Journal: Journal of Advanced Research
Article Title: Dynamic molecular landscape in dorsal root ganglion for peripheral nerve regeneration promoted by tissue engineered nerve graft
doi: 10.1016/j.jare.2025.07.059
Figure Lengend Snippet: The impact of DRG neuron axonal promotion via SKP-SCs. (a) Schematic illustration of DRG neurons culture in vitro. The cultured DRG neurons for 20 h and 40 h (b) showed increased neurite outgrowth. Scale bar: 50 μm. (c) The β-Tubulin III cell immunofluorescent staining (red) showed neurites are undergoing a process of growth in length from 48 h to 96 h. Scale bar: 50 μm. Schematic illustration showed the direct (d) and in-direct (Transwell®) (e) co-culture of DRG neurons and SKP-SCs. (h) After direct co-culture 48 h, GFP-SKP-SCs promoted DRGs neurite outgrowth significantly compared to control group (i). (k) After in-direct co-culture 72 h, GFP-SKP-SCs promoted DRGs neurite outgrowth (j) significantly compared to control group (g). Scale bar: 50 μm for f, g, i, j. (l) Morphological changes under phase contrast microscopy in DRG neurons and SKP-SCs direct co-culture, DRG neurons in separate culture, and in-direct (Transwell®) co-culture of DRG neurons and SKP-SCs after 24 h, 48 h, and 72 h. Scale bar: 100 μm. (m) Schematic illustration of LC-MS/MS analysis to identify the secreted proteins from SKP-SCs cultured in serum free basic medium for 4 h. After concentration, the cleared conditioned medium was collected for LC-MS/MS analysis, database search, and protein identification. The enrichment analysis showed top 20 diseases and functions (n) and canonical pathways (o) according to the IPA database. The PPI network shows the interaction of the proteins involved in the functions highly correlated with regeneration (p). Among them, functions as formation of lamellipodia, extension of neurites, extension of cellular protrusions, and related proteins were highlighted (q). Functions as migration of endothelial cells, development of endothelial tissue, vasculogenesis, development of epithelial tissue, endothelial cell development, and related proteins were highlighted (r). Orange nodes stand for a predicted activation state. Blue nodes stand for a predicted inhibition state. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
Article Snippet: The Ingenuity Pathway Analysis (IPA) software suite (Ingenuity Systems, Redwood City, CA) was employed to identify canonical pathways that are significantly overrepresented.
Techniques: In Vitro, Cell Culture, Staining, Co-Culture Assay, Control, Microscopy, Liquid Chromatography with Mass Spectroscopy, Concentration Assay, Migration, Activation Assay, Inhibition