Aldehyde-functionalized cellulose as reactive sorbents for the capture and retention of polyamine odor molecules associated with chronic wounds.

Aldehyde-functionalized cellulose (AFC) was prepared by oxidizing cellulose with sodium metaperiodate. The reaction was characterized by Schiff's test, FT-IR, and UV-vis study. AFC was evaluated as a reactive sorbent for controlling polyamine-based odor from chronic wounds, and its performance was compared with charcoal, one of the most widely utilized odor-control sorbents through physisorption. Cadaverine was used as the model odor molecule. A liquid chromatography/mass spectrometry (LC/MS) method was established to quantify the compound. AFC was found to rapidly react with cadaverine through the Schiff-base reaction, which was confirmed by FT-IR, visual observation, CHN elemental analysis, and the ninhydrin test. The sorption and desorption behaviors of cadaverine onto AFC were quantified. With clinic-relevant cadaverine concentrations, AFC demonstrated much better sorption performance than charcoal. At even higher cadaverine concentrations charcoal showed higher sorption capacity, probably due to its high surface area. On the other hand, in desorption studies, AFC retained much more of the sorbed cadaverine than charcoal. When AFC and charcoal were combined, the pair demonstrated excellent sorption and desorption behaviors. The XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) assay confirmed that AFC has very good in vitro biocompatibility. These results suggest that AFC-based reactive sorption can be a new strategy to control odors associated with chronic wounds for improved healthcare.

Cyanuric Chloride-Based Reactive Dyes for Use in the Antimicrobial Treatments of Polymeric Materials.

This study reports a simple and practical method to introduce antimicrobial and biofilm-controlling functions into hydroxyl- or amino-containing polymers such as cellulose using compounds derived from widely used reactive dyes. Two dichloro-s-triazine-based dyes, reactive blue 4 and sodium 4-(4,6-dichloro-1,3,5-triazinylamino)-benzenesulfonate (a colorless reactive "dye"), were covalently attached to cellulose at room temperature by replacing one chloride on the dyes with the hydroxyl groups on cellulose followed by hydrolysis under alkaline conditions to transform the remaining chloride into hydroxyl groups. The chemical reactions were confirmed by FT-IR studies, energy-dispersive X-ray spectroscopy, water contact angle measurement, and zeta potential analysis. The resulting cellulose provided powerful antimicrobial activities against Staphylococcus epidermidis (S. epidermidis, ATCC 35984, Gram-positive bacteria), Escherichia coli (E. coli, ATCC 15597, Gram-negative bacteria), and Candida albicans (C. albicans, ATCC 10231, yeast) and effectively prevented the formation of bacterial or fungal biofilms. The minimum inhibition concentrations of the hydrolyzed dyes were similar to that of phenol. In the zone of inhibition studies using phenolic compounds as positive controls, the hydrolyzed dyes and their model compound cyanuric acid demonstrated antimicrobial functions, suggesting that the antimicrobial activities were associated with the phenol-like hydroxyl groups on the triazine rings. Antimicrobial mechanism investigation indicated that the phenol-like structures on the dyed cellulose caused microbial lysis and leakage of intracellular components. The antimicrobial functions were durable upon repeated washing, and the dyed cellulose showed outstanding biocompatibility toward mammalian cells.

Enhanced antimicrobial and antifungal property of two-dimensional fibrous material assembled by N-halamine polymeric electrolytes.

Microbial contamination and biofilm formation cause serious issues in medical, household, industrial and environmental applications. In this study, a series of cationic and anionic N-halamine polymeric precursors, poly (N,N-dimethyl-N-decyl ammonium ethyl methacrylate-co-methacrylamide) (PQDM) and poly (acrylic acid-co-methacrylamide) (PAM), were synthesized and coated onto cotton fabrics through the layer-by-layer (LBL) assembly technique. The coated LBL cotton swatches were characterized by Scanning Electron Microscopy, Fourier transform infrared spectroscopy, and contact angle evaluation. The stability of the LBL samples towards artificial sweat and home laundering was evaluated. The LBL treated fabrics demonstrated effective antimicrobial efficacy and biofilm-controlling against Gram-positive bacteria, Gram-negative bacteria, and Fungi. In vitro cytocompatibility test towards mouse fibroblast cell indicated that the LBL coated cotton fabrics are cytocompatible, pointing to great potentialities of the LBL assembled fabrics for future biomedical applications.

A novel comprehensive efficacy test for textiles intended for use in the healthcare setting.

Soft surfaces, including textiles are found throughout healthcare settings. Pathogens can survive for long periods of time on textiles, and can be transferred to and from the skin. Antimicrobial fabrics are used as an engineering control to prevent infection. Efficacy testing standards have limitations, including single microorganism challenges, multiple fabric plies tested, and lengthy contact times. We developed a novel method that better models in-use conditions through testing standardized mixtures of pathogens and normal skin microorganisms, artificial soils, and a 15-min contact time. Reproducible growth of all microorganisms from frozen stocks was achieved using this method. A novel rechargeable, monitorable N-halamine cotton cellulose fabric, containing 5885 ± 98 ppm of active chlorine, was evaluated with the new method using PBS, artificial sweat, and artificial sweat plus 5% serum as soil. Pathogens tested included Acinetobacter baumannii, Candida albicans, Escherichia coli, vancomycin-resistant Enterococcus faecalis, methicillin-resistant Staphylococcus aureus, methicillin-susceptible Staphylococcus aureus, and Pseudomonas aeruginosa. Each was tested singly and in the presence of a representative normal skin flora mixture, including: Acinetobacter lwoffii, Corynebacterium striatum, Micrococcus luteus, and Staphylococcus epidermidis. When tested singly, all microorganisms were reduced by 3.00 log10 or greater, regardless of artificial soil. In mixture, 4.00 log10 or greater reductions were achieved for all microorganisms. These results suggest that the novel testing method can be used to provide more comprehensive and realistic efficacy information for antimicrobial textiles intended for use in healthcare. Furthermore, the N-halamine fabric demonstrated efficacy against multiple pathogens, singly and in mixtures, regardless of the presence of artificial soils.

Wheat germ agglutinin liposomes with surface grafted cyclodextrins as bioadhesive dual-drug delivery nanocarriers to treat oral cells.

Topical management of oral infection requires combined use of multiple classes of drugs and frequent dosing due to low drug retention rates. The sustained, co-delivery of drugs with different solubilities to cells using nanoparticle drug delivery systems remains a challenge. Here, we developed wheat germ agglutinin (WGA) conjugated liposomes with surface grafted cyclodextrin (WGA-liposome-CD) as bioadhesive dual-drug nanocarriers. We effectively encapsulated two physiochemically different drugs (ciprofloxacin and betamethasone) and demonstrated sustained co-drug release in saliva over a 24 h period in vitro. As proof of therapeutic utility in oral cells, we infected oral keratinocytes with Aggregatibacter actinomycetemcomitans, a bacterial pathogen responsible for chronic periodontal disease. Drug release, resulting from nanocarrier cell binding, produced a significant increase in oral cell survival and synergistically reduced inflammation. These results suggest that WGA-liposome-CD nanocarriers are novel cyto-adhesive candidates for delivering multiple drugs with sustained therapeutic activity for localized drug delivery to oral cells.

Reactions of phenolic compounds with monomeric N-halamines and mesoporous material-supported N-halamines.

The reactions of a monomeric N-halamine, 1-chloro-5,5-dimethylhydantoin (MCDMH), and a mesoporous material-supported N-halamine (MMSNs) with phenol and p-cresol (two common contaminants in water) were investigated. MCDMH reacted rapidly with the phenolic compounds, and pH values had little effects on the reactions. On the contrary, MMSNs reacted with phenol and p-cresol only when the pH values were higher than 10. Phenol showed a lower reaction rate than p-cresol toward MMSNs. GCMS analysis suggested that MMSNs might react with the phenolic compounds through step-wise electrophilic chlorination reactions, and the main product was 2,4,6-trichlorophenol. The reaction kinetics were studied by following the disappearance of phenolic UV absorption bands, and the kinetic parameters were determined.

Demonstration of biofilm removal from type 304 stainless steel using pulsed-waveform electropolishing.

This article describes an electrochemical method to remove bacterial biofilm from a stainless steel (SS) surface using a potential pulse/reverse pulse technique. This technique employs a periodic waveform that consists of anodic and cathodic pulses. The pulses can effectively strip a thin layer of metal off the SS surface, along with the adherent biofilm, in a saline solution. Not only can the pulses effectively remove biofilm from the SS surface, but they also regenerate the original mirror-like shiny surface. The importance of this electrochemical biofilm removal method is its wide applicability for any types of biofilms. That is, instead of directly removing the biofilm, it removes a very thin layer of the metal under the biofilm. Thus, the removal process is independent to the nature of the biofilms. Furthermore, this electrochemical biofilm removal method is rapid (less than 30 s of potential pulse time) and does not require hazardous chemicals.

Silk Fibroin Acts as a Self-Emulsifier to Prepare Hierarchically Porous Silk Fibroin Scaffolds through Emulsion-Ice Dual Templates.

Silk fibroin (SF) has shown enormous potentials in various fields; however, application of SF in emulsion technology is quite limited. Here, we use SF as a self-emulsifier to form an oil-in-water (O/W) emulsion by emulsifying 1-butanol in SF aqueous solution. This showed that SF possessed strong surface activity to stabilize the O/W emulsion without the need for any other surface-active agent until its solidification because of 1-butanol-induced conformational transition of SF to ß-sheet. After freezing the preformed emulsions at -20 °C, robust three-dimensional porous SF scaffolds were prepared without the need for any further post-treatment. The evolution from the O/W emulsion to porous scaffold formation under freezing was tracked, and an emulsion-ice dual template mechanism was proposed for scaffold formation, based on which SF scaffolds with controllable hierarchically porous structures were achieved by tuning the dispersed droplet volume fraction. Furthermore, SF scaffolds with hierarchical porosity showed significantly higher bioactivity toward L929 fibroblasts than that of SF scaffolds with mono macroporosity, highlighting the great asset of this hierarchically porous SF scaffold for broad applications in tissue engineering. Therefore, the strong surface-active characteristic of SF presented here, in addition to its distinct advantages, sheds a bright light on the application of SF in the vast range of emulsion technologies, especially in cosmetic-, food-, and biomedical-related areas.

Understanding the Mechanical Properties and Structure Transition of Antheraea pernyi Silk Fiber Induced by Its Contraction.

Like most major ampullate silks of spider, the length of Antheraea pernyi silkworm silk can shrink to a certain degree when the fiber is in contact with water. However, what happens in terms of molecule chain level and how it correlates to the mechanical properties of the silk during its contraction is not yet fully understood. Here, we investigate the water-induced mechanical property changes as well as the structure transition of two kinds of A. pernyi silk fiber, which are forcibly reeled from two different individuals (silkworm a and silkworm b; the silk fiber from either one represents the lower and upper limit of the distribution of mechanical properties, respectively). The tensile test results present that most of the mechanical parameters except the post-yield modulus and breaking strain for both silk fibers have the same variation trend before and after their water contraction. Synchrotron FTIR and Raman spectra show that the native filament from silkworm a contains more α-helix structures than that in silkworm b filament, and these α-helices are partially converted to ß-sheet structures after the contraction of the fibers, while the order of both ß-sheet and α-helix slightly increase. On the other side, the content and orientation of both secondary structural components in silkworm b fiber keep unchanged, no matter if it is native or contracted. 13C CP/MAS NMR results further indicate that the α-helix/random coil to ß-sheet conformational transition that occurred in the silk of silkworm a corresponds the Ala residues. Based upon these results, the detailed structure transition models of both as-reeled A. pernyi silk fibers during water contraction are proposed finally to interpret their properties transformation.

Salivary polypeptide/hyaluronic acid multilayer coatings act as "fungal repellents" and prevent biofilm formation on biomaterials.

Candida-associated denture stomatitis (CADS) is a common, recurring clinical complication in denture wearers that can lead to serious oral and systemic health problems. Current management strategies are not satisfactory due to their short-acting and ineffective therapeutic effects. Here, we describe a new fungal biofilm controlling strategy using the polyelectrolyte layer-by-layer (LBL) self-assembly technology on denture materials. Conventional poly(methyl methacrylate) (PMMA) denture material discs were functionalized with negatively charged poly(methacrylic acid) (PMAA) via plasma-initiated surface grafting, followed by repetitive alternating coating with the salivary antimicrobial polypeptide histatin 5 (H-5; cationic polymer) and hyaluronic acid (HA; anionic polymer). On the other hand, the H-5/HA LBL coatings (i.e., the outermost layer was H-5) inhibited fungal attachment/adhesion, significantly reduced fungal biofilm formation, and showed synergistic effects with the antifungal drug miconazole. LBL surface hydrophilicity was not the key mechanism in controlling Candida biofilm formation. The current approach demonstrates the utility of a new design principle for fabricating anticandidal denture materials, as well as potentially other related medical devices, for controlling fungal biofilm formation and combating insidious infections.

Porous Three-Dimensional Silk Fibroin Scaffolds for Tracheal Epithelial Regeneration in Vitro and in Vivo.

The regeneration of functional epithelial lining is critical for artificial grafts to repair tracheal defects. Although silk fibroin (SF) scaffolds have been widely studied for biomedical application (e.g., artificial skin), its potential for tracheal substitute and epithelial regeneration is still unknown. In this study, we fabricated porous three-dimensional (3D) silk fibroin scaffolds and cocultured them with primary human tracheobronchial epithelial cells (HBECs) for 21 days in vitro. Examined by scanning electronic microscopy (SEM) and calcein-AM staining with inverted phase contrast microscopy, the SF scaffolds showed excellent properties of promoting cell growth and proliferation for at least 21 days with good viability. In vivo, the porous 3D SF scaffolds (n = 18) were applied to repair a rabbit anterior tracheal defect. In the control group (n = 18), rabbit autologous pedicled trachea wall without epithelium, an ideal tracheal substitute, was implanted in situ. Observing by endoscopy and computed tomography (CT) scan, the repaired airway segment showed no wall collapse, granuloma formation, or stenosis during an 8-week interval in both groups. SEM and histological examination confirmed the airway epithelial growth on the surface of porous SF scaffolds. Both the epithelium repair speed and the epithelial cell differentiation degree in the SF scaffold group were comparable to those in the control group. Neither severe inflammation nor excessive fibrosis occurred in both groups. In summary, the porous 3D SF scaffold is a promising biomaterial for tracheal repair by successfully supporting tracheal wall contour and promoting tracheal epithelial regeneration.

Lectin-Conjugated Liposomes as Biocompatible, Bioadhesive Drug Carriers for the Management of Oral Ulcerative Lesions.

Oral ulcerative lesions are painful and debilitating, particularly for immunosuppressed patients undergoing chemotherapeutic/irradiation treatment. Their clinical management requires multiple drugs to be administered simultaneously. Current formulations available to patients require frequent dosing, leading to poor compliance and suboptimal clinical outcomes. In this study, we prepared wheat germ agglutinin (WGA)-conjugated liposomes (WGA liposomes) to serve as bioadhesive drug carriers that can encapsulate various classes of drugs, rapidly bind to oral epithelial cells within minutes, and stay on the cells to provide sustained, localized drug release for days. Fluorescence binding studies found a significant increase (p < 0.05) in the binding of WGA liposomes to oral cells in as short an incubation time as 1 min compared to that for nonconjugated liposomes. WGA liposomes encapsulating model drug amoxicillin showed sustained in vitro drug release, and the released drugs provided potent antimicrobial activity against Streptococcus mutans in an oral epithelial-bacterial coculture system. Exocytosis studies confirmed that the WGA liposomes stayed within the oral cells for 48 h, after which the cells completely removed the liposomes. Moreover, cell viability studies showed that there was a significant reduction in oral cell damage when the bacterially infected cells were treated with amoxicillin-loaded WGA liposomes compared to the untreated controls. These results point to the great potential of the lectin-conjugated liposomes as cell-binding drug-delivery systems in achieving localized, sustained drug release for the management of oral ulcerative lesions and other related complications.

A simple semi-quantitative approach studying the in vivo degradation of regenerated silk fibroin scaffolds with different pore sizes.

The biocompatibility and in vivo degradation rate of biomaterials represent critical control points in the long-term success of scaffolds for tissue restoration. In this study, new three-dimensional (3D) regenerated silk fibroin scaffolds (RSFs) were prepared by the freezing-defrosting procedure, and then were implanted beneath the dorsal skin of rats. This study aims to develop a kinetic semi-quantitative approach to assess in vivo degradation rate and biocompatibility of this kind of RSFs with different pore sizes for the first time, and to evaluate the relationship between the biodegradation and tissue responses by measuring the thickness of residual scaffolds, fibrous capsules and infiltrated tissues through integrated techniques of histology, optical imaging and image analysis. Our results showed that scaffolds with both pore sizes (74.35±10.84µm and 139.23±44.93µm, respectively) were well tolerated by host animals and pore size was found to be the rate limiting factor to the biodegradation in the subcutaneous implantation model. In addition, the biodegradation of RSFs was inflammation-mediated to a certain degree and fibroblasts may play a critical role in this process. Overall, such semi-quantitative approach was demonstrated to be a simple and effective method to assess the in vivo degradation rate, and the prepared RSFs were presented to have promising potential in tissue engineering applications.

Controlling bacterial fouling with polyurethane/N-halamine semi-interpenetrating polymer networks.

N -halamine-based interpenetrating polymer networks were developed as a simple and effective strategy in the preparation of antimicrobial polymers. An N-halamine monomer, N-chloro-2, 2, 6, 6-tetramethyl-4-piperidyl methacrylate, was incorporated into polyurethane in the presence of a cross-linker and an initiator. Post-polymerization of the monomers led to the formation of polyurethane/N-halamine semi-interpenetrating polymer networks. The presence of N-halamines in the semi-interpenetrating polymer networks was confirmed by attenuated total reflectance infrared, water contact angle, and energy-dispersive X-ray spectroscopy analysis. The N-halamine contents in the semi-interpenetrating polymer networks could be readily controlled by changing reaction conditions. The distribution of active chlorines within the semi-interpenetrating polymer networks was characterized with energy-dispersive X-ray spectroscopy. Contact mode antimicrobial tests, zone of inhibition studies, and scanning electron microscopy observations showed that the semi-interpenetrating polymer networks had potent antimicrobial and antifouling effects against both Gram-positive and Gram-negative bacteria. Release tests demonstrated the outstanding stability of the N-halamine structures in the new semi-interpenetrating polymer networks.

Controlling fungal biofilms with functional drug delivery denture biomaterials.

Candida-associated denture stomatitis (CADS), caused by colonization and biofilm-formation of Candida species on denture surfaces, is a significant clinical concern. We show here that modification of conventional denture materials with functional groups can significantly increase drug binding capacity and control drug release rate of the resulting denture materials for potentially managing CADS. In our approach, poly(methyl methacrylate) (PMMA)-based denture resins were surface grafted with three kinds of polymers, poly(1-vinyl-2-pyrrolidinone) (PNVP), poly(methacrylic acid) (PMAA), and poly(2-hydroxyethyl methacrylate) (PHEMA), through plasma-initiated grafting polymerization. With a grafting yield as low as 2 wt%, the three classes of new functionalized denture materials showed significantly higher drug binding capacities toward miconazole, a widely used antifungal drug, than the original PMMA denture resin control, leading to sustained drug release and potent biofilm-controlling effects against Candida. Among the three classes of functionalized denture materials, PNVP-grafted resin provided the highest miconazole binding capability and the most powerful antifungal and biofilm-controlling activities. Drug binding mechanisms were studied. These results demonstrated the importance of specific interactions between drug molecules and functional groups on biomaterials, shedding lights on future design of CADS-managing denture materials and other related devices for controlled drug delivery.

Functionalized Denture Resins as Drug Delivery Biomaterials to Control Fungal Biofilms.

Colonization and biofilm-formation of Candida species on denture surfaces cause Candida-associated denture stomatitis (CADS), a recurring fungal infection that affects up to 67% of denture wearers. We grafted poly(2-hydroxyethyl methacrylate) (PHEMA) onto poly(methyl methacrylate) (PMMA)-based denture resins through plasma-initiated grafting polymerization. The effects of reaction conditions on grafting and the physical properties of the resulting resins were evaluated. The grafted resins showed significantly increased drug binding capability toward clotrimazole, one of the most widely used antifungal drugs. The mechanisms for the enhancement in drug binding were discussed. The new clotrimazole-containing resins provided sustained drug release for longer than 28 days, and the released drugs demonstrated potent, long-term biofilm-controlling effects against Candida, pointing to an attractive strategy in controlling CADS and related fungal infections.

Fluorinated and Un-fluorinated N-halamines as Antimicrobial and Biofilm-controlling Additives for Polymers.

The objective of this study was to evaluate the effects of fluorination on the antimicrobial and biofilm-controlling activities of N-halamine-based additives for polymers. A fluorinated N-halamine, 1-chloro-3-1H,1H,2H,2H-perflurooctyl-5,5-dimetylhydantoin (Cl-FODMH), and its un-fluorinated counterpart, 1-chloro-3-octyl-5,5-dimethylhydantoin (Cl-ODMH), were synthesized and characterized with FT-IR, 1H-NMR, and DSC studies. Polyurethane (PU) films containing Cl-ODMH and Cl-FODMH as antimicrobial additives were fabricated through solvent casting. With the same additive contents (1wt%-5 wt%), PU films with Cl-FODMH showed higher contact angle values. AFM, SEM and DSC results revealed that while Cl-ODMH distributed evenly within PU, Cl-FODMH aggregated and formed macro-domains in PU. Antimicrobial studies showed that PU films with Cl-ODMH had higher antimicrobial and biofilm-controlling potency against Gram-positive and Gram-negative bacteria than PU samples with Cl-FODMH. These results demonstrated the importance of distribution of additives in polymers on antimicrobial performances, shedding lights on future antimicrobial material design strategies.

N-trimethylchitosan/alginate layer-by-layer self assembly coatings act as "fungal repellents" to prevent biofilm formation on healthcare materials.

Fungal biofilm formation on healthcare materials is a significant clinical concern, often leading to medical-device-related infections, which are difficult to treat. A novel fungal repellent strategy is developed to control fungal biofilm formation. Methylacrylic acid (MAA) is grated onto poly methyl methacrylate (PMMA)-based biomaterials via plasma-initiated grafting polymerization. A cationic polymer, trimethylchitosan (TMC), is synthesized by reacting chitosan with methyl iodide. Sodium alginate (SA) is used as an anionic polymer. TMC/SA multilayers are coated onto the MAA-grafted PMMA via layer-by-layer self-assembly. The TMC/SA multilayer coatings significantly reduce fungal initial adhesion, and effectively prevent fungal biofilm formation. It is concluded that the anti-adhesive property of the surface is due to its hydrophilicity, and that the biofilm-inhibiting action is attributed to the antifungal activity of TMC as well as the chelating function of TMC and SA, which may have acted as fungal repellents. Phosphate buffered saline (PBS)-immersion tests show that the biofilm-modulating effect of the multilayer coatings is stable for more than 4 weeks. Furthermore, the presence of TMC/SA multilayer coatings improves the biocompatibility of the original PMMA, offering a simple, yet effective, strategy for controlling fungal biofilm formation.

Construction of a functional silk-based biomaterial complex with immortalized chondrocytes in vivo.

To explore the feasibility of constructing a functional biomaterial complex with regenerated silk fibroin membrane and immortalized chondrocytes in vivo. Rat auricular chondrocytes (RACs) were transfected with the lentivirus vector pGC-FU-hTERT-3FLAG or pGC-FU-GFP-3FLAG, encoding the human telomerase reverse transcriptase (hTERT) or GFP gene. The effects of regenerated silk fibroin film on the adhesion, growth of immortalized chondrocytes and expression of collagen II in vitro were analyzed with immunofluorescent histochemistry. Immortalized RACs were transformed. Induction by nutrient medium promoted higher expression levels of collagen II in transformed chondrocytes. The regenerated silk fibroin film was not cytotoxic to immortalized chondrocytes and had no adverse influence on their adhesion. Collagen II expression was good in the immortalized chondrocytes in vivo. The construction of a silk-based biomaterial complex with immortalized chondrocytes may provide a feasible kind of functional biomaterial for the repair of cartilage defects in clinical applications.

Facile fabrication of the porous three-dimensional regenerated silk fibroin scaffolds.

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 µm to 350 µm 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 ß-sheet. The porous scaffolds prepared by this facile and energy-saving method with good biocompatibility will have great potential for application in tissue engineering.