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Updated: Oct 3, 2025
A Proinflammatory, Degenerative Organ Culture Model to Simulate Early-Stage Intervertebral Disc Disease.
Published on: February 14, 2021
M2 macrophage-conditioned medium inhibits intervertebral disc degeneration in a tumor necrosis factor-α-rich
Xiao-Chuan Li1,2,3, Shao-Jian Luo3, Wu Fan3
1Postdoctoral Innovation Practice Base of Gaozhou People's Hospital, Gaozhou People's Hospital, Maoming, Guangdong, China.
Abstract:
Inflammation is the primary pathological phenomenon associated with disc degeneration; the inflammatory cytokine tumor necrosis factor (TNF-α) plays a crucial role in this pathology. The anti-inflammatory and regenerative effects of M2 macrophages on nucleus pulposus cells (NPCs) in intervertebral disc degeneration (IDD) progression remain unknown. Here, M2 conditioned medium (M2CM) was harvested and purified from human acute monocytic leukaemia cell line (THP-1) cells and mouse peritoneal macrophages, respectively; it was used for culturing human NPCs and a mouse intervertebral disc (IVD) organ culture model. NPCs and IVD organ models were divided into three groups: group 1 treated with 10% fetal bovine serum (control); group 2 treated with 10 ng/ml TNF-α; and group 3 treated with 10 ng/ml TNF-α and M2CM (coculture group). After 2-14 days, cell proliferation, extracellular matrix synthesis, apoptosis, and NPC senescence were assessed. Cell proliferation was reduced in TNF-α-treated NPCs and inhibited in the M2CM co-culture treatment. Moreover, TNF-α treatment enhanced apoptosis, senescence, and expression of inflammatory factor-related genes, including interleukin-6, MMP-13, ADAMTS-4, and ADAMTS-5, whereas M2CM coculture significantly reversed these effects. In addition, co-culture with M2CM promoted aggrecan and collagen II synthesis, but reduced collagen Iα1 levels in TNF-α treatment groups. Using our established three-dimensional murine IVD organ culture model, we show that M2CM suppressed the inhibitory effect of TNF-α-rich environment. Therefore, co-culture with M2CM promotes cell proliferation and extracellular matrix synthesis and inhibits inflammation, apoptosis, and NPC senescence. This study highlights the therapeutic potential of M2CM for IDD.
Insights
M2 macrophages, via M2 conditioned medium, show potential in treating intervertebral disc degeneration (IDD) by reducing inflammation and promoting cell repair. This study explores their therapeutic effects on nucleus pulposus cells (NPCs) and disc organ models.
Area of Science:
- Biomedical Science
- Regenerative Medicine
- Cell Biology
Background:
- Intervertebral disc degeneration (IDD) is characterized by inflammation, with tumor necrosis factor-alpha (TNF-α) playing a key role.
- The therapeutic potential of M2 macrophages in mitigating IDD progression is not well understood.
Purpose of the Study:
- To investigate the anti-inflammatory and regenerative effects of M2 conditioned medium (M2CM) on nucleus pulposus cells (NPCs) and an intervertebral disc (IVD) organ culture model.
- To assess the impact of M2CM on NPC proliferation, extracellular matrix synthesis, apoptosis, and senescence in the context of TNF-α-induced degeneration.
Main Methods:
- M2CM was derived from human THP-1 cells and mouse peritoneal macrophages.
- Human NPCs and a 3D murine IVD organ culture model were treated with TNF-α alone or in combination with M2CM.
- Assessed outcomes included cell proliferation, apoptosis, senescence, and the expression of key genes related to inflammation and extracellular matrix synthesis.
Main Results:
- TNF-α treatment increased NPC apoptosis, senescence, and inflammatory gene expression while reducing proliferation and extracellular matrix synthesis.
- M2CM co-culture significantly reversed the detrimental effects of TNF-α, promoting NPC proliferation and extracellular matrix synthesis (aggrecan, collagen II) while inhibiting apoptosis, senescence, and inflammation.
- M2CM demonstrated a protective effect in the 3D murine IVD organ culture model against a TNF-α-rich environment.
Conclusions:
- M2CM exhibits significant anti-inflammatory and regenerative properties beneficial for intervertebral disc degeneration.
- M2CM promotes NPC proliferation and extracellular matrix synthesis while inhibiting inflammation, apoptosis, and senescence, highlighting its therapeutic potential for IDD.
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