Chinese Journal of Tissue Engineering Research ›› 2025, Vol. 29 ›› Issue (23): 4924-4930.doi: 10.12307/2025.100

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Ectomesenchymal stem cells-derived extracellular vesicles promote neuronal axonal elongation

Sun Haitao1, Ren Chunpeng1, Yang Yongtao1, Huang Yonghui2, Qin Rujie1, Li Zhen1   

  1. 1First People’s Hospital of Lianyungang, Lianyungang 222000, Jiangsu Province, China; 2Affiliated Hospital of Jiangsu University, Zhenjiang 212000, Jiangsu Province, China
  • Received:2024-03-25 Accepted:2024-06-18 Online:2025-08-18 Published:2024-09-28
  • Contact: Qin Rujie, Master, Chief physician, First People’s Hospital of Lianyungang, Lianyungang 222000, Jiangsu Province, China; Co-corresponding author: Li Zhen, Master, Physician, First People’s Hospital of Lianyungang, Lianyungang 222000, Jiangsu Province, China
  • About author:Sun Haitao, Master, Physician, First People’s Hospital of Lianyungang, Lianyungang 222000, Jiangsu Province, China
  • Supported by:
    Youth Talent Project of First People’s Hospital of Lianyungang, No. QN2205 (to SHT); Scientific Research Project of Zhenjiang City of Jiangsu Province, No. SH2020053 (to HYH)

Abstract: BACKGROUND: The occurrence of neuronal axonal injury can result in neurological dysfunction, and the facilitation of axonal elongation is anticipated to play a pivotal role in the treatment of diseases affecting the nervous system.
OBJECTIVE: To investigate whether ectomesenchymal stem cells-derived extracellular vesicles can promote neuronal axonal elongation.
METHODS: (1) Ectomesenchymal stem cells were obtained from nasal mucosa using the tissue adherence method, and the specific markers of were identified through immunofluorescence. Ectomesenchymal stem cells-derived extracellular vesicles were acquired via ultracentrifugation and identified. (2) Ectomesenchymal stem cells-derived extracellular vesicles (0, 0.5, 1.0, 1.5 mg/mL) were incubated with PC12 cells for 72 hours. The cytotoxicity and proliferation of ectomesenchymal stem cells-derived extracellular vesicles on PC12 cells were assessed using the CCK-8 assay. (3) Ectomesenchymal stem cells-derived extracellular vesicles (1.0 mg/mL) were incubated with PC12 cells or neurons for 72 hours. The changes in axon length were observed using microscopic analysis. The expression levels of axon-related markers β3-tubulin (early stage), growth associated protein 43 (middle stage), and neurofilament 200 (mature stage) were analyzed through real-time fluorescence quantitative PCR and Western blotting. These investigations aimed to explore the potential of ectomesenchymal stem cells-derived extracellular vesicles in promoting neurite elongation within PC12 cells or neurons. 
RESULTS AND CONCLUSION: (1) The majority of the acquired ectomesenchymal stem cells exhibited a spindle-shaped morphology, while a minority displayed irregular shapes, and demonstrated high expression levels of mesenchymal stem cell-specific markers Nestin, CD44, and Vimentin. The obtained ectomesenchymal stem cells-derived extracellular vesicles fulfilled the biological criteria for extracellular vesicles. (2) Within the detected protein concentration range of 0.5 to 1.5 mg/mL, the proliferation of PC12 cells was promoted by ectomesenchymal stem cells-derived extracellular vesicles, and this effect was further enhanced with increasing concentrations. (3) Ectomesenchymal stem cells-derived extracellular vesicles increased the length of axons in PC12 cells and neurons and the expression of axon-related markers β3-tubulin, growth associated protein 43, and neurofilament 200. Above findings suggest that ectomesenchymal stem cells-derived extracellular vesicles have the potential to enhance neuronal axonal elongation. 

Key words: mesenchymal stem cell, neuron, PC12 cell, extracellular vesicle, axonal elongation, β3-tubulin, growth associated protein 43, neurofilament 200

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