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1.

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   

2.
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   

3.
Produced via electrospinning, polyurethane (PU) scaffolds have always attracted the interest of medical applications due of their unique properties such as good adhesion, biocompatibility and excellent mechanical strength. However, the poor hydrophilicity and hemocompatibility of PU presented a problem during PU’s application in the manufacturing of biomedical materials. We hypothesized that the incorporation of polyethylene glycol (PEG) and phosphatidylcholine (PC) into electrospinning solution of PU could improve the cell affinity and hemocompatibility. This research focused on fabricating hybrid PU-PEG and PU-PC random/aligned scaffolds through electrospinning technique and comparing their properties as a potential biocompatible scaffold for vascular tissue engineering. PC was doped into a PU solution in order to prepare an electrospun scaffold through the electrospinning technology while crosslinked electrospun PUPEG hybrid scaffolds were fabricated by photoinduced polymerization. The contact angle dramatically decreased from 122.3±0.8° to 39.1±0.8° with doping of PC in electrospinning solution while it decreased from 122.3±0.8° to 41.6±0.8° with doping of PEG. Furthermore, the mechanical properties of PU scaffolds were altered significantly by the addition of PC. The hemolysis and cytocompatibility assays demonstrated that these composite scaffolds could potentially be used as a smalldiameter vascular graft.  相似文献   

4.
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   

5.
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   

6.
Osteoarthritis (OA) is a multifactorial disease leading to degeneration of articular cartilage, causing morbidity in approximately 8.5 million of the UK population. As the dense extracellular matrix of articular cartilage is primarily composed of collagen, cartilage repair strategies have exploited the biocompatibility and mechanical strength of bovine and porcine collagen to produce robust scaffolds for procedures such as matrix-induced chondrocyte implantation (MACI). However, mammalian sourced collagens pose safety risks such as bovine spongiform encephalopathy, transmissible spongiform encephalopathy and possible transmission of viral vectors. This study characterised a non-mammalian jellyfish (Rhizostoma pulmo) collagen as an alternative, safer source in scaffold production for clinical use. Jellyfish collagen demonstrated comparable scaffold structural properties and stability when compared to mammalian collagen. Jellyfish collagen also displayed comparable immunogenic responses (platelet and leukocyte activation/cell death) and cytokine release profile in comparison to mammalian collagen in vitro. Further histological analysis of jellyfish collagen revealed bovine chondroprogenitor cell invasion and proliferation in the scaffold structures, where the scaffold supported enhanced chondrogenesis in the presence of TGFβ1. This study highlights the potential of jellyfish collagen as a safe and biocompatible biomaterial for both OA repair and further regenerative medicine applications.  相似文献   

7.
This paper reports on the preparation and characterization of nanofibers and nanofiber/film composites fabricated by electrospinning and dip-coating. The polymers in this study consist of polyurethane, nylon-6, and silicone. Scanning electron microscopy (SEM), fiber distribution, X-ray diffraction (XRD) analysis, Fourier transform infrared spectroscopy (FTIR) and tensile tests were conducted. The electrospun nylon-6 nanofiber/dip-coated silicone film (dried for 5 min) showed the optimum tensile strength and strain results, showing an increase in tensile strength of 63 % compared to pure nylon-6 nanofiber alone. XRD and FTIR verified the presence of individual polymers in the composite matrix. The electrospun PU nanofiber produced the biggest fiber diameter, while electrospun nylon-6, and PU/nylon-6 produced uniform fiber diameters, with PU/nylon-6 obtaining very random and curved fiber morphology.  相似文献   

8.
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   

9.
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.  相似文献   

10.
In this study, electrospun wool keratose (WK)/silk fibroin (SF) blend nanofiber was prepared and evaluated as a heavy metal ion adsorbent which can be used in water purification field. The WK, which was a soluble fraction of oxidized wool keratin fiber, was blended with SF in formic acid. The electrospinnability was greatly improved with an increase of SF content. The structure and properties of WK/SF blend nanofibers were investigated by SEM, FTIR, DMTA and tensile test. Among various WK/SF blend ratios, 50/50 blend nanofiber showed an excellent mechanical property. It might be due to some physical interaction between SF and WK molecules although FTIR result did not show any evidence of molecular miscibility. As a result of metal ion adsorption test, WK/SF blend nanofiber mats exhibited high Cu2+ adsorption capacity compared with ordinary wool sliver at pH 8.5. It might be due to large specific surface area of nanofiber mat as well as numerous functional groups of WK. Consequently, the WK/SF blend nanofiber mats can be a promising candidate as metal ion adsorption filter.  相似文献   

11.
As a kind of high-performance fibers, PTFE fiber has been widely used in many fields because of its unique characteristics. In this study, the poly(tetrafloroethylene) (PTFE) nanofibers manufactured by electrospinning method was reported. The gel-spinning solution of poly(tetrafluoroethylene)/poly(vinyl alcohol)/boric acid (PTFE/PVA/BA), which was prepared by the gel process of the mixture of PTFE, PVA, BA and redistilled water, was electrospun to form PTFE/PVA/BA composite nanofibers. After calcinating, the PTFE nanofibers with diameters of 200 nm to 1000 nm were obtained. The fibers before and after calcinating were characterized by scanning electron microscopy (SEM), thermogravimetric analysis (TGA), X-ray diffraction (XRD), FT-IR spectrum analysis and X-ray photoelectron spectroscopy (XPS), respectively, and the mechanical and hydrophobic properties of the fibers were also investigated. The results showed that the PTFE nanofiber membranes could be electrospun effectively used the gel-spinning solution of PTFE/PVA/BA, and may realize the applications in the fields of high-temperature filtration, catalyst supports, battery separator and so on.  相似文献   

12.
As a biomaterial, besides excellent biocompatibility and biodegradability, suitable macropores and pores structure should be provided to guide cell extension and migration. In present study, the silk fibroin (SF) scaffold with uniaxial channels was prepared by directional temperature field freezing technique. The average pore diameter, pore density and porosity of the scaffold with oriented channels are ~128.7 µm, ~158 mm?2 and ~91.4 %, respectively. By controlling of the temperature gradient direction, the oriented multichannels of the scaffolds were formed in longitudinal easily. In process of the scaffolds fabrication, the directional growth of ice crystal could shear and draft to the silk fibroin molecule segments, which resulted in the new crystal nucleus formation in new zone and increase of β-sheet components in the scaffolds. In vitro, L929 cells were seeded on the scaffolds with oriented channels to evaluate the effect on cell behavior. Cell viability, adhesion and morphology were determined by methyl thiazolyl tetrazolium, confocal microscope and scanning electron microscope. The results showed that the cells anchored on the oriented channels, spread along the direction of the channels and hold a higher viability on the scaffolds with oriented channels. These new oriented multichannel scaffold could guide the adhesion and proliferation of L929 cells, which hold a potential in tissue engineering.  相似文献   

13.
Biopolymers, in particular collagen and fibrinogen, are the leading materials for use in tissue engineering. When developing technology for scaffold formation, it is important to understand the properties of the source materials as well as the mechanisms that determine the formation of the scaffold structures. Both factors influence the properties of scaffolds to a great extent. Our present work aimed to identify the features of the molecular characteristics of collagens of different species origin and the changes they undergo during the enzymatic hydrolysis used for the process of scaffold formation. For this study, we used the methods of gel-penetrating chromatography, dynamic light scattering, reading IR spectra, and scanning electron microscopy. It was found that cod collagen (CC) and bovine collagen (BC) have different initial molecular weight parameters, and that, during hydrolysis, the majority of either type of protein is hydrolyzed by the proteolytic enzymes within the first minute. The differently sourced collagen samples were also hydrolyzed with the formation of two low molecular fractions: Mw ~ 10 kDa and ~20 kDa. In the case of CC, the microstructure of the final scaffolds contained denser, closely spaced fibrillar areas, while the BC-sourced scaffolds had narrow, short fibrils composed of unbound fibers of hydrolyzed collagen in their structure.  相似文献   

14.
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   

15.
The present study reports the preparation of a cellulose scaffold for tissue engineering directly from cellulose fiber using ionic liquid (IL) by the NaCl leaching method with bovine serum albumin (BSA), which is well known protein utilized for biomedical applications like degradation of polymer, cell attachment and proliferation on scaffold. The 1-n-allyl-3-methylimidazolium chloride (AmimCl) IL was used as a solvent for cellulose. The morphology of the scaffold was studied by scanning electron microscopy (SEM) and the images showed that the pore sizes of the scaffolds were about 200 µm. In addition, the water uptake (WU) and degree of degradation of the cellulose scaffold were measured. Meanwhile, the biocompatibility and bioactivity of the scaffold were determined via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenytetrazolium bromide (MTT) assay and the Live/Dead viability test. The various results demonstrated the ability of the Mesenchymal stem cells (MSC) to attach to the surface of the scaffolds amplified as percentage of BSA increased in cellulose scaffold.  相似文献   

16.
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   

17.
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   

18.
Easy fabrication, porosity, good mechanical properties, and composition controllable of the electrospun nanofiber mat make this material a promising candidate for wound dressing applications. In the present study, nylon6/gelatin electrospun nanofiber mats are introduced as novel wound dressing materials. The introduced mats were synthesized by electrospinning of nylon6 and gelatin mixtures, three mats containing different gelatin content were prepared; 10, 20 and 30 wt%. Interestingly, addition of the gelatin did not affect the mechanical properties of the nylon 6, moreover the mat containing 10 wt% gelatin revealed higher mechanical properties due to formation of spider-net like structure from very thin nanofibers (~10 nm diameter) bonding the main nanofibers. Biologically study indicates that gelatin incorporation strongly enhances the bioactivity performance as increasing the gelatin content linearly increases the MC3T3-E1 cell attachment. Overall, the obtained results recommend exploiting the introduced mats as wound dressing material.  相似文献   

19.
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   

20.
The formation of thrombosis has limited the applications of small diameter vascular in cardiovascular diseases. In order to improve the anticoagulant activities of scaffolds, this study combined fucoidan with CS/PVA and investigated the complete physicochemical and mechanical characterization of the scaffolds to evaluate the feasibility of Fucoidan/CS/PVA scaffolds used in vascular tissue engineering. The SEM graphs show a well defined and interconnected pore structure and the nanofiber diameters are ranging from 341 nm to 482 nm. After immersing in PBS for 5 days, the tensile strength of the crosslinked scaffolds was 722±38 kPa while the elongation at break was 35.5±1.6 %. Besides, added with fucoidan, the scafflolds showed lower rate of plate adhesion (14.75±2.10 %) and markedly prolonged the APTT and TT. Furthermore, owing to the great water uptake ability, sufficient porosity, enhanced drug release and low cytotoxicity, the Fucoidan/CS/PVA scaffolds might be used for vascular tissue engineering with good prospect.  相似文献   

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