Korean J Pain.  2023 Jan;36(1):72-83. 10.3344/kjp.22279.

The mechanism of human neural stem cell secretomes improves neuropathic pain and locomotor function in spinal cord injury rat models: through antioxidant, anti-inflammatory, anti-matrix degradation, and neurotrophic activities

• Semita IN
• Utomo DN
• Suroto H
• Sudiana IK
• Gandi P

• KMID: 2537614
• DOI: http://doi.org/10.3344/kjp.22279

Abstract

Background
Globally, spinal cord injury (SCI) results in a big burden, including 90% suffering permanent disability, and 60%–69% experiencing neuropathic pain. The main causes are oxidative stress, inflammation, and degeneration. The efficacy of the stem cell secretome is promising, but the role of human neural stem cell (HNSC)-secretome in neuropathic pain is unclear. This study evaluated how the mechanism of HNSC-secretome improves neuropathic pain and locomotor function in SCI rat models through antioxidant, anti-inflammatory, anti-matrix degradation, and neurotrophic activities.
Methods
A proper experimental study investigated 15 Rattus norvegicus divided into normal, control, and treatment groups (30 µL HNSC-secretome, intrathecal in the level of T10, three days post-traumatic SCI). Twentyeight days post-injury, specimens were collected, and matrix metalloproteinase (MMP)-9, F2-Isoprostanes, tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β, and brain derived neurotrophic factor (BDNF) were analyzed. Locomotor recovery was evaluated via Basso, Beattie, and Bresnahan scores. Neuropathic pain was evaluated using the Rat Grimace Scale.
Results
The HNSC-secretome could improve locomotor recovery and neuropathic pain, decrease F2-Isoprostane (antioxidant), decrease MMP-9 and TNF-α (anti-inflammatory), as well as modulate TGF-β and BDNF (neurotrophic factor). Moreover, HNSC-secretomes maintain the extracellular matrix of SCI by reducing the matrix degradation effect of MMP-9 and increasing the collagen formation effect of TGF-β as a resistor of glial scar formation.
Conclusions
The present study demonstrated the mechanism of HNSC-secretome in improving neuropathic pain and locomotor function in SCI through antioxidant, anti-inflammatory, anti-matrix degradation, and neurotrophic activities.

Keyword

Brain-Derived Neurotrophic Factor; F2-Isoprostanes; Gliosis; Matrix Metalloproteinases; Neuralgia; Neural Stem Cells; Oxidative Stress; Secretome; Spinal Cord Injuries; Transforming Growth Factors; Tumor Necrosis Factor-alpha

Figure

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  Fig. 1 Graph of Rat Grimace Scale (RGS) scores (mean value).

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  Fig. 2 Graph of Basso, Beattie, and Bresnahan (BBB) scores locomotors recovery (mean value).

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  Fig. 3 A bar graph representing the comparison of cell counts between groups in matrix metalloproteinase-9 cytokine.

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  Fig. 4 We observed immunohistochemical matrix metalloproteinase (MMP)-9 average value of 10 field of views, every field of view have 625 µ2 with 400× magnification. (A) Treatment group, (B) Control group, (C) Normal group. Microglia (red arrow) are small round cells, solid nuclei and give a positive reaction with anti MMP-9 indicated by brown color. While macrophage (blue arrow) cells are large, vesicular nucleus, and sometimes elongated resembling fibroblasts (macrophages like fibroblast) and give a positive reaction with anti MMP-9 indicated by brown.

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  Fig. 5 A bar graph representing the comparison of cell counts between groups in transforming growth factor (TGF)-β cytokine.

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  Fig. 6 We observed immunohistochemical transforming growth factor (TGF)-β average value of 10 field of views, every field of view have 625 µ2 with 400× magnification. (A) Treatment group, (B) Control group, (C) Normal group. Microglia are small round cells, solid nuclei and give a positive reaction with anti TGF-β indicated by brown color. While macrophage cells are large, vesicular nucleus, and sometimes elongated resembling fibroblasts (macrophages like fibroblast) and give a positive reaction with anti TGF-β indicated by brown.

• 

  Fig. 7 A bar graph representing the comparison of cell counts between groups in brain derived neurotrophic factor (BDNF).

• 

  Fig. 8 We observed immunohistochemical brain derived neurotrophic factor (BDNF) average value of 10 field of views, every field of view have 625 µ2 with 400× magnification. (A) Treatment group, (B) Control group, (C) Normal group. Macrophage cells are large, vesicular nucleus, and sometimes elongated resembling fibroblasts (macrophages like fibroblast) and give a positive reaction with anti BDNF indicated by brown.

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