Blended Silk Fibroin Research Articles

Electrospun silk fibroin/fibrin vascular scaffold with superior mechanical properties and biocompatibility for applications in tissue engineering

Electrospun scaffolds play important roles in the fields of regenerative medicine and vascular tissue engineering. The aim of the research described here was to develop a vascular scaffold that mimics the structural and functional properties of natural vascular scaffolding. The mechanical properties of artificial vascular tissue represent a key issue for successful transplantation in small diameter engineering blood vessels. We blended silk fibroin (SF) and fibrin to fabricate a composite scaffold using electrospinning to overcome the shortcomings of fibrin with respect to its mechanical properties. Subsequently, we then carefully investigated the morphological, mechanical properties, hydrophilicity, hemocompatibility, degradation, cytocompatibility and biocompatibility of the SF/fibrin (0:100), SF/fibrin (15:85), SF/fibrin (25:75), and SF/fibrin (35:65) scaffolds. Based on these in vitro results, we implanted SF/fibrin (25:75) vascular scaffold subcutaneously and analyzed its in vivo degradation and histocompatibility. The fiber structure of the SF/fibrin hybrid scaffold was smooth and uniform, and its fiber diameters were relatively small. Compared with the fibrin scaffold, the SF/fibrin scaffold clearly displayed increased mechanical strength, but the hydrophilicity weakened correspondingly. All of the SF/fibrin scaffolds showed excellent blood compatibility and appropriate biodegradation rates. The SF/fibrin (25:75) scaffold increased the proliferation and adhesion of MSCs. The results of animal experiments confirmed that the degradation of the SF/fibrin (25:75) scaffold was faster than that of the SF scaffold and effectively promoted tissue regeneration and cell infiltration. All in all, the SF/fibrin (25:75) electrospun scaffold displayed balanced and controllable biomechanical properties, degradability, and good cell compatibility. Thus, this scaffold proved to be an ideal candidate material for artificial blood vessels.

Fabrication and drug release behavior of the composite membrane via silk fibroin cross-linked with sodium alginate

ABSTRACT A series of silk fibroin composite membranes were prepared by blending silk fibroin (SF) with sodium alginate (SA) at different ratios. Different contents of polyethylene glycol (PEG) were used as a crosslinking agent to fix the structure of the composite membrane. Water-soluble pazufloxacin mesylate as the model drug was embedded in the composite membrane. The relationship between structure and properties was demonstrated by the sustained-release effect on model drug, and the experimental results revealed that the silk fibroin composite membrane had an excellent function of controlling the release behavior of model drugs, which exhibited pH responsivity to some extent.

Fabrication of antimicrobial poly(lactic‐co‐glycolic acid)/silk fibroin/aloe anthraquinone fibrous membranes for potential application of wound healing

AbstractAntibacterial wound dressings can kill bacteria around the wound and then accelerate the wound healing. To fabricate an antibacterial membrane for potential application of wound healing, poly(lactic‐co‐glycolic acid) (PLGA), silk fibroin (SF), and natural antimicrobial agent aloe anthraquinone (AA) were blended for electrospinning. The electrospun PLGA/SF/AA fibrous membrane consists of smooth and uniform fibers with an average diameter of 583 ± 112 nm. The stress of the PLGA/SF/AA fibrous membrane is 3.88 ± 0.34 MPa, which can ensure its application in skin wound healing. Owing to the good hydrophilicity of blended SF, PLGA/SF/AA fibrous membrane can release 47% of AA at 48 h and 62% of AA in 15 days. PLGA/SF/AA membrane also exhibits good antimicrobial properties and cell compatibility. These results indicate that the fabricated PLGA/SF/AA fibrous membrane in this study has a good application potential for wound healing.

Impacts of Blended Bombyx mori Silk Fibroin and Recombinant Spider Silk Fibroin Hydrogels on Cell Growth.

Binary-blended hydrogels fabricated from Bombyx mori silk fibroin (SF) and recombinant spider silk protein eADF4(C16) were developed and investigated concerning gelation and cellular interactions in vitro. With an increasing concentration of eADF4(C16), the gelation time of SF was shortened from typically one week to less than 48 h depending on the blending ratio. The biological tests with primary cells and two cell lines revealed that the cells cannot adhere and preferably formed cell aggregates on eADF4(C16) hydrogels, due to the polyanionic properties of eADF4(C16). Mixing SF in the blends ameliorated the cellular activities, as the proliferation of L929 fibroblasts and SaOS-2 osteoblast-like cells increased with an increase of SF content. The blended SF:eADF4(C16) hydrogels attained the advantages as well as overcame the limitations of each individual material, underlining the utilization of the hydrogels in several biomedical applications.

Dual-Crystallizable Silk Fibroin/Poly(L-lactic Acid) Biocomposite Films: Effect of Polymer Phases on Protein Structures in Protein-Polymer Blends.

Biopolymer composites based on silk fibroin have shown widespread potential due to their brilliant applications in tissue engineering, medicine and bioelectronics. In our present work, biocomposite nanofilms with different special topologies were obtained through blending silk fibroin with crystallizable poly(L-lactic acid) (PLLA) at various mixture rates using a stirring-reflux condensation blending method. The microstructure, phase components, and miscibility of the blended films were studied through thermal analysis in combination with Fourier-transform infrared spectroscopy and Raman analysis. X-ray diffraction and scanning electron microscope were also used for advanced structural analysis. Furthermore, their conformation transition, interaction mechanism, and thermal stability were also discussed. The results showed that the hydrogen bonds and hydrophobic interactions existed between silk fibroin (SF) and PLLA polymer chains in the blended films. The secondary structures of silk fibroin and phase components of PLLA in composites vary at different ratios of silk to PLLA. The β-sheet content increased with the increase of the silk fibroin content, while the glass transition temperature was raised mainly due to the rigid amorphous phase presence in the blended system. This results in an increase in thermal stability in blended films compared to the pure silk fibroin films. This study provided detailed insights into the influence of synthetic polymer phases (crystalline, rigid amorphous, and mobile amorphous) on protein secondary structures through blending, which has direct applications on the design and fabrication of novel protein–synthetic polymer composites for the biomedical and green chemistry fields.

Silk fibroin/alumina nanoparticle scaffold using for osteogenic differentiation of rabbit adipose-derived stem cells

In the present study, blended silk fibroin (SF)/alumina nanoparticles were prepared to assess the osteogenic differentiation of rabbit adipose-derived stem cells (rADSCs). The structural and biological behaviours of the nanocomposite were evaluated. Themicro-porous structure of SF/alumina nanocomposite had acceptable mechanical strength, water uptake capacity, and biomineralization capability. Increasing the content of alumina nanoparticles in the structure of SF substrate more than 10% enhanced the physiochemical properties of the bulk SF scaffold without any toxicity effect on the proliferation and attachment of rADSCs. More alkaline phosphates activity and calcium phosphate deposition were also observed in nanocomposite with 10% alumina nanoparticle. Compare with other groups, applying the blended scaffold along with osteogenic medium induced rADSCs to express higher osteogenic markers like collagen type I, osteopontin, RUNX2, and osteocalcin. Taken together, SF/alumina nanocomposite serves as good substrates for proliferation and osteogenic differentiation of ADSCs applicable for bone tissue engineering.

Invitro Biodegradability of Silk Fibroin/Xanthan Biopolymeric Composite Scaffolds

Silk fibroin/xanthan scaffolds were prepared by blending silk fibroin and xanthan in the ratios 80SF:20Xa (SFX82), 60SF:40Xa (SFX64), and 50SF:50Xa (SFX55) using freeze drying method. In-vitro degradation behavior of the prepared scaffolds was studied for 37 days in phosphate buffer saline. The degradation rate was the function of silk fibroin, xanthan and β-crystallite contents in the silk fibroin/ xanthan composites. SFX82 degraded extremely slowly whereas SFX55 showed faster degradation rate. Hydrophilic xanthan was the main contributor of weight loss. SFX82 and SFX64 exhibited surface degradation whereas SFX55 showed bulk degradation which indicated that higher silk fibroin ratios favor surface degradation. Due to bulk degradation, SFX55 showed maximum surface roughness among the composite scaffolds. The FTIR spectrum revealed total loss of xanthan from the composites after degradation. The broad and low-intensity peaks in the FTIR spectrum of composite scaffolds confirmed reduction in β-sheet crystallite content during degradation. XRD analysis also confirmed reduction in β-sheet crystals and revealed that degraded composite scaffold had predominantly amorphous structure. The degraded scaffold showed higher porous structure than the non-degraded scaffold. The in vitro degradability testing gives a good approximation of degradation of scaffold in vivo and helps in designing a robust biopolymeric composite scaffold for tissue engineering.

Nature-Derived Aloe Vera Gel Blended Silk Fibroin Film Scaffolds for Cornea Endothelial Cell Regeneration and Transplantation.

The goal of this study was to fabricate an appropriate replacement for cadaveric corneas to overcome a shortage of cadaveric corneas for transplantation. In this study, we fabricated transparent ultrathin film scaffolds with nature-derived aloe vera (AV) gel and silk fibroin (SF) for corneal endothelial cells (CECs). The scaffolds were subjected to analysis of transparency and contact angle using field emission scanning electron microscopy (FESEM) and Fourier transform infrared spectroscopy to determine their physical and chemical properties. FESEM images revealed that the critical morphology of CECs was formed on the AV gel in the blend with SF rather than in the scaffold with SF alone. The cell proliferation, phenotype, and specific gene marker expressions for CECs were determined by MTT assays, immunofluorescence, and reverse transcription polymerase chain reactions. Incorporation of a small amount of AV gel increased the cell viability and maintained its functions well. The scaffolds were easily handled for transplantation into rabbit eyes with small incisions and examined by their transparency after transplantation and histological staining. The scaffolds attached to the surface of the corneal stroma and integrated with surrounding corneal tissue without a significant inflammatory reaction. These results indicate that AV blended SF film scaffolds might be a suitable substitute for alternative corneal grafts for transplantation.

P-025 - Preliminary study of blended silk fibroin for contact lens-based ophthalmic drug delivery

P-025 - Preliminary study of blended silk fibroin for contact lens-based ophthalmic drug delivery

Silk fibroin/copolymer composite hydrogels for the controlled and sustained release of hydrophobic/hydrophilic drugs

In the present study, a composite system for the controlled and sustained release of hydrophobic/hydrophilic drugs is described. Composite hydrogels were prepared by blending silk fibroin (SF) with PLA–PEG–PLA copolymer under mild aqueous condition. Aspirin and indomethacin were incorporated into SF/Copolymer hydrogels as two model drugs with different water-solubility. The degradation of composite hydrogels during the drug release was mainly caused by the hydrolysis of copolymers. SF with stable β-sheet-rich structure was not easily degraded which maintained the mechanical integrity of composite hydrogel. The hydrophobic/hydrophilic interactions of copolymers with model drugs would significantly alter the morphological features of composite hydrogels. Various parameters such as drug load, concentration ratio, and composition of copolymer were considered in vitro drug release. Aspirin as a hydrophilic drug could be controlled release from composite hydrogel at a constant rate for 5 days. Its release was mainly driven by diffusion-based mechanism. Hydrophobic indomethacin could be encapsulated in copolymer nanoparticles distributing in the composite hydrogel. Its sustained release was mainly degradation controlled which could last up to two weeks. SF/Copolymer hydrogel has potential as a useful composite system widely applying for controlled and sustained release of various drugs.

Electrospun nanofibrous SF/P(LLA-CL) membrane: a potential substratum for endothelial keratoplasty

BackgroundCornea transplant technology has progressed markedly in recent decades, allowing surgeons to replace diseased corneal endothelium by a thin lamellar structure. A thin, transparent, biocompatible, tissue-engineered substratum with corneal endothelial cells for endothelial keratoplasty is currently of interest. Electrospinning a nanofibrous structure can simulate the extracellular matrix and have beneficial effects for cell culture. Silk fibroin (SF) has good biocompatibility but poor mechanical properties, while poly(l-lactic acid-co-ε-caprolactone) (P(LLA-CL)) has good mechanical properties but poor biocompatibility. Blending SF with P(LLA-CL) can maintain the advantages of both these materials and overcome their disadvantages. Blended electrospun nanofibrous membranes may be suitable for regeneration of the corneal endothelium. The aim of this study was to produce a tissue-engineered construct suitable for endothelial keratoplasty.MethodsFive scaffolds containing different SF:P(LLA-CL) blended ratios (100:0, 75:25, 50:50, 25:75, 0:100) were manufactured. A human corneal endothelial (B4G12) cell line was cultured on the membranes. Light transmission, speed of cell adherence, cell viability (live-dead test), cell proliferation (Ki-67, BrdU staining), and cell monolayer formation were detected on membranes with the different blended ratios, and expression of some functional genes was also detected by real-time polymerase chain reaction.ResultsDifferent blended ratios of scaffolds had different light transmittance properties. The 25:75 blended ratio membrane had the best transmittance among these scaffolds. All electrospun nanofibrous membranes showed improved speed of cell adherence when compared with the control group, especially when the P(LLA-CL) ratio increased. The 25:75 blended ratio membranes also had the highest cell proliferation. B4G12 cells could form a monolayer on all scaffolds, and most functional genes were also stably expressed on all scaffolds. Only two genes showed changes in expression.ConclusionAll blended ratios of SF:P(LLA-CL) scaffolds were evaluated and showed good biocompatibility for cell adherence and monolayer formation. Among them, the 25:75 blended ratio SF:P(LLA-CL) scaffold had the best transmittance and the highest cell proliferation. These attributes further the potential application of the SF:P(LLA-CL) scaffold for corneal endothelial transplantation.

혼합 실크 피브로인막의 투명도

실크는 천연고분자 단백질로 생체친화적이며, 최근 조직공학 연구가 활성화 되면서 각광받는 생체재료이다. 하지만 실크 단독으로 실크막을 제작하였을 때, 얇은 막은 투명도가 있지만 두께가 두꺼워지게 되면 실크막은 매우 불투명한 막으로 바뀌게 된다. 인체조직에서 사용되는 막중 각막이나 뇌경막 등 투명도가 반드시 필요한 생체막에 실크막을 이용하기 위해서는 실크막의 투명도가 반드시 요구된다. 따라서, 본 연구에서는 실크 피브로인 수용액에 다양한 무기염류와 고분자물질을 혼합하여 혼합 실크막을 제작하고 그 투명도를 분석하였다. 제작된 혼합 실크막은 혼합된 물질에 따라 투명도가 매우 달랐다. 무기 염류가 혼합된 실크막의 경우 고분자물질과 혼합된 실크막에 비해 더 투명도가 높았으며, 특히 염화칼슘이 혼합된 실크막의 투명도는 매우 투명하였다. 따라서, 각막 등 완벽한 투명도를 요구하는 생체막으로 실크막을 사용할 수 있는 가능성을 확인할 수 있었다. 향후 혼합 실크막의 물리적 특성 규명을 통해 좀 더 물성이 개선된다면, 투명한 실크막이 다양한 용도로 이용될 수 있을 것으로 기대된다. Silk fibroin is a natural biomaterial that has the biocompatibility and other many advantages. But as a silk fibroin membrane thickness increases, the transparency becomes more opaque. Because the transparency of membranes tissue such as the cornea and dura mater are necessary, transparent membrane is required to replace these transparent membranes. In this study, we fabricated blending silk fibroin membranes that made by mixing the various inorganic salts or polymer in an aqueous solution of silk fibroin. The transparency of the membranes were analyzed. the transparency of these membranes is very different, depending on the mixed materials. Inorganic salts mixed silk membrane was more transparent than the polymer mixed one. Especially, the silk fibroin membrane with calcium chloride was very transparent. We showed the possibility of blending silk fibroin membrane, which can be used in perfect transparent membrane such as the cornea. In the future, we expect that the transparent blending silk fibroin membrane can be used in various medical applications.

Development of a Porous Scaffold-Manufacturing Method by Blending Silk Fibroin and Agarose Polymer Solutions

Low-melting-temperature agarose gel solution, as a novel porogen was combined with a silk fibroin solution to generate interconnected porous networks. The porosity of the resultant silk fibroin-agarose scaffolds was greater than that of the scaffolds generated with agarose and deionized water. The porosities of silk fibroin scaffolds containing agarose gel at 0.5%, 1.0%, 1.5%, 2.0% [w/v] were 110.9%, 111.7%, 120.9%, and 123.0%, respectively. Lastly, the internal space generated in scaffolds after dissolution of the agarose gel provides a good environment for cell growth and movement within the scaffold.

Copolymer‐induced silk‐based hydrogel with porous and nanofibrous structure

AbstractA novel physical blend method was developed to accelerate the self‐assembly process of silk fibroin (SF) solution into porous and nanofibrous hydrogel by temperature‐sensitive copolymer. Silk‐based hydrogel was firstly achieved through blending SF solution with copolymer aqueous solution and then removed the copolymer from blend solution by heat treatment (50°C) after 24 h hydrogelation. Copolymer molecules would interact with SF molecules resulting in reduction of copolymer micelles, which further affect the hydrogelation of SF solutions. Copolymers could be separated from blend solution by heat treatment under an acceptable temperature (50°C), especially the copolymer2. Fourier transform infrared (FTIR) and X‐ray diffraction showed the blending of copolymer significantly accelerated the self‐assembly of SF into physically crosslinked β‐sheet crystals at room temperature which led to the sol‐gel transition. Results from DTA and X‐ray diffraction showed that the effect of copolymer on crystalline structure of SF in silk‐based hydrogel was very weak. SF molecules transformed from distributed globular nanoparticles to nanofilaments clustered during hydrogelation, resulting in the porous and nanofibrous structure of silk‐based hydrogel. Furthermore, silk‐based hydrogel was prepared in aqueous solution avoiding organic solvents and harsh processing conditions, suggesting that this silk‐based hydrogel could be a potential candidate scaffold for biomedical applications. © 2012 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 2013

Preparation and characterization of blended Bombyx mori silk fibroin scaffolds

The aim of this study was to compare physical, mechanical and biological properties of 3-dimensional scaffolds prepared from Bombyx mori silk fibroin (SF), fibroin blended with collagen (SF/C), and fibroin blended with gelatin (SF/G) using a freeze-drying technique. The prepared scaffolds were sponge-like structure that exhibited homogeneous porosity with highly interconnected pores. Average pore size of these scaffolds ranged from 65–147 µm. All biodegradable scaffolds were capable of water absorption of 90 %. The degradation behavior of these scaffolds could be controlled by varying the amount of blended polymer. The SF/C and SF/G scaffolds showed higher compressive modulus than that of SF scaffolds which could be attributed to the thicker pore wall observed in the blended constructs. The less crystalline SF structure was observed in SF/G scaffolds as compared to SF/C scaffolds. Thus, the highest compressive modulus was observed on SF/C matrix. To investigate the feasibility of the scaffolds for cartilage tissue engineering application, rat articular chondrocytes were seeded onto the scaffolds. The MTT assay demonstrated that blending collagen or gelatin into SF sponge facilitated cell attachment and proliferation better than SF scaffolds. The blended SF scaffolds possessed superior physical, mechanical and biological properties in comparison to SF scaffolds and showed high potential for application in cartilage tissue engineering.

The biocompatibility of silk fibroin films containing sulfonated silk fibroin

Sulfonation reaction may be an effective method for preparation of heparin-like materials. However, no sulfonated polymer based on protein backbone was used for improving the blood compatibility of biomaterials. In this study, the biocompatibility of new kind of composite materials films obtained by blending silk fibroin (SF) with sulfonated silk fibroin (SSF) was evaluated. The anticoagulant activity was characterized with prothrombin time (PT), activated partial thromboplastin time (APTT), and thrombin time (TT), which all increased remarkably, the clot times exceeded the measurement limit of clot detection instrument. Its platelet adhesion was also investigated as another parameter of blood compatibility. The cell compatibility of composite films was evaluated through cell morphologies on the films and cell viability by methyl thiazolyl tetrazolium (MTT) assay. Tensile strength and elongation at break of the composite films reached to 44.6 MPa and 50.3%, respectively. All these results indicated that SF/SSF composite film was a potential material for blood-contact materials and tissue engineering matrix.