Cerium dioxide nanoparticles regulate expression of inflammatory factors in M1 macrophages and affect fibroblast co-culture system

doi:10.12307/2025.899

Abstract

Abstract: BACKGROUND: Macrophage polarization plays a key role in chronic inflammatory joint diseases such as rheumatoid arthritis. Cerium dioxide (CeO2) nanoparticles have a wide range of biomedical applications such as modulating the local inflammatory microenvironment of tissues.
OBJECTIVE: To investigate the role of CeO2 nanoparticles on macrophage polarization and inflammatory factor expression, as well as inflammatory modulation in a co-culture system of macrophages and fibroblasts.
METHODS: (1) CeO2 nanoparticles were dispersed and observed morphologically by transmission electron microscopy. (2) Human leukemia monocytes (THP-1) were induced to differentiate and establish the M1 macrophage pro-inflammatory cell model of rheumatoid arthritis. The cells were divided into M0 group (undifferentiated macrophages), M1 group (successful macrophage modeling), CeO2 nanoparticle treatment group (M1 group with CeO2 nanoparticle treatment), and dexamethasone control group (M1 group with dexamethasone treatment) and incubated for 48 hours. The effects of CeO2 nanoparticles on the expression of inflammatory factors (endogenous nitric oxide synthase, CD86, CD80) in M1 macrophages and M1 macrophage phenotype (CD80, CD206) were detected by RT-qPCR, western blot assay, and flow cytometry. (3) A co-culture system of macrophages and fibroblasts was established, and CeO2 nanoparticles acted on the upper macrophages. The regulation of CeO2 nanoparticles on the expression of inflammatory factors (interleukin-6, tumor necrosis factor-α, cyclooxygenase-2, and endogenous nitric oxide synthase) of fibroblasts in the co-culture system was observed at the mRNA and protein levels.
RESULTS AND CONCLUSION: (1) Transmission electron microscopy showed that the diameter of CeO2 nanoparticles was (19.5±2.0) nm. (2) Compared with the M0 group, the mRNA of endogenous nitric oxide synthase and CD86, and the protein expression of endogenous nitric oxide synthase and CD80 in the M1 group were upregulated. Compared with the M1 group, the mRNA expression of endogenous nitric oxide synthase and CD86, and the protein expression of endogenous nitric oxide synthase and CD80 in the CeO2 nanoparticle treatment group were downregulated. Flow cytometry showed that 20 nm CeO2 nanoparticles downregulated the number of M1 macrophages. (3) Compared with the M1 group, 20 nm CeO2 nanoparticles downregulated the mRNA and protein expression of inflammatory factors (tumor necrosis factor α, interleukin 6, cyclooxygenase 2, and endogenous nitric oxide synthase) in the co-culture system HFL1 cells. (4) The results showed that 20 nm CeO2 nanoparticles can alleviate inflammation in the co-culture system by inhibiting the expression of pro-inflammatory factors in M1 macrophages, providing a new idea for the treatment of inflammatory diseases such as rheumatoid arthritis.

Key words: CeO2 nanoparticle, macrophage polarization, rheumatoid arthritis, synovitis, Transwell co-culture, engineered microenvironment

CLC Number:

Xie Peisen, Guan Zhenpeng, Wei Xianjie, Zhang Keshi, Kang Qingyuan, Xiao Wentao, Guo Xiaoshuai. Cerium dioxide nanoparticles regulate expression of inflammatory factors in M1 macrophages and affect fibroblast co-culture system[J]. Chinese Journal of Tissue Engineering Research, 2026, 30(2): 375-383.

Figures/Tables 4

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