillin-resistant S. aureus can be destroyed by the silk fibroin-silver nanoparticle composite, which meets the demand of clinical application. Therefore, the silk fibroin-silver nanoparticle composite prepared by this clean and facile method is expected to be an effective and economical antimicrobial material in biomedical fields.
The interaction between silk and polar solvents of different molecular size can be an important tool for understanding the structural features of natural silk; in particular, the disordered regions associated with the key property of mechanical toughness. In this work, we investigate the transitions induced in the tensile performance and structure of as-reeled Antheraea pernyi silks from different silkworms by a range of solvents that can only soften the protein chains in the amorphous regions. The results indicate that polar solvents with different molecular sizes affect the silk to different degrees, and silks with slightly different structures display significantly different tensile performance in the same solvent. The solvent molecular size is quantitatively correlated with the accessible volume in the amorphous regions before and after the yield point, which suggests that the volume accessible to the solvent molecules decreases as the solvent radius increases. Moreover, silks with more ordered structure (less free volume) in the amorphous regions are less sensitive to solvents than those with more disordered structures. However, silks with higher free volume have higher toughness due to the greater strain to failure.
In this Communication, we report fabrication of ultrabright water-dispersible silicon nanoparticles (SiNPs) with quantum yields (QYs) up to 75% through a novelly designed chemical surface modification. A simple one-pot surface modification was developed that improves the photoluminescent QYs of SiNPs from 8% to 75% and meanwhile makes SiNPs water-dispersible. Time-correlated single photon counting and femtosecond time-resolved photoluminescence techniques demonstrate the emergence of a single and uncommonly highly emissive recombination channel across the entire NP ensemble induced by surface modification. The extended relatively long fluorescence lifetime (FLT), with a monoexponential decay, makes such surface-modified SiNPs suitable for applications involving lifetime measurements. Experimental results demonstrate that the surface-modified SiNPs can be utilized as an extraordinary nanothermometer through FLT imaging.
The structure-function relationship and mechanism of polycations as gene carriers have attracted considerable research interest in recent years. The present study was to investigate the relationship between polycation chain length and transfection efficiency (RCL-TE), and the corresponding mechanism by O-methyl-free N,N,N-trimethyl chitosans (TMCs) as gene carriers.
Three-dimensional (3D) copper oxide (CuO) nanostructures were synthesized in a regenerated Bombyx mori silk fibroin aqueous solution at room temperature. In the synthesis process, silk fibroin served as the template and helped to form the hierarchical CuO nanostructures by self-assembly. Cu(OH)2 nanowires were formed initially, and then they transformed into almond-like CuO nanostructures with branched edges and a compact middle. The size of the final CuO nanostructures can be tuned by varying the concentration of silk fibroin in the reaction system. A possible mechanism has been proposed based on various characterization techniques, such as scanning and transmission electron microscopy, X-ray diffraction, and thermogravimetric analysis. The synthesized CuO nanostructured material has been evaluated as an anode material for lithium ion batteries, and the result showed that they had a good electrochemical performance. The straightforward energy-saving method developed in this research may provide a useful preparation strategy for other functional inorganic materials through an environmentally friendly process.
Supramolecular polymers can be formed by self-assembly of designed subunits to yield highly ordered materials. In this paper, hierarchically structured materials, from molecules to nanofibers to macroscopical hydrogel, were fabricated by pH-induced assembly of C(12)-GAGAGAGY, a peptide amphiphile (PA) based on silk fibroin. Due to the different acid dissociation constants of the carboxyl and phenolic hydroxyl groups on tyrosine residue (Y), the PAs showed unique pH sensitive assembly and aggregation behaviors. It was found that not only the molecular-scale assemblies of these PAs gradually changed from cylindrical nanofibers to nanoribbons with the decreasing of pH value from 11 to 8 but also most of nanoribbons aggregated into parallel bundles in such a case. Further decrease of pH value resulted in a hierarchically structured robust and plastic hydrogel, of which the rheological moduli reached around 10(5) Pa. Moreover, noodle-like hydrogel fibers with bundles of nanoribbons aggregated parallel along the long axis in them could be steadily prepared under shear force. Taking the pH-sensitive reversible sol-gel transition, high modulus and plasticity into account, the hydrogel is believed to have significant potential applications in tissue engineering or as the biocompatible adhesives.
Synchrotron FTIR (S-FTIR) microspectroscopy was used to monitor both protein secondary structures (conformations) and their orientations in single cocoon silk fibers of the Chinese Tussah silk moth ( Antheraea pernyi ). In addition, to understand further the relationship between structure and properties of single silk fibers, we studied the changes of orientation and content of different secondary structures in single A. pernyi silk fibers when subjected to different strains. The results showed that the content and orientation of ?-sheet was almost unchanged for strains from 0 to 0.3. However, the orientation of ?-helix and random coil improved progressively with increasing strain, with a parallel decrease in ?-helix content and an increase in random coil. This clearly indicates that most of the deformation upon stretching of the single fiber is due to the change of orientation in the amorphous regions coupled with a conversion of some of the ?-helix to random coil. These observations provide an explanation for the supercontraction behavior of certain animal silks and are likely to facilitate understanding and optimization of postdrawing used in the conjunction with the wet-spinning of silk fibers from regenerated silk solutions. Thus, our work demonstrates the power of S-FTIR microspectroscopy for studying biopolymers.
In the present work, we report a new facile method to fabricate porous three-dimensional regenerated silk fibroin (RSF) scaffolds through n-butanol- and freezing-induced conformation transition and phase separation. The effects of RSF concentration, freezing temperature and n-butanol addition on the microstructure, the secondary structures of silk fibroin and apparent mechanical properties of the RSF scaffolds were investigated by SEM, (13)C CP-MAS NMR spectra and mechanical testing, respectively. By adjusting the RSF concentration and n-butanol addition, the pore size of the scaffold could be controlled in the range from of 10 ?m to 350 ?m with 84%-98% of porosity. The tensile strength of the wet scaffold reached the maximum of 755.2±33.6 kPa when the concentration of RSF solution was increased to 15% w/w. Moreover, post-treatment with ethanol further induced conformation transition of RSF from random coil or helix to ?-sheet. The porous scaffolds prepared by this facile and energy-saving method with good biocompatibility will have great potential for application in tissue engineering.
A well-defined amphiphilic polypeptide, poly(glutamic acid)22 -block-poly(alanine)8 (PGlu22 -b-PAla8 ), which plays the roles of both soluble (functional) additive and insoluble (structural) matrix, is employed to mediate the mineralization of CaCO3 at the air/water interface. X-ray diffraction (XRD) and Raman spectroscopy, for example, show that the polymorph of CaCO3 particles obtained is calcite. The observations from SEM and TEM suggest that PGlu22 -b-PAla8 initiates the amorphous precursor phase and heterogeneous nucleation of CaCO3 at the air/water interface, while temporarily stabilizes the gelatinous precursors as a process-directing agent; nevertheless, the initial concentration of Ca(2+) controls the procedure of crystallization and the final morphology of CaCO3 particles. Such "bifunctional" amphiphilic-polypeptide-regulated mineralization at the air/water interface may be applied to the synthesis of many kinds of symmetrical inorganic/organic hybrids.
An octapeptide, GAGAGAGY, was obtained by a novel method, i.e. hydrolysing Bombyx mori silk fibroin. Afterward, a dodecanoic acid-peptide conjugation was synthesized. This amphiphile assembled into cylindrical nanofibers of planar ?-sheets at pH 9 and twisted ?-sheets at pH 4.
Synchrotron FTIR (S-FTIR) microspectroscopy was used to monitor the silk protein conformation in a range of single natural silk fibers (domestic and wild silkworm and spider dragline silk). With the selection of suitable aperture size, we obtained high-resolution S-FTIR spectra capable of semiquantitative analysis of protein secondary structures. For the first time, we have determined from S-FTIR the ?-sheet content in a range of natural single silk fibers, 28 ± 4, 23 ± 2, and 17 ± 4% in Bombyx mori, Antheraea pernyi, and Nephila edulis silks, respectively. The trend of ?-sheet content in different silk fibers from the current study accords quite well with published data determined by XRD, Raman, and (13)C NMR. Our results indicate that the S-FTIR microspectroscopy method has considerable potential for the study of single natural silk fibers.
Silk fibroin is a very promising biomedical material because of its renewability, nontoxicity, biocompatibility, and biodegradability. On the basis of a simple and mild method for the preparation of silk fibroin nanospheres with controllable size, the authors developed earlier, anti-cancer drug paclitaxel (PTX)-loaded silk fibroin nanospheres ranging from 270 to 520 nm were produced accordingly. The drug loading, encapsulation efficiency, and released property of PTX-loaded silk fibroin nanospheres are depended on the silk fibroin concentration and initial PTX-loading capacity. The maximum drug loading is about 6.9% and the release time of such a kind of nanospheres is over 9 days. The release time of PTX-loaded silk fibroin nanospheres can be as long as 2 weeks when the drug loading is about 3.0%. All these results imply that such a kind of biomacromolecule-based anti-cancer drug nanocarrier has a great potential for chemotherapy in clinical applications.
The crystallization of calcium carbonate (CaCO3) was investigated using a mineralization system composed of a chitosan membrane and regenerated silk fibroin (RSF). Such a system may resemble the mineralization in molluscs, where chitosan is a derivative of chitin and RSF an analogue of nacreous protein. It was found that the vaterite disks generally formed on the chitosan membrane while the aragonite disks also appeared with changes of pH value or temperature of the solution. The crystallization of CaCO3 in the vicinity of the chitosan membrane was much more affected by the environment of crystallization, compared to that in bulk solution. Detailed observation from high-resolution scanning and transmission electron microscopy (HRSEM and TEM) showed that these disks consisted of nanoparticles about 20 nm in size, thus suggesting that the accumulation of hybrid CaCO3/RSF nanoparticles induced the formation of crystalline disks on the chitosan membrane.
The treatment of osteomyelitis remains a challenge for orthopaedic surgeons. Controlled release of vancomycin from biodegradable microspheres is a promising method for eliminating infection. However, the large initial burst release may make it difficult to maintain the local vancomycin concentration superior to minimum inhibitory concentration for several weeks. The aims of this study were to explore applications of the silk fibroin (SF) as an aqueous coating material for vancomycin-loaded poly(?-caprolactone) (PCL) microspheres, and investigate the effects of silk coating on in vitro drug release. Examinations of particle size analyses, vancomycin content, Fourier transform infrared spectroscopy, differential scanning calorimetry, scanning electron microscopy and in vitro drug release were performed. The results showed that silk coating could reduce the large initial burst release and retard the vancomycin release. Therefore, we suggest that the SF could be used as an aqueous coating material for vancomycin-loaded PCL microspheres and prolonged the drug release. SF coating on vancomycin-loaded PCL microspheres may be considered as an effective approach to prolong the drug release and improve the anti-infection effects.
In this work, we developed a simple and flexible method to manufacture a 3D porous scaffold based on the blend of regenerated silk fibroin (RSF) and chitosan (CS). No crosslinker or other toxic reagents were used in this method. The pores of resulted 3D scaffolds were connected with each other, and their sizes could be easily controlled by the concentration of the mixed solution. Compared with pure RSF scaffolds, the water absorptivities of these RSF/CS blend scaffolds with significantly enhanced mechanical properties were greatly increased. The results of MTT and RT-PCR tests indicated that the chondrocytes grew very well in these blend RSF/CS porous scaffolds. This suggested that the RSF/CS blend scaffold prepared by this new method could be a promising candidate for applications in tissue engineering.
A natural electroactive protein hydrogel was prepared from soy protein isolate (SPI) solution by cross-linking with epichlorohydrin. Under electrical stimulus, such SPI hydrogel quickly bends toward one electrode, showing a good electroactivity. Because of its amphoteric nature, the SPI hydrogel bends either toward the anode (pH < 6) or cathode (pH > 6), depending on the pH of the electrolyte solution. Other factors, such as electric field strength, ionic strength and gel thickness also influence the electromechanical behavior of the SPI hydrogels. Moreover, this SPI hydrogel exhibits a good electroactive behavior under strong acidic (pH = 2 - 3) or basic (pH = 11 - 12) solutions, which is a significant improvement over two other kinds of natural electroactive hydrogels, i.e., chitosan/carboxymethylcellulose and chitosan/carboxymethylchitosan hydrogel, which we reported previously. The wide pH range and good electroactivity of this natural protein hydrogel suggests its great potential for microsensor and actuator applications, especially in the biomedical field, and also to increase the scope of natural polymer-based electroactive hydrogels.
Films of regenerated silk fibroin (RSF) are usually brittle and weak, which prevents its wide application as a structural material. To improve the mechanical properties of RSF film, uniaxial extension under swollen conditions was employed to introduce preferred orientation of molecular chains of silk fibroin. Such a prestretching treatment resulted in the strain at break, ultimate stress, Youngs modulus, and energy to break along the predrawn direction of the RSF film increasing from approximate 5%, 90 MPa, 2.7 GPa, and 2.1 kJ/kg to 35%, 169 MPa, 3.5 GPa, and 38.9 kJ/kg, respectively, which is an attractive combination of strength and toughness. The mechanism of these property enhancements was investigated using techniques such as small-angle X-ray scattering, wide-angle X-ray diffraction, atomic force microscopy, and dynamic mechanical analysis.
Polysaccharides were believed to play an important role in the mineralization process of many organisms. As the source of continuously and uniformly releasing alginate molecules and Ca(2+), alginate/Ca nanospherical gel was employed in the solution to induce the nucleation and growth of CaCO(3). Time-resolved transmission electron microscopy (TEM) was applied to study the crystallization at a very early stage. It was found that the initially formed lens-like vaterite particles gradually dissolved from the middle of the particle and released alginate molecules and Ca(2+) back into the system. As reaction time increased, the released substances were involved in the next stage of crystallization of CaCO(3), in the form of needle-like and shuttle-like aragonite particles sequentially depending on the concentration of alginate molecules and Ca(2+). "Egg-box" conformation of alginate and Ca(2+) was considered a skeleton for the growth of such aragonite particles. Notably, shuttle-like aragonite particles were composed of "bricks" of several hundred nanometers in size, which were very similar to biogenetic nacreous layers in shells.
Chitosan (CS) can mediate the formation of spherical, tabulate, and unique starfruit-like silica in the presence of phosphate ions (Pi). CryoTEM and cryoET were used to examine the CS aggregates in the hydrated state. 3D starfruit-like CS/Pi aggregates were reconstructed, which unambiguously confirmed the templating effect of CS/Pi in biomimetic silicification.
This study showed that Bombyx mori silk protein could be selectively induced to fold into fibrils dominated by either cross- or parallel-beta-sheet structure, where the beta-strands arrange perpendicular or parallel to the long fibril axis, incubated in ethanol-water quiescently or in water under shear.
The ability to control the processing of artificial silk is key to the successful application of this important and high performance biopolymer. Understanding where our current reconstitution process can be improved will not only aid us in the creation of better materials, but will also provide insight into the natural material along the way. This study aims to understand what proportion of reconstituted silk contributes to its rheological properties and what conformational state the silk proteins are in. It shows, for the first time, that a change in rheological properties can be related to a change in silk structures present in solution and reveals a low concentration gel state for silk that may have important implications for future successful artificial processing of silk.
This feature article reviews recent progress in the understanding of the hierarchically organized structures, the perfectly balanced mechanical properties and the structure-property relationship of the natural animal silk fibres, as well as the experimental attempts to fabricate man-made silk fibres by means of wet spinning, dry spinning, electrospinning and transgenosis.
Common cotton textiles are hydrophilic and oleophilic in nature. Superhydrophobic cotton textiles have the potential to be used as self-cleaning fabrics, but they typically are not super oil-repellent. Poor oil repellency may easily compromise the self-cleaning property of these fabrics. Here, we report on the preparation of superoleophobic cotton textiles based on a multilength-scale structure, as demonstrated by a high hexadecane contact angle (153 degrees for 5 microL droplets) and low roll-off angle (9 degrees for 20 microL droplets). The multilength-scale roughness was based on the woven structure, with additional two layers of silica particles (microparticles and nanoparticles, respectively) covalently bonded to the fiber. Superoleophobicity was successfully obtained by incorporating perfluoroalkyl groups onto the surface of the modified cotton. It proved to be essential to add the nanoparticle layer in achieving superoleophobicity, especially in terms of low roll-off angles for hexadecane.
Silk fibroin (SF) has played a curial role for the surface modification of conventional materials to improve the biocompatibility, and SF modified poly(ethylene terephthalate) (PET) materials have potential applications on tissue engineering such as artificial ligament, artificial vessel, artificial heart valve sewing cuffs dacron and surgical mesh engineering. In this work, SF was immobilized onto PET film via two different methods: 1) plasma pretreatment followed by SF dip coating (PET-SF) and 2) plasma-induce acrylic acid graft polymerization and subsequent covalent immobilization of SF on PET film (PET-PAA-SF). It could be found that plasma treatment provided higher surface roughness which was suitable for further SF dip coating, while grafted poly(acrylic acid) (PAA) promised the covalent bonding between SF and PAA. ATR-FTIR adsorption band at 3284 cm(-1), 1623 cm(-1) and 1520 cm(-1) suggested the successful introduction of SF onto PET surface, while the amount of immobilized SF of PET-SF was higher than PET-PAA-SF according to XPS investigation (0.29 vs 0.23 for N/C ratio). Surface modified PET film was used as substrate for mesenchymal stem cells (MSCs) culture, the cells on PET-SF surface exhibited optimum density compared to PET-PAA-SF according to CCK-8 assays, which indicated that plasma pretreatment followed by SF dip coating was a simple and effective way to prepare biocompatible PET surface.
A novel hydroxyapatite/regenerated silk fibroin scaffold was prepared and investigated for its potential to enhance both osteoinductivity and osteoconductivity of bone marrow-derived mesenchymal stromal cells in vitro. Approx. 12.4 ± 0.06 % (w/w) hydroxyapatite was deposited onto the scaffold, and cell viability and DNA content were significantly increased (18.5 ± 0.6 and 33 ± 1.2 %, respectively) compared with the hydroxyapatite scaffold after 14 days. Furthermore, alkaline phosphatase activity in the novel scaffold increased 41 ± 2.5 % after 14 days compared with the hydroxyapatite scaffold. The data indicate that this novel hydroxyapatite/regenerated silk fibroin scaffold has a positive effect on osteoinductivity and osteoconductivity, and may be useful for bone tissue engineering.
Silk fibroin (SF) shows promise for tissue engineering and other biomedical applications due to its excellent biocompatibility, unique biomechanical properties, and controllable biodegradability. The particulate form of SF materials may have many potential uses, including the use as a filler for tissue defects or as a controlled-release agent for drug delivery. However, many past in vivo and in vitro studies evaluating the biocompatibility and biodegradability of SF have involved bulk implants. It is essential to evaluate the inflammatory effects of SF particles before further use. In this study, two different sizes of SF particles were evaluated to assess their impact on the release of tumor necrosis factor (TNF)-?, interleukin (IL)-1?, and IL-6, in comparison with lipopolysaccharide positive control stimulation. The inflammatory processes were characterized using real-time reverse transcription polymerase chain reaction, enzyme-linked immunosorbent assay, and light microscopy evaluations. The results indicated that small silk fibroin particles and large silk fibroin particles, in culture with RAW 264.7 murine macrophage cells for 24 h, caused up-regulation of mRNA coding for TNF-?, which indicated that both size of particles have potential inflammatory effects. There was a statistically significant increase in this up-regulation under small silk fibroin stimulation. However, the immunosorbent assay suggested that there was virtually no observed release of IL-1?, IL-6, or TNF-?, relative to the control group. The results suggest that SF particles of the chosen dimensions may have good biocompatibility in culture with RAW 264.7 murine macrophages.
A chitosan-based membrane chromatography was set up by using natural chitosan/carboxymethylchitosan (CS/CMCS) blend membrane as the matrix. The dynamic adsorption property for protein (lysozyme as model protein) was detailed discussed with the change in pore size of the membrane, the flow rate and the initial concentration of the feed solution, and the layer of membrane in membrane stack. The best dynamic adsorption capacity of lysozyme on the CS/CMCS membrane chromatography was found to be 15.3mg/mL under the optimal flow conditions. Moreover, the CS/CMCS membrane chromatography exhibited good repeatability and reusability with the desorption efficiency of ~90%. As an application, lysozyme and ovalbumin were successfully separated from their binary mixture through the CS/CMCS membrane chromatography. This implies that such a natural chitosan-based membrane chromatography may have great potential on the bioseparation field in the future.
The orthopedic infection, such as osteomyelitis, especially those caused by Methicillin-resistant Staphylococcus aureus (MRSA), remains a major complication of open fractures. Local vancomycin delivery is considered to provide better methods when avascular zones prevent the delivery of drugs from conventional routes of administration. Chitosan (CS) delivery system has been developed with the disadvantages, such as mechanically weakness, lacking osteoconductivity, and the initial burst of antibiotics into the environment. The aim of this study was to confirm that the prepared CS/?-tricalcium phosphate (?-TCP) composites coated with poly (?-caprolactone) (PCL), similar to natural bone in components, had a three-dimensional porous structure and could be used as drug carriers to deliver vancomycin in a sustained and controlled manner effectively for 6 weeks at levels to inhibit MRSA proliferation. We prepared porous CS/?-TCP composites by incorporating ?-TCP into the system, and coated the composites with PCL of three different concentrations. The morphological structure of composites, including pore size and porosity, was examined. The result showed that CS/?-TCP coated with 2.5w/v% PCL solution had the best coating effect and it retarded the release of vancomycin in a near zero-order mechanism from 0 to 14 days. The drug delivery was significantly delayed after coated with 2.5w/v% PCL. The quantitative release of vancomycin was extended to 42 days. Therefore PCL coating could be used to retard the release of vancomycin from CS/?-TCP composites in a sustained and controlled manner. Porous CS/?-TCP coated with PCL might be one of the candidate vancomycin carriers for treating MRSA-related osteomyelitis.
The conformation transition from random coil and/or helix to ?-sheet of silk protein is the most important step in the formation of silk fiber in nature as well as by artificial spinning. Time-dependent Fourier transform infrared (FT-IR) spectroscopy was used in this research to monitor such a conformation transition process induced by the organic solvents methanol, ethanol, propanol, isopropanol, and acetone. The kinetics of ?-sheet formation of regenerated Bombyx mori silk fibroin in these organic solvents was obtained by the ?absorbance-time curve from the time-dependent difference infrared spectra. The results showed that the conformation transition rate of silk fibroin was methanol > ethanol > acetone > propanol > isopropanol, which is in accordance with the polarity of these organic solvents. In connection with the mechanical properties and morphologies of regenerated silk fibers using these organic solvents as coagulation bath reported in the literature, we may conclude that the conformation transition rate of silk protein in the organic solvent is very important in wet-spinning to produce high-performance regenerated silk fibers.
The conformation and eventual morphology of silk fibroin (SF) chains are crucial for the mechanical properties of SF materials, and are strongly related to the solvation step as a key stage in their processing conditions. In this work, a novel SF/AmimCl (1-allyl-3-methylimidazolium chloride) solution with unique properties is reported and compared with conventional regenerated SF aqueous solutions, based on an investigation of its rheological properties. The steady shearing behavior suggested that AmimCl is a good solvent for SF molecules, and shear thinning of semidiluted SF/AmimCl solution at high shear rates showed behavior similar to that in native spinning, which is due to the rearrangement and orientation of SF molecular chains. Fitting of experimental dynamic viscoelastic data to the Rouse model provided an effective method to estimate the molecular weight of SF. We believe that this work not only provides a better understanding of the relationship between properties of silk protein and aggregation states of their molecular chains, but also provides tools to fabricate high-performance SF-based materials.
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