Establishment of chondrocyte model of iron overload and the mechanism of injury

doi:10.12307/2023.590

Abstract

Abstract: BACKGROUND: It is unclear how excessive iron is involved in the development of arthritis, and investigating the potential mechanisms by which iron overload is involved in the development of arthritis could provide new directions for the treatment of arthritis.
OBJECTIVE: To establish an iron overload model of chondrocytes by co-culture of ferric ammonium citrate and chondrocytes, and to observe the effects of iron overload on chondrocyte survival, iron deposition, oxidative stress, and mitochondrial damage.
METHODS: The chondrocytes of C57BL/6J mouse knee joint were extracted by enzyme digestion and cultured with ferric ammonium citrate (100, 250, 500, 1 000 and 2 000 μmol/L) chondrocytes for 24, 48 and 72 hours to establish chondrocyte models of iron overload. Cell viability was measured by cell counting kit-8. Content of iron was determined by Calcein-AM, and ferric ammonium citrate concentration was screened for subsequent experiments. Then, the level of reactive oxygen species was detected by fluorescence probe DCFH-DA and the change of mitochondrial membrane potential was detected by JC-1. Real-time quantitative PCR and western blot assays were used to detect the expression levels of type II collagen, matrix metalloproteinases 3 and 13.
RESULTS AND CONCLUSION: The iron overload model could be successfully established in chondrocytes co-cultured with ferric ammonium citrate at the concentration of 500 and 1 000 μmol/L for 48 hours. Ferric ammonium citrate could induce iron overload in chondrocytes, and iron overload subsequently destroy mitochondrial function, increase the level of intracellular reactive oxygen species, impair the functional activity of chondrocytes, and decrease the cell ability to secrete type II collagen and aggrecan, which further increases the difficulty of articular cartilage self-repair. It lays a foundation for further research on the mechanism of chondrocyte injury caused by iron overload and the exploration of effective therapeutic drugs for the treatment of iron overload-induced arthritis.

Key words: iron overload, chondrocyte, oxidative stress, mitochondrial function

CLC Number:

He Qi, Pan Zhaofeng, Chen Baihao, Yang Junzheng, Li Shaocong, Zeng Jiaxu, Zhou Chi, Wang Haibin. Establishment of chondrocyte model of iron overload and the mechanism of injury[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(32): 5126-5132.

Figures/Tables 7

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