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Updated: Feb 27, 2026
Isolation and In vitro Culture of Bone Marrow-Derived Macrophages for the Study of NO-Redox Biology
Published on: May 31, 2022
[Effects of paeonol on the function of bone marrow-derived macrophage from Porphyromonas gingivalis-induced mice]
1Dept. of Conservative Dentistry and Endodontics, Guiyang Hospital of Stomatology, Affiliated Guiyang Hospital of Stomatology, Zunyi Medical University, Guiyang 550002, China.
Objective:
This work aims to examine the effects of paeonol treatment on the ability of bone marrow-derived macrophage (BMM) to excrete inflammatory factors and to differentiate into osteoclasts upon induction with Porphyromonas gingivalis (P. gingivalis). This work also aims to investigate the underlying mechanisms of these abilities.
Methods:
BMM culture was treated with different paeonol concentrations at for 1 h and then stimulated with P. gingivalis for 24 h before programmed death-ligand 1 (PD-L1) was quantified with flow cytometry. Tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6 were detected by enzyme-linked immunosorbent assay (ELISA). The BMM culture was treated with the receptor activator for nuclear factor-κB ligand (RANKL) and macrophage colony-stimulating factor (M-CSF), and then with paeonol for 1 h prior to induction with P. gingivalis. Then, osteoclast formation was assessed using tartrate resistant acid phosphatase (TRAP) staining. The osteoclast-related proteins TRAP and receptor activator of nuclear factor-κB (RANK) were quantified by Western blotting.
Results:
Paeonol was nontoxic to BMM within a range of 10-50 μmol·L⁻¹. Flow cytometry showed that paeonol inhibited PD-L1 expression in P. gingivalis-induced BMM in a dose-dependent manner. ELISA indicated that paeonol dose-dependently inhibited the excretion of TNF-α, IL-1β, and IL-6 by P. gingivalis-induced BMM (P<0.01). TRAP staining revealed that paenol treatment inhibited the differentiation of P. gingivalis-induced BMM into osteoclasts. Western blot results suggested that paeonol decreased the expression of TRAP and RANK in BMM.
Conclusions:
Paeonol dose-dependently inhibited the excretion of the inflammatory factors TNF-α, IL-1β, and IL-6 by P. gingivalis-induced BMM in a dose-dependent manner. Moreover, paenol treatment prevented the differentiation of P. gingivalis-induced BMM differentiation into osteoclasts. .
Insights
Paeonol effectively reduces inflammatory factors like TNF-α, IL-1β, and IL-6 in bone marrow-derived macrophages (BMM) stimulated by P. gingivalis. It also inhibits BMM differentiation into osteoclasts, offering potential therapeutic benefits.
Area of Science:
- Immunology
- Cell Biology
- Pharmacology
Background:
- Periodontitis is often associated with Porphyromonas gingivalis (P. gingivalis) infection.
- Bone marrow-derived macrophages (BMM) play a crucial role in inflammatory responses and osteoclastogenesis.
- P. gingivalis can induce inflammatory factor excretion and osteoclast differentiation in BMM.
Purpose of the Study:
- To investigate the effects of paeonol on inflammatory factor excretion by P. gingivalis-induced BMM.
- To determine paeonol's impact on osteoclast differentiation from BMM.
- To explore the underlying mechanisms of paeonol's actions.
Main Methods:
- BMM were treated with varying concentrations of paeonol and stimulated with P. gingivalis.
- Programmed death-ligand 1 (PD-L1) expression was measured by flow cytometry.
- Tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6 were quantified using ELISA.
- Osteoclast differentiation was assessed via TRAP staining and Western blotting for TRAP and RANK expression.
Main Results:
- Paeonol demonstrated no toxicity to BMM within the tested concentration range (10-50 μmol·L⁻¹).
- Paeonol dose-dependently inhibited PD-L1 expression, TNF-α, IL-1β, and IL-6 excretion in P. gingivalis-stimulated BMM.
- Paeonol treatment significantly suppressed osteoclast formation and reduced the expression of TRAP and RANK proteins.
Conclusions:
- Paeonol exhibits potent anti-inflammatory effects by inhibiting key inflammatory mediators in BMM.
- Paeonol effectively prevents the differentiation of BMM into osteoclasts, suggesting a role in modulating bone resorption.
- These findings highlight paeonol's potential as a therapeutic agent against inflammatory conditions linked to P. gingivalis and osteoclast activity.
