Investigation and optimization the effect of electrical stimulation parameters on the differentiation of human adipose mesenchymal stem cells into neurons-like cells on carbon nanofibers

Abstract

BackgroundNeurodegenerative diseases are among the most challenging diseases because neuron cells are not able to regenerate spontaneously. Tissue engineering is one of the most promising stem cell-based therapies. Controlling stem cell differentiation is a very crucial aspect of tissue engineering. MethodsIn this study, carbon nanofibers with an average diameter of 181{+/-}45 nm were prepared as a conductive scaffold based on the electrospinning method and subsequent thermal processing. Scaffold structure characterization were performed with XRD, Raman and Electrical conductivity tests. A homemade device was prepared to transmit electrical current to cells seeded on the scaffold in a culture plate. Various current parameters such as current intensity, frequency, waveform, daily shock duration, and shock period on adipose mesenchymal stem cells were examined for differentiation into neuronal cells. SPSS software and the one-way analysis of variance (ANOVA) was used as statistical analysis. ResultsCharacterization tests confirmed the formation of the carbon and crystallite structure with the electrical conductivity . Current with 1500 uA intensity, 500Hz frequency, and square waveform were selected as the optimal current parameters. It was found that the daily and periodic increase in shock time leads to an increase in the expression of neural and glial genes. A comparison of groups with real-time PCR and immunofluorescence of nestin, Map2, TubB3, and GFAPgenes was evaluated. ConclusionsThere are a variety of chemical and physical methods to control cell behavior, one of which is electrical stimulation. Conductive scaffolding is required for direct electrical stimulation of cells. The results showed that the method based on electrical stimulation can well cause neural differentiation, and considering the problems in preparing and maintaining chemical differentiation agents, it can be used practically. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=126 SRC="FIGDIR/small/593090v1_ufig1.gif" ALT="Figure 1"> View larger version (32K): org.highwire.dtl.DTLVardef@159d415org.highwire.dtl.DTLVardef@72bbb3org.highwire.dtl.DTLVardef@811222org.highwire.dtl.DTLVardef@1cda185_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOGraphical abstract:C_FLOATNO stages of scaffold preparation and electrical stimulation of stem cells C_FIG