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Global burden of chronic wounds has increased drasticallyas they are vulnerable to bacterial infections that causes inflammation, thereby leads to a delay in the healing process. Furthermore, wound care and dressing industry is subjected to a global market of $30.4 billion by 2024. Our work entails fabrication of polymeric electrospun nanofibers loaded with different concentration of the amoxicillin (AMX) antibiotic. Biodegradable and biocompatible poly (vinyl) alcohol (PVA)/poly(meth)(methacrylate)(PMMA) polymerswere blended with different AMX concentration (100, 150, 200 and 250 mg) and fabricated by electrospinning technique. Morphology, structural properties and drug release from electrospun nanofibers depend on the different concentrations of drug incorporated in PVA:PMMA blend of polymer. Furthermore, these studies revealed drug-excipient compatibility and drug encapsulation within the nanofiber. In-vitro release study showed the AMX release time from PVA: PMMA: AMX was extended up to 7 days for AMX-250 with an initial burst release of 70% and further sustained drug release. Electrospun nanofibers of PVA:PMMA:AMX showed greater zone of inhibition of S. aureus as 2.1±0.4 cm for 100-AMX, 2.3±0.5 cm for 150-AMX, 2.4±0.1 for 200-AMX and 3.4±0.3 cm for 250-AMX. These results demonstrate that AMX retains the anti-bacterial activity and hence can be used as a potential wound dressing candidate.
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The study aimed to evaluate the safety and function of poly(lactic-acid-co-ε-caprolactone) (PLCL)/fibrinogen nanofibers (P/F-Ns), and provide theoretical basis for the clinical application. The surface morphology, mechanical properties, the hydrophilicity and the fibrinogen content of P/F-Ns were tested by scanning electron microscope, the material testing machine, the contact angle meter and the microplate reader, respectively. The cell adhesion, proliferation and ligament remodeling genes expression of Hig-82 cells on P/F-Ns were conducted through cell counting kit-8 (CCK-8) and real-time quantitative PCR analyses, respectively. The results showed that with the increase of the fibrinogen content, the pore sizes and hydrophilicity of three P/F-Ns increased, but the mechanical properties decreased. Cell adhesion and proliferation tests showed that P/F-N-2 held the best ability to promote cell adhesion and proliferation. The ligament remodeling genes expressions of Hig-82 cells on P/F-N-1, P/F-N-2 and P/F-N-3 were all up-regulated compared to P/F-N-0 on days 3 and 7. All the three P/F-Ns containing fibrinogen (P/F-N-1, P/F-N-2 and P/F-N-3) had better biocompatibility compared to P/F-N-0, and could be efficiently applied to the reconstruction of anterior cruciate ligament.
Subject(s)
Anterior Cruciate Ligament Reconstruction , Cell Adhesion , Fibrinogen , Materials Testing , NanofibersABSTRACT
Objective:For severe skin defects which are deep to dermis, engineered skin with epidermis and dermis (bilayered) is required. Based on the success of engineering epidermis with GT/PCL electrospun membranes, our study was to investigate whether this membrane could be also used for engineering bilayered skin graft.Methods:From 2013 to 2019, we first prepared three GT/PCL electrospun membranes with different proportion (70∶30; 50∶50; 30∶70) in our laboratory; the biocompatibility of the membrane was evaluated in vitro by seeding fibroblasts or keratinocytes on the membranes. Then the outcome of GT/PCL membranes repairing skin defects in the nude mouse was investigated.Results:Cell attachment and proliferation were significantly improved with increase of gelatin. Histological analyses showed that bilayered skin engineered with GT/PCL (70∶30) group could form relatively better structure after 3 weeks of cultivation in vitro. Further in vivo transplantation studies revealed that scaffolds were not degraded in all three groups, indicating that these materials were not suitable for engineering bilayered skin although they had good biocompatibility.Conclusions:The higher gelatin membranes possess better biocompatibility. Further in vivo transplantation studies reveal that bilayered skin engineered with GT/PCL membranes is able to repair skin defects in the nude mouse.
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The utilization of electrospinning in drug delivery has thrived in recent years, with the ability to incorporate drugsand enhance dissolution; this technique is employed to improve the dissolution of poorly water-soluble selectivephosphodiesterase-5 inhibitor, tadalafil. The strategy involved direct electrospinning of tadalafil/polyvinylpyrrolidoneand polyethylene oxide (PEO) solution. The optimization process included a 32 full factorial design based on theinfluence of polymers concentration as independent variables on the electrospun yield, loading efficiency, nanofibersdiameter, number of beads, and in vitro release. Optimization studies revealed the negative influence of bothpolymers on the electrospun yield, while the loading efficiency and in vitro dissolution rate were reduced by the PEOconcentration solely. The higher polymer concentrations were favorable for the declination of beads number, and adriving factor for fiber diameter reduction. Further physicochemical characterization of the optimized formulationrevealed the presence of the drug in an amorphous state or molecular dispersion within the polymer matrix. In vitrodissolution studies revealed about 81.5% ± 8.34% release in less than 2 minutes compared to a negligible dissolutionof free drug. From the derived outcomes, the electrohydrodynamic spun tadalafil-loaded nanofibers pave the way fordissolution enhancement for insoluble low bioavailability class II drugs.
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We report fabrication of electro-spun polycaprolactone (PCL), gelatin and PCL–gelatin scaffolds in tissue engineering based applications. The structural characterizations were investigated by fourier transform infrared spectroscopy (FT-IR) and scanning electron microscopy (SEM) for the fabricated scaffolds. Pure PCL and gelatin scaffolds showed 3D nano−fibrous porous structures at 20 wt. % and 30 wt. %, respectively. PCL−gelatin scaffold at 70:30 vol. % showed good fibrous morphology without any structural discontinuities such as polymer beads, droplet formation and uneven fibers. Cellular activities such as cell adhesion and growth on scaffold surfaces were performed with F11 human fibroblast cells via MTT assay. Cell viability study showed that PCL−gelatin scaffold significantly improved the cell adhesion and proliferation when compared to pure gelatin and PCL scaffolds. Thus, the PCL−gelatin scaffold reported in our work has major prospective in the area of wound dressings, treatment of diabetic foot ulcers, implants and transdermal drug delivery.
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Open abdomen therapy (OA) is a breakthrough in treating abdominal trauma and infections. Enterorrhexis and entero-atmospheric fistula are both severe complications of OA and impact the therapeutic effect of OA seriously. To solve this problem, the pathophysiology of the OA wound formation and development, and the early protection measures of the OA wound are studied. Many biomaterials were developed, including autologous fibrin glue, electrospinning nanofiber membrane and hydrogel, to realize the target of early protection of the OA wound through hemostasis, anti-infection, antioxidation, and proliferation promoting.
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Objective · To investigate the effect of poly (L-lactic acid caprolactone) (PLCL)/gelatin electrospinning on the angiogenesis differentiation of endothelial progenitor cells (EPCs).Methods· Rat bone marrow-derived EPCs were isolated and cultured,then identification was performed.After preparation of PLCL/gelatin blend electrospun scaffold,scanning electron microscopy and water contact angle test were carried out.EPCs were grown on PLCL/gelatin electrospinning and CCK8 was used to detect cell proliferation.The expression of vascular endothelial growth factor (Vegf) and kinases insert region receptor (Kdr) was observed by RT-PCR and the expression of VEGF protein was observed by Western blotting.Results· The density gradient centrifugation combined with differential adherence method could effectively isolate EPCs.PLCL/gelatin electrospun nanofibers were porous,and the hydrophilic properties were favorable for cell adhesion,and EPCs grew well on the scaffold.The expression of Vegfand Kdr gene in PLCL/gelatin group was higher than that in control group (P=0.000),and the expression of VEGF protein was also increased (P=0.000).Conclusion · PLCL/gelatin is an ideal scaffold for tissue engineering,and it can promote the angiogenesis differentiation of EPCs.
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Cell adhesion in three-dimensional scaffolds plays a key role in tissue development. However, stem cell behavior in electrospun scaffolds under perfusion is not fully understood. Thus, an investigation was made on the effect of flow rate and shear stress, adhesion time, and seeding density under direct perfusion in polycaprolactone electrospun scaffolds on human dental pulp stem cell detachment. Polycaprolactone scaffolds were electrospun using a solvent mixture of chloroform and methanol. The viable cell number was determined at each tested condition. Cell morphology was analyzed by confocal microscopy after various incubation times for static cell adhesion with a high seeding density. Scanning electron microscopy images were obtained before and after perfusion for the highest flow rate tested. The wall pore shear stress was calculated for all tested flow rates (0.005-3 mL/min). An inversely proportional relationship between adhesion time with cell detachment under perfusion was observed. Lower flow rates and lower seeding densities reduced the drag of cells by shear stress. However, there was an operational limit for the lowest flow rate that can be used without compromising cell viability, indicating that a flow rate of 0.05 mL/min might be more suitable for the tested cell culture in electrospun scaffolds under direct perfusion.
Subject(s)
Humans , Dental Pulp/cytology , Perfusion , Polyesters , Stem Cells/cytology , Tissue Scaffolds , Cell Adhesion , Cell Culture TechniquesABSTRACT
Objective: To investigate the effect of poly (L-lactic acid caprolactone) (PLCL) /gelatin electrospinning on the angiogenesis differentiation of endothelial progenitor cells (EPCs). Methods: Rat bone marrow-derived EPCs were isolated and cultured, then identification was performed. After preparation of PLCL/gelatin blend electrospun scaffold, scanning electron microscopy and water contact angle test were carried out. EPCs were grown on PLCL/gelatin electrospinning and CCK8 was used to detect cell proliferation. The expression of vascular endothelial growth factor (Vegf ) and kinases insert region receptor (Kdr) was observed by RT-PCR and the expression of VEGF protein was observed by Western blotting. Results: The density gradient centrifugation combined with differential adherence method could effectively isolate EPCs. PLCL/gelatin electrospun nanofibers were porous, and the hydrophilic properties were favorable for cell adhesion, and EPCs grew well on the scaffold. The expression of Vegf and Kdr gene in PLCL/gelatin group was higher than that in control group (P=0.000), and the expression of VEGF protein was also increased (P=0.000). Conclusion: PLCL/gelatin is an ideal scaffold for tissue engineering, and it can promote the angiogenesis differentiation of EPCs.
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Objective·To construct C-shaped cartilage rings by rabbit auricular cartilage-derived chondrocytes combing with both electrospun gelatin/ polycaprolactone(GT/PCL) nanofibrous membranes and 3D printed supporters for repairing tracheal cartilage defects.Methods·Primary chondrocytes were isolated from rabbit auricular cartilage with methods of trypsin enzyme digestion and collagenase enzyme digestion.After proliferation in vitro,the chondrocytes of passage 2 were harvested for further experiments.Ultrafine composite fibers of GT/PCL were fabricated via electrospinning.The electrospun GT/PCL membranes were tailored into rectangle shape,the length of which is 12 cm and the width is 2.5 cm.Chondrocytes were seeded on membrane at a density of 1 × 108 cells/mL.Then the membrane were rolled onto a 3D printed supporter of poly(DL-lactide-ε-caprolactone) (PLCL) material to construct a C-shaped cartilage-like complex.After 8 weeks of subcutaneous incubation in vivo,gross inspection and paraffin section staining were applied for evaluation.Results·After 8 weeks of culture in vivo,mature cartilage-like tissue were formed with open-cylindrical bellow appearance and pecific mechanical property.C-shaped rings arranged at regular intervals on the inner surface of tissue,which were similar to the normal structure of tracheal cartilages.Histological and immunohistological staining showed a large number of typical lacunar structures and extracellular matrix secretions.Conclusion·It is feasible to construct tissue engineered C-shaped cartilage tissue by combing chondrocytes with GT/PCL membrane and 3D printed PLCL supporter for tracheal cartilage repair.
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Increasing bioactivity and mechanical properties of polymers to produce more suitable scaffold for tissue engineering is a recurrent goal in the development of new biomedical materials. In this study, collagen-functionalized poly (lactic acid), PLA, was obtained by means of a simple grafting route, and electrospun scaffolds were produced to grow cells in vitro; their bioactivity was compared with scaffolds made of physical blends of PLA and collagen. Grafting was verified via nuclear magnetic resonance, attenuated total reflection-Fourier transform infrared and X-ray photoelectron spectroscopy. The cell adhesion performance of the scaffolds was studied using macrophages. Elastic modulus (74.7 megapascals) and tensile strength (3.0 megapascals) of the scaffold made from PLA grafted with collagen were substantially higher than the scaffolds made from physical blends of collagen and PLA: 32 and 2.16 megapascals, respectively, implying a more resistant material because of the chemical bond of the polypeptide to PLA. Besides, the fibers had more uniform diameter without defects. Scaffolds made from PLA grafted with collagen presented four-fold increase in cell adhesion than those of PLA blended with collagen. Furthermore, cell spreading within the scaffolds occurred only when collagen-functionalized poly (lactic acid) was used. These results open a new option for the easy tailoring of nanofiber-based scaffolds in three dimensions for tissue engineering.
Subject(s)
Cell Adhesion , Collagen , Elastic Modulus , In Vitro Techniques , Macrophages , Magnetic Resonance Spectroscopy , Photoelectron Spectroscopy , Polymers , Tensile Strength , Tissue Engineering , TransplantsABSTRACT
<p><b>OBJECTIVE</b>To evaluate the biomineralization of the tissue-engineering electrospun polycaprolactone (PCL) scaffold and its potential use for guided bone regeneration (GBR) membranes.</p><p><b>METHODS</b>PCL ultrafinefiber scaffolds were fabricated by electrospinning and then immersed in supersaturated calcification solution (SCS) for biomineralization investigation. The electrospun PCL scaffolds and the calcium phosphate coating were identified by scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). Water-contact angles were also measured to evaluate the hydrophilicity of the modified surface. The biocompatibility of the composite was investigated by culturing osteoblasts on the scaffolds. Cell behavior was observed by SEM.</p><p><b>RESULTS</b>The electrospun PCL scaffold was composed of ultrafine fibers and well-interconnected pores. The deposits on the fibers grew in number and size as the biomineralization time increased. Then, the covering of the whole PCL film was identified as dicalcium phosphate dehydrate and apatite. Good cell attachment and proliferation behavior were observed on the membranes.</p><p><b>CONCLUSIONS</b>The quick apatite formation on the surface of the PCL electrospun scaffold indicated that SCS biomineralization may be an effective approach for obtaining PCL/calcium phosphate composites. The cellular biocompatibility of the composite scaffold was preliminarily confirmed by the in vitro culture of osteoblasts on the scaffold. As such, the composite scaffold is a promising biomimetic extracellular matrix biomaterial for bone tissue engineering and GBR membranes.</p>
Subject(s)
Biocompatible Materials , Bone Regeneration , Calcium Phosphates , Microscopy, Electron, Scanning , Osteoblasts , Polyesters , Tissue Scaffolds , X-Ray DiffractionABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of electronspun PLGA/HAp/Zein scaffolds on the repair of cartilage defects.</p><p><b>METHODS</b>The PLGA/HAp/Zein composite scaffolds were fabricated by electrospinning method. The physiochemical properties and biocompatibility of the scaffolds were separately characterized by scanning electron microscope (SEM), transmission electron microscope (TEM), and fourier transform infrared spectroscopy (FTIR), human umbilical cord mesenchymal stem cells (hUC-MSCs) culture and animal experiments.</p><p><b>RESULTS</b>The prepared PLGA/HAp/Zein scaffolds showed fibrous structure with homogenous distribution. hUC-MSCs could attach to and grow well on PLGA/HAp/Zein scaffolds, and there was no significant difference between cell proliferation on scaffolds and that without scaffolds (P>0.05). The PLGA/HAp/Zein scaffolds possessed excellent ability to promote in vivo cartilage formation. Moreover, there was a large amount of immature chondrocytes and matrix with cartilage lacuna on PLGA/HAp/Zein scaffolds.</p><p><b>CONCLUSION</b>The data suggest that the PLGA/HAp/Zein scaffolds possess good biocompatibility, which are anticipated to be potentially applied in cartilage tissue engineering and reconstruction.</p>
Subject(s)
Animals , Female , Humans , Male , Young Adult , Biocompatible Materials , Bone Development , Physiology , Cartilage , Cells, Cultured , Durapatite , Chemistry , Lactic Acid , Chemistry , Mesenchymal Stem Cells , Physiology , Polyglycolic Acid , Chemistry , Regeneration , Physiology , Tissue Scaffolds , Chemistry , Zein , ChemistryABSTRACT
One of the key contents in tissue engineering trachea replacement research is the scaffold selection.This review summarizes the latest original literatures and investigations about electrospun technique as well as recent progress.To discuss the advantages and disadvantages of natural,synthetic and combined electrospun scaffolds,the versatility in material choosing and production methods is the unique superiority.For specific experimental or clinical objects,the further research is to choose a suitable polymer,to improve surface modification techniques and to control the dimension and arrangement of the fibrous structure of electrospun fibers,which can offer versatility and tissue specificity,and therefore provide a bright prospect for tissue engineering trachea.
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OBJECTIVE: To fabricate moxifloxacin hydrochloride-loaded polyvinyl alcohol (PVA)-sodium alginate (SA) nanofiber using the electrospinning technique and investigate the drug release in vitro. METHODS: Central composite design-response surface methodology was used to investigate the influences of various factors, i.e., PVA content, SA content, the ratio of PVA-SA, on the accumulated release rate of moxifloxacin hydrochloride from the nanofibers. The scaffolds were cross-linked by CaCl2 ethanol solution. Fiber morphology was characterized using optical microscope and scanning electron microscopy (SEM). Drug entrapment efficiency was determined, and drug release profiles were tested. RESULTS: Moxifloxacin hydrochloride entrapment efficiencies were well above 65%. Drug was released from the nanofibers as a linear function of the square root of time, suggesting accordance with Fickian kinetics. The relationship between the independent variables and the dependent variables was in accordance with quadratic model. CONCLUSION: The PVA-SA nanofiber presents good release characteristics, and has a good prospect as drug carrier.