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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   

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3.
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   

4.
In this study, a three-dimensional (3D) poly(lactide-co-glycolide) (PLGA) microfibrous scaffold with high porosity (ca. 90 % porosity) was developed for evaluating its performance in tissue engineering application. A dope solution of PLGA/polyethylene oxide (PEO) blend was electrospun into a methanol coagulation bath for fabricating highly porous 3D PLGA scaffold and a salt leaching method was used for making interconnected pores of 100?C200 ??m size inside the scaffold. The morphological structure, pore size and porosity of the microfibrous scaffold were determined, and compared with twodimensional (2D) mat-type and 3D sponge-type of PLGA scaffold. Also, swelling ratio, water uptake and compressive strength were compared in order to elucidate the structure-property relationships of different types of the scaffolds, especially in a wet condition. As a result of scanning electron microscopy (SEM) observation, normal human dermal fibroblasts (nHDF) were migrated, attached, and proliferated well inside the 3D scaffold. MTT assay confirmed that the highly porous 3D PLGA microfibrous scaffold had superior cell adhesion and proliferation abilities due to fibrous structure of large specific surface area, and interconnected pore structure. Therefore, this high performance 3D PLGA scaffold can have a high potentiality for application in tissue engineering in comparison with conventional PLGA scaffolds.  相似文献   

5.
In bone tissue regeneration, extracellular matrix (ECM) and bioceramics are important factors, because of their osteogenic potential and cell–matrix interactions. Surface modifications with hydrophilic material including proteins show significant potential in tissue engineering applications, because scaffolds are generally fabricated using synthetic polymers and bioceramics. In the present study, carbonated hydroxyapatite (CHA) and marine atelocollagen (MC) were extracted from the bones and skins, respectively, of Paralichthys olivaceus. The extracted CHA was characterized using Fourier transform infrared (FTIR) spectroscopy and X-ray diffraction (XRD) analysis, while MC was characterized using FTIR spectroscopy and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The scaffolds consisting of polycaprolactone (PCL), and different compositions of CHA (2.5%, 5%, and 10%) were fabricated using a three-axis plotting system and coated with 2% MC. Then, the MC3T3-E1 cells were seeded on the scaffolds to evaluate the osteogenic differentiation in vitro, and in vivo calvarial implantation of the scaffolds was performed to study bone tissue regeneration. The results of mineralization confirmed that the MC/PCL, 2.5% CHA/MC/PCL, 5% CHA/MC/PCL, and 10% CHA/MC/PCL scaffolds increased osteogenic differentiation by 302%, 858%, 970%, and 1044%, respectively, compared with pure PCL scaffolds. Consequently, these results suggest that CHA and MC obtained from byproducts of P. olivaceus are superior alternatives for land animal-derived substances.  相似文献   

6.
Particular attention has been given to axonal outgrowth of neurons to understand how topographical surface cues influence attachment and subsequent directional migration and growth. In present study, the silk fibroin (SF) scaffold with uniaxial channels was prepared by directional freeze-drying processes. The average pore diameter, the porosity, and pore density of the scaffold are 120 µm, 88 %, and 203 mm?2, respectively. Further, hippocampal neurons were seeded onto the scaffold and the hippocampal neurons morphology was investigated. Cell-cell networks and cell-matrix interactions had been established by newly formed axons and the diversity of neurons was much higher after culturing 7 days. The neurons expressed β-III-tubulin and nerve filament, while glial fibrillary acidic protein immunofluorescence was barely above background. These results indicated that the SF scaffolds with uniaxial multichannels could be guided axons of neurons spread along the channels. SF scaffolds with oriented pores have a potential for nerve tissue regeneration.  相似文献   

7.

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   

8.
The PLGA ultrafine fibrous scaffold was successfully fabricated by electrospinning. The morphology and properties of the PLGA vascular scaffolds were examined. In particular, the in vitro degradation behavior of the electrospun PLGA vascular scaffolds was investigated by means of morphology, microstructure, mass loss, Mw, and breaking strength characterization. The results showed that electrospun scaffold possessed ultrafine fibrous and porous structure, and had adequate mechanical properties to be developed as a substitute for native blood vessels. In vitro degradation study showed that the PLGA ultrafine fibrous scaffold could biodegrade in the PBS solution, and the mass loss, Mw, and breaking strength studies indicated that degradation rate of the electrospun PLGA nanofibers was greater in the first 2 weeks. After the degradation of 2 weeks, the degradation slowed down. Furthermore, with the extension of the degradation time, the thermal decomposition temperature of the PLGA scaffold decreased gradually. The results indicated that the electrospun PLGA vascular scaffold could be considered as an ideal candidate for tissue-engineered blood vessel.  相似文献   

9.
Background: Spinal cord has a limited capacity to repair; therefore, medical interventions are necessary for treatment of injuries. Transplantation of Schwann cells has shown a great promising result for spinal cord injury (SCI). However, harvesting Schwann cell has been limited due to donor morbidity and limited expansion capacity. Furthermore, accessible sources such as bone marrow stem cells have drawn attentions to themselves. Therefore, this study was designed to evaluate the effect of bone marrow-derived Schwann cell on functional recovery in adult rats after injury. Methods: Mesenchymal stem cells were cultured from adult rats’ bone marrow and induced into Schwann cells in vitro. Differentiation was confirmed by immunocytochemistry and RT-PCR. Next, Schwann cells were seeded into collagen scaffolds and engrafted in 3 mm lateral hemisection defects. For 8 weeks, motor and sensory improvements were assessed by open field locomotor scale, narrow beam, and tail flick tests. Afterwards, lesioned spinal cord was evaluated by conventional histology and immunohistochemistry. Results: In vitro observations showed that differentiated cells had Schwann cell morphology and markers. In this study, we had four groups (n = 10 each): laminectomy, control, scaffold and scaffold + Schwann cells. Locomotor and sensory scores of cell grafted group were significantly better than control and scaffold groups. In histology, axonal regeneration and remyelination were better than control and scaffold groups. Conclusion: This study demonstrates that bone marrow-derived Schwann cells can be considered as a cell source for Schwann cells in SCI treatment. Key Words: Rats, Spinal cord injuries (SCI), Bone marrow, Schwann cells, Cell transdifferentiation  相似文献   

10.
Background:Biomaterials used as cell growth stimulants should be able to provide adequate cell adhesion with no alteration in cell function. In this work, we developed a 3D model of mouse spinal cord motoneurons on scaffolds composed of electrospun PLA fibers and plasma-polymerized PPy-coated PLA fibers. Methods:The functionality of the cultured motoneurons was assessed by evaluating both the electrophysiological response (i.e., the whole-cell Na+ and K+ currents and the firing of action potentials) and also the expression of the VAChaT by immunostaining techniques. While the expression of the VAChaT was confirmed on motoneurons cultured on the fibrous scaffolds, the electrophysiological responses indicated Na+ and K+ currents with lower amplitude and slower action potentials when compared to the response recorded from spinal cord motoneurons cultured on Poly-DL-Ornithine/Laminin- and plasma-polymerized PPy-coated coverslips.Results:From a morphological viewpoint, motoneurons cultured on PLA and PPy-coated PLA scaffolds did not show the development of dendritic and/or axonal processes, which were satisfactorily observed in the bidimensional cultures.Conclusion:We hypothesize that the apparently limited development of dendritic and/or axonal processes could produce a deleterious effect on the electrophysiological response of the cells, which might be due to the limited growth surface available in the fibrous scaffolds and/or to an undesired effect of the purification process.Key Words: Electrophysiology, Pyrrol, Scaffolds  相似文献   

11.
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   

12.
Cartilage repair is a challenge in bone tissue reconstruction. In this study, silk fibroin (SF), chondroitin sulfate (CS) and hyaluronic acid (HA) were employed to fabricate scaffolds for tissue engineered cartilage by freeze drying technique. The secondary pores were formed in the main pores of SF/CS/HA scaffold which improved the pore connectivity and equilibrium swelling of the scaffold. Furthermore, rat bone marrow mesenchymal stem cells were seeded on the scaffolds to evaluate the cell adhesion and proliferation. Results of hematoxylin/eosin staining and cell counting kit-8 assay showed that the cells migration and differentiation of SF/CS/HA (80/15/5) scaffold were better than that of SF/CS/HA scaffolds with different ratios after 7 days culture. Moreover, immunohistochemistry and scanning electron microscope demonstrated that large amounts of collagen II and proteoglycans of the cells were expressed in the SF/CS/HA 3D scaffold, while the expression of collagen I was barely visible by immunohistochemistry. Abound of extracellular matrix was formed to morphologically round and distributed uniformly throughout the scaffolds. The 3D ternary scaffold could promote the cells chondrogenic differentiation without using any inductive agent and offer potential for cartilage tissue regeneration.  相似文献   

13.
Background:One of the main challenges with conventional scaffold fabrication methods is the inability to control scaffold architecture. Recently, scaffolds with controlled shape and architecture have been fabricated using 3D-printing. Herein, we aimed to determine whether the much tighter control of microstructure of 3DP PLGA/β-TCP scaffolds is more effective in promoting osteogenesis than porous scaffolds produced by solvent casting/porogen leaching. Methods:Physical and mechanical properties of porous and 3DP scaffolds were studied. The response of pre-osteoblasts to the scaffolds was analyzed after 14 days. Results:The 3DP scaffolds had a smoother surface (Ra: 22 ± 3 µm) relative to the highly rough surface of porous scaffolds (Ra: 110 ± 15 µm). Water contact angle was 112 ± 4° on porous and 76 ± 6° on 3DP scaffolds. Porous and 3DP scaffolds had the pore size of 408 ± 90 and 315 ± 17 µm and porosity of 85 ± 5% and 39 ± 7%, respectively. Compressive strength of 3DP scaffolds (4.0 ± 0.3 MPa) was higher than porous scaffolds (1.7 ± 0.2 MPa). Collagenous matrix deposition was similar on both scaffolds. Cells proliferated from day 1 to day 14 by fourfold in porous and by 3.8-fold in 3DP scaffolds. ALP activity was 21-fold higher in 3DP scaffolds than porous scaffolds. Conclusion:The 3DP scaffolds show enhanced mechanical properties and ALP activity compared to porous scaffolds in vitro, suggesting that 3DP PLGA/β-TCP scaffolds are possibly more favorable for bone formation. Key Words: Alkaline phosphatase, β-tricalcium phosphate, Poly(lactic-co-glycolic) acid copolymer  相似文献   

14.
Biomaterial bridges constructed from electrospun fibers offer a promising alternative to traditional nerve tissue regeneration substrates. Aligned and unaligned polycaprolactone (PCL) electrospun fibers were prepared and functionalized with the extracellular matrix proteins collagen and laminin using covalent and physical adsorption attachment chemistries. The effect of the protein modified and native PCL nanofiber scaffolds on cell proliferation, neurite outgrowth rate, and orientation was examined with neuronlike PC12 cells. All protein modified scaffolds showed enhanced cellular adhesion and neurite outgrowth compared to unmodified PCL scaffolds. Neurite orientation was found to be in near perfect alignment with the fiber axis for cells grown on aligned fibers, with difference angles of less than 7° from the fiber axis, regardless of the surface chemistry. The bioavailability of PCL fibers with covalently attached laminin was found to be identical to that of PCL fibers with physically adsorbed laminin, indicating that the covalent chemistry did not change the protein conformation into a less active form and the covalent attachment of protein is a suitable method for enhancing the biocompatibility of tissue engineering scaffolds.  相似文献   

15.
The main objective of this work was chemically bioactivation of the conducting polyanizidine (PANIZ) by incorporating a polyester such as polycaprolactone (PCL). Modified PANIZ nanocomposites were synthesized through ring opening and chemical oxidation polymerizations. A four-point probe was applied to measure the conductivity of newly synthesized star-like block copolymer (S-PCL-PANIZ) nanocomposite, which was about 0.44 S cm-1. Conductive biodegradable nanofibers were prepared by electrospinning with 25 and 75 % (wt/wt) S-PCL-PANIZ to PCL. The contact angle of each prepared nanofiber was 87±3°, supporting their usefulness for cell culture. The cultured mouse osteoblast MG63 cells demonstrated normal morphology and significantly higher adhesion and spreading on the nanofiber. The bioactivated PANIZ based nanocomposite may be fruitful in tissue engineering to fabricate conducting biodegradable scaffolds with improved cell adhesion properties for various cell cultures.  相似文献   

16.
Background:Solvent casting/particulate leaching is one of the most conventional methods for fabricating polymer/ceramic composite scaffolds. In this method, the solvent generally affects resulting scaffold properties, including porosity and degradation rate. Methods:Herein, composite scaffolds of PLGA/nHA with different percentages of nHA (25, 35, and 45 wt. %) were prepared by the solvent casting/particle leaching combined with freeze drying. The effects of two different solvents, DIO and NMP, on morphology, porosity, bioactivity, degradation rate, and biocompatibility of the resulting scaffolds were investigated. Results:The results revealed that increasing the nHA percentages had no significant effect on the porosity and interconectivity of scaffolds (p > 0.05), whereas altering the solvent from DIO into NMP decreased the porosity from about 87% into 71%, respectively. Moreover, scaffolds of DIO illustrated the high results of cell proliferation compared to those of NMP; the cell viability of GD25 decreased from 85% to 65% for GN25. The findings also indicated that scaffolds prepared by NMP had a higher rate of losing weight in comparison to DIO. Adding nHA to PLGA had a significant effect on the bioactivity of scaffolds (p < 0.05), composite scaffolds with 45 wt % nHA had at least 30% more weight gain compared to the neat polymer scaffolds. Conclusion:The DIO scaffolds have higher rates of porosity, interconnectivity, bioactivity, and biocompatibility than NMP scaffolds due to its high evaporation rate. Key Words: Freeze drying, Porosity, Solvents  相似文献   

17.
The study aims at performing a comparative assessment of two types of burn wound treatment. The present study was designed to prepare crosslinked and blended two natural polymers nanofiber scaffolds using gelatin (GE) and hyaluronic acid (HA). The GE/HA composite nanofibrous membranes with varied GE/HA weight ratio have also been successfully fabricated by an electrospinning method. The average diameter of GE/HA fibers was in the range of 20 to 150 nm. In vivo efficacy was also investigated based on a deep second degree burns model for Wistar rats. At 14 days post-operation, the dermal defect basically recovered its normal condition. A percentage of wound closure of GE/HA composite nanofibrous membranes and ChitoHeal gel reached up to 81.9 % and 77.8 % respectively, compared with 65 % of the untreated control (p<0.05). Also, histological parameters were assessed on postoperative day 7 and 14. The results of in vivo experiments showed that more epidermis was formed in the gel and scaffold groups compared to the control group. The numbers of inflammatory cells in these two groups were also smaller as compared with the control group, which could well be the reason for the delayed healing in the control group.  相似文献   

18.
Electrical conductive nanocomposite fibers were prepared with polyaniline (PANI), polyacrylonitrile (PAN) and multi-walled carbon nanotubes (MWCNTs) via electrospinning. The morphology and electrical conductivity of the PANI/PAN/MWCNTs nanocomposite fibers were characterized by scanning electron microscope (SEM) and Van De Pauw method. Electrical conductivity of nanocomposite fibers increased from 1.79 S·m?1 to 7.97 S·m?1 with increasing the MWCNTs content from 3.0 wt% to 7.0 wt%. Compared with PANI/PAN membranes, the mechanical property of PANI/PAN/MWCNTs nanocomposites fiber membranes decreased. The microwave absorption performance of composite films was analyzed using waveguide tube, which indicated that with the thickness increasing the value of RL reduced from ?4.6 to ?5.9 dB.  相似文献   

19.
Background:Freeze dried bone allograft nanoparticles on a nanofiber membrane may serve as an ideal scaffold for bone regeneration. This study aimed to assess the biological behavior of human MSCs in terms of proliferation and adhesion to nanoparticulate and microparticulate FDBA scaffolds on PLLA nanofiber membrane. Methods:In this experimental study, PLLA nanofiber scaffolds were synthesized by the electrospinning method. The FDBA nanoparticles were synthesized mechanically. The FDBA nanoparticles and microparticles were loaded on the surface of PLLA nanofiber membrane. A total of 64 scaffold samples in four groups of n-FDBA/PLLA, FDBA/PLLA, PLLA and control were placed in 24-well polystyrene tissue culture plates; 16 wells were allocated to each group. Data were analyzed using one-way ANOVA and Bonferroni test. Results:The proliferation rate of MSCs was significantly higher in the nanoparticulate group compared to the microparticulate group at five days (p = 0.034). Assessment of cell morphology at 24 hours revealed spindle-shaped cells with a higher number of appendages in the nanoparticulate group compared to other groups. Conclusion:MSCs on n-FDBA/PLLA scaffold were morphologically more active and flatter with a higher number of cellular appendages, as compared to FDBA/PLLA. It seems that the nanoparticulate scaffold is superior to the microparticulate scaffold in terms of proliferation, attachment, and morphology of MSCs in vitro.Key Words: Allografts, Bone regeneration, Mesenchymal stem cells, Nanofibers  相似文献   

20.
In this study, biodegradable poly(D,L-lactic-co-glycolic acid) (PLGA) fibers were prepared by a melt-electrospinning and treated with plasma in the presence of either oxygen or ammonia gas to modify the surface of the fibers. The effects of processing parameters on the melt-electrospinning of PLGA were examined in terms of fiber morphology and diameter. Among the processing parameters, the spinning temperature and mass flow rate had a significant effect on the average fiber diameter and its distribution. The water contact angle of melt-electrospun PLGA fibers decreased significantly from 123 ° to 55 ° (oxygen plasma treatment) or to 0 ° (ammonia plasma treatment) by plasma treatment for 180 sec, while their water content increased significantly from 2.4 % to 123 % (oxygen plasma treatment) or to 189 % (ammonia plasma treatment). Ammonia gas-plasma enhanced the surface hydrophilicity of PLGA fibers more effectively compared to oxygen gas-plasma. X-ray photoelectron spectroscopy analysis supported that the number of polar groups, such as hydroxyl and amino groups, on the surface of PLGA fibers increased after plasma treatment. Overall, the microfibrous PLGA scaffolds with appropriate surface hydrophilicity and fiber diameter could be fabricated by melt electrospinning and subsequent plasma treatment, without a significant deterioration of fiber structure and dimensional stability. This approach of controlling the surface properties and structures of fibers could be useful in the design and tailoring of novel scaffolds for tissue engineering.  相似文献   

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