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BACKGROUND: Fibrin as an extracellular matrix feature like biocompatibility, creates a favorable environment for proliferation and migration of cells and it can act as a reservoir for storage and release of growth factors in tissue engineering. METHODS: In this study, the inner surface of electrospun poly (lactic-co-glycolic acid) (PLGA) nanofibrous conduit was biofunctionalized with laminin containing brain derived neurotrophic factor (BDNF) and gold nanoparticles in chitosan nanoparticle. The rats were randomly divided into five groups, including autograft group as the positive control, PLGA conduit coated by laminin and filled with DMEM/F12, PLGA conduit coated by laminin and filled with rat-adipose derived stem cells (r-ADSCs), PLGA conduit coated by laminin containing gold-chitosan nanoparticles (AuNPs-CNPs), BDNF-chitosan nanoparticles (BDNF-CNPs) and filled with r-ADSCs or filled with r-ADSCs suspended in fibrin matrix, and they were implanted into a 10 mm rat sciatic nerve gap. Eventually, axonal regeneration and functional recovery were assessed after 12 weeks. RESULTS: After 3 months post-surgery period, the results showed that in the PLGA conduit filled with r-ADSCs without fibrin matrix group, positive effects were obtained as compared to other implanted groups by increasing the sciatic functional index significantly (p < 0.05). In addition, the diameter nerve fibers had a significant difference mean in the PLGA conduit coated by laminin and conduit filled with r-ADSCs in fibrin matrix groups relative to the autograft group (p < 0.001). However, G-ratio and amplitude (AMP) results showed that fibrin matrix might have beneficial effects on nerve regeneration but, immunohistochemistry and real-time RT-PCR outcomes indicated that the implanted conduit which filled with r-ADSCs, with or without BDNF-CNPs and AuNPs-CNPs had significantly higher expression of S100 and MBP markers than other conduit implanted groups (p < 0.05). CONCLUSIONS: It seems, in this study differential effects of fibrin matrix, could be interfered it with other factors thereby and further studies are required to determine the distinctive effects of fibrin matrix combination with other exogenous factors in peripheral nerve regeneration.
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Fator Neurotrófico Derivado do Encéfalo/administração & dosagem , Ouro/administração & dosagem , Células-Tronco Mesenquimais , Nanopartículas Metálicas/administração & dosagem , Regeneração Nervosa/fisiologia , Neuropatia Ciática/terapia , Animais , Terapia Combinada , Sistemas de Liberação de Medicamentos/métodos , Quimioterapia Combinada , Fibrina/administração & dosagem , Masculino , Regeneração Nervosa/efeitos dos fármacos , Ratos , Ratos Wistar , Neuropatia Ciática/patologia , Neuropatia Ciática/fisiopatologiaRESUMO
Tissue engineering techniques particularly using electrospun scaffolds have been intensively used in recent years for the development of small diameter vascular grafts. However, the development of a completely successful scaffold that fulfills multiple requirements to guarantee complete vascular regeneration remains challenging. In this study, a hydrophilic and compliant polyurethane namely Tecophilic (TP) blended with gelatin (gel) at a weight ratio of 70:30 (TP(70)/gel(30)) was electrospun to fabricate a tubular composite scaffold with biomechanical properties closely simulating those of native blood vessels. Hydrophilic properties of the composite scaffold induced non-thrombogenicity while the incorporation of gelatin molecules within the scaffold greatly improved the capacity of the scaffold to serve as an adhesive substrate for vascular smooth muscle cells (SMCs), in comparison to pure TP. Preservation of the contractile phenotype of SMCs seeded on electrospun TP(70)/gel(30) was yet another promising feature of this scaffold. The nanostructured TP(70)/gel(30) demonstrated potential feasibility toward functioning as a vascular graft.
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Prótese Vascular , Gelatina/farmacologia , Nanofibras/química , Poliuretanos/farmacologia , Engenharia Tecidual/métodos , Animais , Aorta/citologia , Proliferação de Células/efeitos dos fármacos , Forma Celular/efeitos dos fármacos , Células Cultivadas , Hemólise/efeitos dos fármacos , Humanos , Imageamento Tridimensional , Imuno-Histoquímica , Teste de Materiais , Fenômenos Mecânicos , Miócitos de Músculo Liso/citologia , Miócitos de Músculo Liso/efeitos dos fármacos , Nanofibras/ultraestrutura , Sus scrofa , Alicerces Teciduais/química , Água/químicaRESUMO
Nerve regeneration following the injury of nerve tissue remains a major issue in the therapeutic medical field. Various bio-mimetic strategies are employed to direct the nerve growth in vitro, among which the chemical and topographical cues elicited by the scaffolds are crucial parameters that is primarily responsible for the axon growth and neurite extension involved in nerve regeneration. We carried out electrospinning for the first time, to fabricate both random and aligned nanofibers of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate; PHBV) and composite PHBV/collagen nanofibers with fiber diameters in the range of 386-472 nm and 205-266 nm, respectively. To evaluate the potential of electrospun aligned nanofibers of PHBV and composite scaffolds as a substrate for nerve regeneration, we cultured nerve cells (PC12) and studied the biocompatibility effect along with neurite extension by immunostaining studies. Cell proliferation assays showed 40.01% and 5.48% higher proliferation of nerve cells on aligned PHBV/Coll50:50 nanofibers compared to cell proliferation on aligned PHBV and PHBV/Col75:25 nanofibers, respectively. Aligned nanofibers of PHBV/Coll provided contact guidance to direct the orientation of nerve cells along the direction of the fibers, thus endowing elongated cell morphology, with bi-polar neurite extensions required for nerve regeneration. Results showed that aligned PHBV/Col nanofibers are promising substrates than the random PHBV/Col nanofibers for application as bioengineered grafts for nerve tissue regeneration.
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Colágeno/química , Nanofibras/química , Neurônios/fisiologia , Poliésteres/química , Engenharia Tecidual , Alicerces Teciduais/química , Animais , Proliferação de Células/efeitos dos fármacos , Colágeno/farmacologia , Módulo de Elasticidade , Técnicas Eletroquímicas , Teste de Materiais , Neurônios/citologia , Neurônios/efeitos dos fármacos , Células PC12 , Poliésteres/farmacologia , Ratos , Estresse Mecânico , Engenharia Tecidual/instrumentação , Engenharia Tecidual/métodosRESUMO
The presence of cyclic organic compounds, including phenol, in the wastewater of many industries has made phenol removal an important issue. Meanwhile, the biological methods of removing phenol have attracted the attention of researchers in recent years. Recently, the use of immobilized microbial cells is proposed as a new approach in industrial wastewater treatment. In this research, the aim is to study the effect of immobilized beads size on the phenol biodegradation efficiency and specific microbial growth rate. For this purpose, electrospray technique was used to immobilize activated sludge in hybrid matrix of alginate and polyvinyl alcohol (PVA). The fabricated alginate/PVA beads were characterized using Fourier transform infrared spectroscopy (FTIR). Evaluation of the results related to the free and immobilized cell systems in the shake flask experiments showed that at low phenol concentrations the immobilized cell system had the same performance as the free cell system, while the immobilized cell system at higher concentrations had a better performance in removing phenol so that at a concentration of 2000 mg/L, removal percentage has increased from 15% to 25-34%. On the other hand, in this survey, the kinetic behavior of activated sludge was in good agreement with Haldane's equation. Moreover, the maximum specific growth rate was measured 0.033 and 0.041 (h-1) beside 544 and 636 mg/L substrate inhibition constant, for free and immobilized cell systems, respectively. This result shows that the phenol biodegradation has been improved by using the cell immobilization technique especially with applying the smaller beads, which is due to improved mass transfer and microbial cell protection from harsh environments.
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Tissue-engineered vascular grafts (TEVGs) are promising alternatives to existing prosthetic grafts. The objective of this study is to evaluate the clinical feasibility of a novel multi-layered small-diameter vascular graft that has a hierarchical structure. Vascular grafts with elaborately designed composition and architecture were prepared by 3D printing and electrospinning and were implanted into the femoral artery of 5 dogs. The patency of the grafts was assessed using Doppler ultrasonography. After 6 months, the grafts were retrieved and histological and SEM examinations were conducted. During implantation, the grafts exhibited resistance to kinking and no blood seepage thanks to the helical structure of the innermost and outermost layers. The grafts showed a high patency rate and remodelling ability. At 6 months post-implantation, the lumen was endothelialized and middle layers were regenerated by infiltration of smooth muscle cells (SMCs) and deposition of extracellular matrix (ECM). These results suggest that the multi-layered vascular graft may be a promising candidate for small-diameter blood vessel tissue engineering in clinical practice.
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The purpose of the present article is to study the bending strength of glulam prepared by plane tree (Platanus Orientalis-L) wood layers adhered by UF resin with different formaldehyde to urea molar ratios containing the modified starch adhesive with different NaOCl concentrations. Artificial neural network (ANN) as a modern tool was used to predict this response, too. The multilayer perceptron (MLP) models were used to predict the modulus of rapture (MOR) and the statistics, including the determination coefficient (R2), root mean square error (RMSE), and mean absolute percentage error (MAPE) were used to validate the prediction. Combining the ANN and the genetic algorithm by using the multiple objective and nonlinear constraint functions, the optimum point was determined based on the experimental and estimated data, respectively. The characterization analysis, performed by FTIR and XRD, was used to describe the effect of the inputs on the output. The results indicated that the statistics obtained show excellent MOR predictions by the feed-forward neural network using Levenberg-Marquardt algorithms. The comparison of the optimal output of the actual values obtained by the genetic algorithm resulting from the multi-objective function and the optimal output of the values estimated by the nonlinear constraint function indicates a minimum difference between both functions.
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Triboelectric nanogenerators (TENGs) provide promising potential to sustainably power modern portable electronic devices through harvesting mechanical energy from the environment. There is a need to further improve the sustainability credentials of TENGs by reducing the usage of non-renewable materials. Nanocellulose possessing outstanding properties, can partially replace non-renewable materials used in TENGs. Here, main sources and types of nanocellulose and methods of forming nanocellulose-based 2D/3D configurations used as triboelectric layers of TENGs are introduced. Cellulose has relatively weak tribopositivity and this review describes effective strategies of amplifying the output performance of TENGs possessing a nanocellulose-based tribopositive layer. Furthermore, efforts made to change the polarity of nanocellulose structures in order to use them as both positive and negative triboelectric layers are reviewed and their output performance is compared to that of all-synthetic polymer-based TENGs. Finally, requirements expected to be met in near future for development of commercial green nanocellulose-based TENGs are highlighted.
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Superabsorbent polymers currently used in health and agricultural sectors are based on petroleum-based materials which led to serious concerns in the society. Here, superabsorbent fibers (SAFs) based on electrospun cellulose nanofibers (ECNFs) were prepared. Firstly, cellulose was removed from wheat straw, pre-treated with the TEMPO-mediated oxidation and subsequently dissolved into Trifluoroacetic acid for production of ECNFs through the electrospinning approach. The maximum swelling ratios of 225 g/g and 208 g/g in distilled water and 0.9 wt% NaCl solution were measured for ESAFs composed of oxidized ECNFs containing 15 % poly (sodium acrylate), respectively. The ESAFs were characterized using Fourier transform infrared spectroscopy and field emission scanning electron microscopy analysis. The FESEM showed that ESAFs formed high strength three-dimensional architecture networks. Also, the results showed that the ionic sensitivity of this ECNFs were low. The prepared ESAFs are attractive renewable alternatives for different applications, contributing to a reduction of plastic microspheres.
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Celulose Oxidada/química , Celulose/química , Eletroquímica/métodos , Nanofibras/química , Polímeros/química , Triticum/química , Água/química , Celulose/isolamento & purificação , Microscopia Eletrônica de Varredura/métodos , Espectroscopia de Infravermelho com Transformada de Fourier/métodosRESUMO
In this paper, a multifunctional nanofibrous cellulose acetate/gelatin/Zataria multiflora-nanoemulsion (CA/Gel/ZM-nano) wound dressing was fabricated, in which nanoemulsion of a natural active antibacterial plant, by the scientific name of Zataria multiflora (ZM) was loaded into the nanofibrous mat. To fabricate the wound dressing, different weight ratios of CA/Gel (100, 0, 75, 25, 50, 50 and 25, 75) were selected, and solutions with concentrations of 16, 15, 14 and 12% w/v were prepared for each ratio, respectively to achieve smooth and uniform fibers by electrospinning. In vitro and in vivo analysis was taken for the samples. Nanofibrous mats with a lower ratio of CA/Gel and incorporated with ZM-nano promoted the adhesion and proliferation of L929 fibroblast cells significantly. Also, by lowering the ratio of CA/Gel, nanoemulsion drug-loading into nanofibers increased considerably, as the amount of ZM-nano loaded into CA/Gelâ¯=â¯50:50 was found to be 2.4-fold higher than CA/Gelâ¯=â¯100:0. Moreover, the rat model experiment in our study revealed that the nanofibrous samples incorporated with nanoemulsion drug (ZM-nano) accelerated the wound healing process so that the relative wound area for the nanoemulsion-loaded dressings was much smaller than the other samples after 22â¯days. Therefore, this multifunctional CA/Gel/ZM-nano wound dressing could be a promising and potential candidate for wound healing applications.
Assuntos
Antibacterianos/farmacologia , Bandagens , Gelatina , Lamiaceae/química , Nanofibras/química , Preparações de Plantas , Animais , Linhagem Celular , Celulose/análogos & derivados , Celulose/química , Celulose/farmacologia , Portadores de Fármacos/química , Portadores de Fármacos/farmacologia , Emulsões/química , Emulsões/farmacologia , Fibroblastos , Gelatina/química , Gelatina/farmacologia , Masculino , Camundongos , Preparações de Plantas/química , Preparações de Plantas/farmacologia , Ratos , Ratos Wistar , Cicatrização/efeitos dos fármacosRESUMO
Alginate spherical hydrogel beads have several applications in biomedical and biological processes in which the bead size and sphericity are critical factors affecting mass transfer phenomena. Electrospraying technology facilitates generation of small and almost uniform beads with higher diffusion rate resulting in process performance improvement. There are several key factors affecting particle size and shape behavior of electrosprayed alginate beads meanwhile interactions between these factors introduce complexity in determining appropriate conditions to produce spherical beads with the size of interest. Thus, the need to achieve reliable products has put growing emphasis on the use of modeling methodology to establish correlations between particle size and affecting variables as well as sphericity coefficient and meaningful factors. Obviously, a more applicable model based on intentionally manipulatable factors would spark a great deal of interest for practical engineering applications. In this regard we employed a central composite design (CCD) and response surface methodology (RSM) to model the diameter and sphericity coefficient of electrosprayed alginate beads for the first time. Two quadratic models were obtained in which the effectiveness order of the variables were found. We could benefit from this RSM-based empirical model not only for better understanding the complex physics of the electrospraying process, but also for selection of factors and their levels to produce alginate micro-beads with appropriate size and sphericity. The results indicate that the alginate concentration, voltage and needle size have the strongest influence on both response variables. The quite spherical beads with a minimum size of 130 µm can be obtained at alginate concentration of 1.5%, voltage of 11 kV, and needle size of 26 G.
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This study seeks to develop a nanofibrous matrix containing rosmarinic acid (RosA), an herbal non-steroidal anti-inflammatory and antioxidant drug with low water solubility, for drug delivery applications. Neat and two types of RosA-loaded cellulose acetate (CA) mats varying in the initial content of RosA were electrospun. Microstructure of nanofibers, chemistry and physical state of RosA in nanofibers, RosA loading efficiency and RosA release in acetate buffer were investigated. To evaluate bioactivity of RosA-loaded nanofibers, their ability to inhibit protein denaturation was assayed as an indicator of anti-inflammatory properties and their antioxidant activity was determined by radical scavenging assay. The indirect cytotoxicity assay was used to find if there is a cytotoxic response to nanofibers. The homogeneous distribution of the drug within nanofibers through electrospinning led to high loading efficiency, low burst release and prolonged release of a large percentage of RosA over a period of 64h following Fickian diffusion mechanism. Nanofibers with higher RosA content exhibited anti-inflammatory activity comparable to ibuprofen, and higher antioxidant activity compared to nanofibers with lower RosA content. Additionally, extracts from nanofibers did not give any major harmful effect on cells. Sustained release of RosA, and bioactivity of RosA-loaded nanofibers confirmed the potential of the produced matrix as a drug delivery system.
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Physiological functionality of a tissue engineered vascular construct depends on the phenotype of smooth muscle cells (SMCs) cultured into the scaffold and mechanical robust of the construct relies on two simultaneous mechanisms including scaffold biodegradation and de novo matrix synthesis by SMCs which both can be influenced by scaffold properties and culture condition. Our focus in this study was to provide an appropriate environmental condition within tissue engineering context to meet foregoing requisites for a successful vascular regeneration. To this end, SMCs seeded onto electrospun Tecophilic/gelatin (TP(70)/gel(30)) scaffolds were subjected to orbital shear stress. Given the improvement in mechanical properties of dynamically stimulated cell-seeded constructs after a span of 10days, effect of fluctuating shear stress on scaffold biodegradation and SMC behavior was investigated. Compared to static condition, SMCs proliferated more rapidly and concomitantly built up greater collagen content in response to dynamic culture, suggesting a reasonable balance between scaffold biodegradation and matrix turnover for maintaining the structural integrity and mechanical support to seeded cells during early phase of vascular tissue engineering. Despite higher proliferation of SMCs under dynamic condition, cells preserved nearly spindle like morphology and contractile protein expression likely thanks to composition of the scaffold.
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Biomimética , Proliferação de Células , Células Cultivadas , Miócitos de Músculo Liso , Engenharia Tecidual , Alicerces TeciduaisRESUMO
Aiming to mimic a blood vessel structurally, morphologically, and mechanically, a sequential electrospinning technique using a small diameter mandrel collector was performed and a three-layered tubular scaffold composed of nanofibers of polycaprolactone, collagen, and poly(l-lactic acid) as inner, intermediate, and outer layers, respectively, was developed. Biological performances of the scaffold in terms of compatibility with blood and endothelial cells were assessed to get some insights into its potential use as a tissue engineered small-diameter vascular replacement compared to an expanded polytetrafluoroethylene vascular graft. Due to direct contact of the blood and endothelial cells with inner surface of the scaffold, polycaprolactone fibers were characterized using SEM, water contact angle measurement, and ATR-FTIR. Despite similar surface wettability of the electrospun scaffold and the expanded polytetrafluoroethylene graft, the three-layered scaffold significantly reduced platelet adhesion and hemolysis ratio compared to expanded polytetrafluoroethylene graft while comparable blood clotting profiles were observed for both electrospun scaffold and expanded polytetrafluoroethylene graft. However, inflammatory response to nanofibrous surface of the scaffold was reduced compared to expanded polytetrafluoroethylene graft. The electrospun scaffold also presented a significantly more supportive substrate for endothelialization than the expanded polytetrafluoroethylene graft. The results described herein suggested that the three-layered scaffold has superior biological properties compared to an expanded polytetrafluoroethylene graft for vascular tissue engineering.
Assuntos
Prótese Vascular , Colágeno/química , Células Endoteliais/fisiologia , Nanofibras/química , Poliésteres/química , Alicerces Teciduais , Materiais Biocompatíveis/administração & dosagem , Materiais Biocompatíveis/síntese química , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Colágeno/administração & dosagem , Células Endoteliais/efeitos dos fármacos , Desenho de Equipamento , Humanos , Teste de Materiais , Nanofibras/administração & dosagem , Nanofibras/ultraestrutura , Tamanho da Partícula , Poliésteres/administração & dosagem , RotaçãoRESUMO
The major component of fibrous extracellular matrix of dermis is composed of a complex combination of proteins and polysaccharides. Electrospun cellulose acetate/gelatin might be an effective simulator of the structure and composition of native skin and during this study, we electrospun cellulose acetate/gelatin membranes in various compositions and their performance as a scaffold for either skin tissue engineering or as a wound dressing was evaluated. Skin treatment products, whether tissue-engineered scaffolds or wound dressings, should be sufficiently hydrophilic to allow for gas and fluid exchange and absorb excess exudates while controlling the fluid loss. However, a wound dressing should be easily removable without causing tissue damage and a tissue-engineered scaffold should be able to adhere to the wound, and support cell proliferation during skin regeneration. We showed that these distinct adherency features are feasible just by changing the composition of cellulose acetate and gelatin in composite cellulose acetate/gelatin scaffolds. High proliferation of human dermal fibroblasts on electrospun cellulose acetate/gelatin 25:75 confirmed the capability of cellulose acetate/gelatin 25:75 nanofibers as a tissue-engineered scaffold, while the electrospun cellulose acetate/gelatin 75:25 can be a potential low-adherent wound dressing.
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Celulose/análogos & derivados , Gelatina , Nanofibras , Pele Artificial , Ferimentos e Lesões/terapia , Proliferação de Células , Células Cultivadas , Colágeno/metabolismo , Humanos , Espectroscopia de Infravermelho com Transformada de Fourier , Engenharia Tecidual , MolhabilidadeRESUMO
Scaffolds for tissue engineering (TE) require the consideration of multiple aspects, including polymeric composition and the structure and mechanical properties of the scaffolds, in order to mimic the native extracellular matrix of the tissue. Electrospun fibers are frequently utilized in TE due to their tunable physical, chemical, and mechanical properties and porosity. The mechanical properties of electrospun scaffolds made from specific polymers are highly dependent on the processing parameters, which can therefore be tuned for particular applications. Fiber diameter and orientation along with polymeric composition are the major factors that determine the elastic modulus of electrospun nano- and microfibers. Here we have developed a neural network model to investigate the simultaneous effects of composition, fiber diameter and fiber orientation of electrospun polycaprolactone/gelatin mats on the elastic modulus of the scaffolds under ambient and simulated physiological conditions. The model generated might assist bioengineers to fabricate electrospun scaffolds with defined fiber diameters, orientations and constituents, thereby replicating the mechanical properties of the native target tissue.
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Módulo de Elasticidade , Gelatina/química , Redes Neurais de Computação , Poliésteres/química , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Algoritmos , Animais , Polímeros/química , Análise de Regressão , Sus scrofaRESUMO
The ability of mature smooth muscle cells (SMCs) to modulate their phenotype in response to environmental cues is a critical issue related to vascular diseases. A tissue engineered vascular graft shall promote the contractile phenotype of vascular SMCs. To this aim, Tecophilic/gelatin (TP/gel) was electrospun at different weight ratios of TP/gelatin (100:0, 70:30, 50:50, 30:70), leading to differences in biochemical and mechanical properties of the nanofibers which in turn influenced the phenotype of SMCs. Results indicated that both the substrate with higher ligand density and lower stiffness could enhance SMC contractility and reduce cell proliferation. However, observing the highest SMCs contractility on electrospun TP(70)/gel(30) among the composite scaffolds demonstrated stiffness as the most critical parameter. Due to conflicting effects of softness versus minor fraction of gelatin (reduced ligand density) within TP(70)/gel(30) fibers, a relatively high proliferation of SMCs was still observed on TP(70)/gel(30) scaffold. The surface of TP(70)/gel(30) scaffold was further modified through physical adsorption of gelatin molecules so as to increase the ligand density on its surface, whereby a functional vascular construct that promotes the contractile behavior of SMCs with low cell proliferation was developed.