Background: Researchers are trying to study the mechanism of neural stem cells (NSCs) differentiation to oligodendrocyte-like cells (OLCs) as well as to enhance the selective differentiation of NSCs to oligodendrocytes. However, the limitation in nerve tissue accessibility to isolate the NSCs as well as their differentiation toward oligodendrocytes is still challenging. Purpose: In the present study, a hybrid polycaprolactone (PCL)-gelatin nanofiber scaffold mimicking the native extracellular matrix and axon morphology to direct the differentiation of bone marrow-derived NSCs to OLCs was introduced. Materials and Methods: In order to achieve a sustained release of T3, this factor was encapsulated within chitosan nanoparticles and chitosan-loaded T3 was incorporated within PCL nanofibers. Polyaniline graphene (PAG) nanocomposite was incorporated within gelatin nanofibers to endow the scaffold with conductive properties, which resemble the conductive behavior of axons. Biodegradation, water contact angle measurements, and scanning electron microscopy (SEM) observations as well as conductivity tests were used to evaluate the properties of the prepared scaffold. The concentration of PAG and T3-loaded chitosan NPs in nanofibers were optimized by examining the proliferation of cultured bone marrow-derived mesenchymal stem cells (BMSCs) on the scaffolds. The differentiation of BMSCs-derived NSCs cultured on the fabricated scaffolds into OLCs was analyzed by evaluating the expression of oligodendrocyte markers using immunofluorescence (ICC), RT-PCR and flowcytometric assays. Results: Incorporating 2% PAG proved to have superior cell support and proliferation while guaranteeing electrical conductivity of 10.8 × 10 −5 S/cm. Moreover, the scaffold containing 2% of T3-loaded chitosan NPs was considered to be the most biocompatible samples. Result of ICC, RT-PCR and flow cytometry showed high expression of O4, Olig2, platelet-derived growth factor receptor-alpha (PDGFR-α), O1, myelin/oligodendrocyte glycoprotein (MOG) and myelin basic protein (MBP) high expressed but low expression of glial fibrillary acidic protein (GFAP). Conclusion: Considering surface topography, biocompatibility, electrical conductivity and gene expression, the hybrid PCL/gelatin scaffold with the controlled release of T3 may be considered as a promising candidate to be used as an in vitro model to study patient-derived oligodendrocytes by isolating patient's BMSCs in pathological conditions such as diseases or injuries. Moreover, the resulted oligodendrocytes can be used as a desirable source for transplanting in patients.
Peripheral nervous system in contrary to central one has the potential for regeneration, but its regrowth requires proper environmental conditions and supporting growth factors. The aim of this study is to design and fabricate a conductive polyaniline/graphene nanoparticles incorporated gelatin nanofibrous scaffolds suitable for peripheral nervous system regeneration. The scaffolds were fabricated with electrospinning and the fabrication process was designed with Design-Expert software via response surface methodology. The effect of process parameters including applied voltage (kV), syringe pump flow rate (cm 3 /h), and PAG concentration (wt%), on the scaffold conductivity, nanofibers diameter, and cell viability were investigated. The obtained results showed that the scaffold conductivity and cell viability are affected by polyaniline/graphene concentration while nanofiber diameter is more affected by the applied voltage and syringe pump flow rate. Optimum scaffold with maximum conductivity (0.031 AE 0.0013 S/cm) and cell compatibility and suitable diameter were electrospun according to the software introduced values for the process parameters (voltage of 13 kV, flow rate of 0.1 cm 3 /h, and PAG wt.% of 1.3) and its morphology, cell compatibility, and biodegradability were further investigated, which showed its potential for applying in peripheral nervous system injury regeneration.
Electrospinning is an economical and alluring method to fabricate wound dressing mats from a variety of natural and synthetic materials. In this study, polyvinylidene fluoride (PVDF) and starch composite mats containing ciprofloxacin (CIP) loaded on titanium dioxide nanoparticles (TiO 2 ) were prepared. Fourier Transform Infrared spectra of CIP, synthesized TiO 2 NPs, TiO 2 /CIP, and PVDF/starch composite mats are analyzed. Scanning electron microscopy images revealed that the fiber diameter of PVDF nanofibers thickens by increasing starch, which varies between 180 nm and 550 nm. Strain at break of PVDF, starch, PVDF/starch (2:1 w:w; P2S1), PVDF/starch (1:1 w:w; P1S1), PVDF/starch (1:2 w:w; P1S2), and nanofibers were 103 AE 11, 3 AE 0.6, 27 AE 4, 52 AE 5.2, 7.7 AE 1%, respectively. Based on strain at break and young modulus, P2S1 was selected as a suitable candidate for wound dressing to which load TiO 2 / CIP as a bioactive agent. The release rate of CIP showed that about 40% of the drug is released in the first 2 days. Furthermore, the antibacterial activity of dressings was investigated using Escherichia coli and Staphylococcus aureus microorganisms and zones of clearance were obvious around the specimen on the agar plate.
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