Electrospun Silk Fibroin Nanofibers Research Articles

Electrospun Silk Fibroin and Collagen Composite Nanofiber Incorporated with Palladium and Platinum Nanoparticles for Wound Dressing Applications

Electrospun Silk Fibroin and Collagen Composite Nanofiber Incorporated with Palladium and Platinum Nanoparticles for Wound Dressing Applications

Silk Fibroin Combined with Electrospinning as a Promising Strategy for Tissue Regeneration.

The development of tissue engineering scaffolds is of great significance for the repair and regeneration of damaged tissues and organs. Silk fibroin (SF) is a natural protein polymer with good biocompatibility, biodegradability, excellent physical and mechanical properties and processability, making it an ideal universal tissue engineering scaffold material. Nanofibers prepared by electrospinning have attracted extensive attention in the field of tissue engineering due to their excellent mechanical properties, high specific surface area, and similar morphology as to extracellular matrix (ECM). The combination of silk fibroin and electrospinning is a promising strategy for the preparation of tissue engineering scaffolds. In this review, the research progress of electrospun silk fibroin nanofibers in the regeneration of skin, vascular, bone, neural, tendons, cardiac, periodontal, ocular and other tissues is discussed in detail.

Electrospun silk fibroin and gelatin blended nanofibers functionalized with noble metal nanoparticles for enhanced biomedical applications

The combination of highly anticancer active platinum group metals and biocompatible natural fibers exhibits significant anticancer activity while also allowing cell regeneration and repair to the damaged region. In this research, platinum and palladium nanoparticles anchored silk fibroin and gelatin (SF/GL/Pd-Pt) hybrid composite nanofiber was fabricated through the electrospinning method. The amino groups in SF and GL act as a reducingd stabilizing agent for the resulting Pd and Pt nanoparticles. Morphological and functional studies confirmed that crystalline Pd and Pt nanoparticles with average sizes of 5.11 and 3.03 nm are anchored on a 154.93 ± 34.19 nm SF/GL nanofiber matrix. The atomic force microscopic (AFM) images show that the SF/GL/Pd-Pt composite nanofiber has a thickness of 4.74 nm and a roughness area (Ra) of 258.5 nm. X-ray photoelectron spectroscopic (XPS) results confirmed the presence of C, N, O, Pt, and Pd in the composite fiber with an atomic percentage of 67.56%, 11.05%, 17.72%, 2.01% and 1.66%. respectively. This SF/GL/Pd-Pt composite nanofiber is more effective than its counterparts in terms of anticancer activity (IC50 value of 28.82 µg/mL−1) against A549 human lung carcinoma cells. Furthermore, the β-actin and Hsp90 gene expression in A549 cells confirmed its superior anticancer activity.

The remarkable effect of menstrual blood stem cells seeded on bilayer scaffold composed of amniotic membrane and silk fibroin aiming to promote wound healing in diabetic mice

IntroductionImpaired chronic wound healing frequently occurs in diabetic patients. We hypothesized that menstrual blood-derived mesenchymal stem cells (MenSCs) in combination with bilayer scaffold consisted of human amniotic membrane (AM) and electrospun silk fibroin nanofibers could potentially promote wound healing in diabetic mice. Methods & MethodsTwo bilateral full-thickness wounds were created on dorsal skin of type-1 diabetic mice model and animals were equally divided in four groups including: no-treatment group (NT), amniotic membrane treated group (AM), bilayer scaffold treated group (bSC), and MenSCs-seeded bilayer scaffold treated group (bSC + MenSCs). Wound healing evaluations were performed at 3, 7, and 14 days after their treatment. The wound healing was analyzed by macroscopic and microscopic evaluations, and immunofluorescence staining of involucrin (IVL), type III collagen, CD31/ von Willebrand factor (vWF), and PGP9.5 were performed. Furthermore, number of neutrophils and macrophages and subpopulation of macrophages were assessed. In addition, the expression of Egr2, Mmp9, CXCL12, IDO1, Ptgs2 and VEGFA transcripts involved in wound repair were also analyzed. ResultsAfter 14 days, the best epidermal and dermal regeneration belonged to the cases received bSC + MenSCs as wound dressing. Moreover, the wound healing was typically faster in this group compared to other groups. Immunofluorescence evaluation represented higher levels of CD31 and VWF, higher ratio of M2/M1 macrophages, greater expression of IVL, and higher levels of the PGP9.5 in the bSC + MenSCs group in comparison with other groups. Expression analysis of assessed genes also supported assumption of more regeneration and healing in the bSC + MenSCs group versus other groups. ConclusionThese results indicate that enhanced immunomodulatory and reparative properties of MenSCs in conjunction with bilayer scaffold specified this cellular skin substitute for modulating wound chronicity and contribution to resolution of wound healing process in diabetic ulcer.

Crystallinity modulated silk fibroin electrospun nanofibers based floating scaffold as a candidate for controlled release of felodipine

Floating gastro-retentive delivery approach provides a significant pathway for controlled release of drug with increase gastric residence. In this study, we report crystallinity modulated electrospun silk fibroin nanofibers (SF-NFs) floating scaffolds for the controlled release of felodipine (FD). The alteration in the crystallinity behavior due to changes in the structural conformation of SF helps to customize the release kinetics of FD-loaded SF-NFs scaffolds. Additionally, FD-loaded SF scaffolds system having a density less than the acidic gastric fluid explore as a new tactic for floating drug delivery system. The prepared FD-loaded SF nanofibers (FD-loaded SF-NFs) were characterized by spectral, thermal, and diffractometric techniques, scanning electron microscopy, floating profile, in-vitro degradation, mucoadhesion, and in-vitro dissolution studies, etc. The optimized batch had the least porosity and swelling, was annealed with ethanol and water for crystallinity modulation of SF-NFs to get controlled release of FD. Spectral, thermal, and diffractometric analyses could unveil the molecular dispersion of FD, coupled with amorphous form stabilization in NF. Excellent floating profile and satisfactory mucoadhesion of FD-SF-NFs also endorsed the formation of a novel floating drug delivery system. Temporal control over FD release was elucidated by in-vitro dissolution, demonstrating controlled release due to crystallinity modulation of SF-NFs. In conclusion, crystallinity-modulated electrospun NFs fabricated from SF waste could be used as a customizable carrier for drug delivery to the gastric region.

Green Electrospun Silk Fibroin Nanofibers Loaded with Cationic Ethosomes for Transdermal Drug Delivery

Transdermal drug delivery system(TDDS) facilitates the controlled release of active ingredients penetrating through the skin, avoiding the liver first pass effect. Electrospinning is a simple process to fabricate ultrafine fibers with a higher specific surface area, making them excellent candidates for drug delivery. In current work, a novel silk fibroin(SF) nanofiber loaded with cationic ethosomes(CEs) was prepared via green electrospinning. The data of Fourier transform infrared spectroscopy(FTIR) and laser scanning confocal microscopy(LSCM) confirmed the existence of CEs in the SF nanofibers. The morphology of the nanofibers was not significantly affected by the incorporation of CEs as shown by scanning electron microscopy(SEM) images. The CEs-loaded SF nanofibrous patch (CEs-SFnP) showed good cytocompatibility as proved by both cell counting Kit-8(CCK-8) assay and SEM. Using doxorubicin hydrochloride(Dox) as a model drug, the transdermal performance of CEs-SFnP was evaluated through Franz diffusion cell against mouse skin. The results indicated that CEs-SFnP can effectively deliver drug into the skin, with a much higher permeation rate than the normal nanofibers without CEs. The as-spun CEs-SFnP in this study could find promising applications in TDDS.

Biofunctionalized silk fibroin nanofibers for directional and long neurite outgrowth.

Engineered scaffolds simultaneously exhibiting multiple cues are highly desirable for neural tissue regeneration. Silk fibroin is a promising natural protein material for nerve repair. However, the lack of specific bioactive cues significantly hinders its application. In this study, the electrospun silk fibroin nanofibers with both biochemical and topographical cues were prepared. The alignment of electrospun nanofibers was optimized by controlling the surface linear velocity of a rotating drum. The silk fibroin nanofibers were further functionalized with laminin through covalent binding, confirmed by immunostaining observation. Cell proliferation and neurite outgrowth assays confirmed that the functionalized aligned nanofibers significantly enhanced directional axonal extensions, providing physical and bioactive cues for neurite outgrowth. Furthermore, the tubular scaffolds with longitudinally aligned microchannels were designed by rolling the functionalized silk fibroin nanofibers. The neurite extension across the lumen of the conduit along the direction of the aligned fibers is apparent. These results highlight the ability of laminin-immobilized silk fibroin nanofibers to enhance neurite outgrowth and to control directional neurite extension, providing a useful approach to construct a regenerative microenvironment for nerve repair materials.

Efficient Wound Healing Using a Synthetic Nanofibrous Bilayer Skin Substitute in Murine Model

Treatment of full-thickness skin wounds with minimal scarring and complete restoration of native tissue properties still exists as a clinical challenge. A bilayer skin substitute was fabricated by coating human amniotic membrane (AM) with electrospun silk fibroin nanofibers, and its in vivo biological behavior was studied using murine full-thickness skin wound model. Donut-shaped silicon splints were utilized to prevent wound contraction in mouse skin and simulate re-epithelialization, which is the normal path of human wound healing. Skin regeneration using the bilayer scaffold was compared with AM and untreated defect after 30 d. Tissue samples were taken from healed wound areas and investigated through histopathological and immunohistochemical staining to visualize involucrin (IVL), P63, collagen I, CD31, and vascular endothelial growth factor. In addition, mRNA expression of IVL, P63, interleukin-6, and cyclooxygenase-2 was studied. The application of bilayer scaffold resulted in the best epidermal and dermal regeneration, demonstrated by histopathological examination and molecular analysis. In regenerated wounds of the bilayer scaffold group, the mRNA expression levels of inflammatory markers (interleukin-6 and cyclooxygenase-2) were downregulated, and the expression pattern of keratinocyte markers (IVL and P63) at both mRNA and protein levels was more similar to native tissue in comparison with AM and no-treatment groups. There was no significant difference in the expression level of collagen I, CD31, and vascular endothelial growth factor among different groups. Conclusively, these promising results serve as a supporting evidence for proceeding to clinical phase to examine the capacity of this bilayer scaffold for human skin regeneration.

Magnetic silk fibroin composite nanofibers for biomedical applications: Fabrication and evaluation of the chemical, thermal, mechanical, and in vitro biological properties

ABSTRACTMagnetically responsive polymer composites have great potential for use in diverse biomedical applications. In this study, composite biomaterials consisting of silk fibroin (SF) and superparamagnetic iron oxide nanoparticles (SPIONs) were fabricated by the electrospinning method. Two different methods were employed to incorporate the SPIONs into the SF nanofibers. In the first encapsulation method (M1), SPIONs (1.0, 3.0, and 5.0 wt%) were initially included in the electrospinning solution. In the second dip‐coating method (M2), electrospun SF nanofiber mats were immersed in the aqueous suspensions of SPIONs (10, 30 and 50% v/v). Then, the pure and composite silk fibroin composite mats were comparatively evaluated for their morphological, chemical, magnetic, mechanical and in vitro biological properties, by using a number of methods including SEM, TEM, FTIR, XRD, EDS, VSM, TGA, mechanical tensile tests, as well as by indirect in vitro cytotoxicity and in vitro hemocompatibility analyses. Overall findings suggested that, while M1 nanofiber mats could be a suitable candidate for use in tissue engineering as a magnetically responsive cytocompatible scaffold, the M2 nanofiber mats perhaps could be more appropriate as an interface for triggering the in vitro stem cell differentiation and/or biosensor applications. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019, 136, 48040.

Modulation of myelin formation by combined high affinity with extracellular matrix structure of electrospun silk fibroin nanoscaffolds

Remyelination is a major therapeutic goal in peripheral nerve regeneration, serving to restore function of demyelinated axons and provide neuroprotection. In order to apply myelin biogenesis strategies to peripheral nerve defects, the tissue engineered substitutes might be amenable to the promotion of this repair process. Electrospun nanofibers are considered as promising scaffolds for tissue engineering due to extracellular matrix mimicking factor and enhanced electrostatic interaction resulting in a controllable 3 D nanofibrous membrane. In order to explore the role of electrospun silk fibroin (SF) membrane in myelination, co-culture of dorsal root ganglion (DRG) neurons and Schwann cells (SCs) in vitro was established and observed. Scanning electron microscopy was used to observe DRG adhesion to the membranes, the electrospinning SF membrane is more favorable to the adhesion of DRG. The immunofluorescence staining of MAG and NF showed considerable amount of myelin were formed, and the myelin was tightly wrapped around the axons of the neurons, which was confirmed under the scanning electron microscope observation. Real-time quantitative PCR technique was used to determine the gene expression level of DRG neurons cultured at different time points. The results showed that the mRNA levels of N-cadherin, laminin, fibronectin were higher than those in the control group. Our results showed that the electrospun SF nanofibers can provide topographical and chemical cues that mimic (to a certain extent) the extracellular matrix.

Modification of electrospun silk fibroin nanofiber mats: using an EDC/NHS ethanol solvent

The application of electrospun silk fibroin nanofiber mats ware restricted for its poor mechanical property. The goal of the present study was to improve the mechanical property of silk fibroin nanofiber mats. EDC/NHS ethanol system was employed to treat silk fibroin nanofiber mats to modify the property of silk fibroin nanofiber mats. The as-spun, ethanol treated, and EDC/NHS ethanol treated silk fibroin nanofiber mats were characterized by scanning electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction, thermogravimetry-Differential thermal analysis, and Instron tester. After ethanol and EDC/NHS treatments, the conformation transition of silk fibroin was detected by FTIR and WAXD. The crosslinker EDC/NHS will form abundant “amide” bonds within silk fibroin which was confirmed by FTIR and tensile mechanical test. Compared to as-spun and ethanol treated silk fibroin nanofiber mats, the EDC/NHS ethanol treated one demonstrated significantly enhanced breaking elongation by 136%. The significant increase in breaking elongation made the electrospun silk fibroin nanofiber mats more promising scaffold materials for practical tissue engineering application.

Three-dimensional electrospun silk-fibroin nanofiber for skin tissue engineering

Tissue-engineered skin substitutes may offer an effective therapeutic option for the treatment of patients with skin damages. In this study, a novel three-dimensional (3D) scaffold composed of electrospun silk fibroin (SF) nanofiber was fabricated using electrospinning with the addition of NaCl crystals. It has well known that the electrospun SF nanofibers were excellent scaffold for tissue. However, it is generally difficult for cells to infiltrate the electrospun silk fibroin due to its small pore size. To resolve this problem, we dropped the NaCl crystals above the rotating collector, which become incorporated into the nanofibers. Three methods (freeze-drying, salt-leaching, and electrospinning with NaCl) for fabrication of SF scaffolds were compared to the difference of their characteristics using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), mechanical strength, porosity, swelling abilities, and cell proliferation. Additionally, using air-liquid culture system, keratinocytes were co-cultured with fibroblasts in each type of SF scaffolds to construct an artificial bilayer skin in vitro. In our experimental results, histologic findings in only electrospun SF scaffolds showed more proliferation of fibroblasts in deep layer and more differentiation of keratinocytes in superficial layer. The present study suggests that 3D electrospun SF scaffolds might be a suitable for skin tissue engineering.

Multi-biofunction of antimicrobial peptide-immobilized silk fibroin nanofiber membrane: Implications for wound healing

An antimicrobial peptide motif (Cys-KR12) originating from human cathelicidin peptide (LL37) was immobilized onto electrospun SF nanofiber membranes using EDC/NHS and thiol-maleimide click chemistry to confer the various bioactivities of LL37 onto the membrane for wound care purposes. Surface characterizations revealed that the immobilization density of Cys-KR12 on SF nanofibers could be precisely controlled with a high reaction yield. The Cys-KR12-immobilized SF nanofiber membrane exhibited antimicrobial activity against four pathogenic bacterial strains (Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas aeruginosa) without biofilm formation on the membrane surface. It also facilitated the proliferation of keratinocytes and fibroblasts and promoted the differentiation of keratinocytes with enhanced cell-cell attachment. In addition, immobilized Cys-KR12 significantly suppressed the LPS-induced TNF-α expression of monocytes (Raw264.7) cultured on the membrane. These results suggest that a Cys-KR12-immobilized SF nanofiber membrane, which has multiple biological activities, would be a promising candidate as a wound dressing material. Statement of SignificanceThis research article reports various bioactivities of an antimicrobial peptide on electrospun silk fibroin nanofiber membrane. Recently, human cathelicidin peptide LL37 has been extensively explored as an alternative antibiotic material. It has not only a great antimicrobial activity but also a wide variety of bioactivities which can facilitate wound healing process. Especially, many studies on immobilization of LL37 or its analogues have shown the immobilization technique could improve performance of wound dressing materials or tissue culture matrices. Nevertheless, so far studies have only focused on the bactericidal effect of immobilized peptide on material surface. On the other hand, we tried to evaluate multi-biofunction of immobilized antimicrobial peptide Cys-KR12, which is the shortest peptide motif as an analogue of LL37. We fabricated silk fibroin nanofiber membrane as a model wound dressing by electrospinning and immobilized the antimicrobial peptide. As a result, we confirmed that the immobilized peptide can play multi-role in wound healing process, such as antimicrobial activity, facilitation of cell proliferation and keratinocyte differentiation, and inhibition of inflammatory cytokine expression. These findings have not been reported and can give an inspiration in wound-care application.

Electrospun silk-based nanofibrous scaffolds: fiber diameter and oxygen transfer.

In this study, silk fibroin was extracted from cocoons of silkworms and fabricated into nonwoven mats by electrospinning method. A new model based on the group method of data handling (GMDH) and artificial neural network (ANN) was developed for estimation of the average diameter of electrospun silk fibroin nanofibers. In this regard, concentration, flow rate, voltage, distance, and speed of collector were used as input parameters and average diameter of the fibers was considered as output parameter. Two models were capable to estimate average diameter of fibers with good accuracy. The average absolute relative deviation for GMDH and ANN models was equal to 3.56 and 2.28 %, respectively. Furthermore, due to importance of oxygen delivery to site of injury to promote wound healing, continuity equation for mass transport was employed for prediction of oxygen profile in the system containing wound dressing and skin. The result showed that our prepared wound dressing is capable to pass the oxygen completely to the skin layer and is not acting as a barrier for oxygen delivery to wound site. Since average nanofibers diameter can influence the mat physical, mechanical and biological properties then this model may serve as a useful guide to obtain tailor made and uniform silk nanofibers at various combinations of process variables.

Development of novel electrospun nanofibrous scaffold from P. Ricini And A. Mylitta silk fibroin blend with improved surface and biological properties.

Biomaterials that stimulate cell attachment and proliferation without any surface modification (e.g. RGD coating) provide potent and cost effective scaffold for regenerative medicine. This study assessed the physico-chemical properties and cell supportive potential of a silk fibroin blend scaffold derived from eri (Philosamia ricini) and tasar (Antheraea mylitta) silk (ET) respectively by electrospinning process. The scanning electron microscopy and transmission electron microscopy study found that the fiber diameters are in 200 to 800nm range with flat morphology. The porosity of ET scaffold is found to be 79±5% with majority of pore diameter between 2.5 to 5nm. Similarly, Bombyx mori (BM) silk fibroin and gelatin nanofibrous scaffolds were prepared and taken as control. The ultimate tensile strength of the ET and BM scaffold are found to be 1.83±0.13MPa and 1.47±0.10MPa respectively. The measured contact angle (a measure of hydrophilicity) for ET (54.7°±1.8°) is found to be lower than BM (62°±2.3°). The ability to deposit apatite over ET is comparable to that of BM nanofibers. All the scaffolds were seeded with cord blood derived mesenchymal stem cells (hMSCs) and cultured for 14days in vitro. The immunofluorescence study reveals enhanced cell attachment with higher metabolic activity for MSCs grown over ET than BM and gelatin. The ET scaffold also demonstrated expression of higher amount cell adhesion molecules (CD29/CD44) and higher proliferation rate than BM and gelatin as confirmed by MTT assay, DNA content estimation assay, flow cytometry study and SEM study. Overall, it may be concluded that ET scaffold may have potential in developing bone tissue grafts for clinical applications in the future.

Preparation of electrospun silk fibroin nanofibers from solutions containing native silk fibrils

ABSTRACTElectrospinning solutions containing native silk fibrils with varied diameter and length were firstly achieved by dissolving silk in CaCl2/Formic acid solvents. The structure of nanofibrils significantly improved the spinnability of electrospinning solution. The diameter of electrospun silk fibroin (SF) nanofibers increased from 40 nm to 1.8 μm, which could be achieved through increasing the solution concentration from 2 to 10%, implying a good size control over a wide range in this process. The structure of SF nanofibers transferred from random coil to beta‐sheet, before and after ethanol treatment, respectively. The mechanical properties of the SF nanofibers were improved significantly with stress and strain at break of 11.15 MPa and 7.66% in dry state, and 3.32 MPa and 174.0% in wet state. The strategy for preparing SF nanofibers with improved mechanical properties and fiber diameter control over a wide range provides benefits for the application of this material. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015, 132, 41236.

Development of artificial dermis using 3D electrospun silk fibroin nanofiber matrix.

Several auto- or alloplastic dermal substitutes have been used for full thickness skin defects. However, there is no ideal dermal substitute that is biocompatible and offers adequate mechanical properties and sufficient thickness and volume. Silk fibroin can be considered a possible alternative material for the construction of dermal substitutes. Nanofibers fabricated by electrospinning silk fibroin were considered an excellent candidate due to their desirable biocompatibility. However, it is generally difficult for cells to infiltrate the electrospun silk fibroin due to its small pore size. To overcome this challenge, a key parameter is the control of the pore size of silk fibroin electrospun such that cells can easily infiltrate into the nanofibers and proliferate internally. In this work, a novel approach is introduced by dropping two different size NaCl crystals above the rotating collector, which become incorporated into the nanofibers. The structures and properties of the, resulting electrospun silk fibroin were characterized using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and its wound healing effect for a full thickness skin defect was compared with Matriderm. Although the dermal regenerative effect of the 3D-SF (3D electrospun silk fibroin nanofiber matrix) was similar to that of Matriderm, the 3D-SF was almost completely degraded and did not induce wound contracture like Matriderm. We suggest that 3D-SF can be applied to the treatment of full thickness skin defects.

Effect of Sodium Carbonate Concentrations on the Formation and Mechanism of Regenerated Silk Fibroin Nanofibers by Electrospinning

Degumming is the first process for the preparation of all silk-based products. In this paper, effect of sodium carbonate concentrations for silk degumming on the formation of electrospun silk fibroin nanofibers was investigated and the reason for the silk electrospinning process was explained for the first time by differences from the microstructure of regenerated silk fibroin. With increasing the sodium carbonate concentration, microstructure both in the aqueous solutions and in the electrospinning solutions transformed from nanofibrils to nanoparticles, leading to obvious changes on rheological property; electrospinning solutions with nanofibrils behaved like the native silk dope and owned remarkably higher viscosity than the solutions with nanoparticles showing very low viscosity. More interestingly, nanofibrils favored the formation of silk nanofibers with ease, and even nanofibers could be electrospun at concentration 2%. However, nanoparticles were completely unable to generate nanofibers at high spinning concentration 8%. Importance of sodium carbonate concentrations is heavily emphasized for impacting the microstructure types and further influencing the electrospinning performance of regenerated silk. Hence, sodium carbonate concentrations provide a controllable choice for the preparation of silk-based electrospun biomaterials with desired properties.

Facile and highly efficient approach for the fabrication of multifunctional silk nanofibers containing hydroxyapatite and silver nanoparticles

In this study, a good combination consisting of electrospun silk fibroin nanofibers incorporated with high-purity hydroxyapatite (HAp) nanoparticles (NPs) and silver NPs is introduced as antimicrobial for tissue engineering applications. The variable pressure field emission scanning electron microscope results confirmed randomly placed nanofibers are produced with highly dispersed HAp and silver NPs in nanofibers after electrospinning. The X-ray diffraction results demonstrated crystalline features of each of the three components used for electrospinning. Moreover, the TEM-EDS analysis confirmed the presence and chemical nature of each component over individual silk nanofiber. The FT-IR analyses was used confirm the different vibration modes caused due to functional groups present in silk fibroin, Hap, and silver NPs. The obtained nanofibers were checked for antimicrobial activity by using two model organisms Escherichia coli and Staphylococcus aureus. Subsequently, the antimicrobial tests have indicated that prepared nanofibers do possess good bactericidal activity. The ability of N,N-dimethylformamide and silk fibroin used to reduce silver nitrate into silver metal was evaluated using MTT assay. The nanofibers were grown in presence of NIH 3T3 fibroblasts, which revealed toxic behavior to fibroblasts at higher concentrations of silver nitrate used in this study. Furthermore, cell attachment studies on nanofibers for 3 and 12 days of incubation time were minutely observed and correlated with the results of MTT assay. The reported results confirmed the high amounts of silver nitrate can lead to toxic effects on viability of fibroblasts and had bad effect in cell attachment.

Electrospun silk fibroin nanofibers in different diameters support neurite outgrowth and promote astrocyte migration

Nerve tissue engineering has been one of the promising strategies for regenerative treatment in patients suffering from neural tissue loss, but considerable challenges remain before it is able to progress toward clinical application. It has been demonstrated that transplantation of cells in combination with physically or chemically modified biomaterials provides better environments for neurite outgrowth and further promotes axonal regeneration in animal models of spinal cord injury. In this study, neurons and astrocytes were incorporated into 400-nm, 800-nm, and 1200-nm electrospun Bombyx mori silk fibroin (SF) materials to investigate the effects of scaffold-diameter in regulating and directing cell behaviors. β-III-tubulin immunofluorescence analyses reveal that SF nanofibers with smaller diameters are more favorable to the development and maturation of subventricular zone-derived neurons than 1200-nm SF scaffolds. In addition, astrocytes exhibited well-arranged glial fibrillary acidic protein (GFAP) expression on SF scaffolds, and a significant increase in cell-spreading area was observed on 400-nm but not 1200-nm SF scaffolds. Moreover, a significantly enhanced migration efficiency of astrocytes grown on SF scaffolds was verified, which highlights the guiding roles of SF nanofibers to the migratory cells. Overall, our results may provide valuable information to develop effective tissue remodeling substrates and to optimize existing biomaterials for neural tissue engineering applications.