Alleviation of oxidative stress damage in chondrocytes by a new Mn-containing bioceramic powder

doi:10.12307/2025.434

Abstract

Abstract: BACKGROUND: Mn can participate in oxidation-reduction reactions in various organisms. For example, as a metal-assisted group in superoxide dismutase 2, Mn plays a role in helping to remove reactive oxygen species. Therefore, the development of novel anti-oxidative stress materials containing Mn has become a research focus in recent years.
OBJECTIVE: To investigate the protective effect of Mn bioceramic powder material on oxidative stress damage to chondrocytes by reducing the reactive oxygen species pathway.
METHODS: Bioceramic powders containing Mn were prepared by molten salt method. Primary mouse chondrocytes were isolated and cultured. Bioceramic powder containing 0, 0.15, and 0.30 mg/mL of Mn was added into H2O2 solution. The H2O2 clearance rate was detected after incubation without light. The passage 2 to passage 4 chondrocytes were co-cultured with complete media containing Mn-containing bioceramic powder with different mass concentrations (0, 0.15, and 0.30 mg/mL). Cell viability was detected by cell live/dead staining. The passage 2-4 chondrocytes (or cartilage tissue) were divided into four groups for intervention: Complete culture medium was added to the blank control group. The H2O2 group was added and cultured with complete medium containing H2O2. H2O2+low mass concentration Mn powder group was cultured by adding H2O2+0.15 mg /mL Mn-containing bioceramic powder. The complete medium containing H2O2+0.30 mg/mL Mn-containing bioceramic powder was added to the H2O2+high mass concentration Mn powder group. Viability of chondrocytes was detected by CCK-8 assay. Generation of reactive oxygen species of chondrocytes was detected by 2,7-dichlorofluorescein diacetate probe. Expression of chondrocyte-related factors was detected by qRT-PCR. The tissue structure and function of cartilage were detected by toluidine blue staining.
RESULTS AND CONCLUSION: (1) Both doses of Mn-containing bioceramic powders could significantly remove H2O2 in vitro, and they were concentration dependent. The results of cell live/death staining showed that 0.15 mg/mL bioceramic powder containing Mn had chondrocyte safety, and 0.30 mg/mL bioceramic powder containing Mn had chondrocytotoxicity. (2) The results of CCK-8 assay showed that the two mass concentrations of Mn-containing bioceramic powders could significantly reduce the inhibitory effect of H2O2 on chondrocyte viability, and inhibit the generation of reactive oxygen species induced by H2O2 in chondrocytes in a mass concentration dependent manner. Both kinds of Mn-containing bioceramic powders could reverse the H2O2-induced increase of mRNA expression of a disintegrin and metalloproteinase with thrombospondin motifs-5 and decrease of proteoglycan mRNA expression in chondrocytes. (3) Toluidine blue staining results showed that both concentrations of Mn-containing bioceramic powder could protect the integrity of cartilage tissue structure under oxidative stress, and 0.30 mg/mL of Mn-containing bioceramic powder could also reduce the functional damage of cartilage tissue. (4) The results indicate that the Mn-containing bioceramic powder can protect chondrocytes under oxidative stress by clearing reactive oxygen species, maintaining the extracellular matrix homeostasis. However, 0.30 mg/mL Mn-containing bioceramic powder has certain chondrocytotoxicity, so 0.15 mg/mL Mn-containing bioceramic powder is preferred for follow-up studies.

Key words: bioceramic, Mn powder material, cartilage injury, oxidative stress, reactive oxygen species, osteoarthritis, extracellular matrix, biosafety

CLC Number:

hang Zining, Deng Ronghui, Yu Jiakuo. Alleviation of oxidative stress damage in chondrocytes by a new Mn-containing bioceramic powder[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(16): 3335-3342.

Figures/Tables 6

References

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