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1.
Application of electrospun nanofibrous scaffolds has received immense attention in tissue engineering. Fabrication of scaffolds with appropriate electrical properties plays a key role in neural tissue engineering. Since fibers orientation in the scaffolds affects the growth and proliferation of the cells, this study aimed to prepare aligned electrospun conductive nanofibers by mixing 1 %, 10 % and 18 % (w/v) doped polyaniline (PANI) with polycaprolactone (PCL)/poly lactic-coglycolic acid (PLGA) (25/75) solution through the electrospinning process. The fibers diameter, hydrophilicity and conductivity were measured. In addition, the shape and proliferation of the nerve cells seeded on fibers were evaluated by MTT cytotoxicity assay and scanning electron microscopy. The results revealed that the conductive nanofibrous scaffolds were appropriate substrates for the attachment and proliferation of nerve cells. The electrical stimulation enhanced neurite outgrowth compared to those PLGA/PCL/PANI scaffolds that were not subjected to electrical stimulation. As polyaniline ratio increases, electric stimulation through nanofibrous PLGA/PCL/PANI scaffolds results in cell proliferation enhancement. However, a raise more than 10 % in polyaniline will result in cell toxicity. It was concluded that conductive scaffolds with appropriate ratio of PANI along with electrical stimulation have potential applications in treatment of spinal cord injuries.  相似文献   

2.
In this study, the in vitro degradation of yarns made from PCL electrospun fibers containing various concentrations of ampicillin sodium salt was investigated. PCL fibers were electrospun and collected as well-aligned fiber bundles and then twisted into yarns. Yarn weight loss, morphology changes in the yarns, and morphology changes in the fiber microstructure with degradation were evaluated. Results showed that the electrospun PCL yarns degraded slowly with a weight loss less than 3 % in 12 weeks. The addition of various concentrations of ampicillin salt increased the degradation rate slightly. The morphological changes observed in fiber microstructure suggested that the degradation underwent bulk erosion and the degradation began with the amorphous regions. Revealed by the fiber morphological changes with degradation, the microstructure of electrospun PCL fibers followed the fringed fibril fiber model. The fiber arrangement in the yarn was impaired by the in vitro degradation environment as well.  相似文献   

3.

Background

The aim of this study was to fabricate the poly caprolactone (PCL) aligned nanofiber scaffold and to evaluate the survival, adhesion, proliferation, and differentiation of rat hair follicle stem cells (HFSC) in the graft material using electrospun PCL nanofiber scaffold for tissue engineering applications.

Methods

The bulge region of rat whisker was isolated and cultured in DMEM: nutrient mixture F-12 supplemented with epidermal growth factor. The morphological and biological features of cultured bulge cells were observed by light microscopy using immunocytochemistry methods. Electrospinning was used for production of PCL nanofiber scaffolds. Scanning electron microscopy (SEM), 3-(4, 5-di-methylthiazol- 2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, and histology analysis were used to investigate the cell morphology, viability, attachment and infiltration of the HFSC on the PCL nanofiber scaffolds.

Results

The results of the MTT assay showed cell viability and cell proliferation of the HFSC on PCL nanofiber scaffolds. SEM microscopy images indicated that HFSC are attached, proliferated and spread on PCL nanofiber scaffolds. Also, immunocytochemical analysis showed cell infiltration and cell differentiation on the scaffolds.

Conclusion

The results of this study reveal that PCL nanofiber scaffolds are suitable for cell culture, proliferation, differentiation and attachment. Furthermore, HFSC are attached and proliferated on PCL nanofiber scaffolds.Key Words: Nanofiber, Electrospinning, Stem cells, Tissue engineering  相似文献   

4.
Tissue engineering is an interdisciplinary field which combines the basic principles of life sciences and engineering. One promising idea is the combination of scaffolds and living cells in order to produce new functional tissue. The scaffolds play the role of a microenvironment that guides the cells towards tissue formation and regeneration. One of the most frequently used techniques to produce scaffolds is electrospinning. Tissue engineered constructs have to exhibit physiological and mechanical properties comparable to the native tissue they are intended to replace. To create polymeric fibers with controlled orientation, a cylindrical collector that rotates at a certain speed could be used, creating fibers that run longitudinally. The process of gap-spinning enables the production of specifically aligned fibers. Aim of this study was to develop a novel setup capable of producing multilayered structures with controlled fiber angle. The structural, morphological and mechanical characteristics of the fibers were accessed using scanning electron microscopy and uniaxial tensile tests. Longer pre-stretching led to thinner (in the sub-micron scale), more brittle and less elastic fibers. In a nutshell, the results indicated that fiber mats of desired orientation, fiber diameter and mechanical properties could be produced by controlled gap-spinning with a translational collector.  相似文献   

5.
Electrospun fiber scaffolds crafted from polyesters are studied extensively for potential tissue engineering applications. For translation of electrospun fibers into the clinic, the FDA requires analysis and quantification of any organic solvent that may be retained in the fibers since many organic solvents can negatively affect cells and tissues. If a significant amount of solvent is retained, then developing procedures for efficient solvent removal may enhance the clinical potential of these materials. In this study we use fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), and nuclear magnetic resonance spectroscopy (NMR) to analyze solvent retention. A correlative analysis shows that both FTIR and TGA accurately predicted retention of two different solvents (HFP and chloroform) in our electrospun PLLA scaffolds, thus validating these procedures. We also assess the efficacy of various fiber treatment methods to facilitate organic solvent removal and conclude that submersion in 70 % ethanol and heat treatment at 100 °C were the most efficient methods of removing solvent from electrospun PLLA fibers.  相似文献   

6.
This article focuses on elucidating the key presentation features of neurotrophic ligands at polymer interfaces. Different biointerfacial configurations of the human neural cell adhesion molecule L1 were established on two-dimensional films and three-dimensional fibrous scaffolds of synthetic tyrosine-derived polycarbonate polymers and probed for surface concentrations, microscale organization, and effects on cultured primary neurons and neural stem cells. Underlying polymer substrates were modified with varying combinations of protein A and poly-D-lysine to modulate the immobilization and presentation of the Fc fusion fragment of the extracellular domain of L1 (L1-Fc). When presented as an oriented and multimeric configuration from protein A-pretreated polymers, L1-Fc significantly increased neurite outgrowth of rodent spinal cord neurons and cerebellar neurons as early as 24 h compared to the traditional presentation via adsorption onto surfaces treated with poly-D-lysine. Cultures of human neural progenitor cells screened on the L1-Fc/polymer biointerfaces showed significantly enhanced neuronal differentiation and neuritogenesis on all protein A oriented substrates. Notably, the highest degree of βIII-tubulin expression for cells in 3-D fibrous scaffolds were observed in protein A oriented substrates with PDL pretreatment, suggesting combined effects of cell attachment to polycationic charged substrates with subcellular topography along with L1-mediated adhesion mediating neuronal differentiation. Together, these findings highlight the promise of displays of multimeric neural adhesion ligands via biointerfacially engineered substrates to "cooperatively" enhance neuronal phenotypes on polymers of relevance to tissue engineering.  相似文献   

7.
The complex nature of spinal cord injuries has provided much inspiration for the design of novel biomaterials and scaffolds which are capable of stimulating neural tissue repair strategies. Recently, conductive polymers have gained much attention for improving the nerve regeneration. In our previous study, a three-dimensional (3D) structure with reliable performance was achieved for electrospun scaffolds. The main purpose in the current study is formation of electrical excitable 3D scaffolds by appending polyaniline (PANI) to biocompatible polymers. In this paper, an attempt was made to develop conductive nanofibrous scaffolds, which can simultaneously present both electrical and topographical cues to cells. By using a proper 3D structure, two kinds of conductive scaffolds are compared with a non-conductive scaffold. The 3D nanofibrous core-sheath scaffolds, which are conductive, were prepared with nanorough sheath and aligned core. Two different sheath polymers, including poly(lactic-co-glycolic acid) PLGA and PLGA/PANI, with identical PCL/PANI cores were fabricated. Nanofibers of PCL and PLGA blends with PANI have fiber diameters of 234±60.8 nm and 770±166.6 nm, and conductivity of 3.17×10-5 S/cm and 4.29×10-5 S/cm, respectively. The cell proliferation evaluation of nerve cells on these two conductive scaffolds and previous non-conductive scaffolds (PLGA) indicate that the first conductive scaffold (PCL/ PANI-PLGA) could be more effective for nerve tissue regeneration. Locomotor scores of grafted animals by developed scaffolds showed significant performance of non-conductive 3D scaffolds. Moreover, the animal studies indicated the ability of two new types of conductive scaffolds as spinal cord regeneration candidates.  相似文献   

8.
Flexible tubular structures fabricated from solution electrospun fibers are finding increasing use in tissue engineering applications. However it is difficult to control the deposition of fibers due to the chaotic nature of the solution electrospinning jet. By using non-conductive polymer melts instead of polymer solutions the path and collection of the fiber becomes predictable. In this work we demonstrate the melt electrospinning of polycaprolactone in a direct writing mode onto a rotating cylinder. This allows the design and fabrication of tubes using 20 μm diameter fibers with controllable micropatterns and mechanical properties. A key design parameter is the fiber winding angle, where it allows control over scaffold pore morphology (e.g. size, shape, number and porosity). Furthermore, the establishment of a finite element model as a predictive design tool is validated against mechanical testing results of melt electrospun tubes to show that a lesser winding angle provides improved mechanical response to uniaxial tension and compression. In addition, we show that melt electrospun tubes support the growth of three different cell types in vitro and are therefore promising scaffolds for tissue engineering applications.  相似文献   

9.
In this study, electrospinning of poly(ε-caprolactone) (PCL) and its optimum preparation conditions were examined in detail using various solvent systems, such as formic acid, dichloromethane/dimethyl formamide (DMF), chloroform/DMF, and dichloroethane. The average fiber diameter of the electrospun PCL mat was controlled by the solvent used with a proper concentration of PCL dope solution. Different fiber sizes (0.1, 0.8, 1.9, and 3.4 μm) of uniform PCL mats were fabricated and the effects of fiber size on surface morphology, tensile properties and cell behavior were investigated. Here, we manipulated much broader range of average fiber diameter of the mats, from nano to several micron size of fiber. It was found that the fiber diameter greatly affected topology (surface roughness) and mechanical properties of the electrospun PCL mat and consequently, they influenced the cell behavior (adhesion and proliferation) significantly. We expect that these results will provide more feasible application of electrospun PCL scaffold in tissue engineering through the co-relations in structure and property of PCL fiber mat on cell behavior.  相似文献   

10.
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11.
A major goal of biomimetics is the development of chemical compositions and structures that simulate the extracellular matrix. In this study, gelatin-based electrospun composite fibrous membranes were prepared by electrospinning to generate bone scaffold materials. The gelatin-based multicomponent composite fibers were fabricated using co-electrospinning, and the composite fibers of chitosan (CS), gelatin (Gel), hydroxyapatite (HA), and graphene oxide (GO) were successfully fabricated for multi-function characteristics of biomimetic scaffolds. The effect of component concentration on composite fiber morphology, antibacterial properties, and protein adsorption were investigated. Composite fibers exhibited effective antibacterial activity against Staphylococcus aureus and Escherichia coli. The study observed that the composite fibers have higher adsorption capacities of bovine serum albumin (BSA) at pH 5.32–6.00 than at pH 3.90–4.50 or 7.35. The protein adsorption on the surface of the composite fiber increased as the initial BSA concentration increased. The surface of the composite reached adsorption equilibrium at 20 min. These results have specific applications for the development of bone scaffold materials, and broad implications in the field of tissue engineering.  相似文献   

12.
Artificial keratoprostheses are indispensable for visual rehabilitation in patients with end-stage corneal blindness. This study aimed to assess the biocompatibility of polyethylene terephthalate nanofibrous mats and its potential as a novel synthetic keratoprosthesis skirt material for corneal tissue engineering. Nanofibrous mats were prepared by an electrospinning method and were first treated with the CO2 plasma to yield carboxylic groups on the surface; finally, the modified PET mat was cross linked with collagen using water-soluble carbodiimide as a coupling agent. The samples were evaluated by ATR-FTIR, scanning electron microscope (SEM), contact angle, and cell culture. The cross-linking of collagen on PET surface was confirmed by ATR-FTIR spectroscopy and SEM images The 79° difference was obtained in the contact angle analysis, obtained for the collagen-cross-linked nanofibrous mat than the non-modified nanofibrous mat. Cellular investigation showed limbal epithelial progenitor cells (LEPCs) has been better adhesion, cell growth, and proliferation of collagen-crosslinked nanofibrous samples than other samples. The bioavailability of PET fibers with covalently attached collagen was found to be identical to that of PET fibers with covalent attachment is a suitable method for enhancing the biocompatibility of scaffolds special as a good skirt in keratoprosthesis designs.  相似文献   

13.
Electrospinning has been recognized as an efficient technique for the fabrication of neural tissue engineering scaffolds. Many approaches have been developed on material optimization, electrospinning techniques, and physical properties of scaffolds to produce a suitable scaffold for tissue engineering aspects. In this study, structural properties of scaffolds were promoted by controlling the speed of fiber collection without any post-processing. PLGA scaffolds, in two significantly different solution concentrations, were fabricated by the electrospinning process to produce scaffolds with the optimum nerve cell growth in a desired direction. The minimum, intermediate and maximum rate of fiber collection (0.4, 2.4, 4.8 m/s) formed Random, Aligned and Drown-aligned fibers, with various porosities and hydrophilicities. The scaffolds were characterized by fiber diameter, porosity, water contact angle and morphology. Human nerve cells were cultured on fiber substrates for seven days to study the effects of different scaffold structures on cell morphology and proliferation, simultaneously. The results of MTT assay, the morphology of cells and scaffold characterization recommend that the best structure to promote cell direction, morphology and proliferation is accessible in an optimized hydrophilicity and porosity of scaffolds, which was obtained at the collector linear speed of 2.4 m/s.  相似文献   

14.
Electrospinning is a straightforward method to produce sub-micrometer or nanoscale fiber. Polycaprolactone (PCL), an important polymer in biomedical applications, has been electrospun in several solvent systems. N,Ndimethylformamide (DMF) is often used as an additive in the solvent system to prepare PCL nanofibers. The adding of the DMF changes the physical properties of the solution. To trace and understand the influence of these changes on the jet formation as well as the resultant fibers morphologies, a model of jet perturbation based on the Plateau-Rayleigh Instability theory was established to explicate the formation of the particles/fibers and some experiments for testing the solution properties and fibers morphologies were carried out. With the adding of DMF in dichloromethane (DCM)/DMF mixed solvents, the solution surface tensions increase while solution viscosities decrease, which triggers the change of electrospinning to electrospraying in general. However, according to the obtained results, the addition of the DMF makes it easier to induce the transformation of particles electrospraying to fibers electrospinning with smaller diameter. This is attributed to the higher dielectric constant, lower vapor pressure, and higher electric conductivity of DMF. The theoretical model and experimental results strengthen the relations of solution properties, jet moving behaviors, and the resultant fiber morphologies.  相似文献   

15.
Electrospun atactic polypropylene (PP) fibers are thicker than those obtained from isotactic PP, although the viscosity of molten PPs is almost same. Thus we focused on the effect of tacticity of PP on fiber diameters. The PP samples with various tacticity were prepared by changing the blend ratio of isotactic PP and atactic PP. Melt-electrospinning is performed by using blended samples, and then electrospun fibers were observed by scanning electron microscope to evaluate fiber diameter of obtained fibers. It is clear that the diameter of electrospun PP fibers decreases as high tacticity content of PP increases. This result suggests that tacticity of samples is an important factor to control the electrospun fiber diameter.  相似文献   

16.
In this study, to improve the cellular biocompatibility of PVP-PCL micro- and nanofiber scaffold, a novel electrospun collagen/PVP-PCL micro- and nanofiber scaffold was sucessfully prepared assisted by ultrasonic irradiation using chloroform/ethanol mixtures as solvent. The micro- and nanofibers of the electrospun PCL-PVP scaffolds still presented compact inter-fiber entanglement and three-dimensional netlike network with some certain range of pore space after introducing collagen. The incorporated collagen phase was dispersed as inclusions within the electrospun fibers, and then could be easily released by immersing the scaffold in Hanks simulated body fluid. Meanwhile, the integral triple helix structure of collagen could be maintained after blending with the PVP-PCL mixture due to the weak intermolecular interactions. Furthermore, the suitable mechanical and degradation properties of the PVP-PCL scaffold were still reserved after introducing collagen, and the introduction of collagen could further promote the thermostability of the PVP-PCL scaffold. Above all, the collagen/PVP-PCL scaffold showed no cytotoxicity, better cell proliferation, and improved viability of primary fibroblasts than the PVP-PCL scaffold. In conclusion, blending collagen with the PVP-PCL mixture in this study has potential for promoting the biocompatibility of PVP-PCL micro- and nanofiber scaffolds for tissue engineering.  相似文献   

17.
Photoluminescence electrospun fibers were prepared from poly(aryl ether)s solutions. The porosity and wrinkle fibers could be observed by scanning electron microscopy (SEM). The effect of solution properties on fiber surface morphologies was studied. Meanwhile, the rough fiber surfaces could make the electrospun membranes possess water repellency. The contact angles of electrospun membranes for water were around 140°. The emission spectra of these membranes indicated that the fibers exhibited multi-color including sapphire blue, olive green and rose red. It could provide a proposal for improving flexible optoelectronic devices based on electrospun membranes of conjugated polymers.  相似文献   

18.
While electrospun chitosan membranes modified to retain nanofibrous morphology have shown promise for use in guided bone regeneration applications in in vitro and in vivo studies, their mechanical tear strengths are lower than commercial collagen membranes. Elastin, a natural component of the extracellular matrix, is a protein with extensive elastic property. This work examined the incorporation of elastin into electrospun chitosan membranes to improve their mechanical tear strengths and to further mimic the native extracellular composition for guided bone regeneration (GBR) applications. In this work, hydrolyzed elastin (ES12, Elastin Products Company, USA) was added to a chitosan spinning solution from 0 to 4 wt% of chitosan. The chitosan–elastin (CE) membranes were examined for fiber morphology using SEM, hydrophobicity using water contact angle measurements, the mechanical tear strength under simulated surgical tacking, and compositions using Fourier-transform infrared spectroscopy (FTIR) and post-spinning protein extraction. In vitro experiments were conducted to evaluate the degradation in a lysozyme solution based on the mass loss and growth of fibroblastic cells. Chitosan membranes with elastin showed significantly thicker fiber diameters, lower water contact angles, up to 33% faster degradation rates, and up to seven times higher mechanical strengths than the chitosan membrane. The FTIR spectra showed stronger amide peaks at 1535 cm−1 and 1655 cm−1 in membranes with higher concentrated elastin, indicating the incorporation of elastin into electrospun fibers. The bicinchoninic acid (BCA) assay demonstrated an increase in protein concentration in proportion to the amount of elastin added to the CE membranes. In addition, all the CE membranes showed in vitro biocompatibility with the fibroblasts.  相似文献   

19.
In recent decades, tremendous research has focused on the production of nanoscale fibers using synthetic polymers, with the goal of fabricating nanofibrous scaffolds for wound healing. However, the hydrophobicity of such polymers typically hinders attachment and proliferation of the cells. In this study, we combined poly-d,l-lactide-co-glycolide (PLGA) and small intestine submucosa (SIS) to fabricate blended nanofibers for wound healing by electrospinning. PLGA and SIS were dissolved in 1,1,1,3,3,3-hexafluoro isopropanol to produce different weight ratios of PLGA/SIS-blended nanofibrous membranes (NFM). Physicochemical characterization of the electrospun NFM was performed by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy, water contact angle analysis, degradation test and tensile testing. The PLGA/SIS-blended NFM showed improved hydrophilicity and tensile strength. Better infiltration, attachment and proliferation of rat granulation fibroblasts of PLGA/SIS-blended NFMs compared to PLGA NFMs were identified by morphological differences determined by SEM and a water-soluble tetrazolium salt assay kit. Based on our results, the PLGA/SIS blended NFMs were found to be suitable for use as a potential material for wound dressing.  相似文献   

20.
The development of biomaterials with controllable interfacial features which have the capability to instruct cellular behavior are required to produce functional scaffolds for the treatment of spinal cord injury (SCI). Here, poly-?-caprolactone surfaces were biofunctionalized via layer-by-layer (LbL) deposition. The polyelectrolytes employed in this LbL technique were heparin and poly-L-lysine (PLL), the latter being chosen to improve cell adhesion and the subsequent cellular function of in vitrocultured neural progenitor cells. Material characterization results confirmed the deposition of well structured multilayers. Cell culture studies revealed significant differences in the cellular response to these adhesive/nonadhesive (PLL/heparin) polyelectrolyte multilayer (PEM)surfaces, with neurite outgrowth being significantly promoted on the PLL terminating layers. In addition, brain derived neurotrophic factor (BDNF) was adsorbed onto the LbL surfaces. This combined chemical and biological effect was then characterized in terms of neurite length along with the full length/truncated isoform 1 tyrosine kinase receptor (TrkB-FL/TrkB-T1) and growth associated protein-43 mRNA levels. Here, the authors report the differential effect of adsorbed and soluble BDNF of different concentrations. Adsorbed BDNF promoted neurite outgrowth and led to elevated, sustained TrkB mRNA levels. These findings highlight the potential of PEM biofunctionalized surfaces with integrated chemical and neurotrophin supportive cues to overcome SCI inhibitory environments and to promote regeneration.  相似文献   

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