Metformin affects changes of cartilage and subchondral bone in mouse models of early osteoarthritis

doi:10.3969/j.issn.2095-4344.1249

Abstract

Abstract:

BACKGROUND: Metformin has been shown to hold anti-inflammatory effect, inhibit the expression of interleukin1β and tumor necrosis factor α, and affect bone metabolism by activating AMPK signaling pathway. However, the effects of metformin on cartilage and subchondral bone remain obscure.
OBJECTIVE: To investigate the effects of metformin on articular cartilage injury and pathological changes of subchondral bone in early osteoarthritis in mice.
METHODS: C57 mice, SPF grade and postnatal 3 day neonate mice (used for primary chondrocyte culture) were provided by Laboratory Animal Center of Southern Medical University, and the experimental protocol was approved by the Animal Ethics Committee of the Third Affiliated Hospital of Southern Medical University. (1) Modeling: Eight-week-old C57 male mice were randomly divided into control and experimental groups. The traumatic osteoarthritis model was established in all mice, the experimental group was treated with 200 mg/(kg•d) metformin by intragastric administration, and the control group the same volume of normal saline. The knee joint specimens were taken for histological analysis after 5 weeks of treatment. Safranin O fast green staining and Osteoarthritis Research Society International score were used to evaluate the damage of articular cartilage and the degree of subchondral osteosclerosis. The expression of matrix metalloproteinase 13, collagen type II, Runt-related transcription factor 2 and osteocalcin protein was detected by immunohistochemical staining. (2) Cytological experiment: Primary chondrocytes were isolated from neonate mice born within three days, induced by 10 μg/L interleukin 1β, and co-cultured with metformin (5, 10 mmol/L). The expression levels of collagen type II, Runt-related transcription factor 2 and osteocalcin protein in chondrocytes were detected by western blot assay.
RESULTS AND CONCLUSION: (1) The results of safranin O fast green staining and Osteoarthritis Research Society International score showed that the degree of articular cartilage damage and subchondral osteosclerosis in the control group was more serious than that in the experimental group. (2) The immunohistochemical staining results revealed that the expression level of matrix metalloproteinase 13 in the control group was significantly higher than that in the experimental group. In the experimental group, the level of collagen type II was on a rise, and the levels of Runt-related transcription factor 2 and osteocalcin proteins were on a decline (P < 0.05). (3) The expression levels of matrix metalloproteinase 13 and Runt-related transcription factor 2 were significantly increased after interleukin 1β induction, and the expression level of collagen type II was significantly decreased. After metformin added, the expression levels of matrix metalloproteinase 13 and Runt-related transcription factor 2 were significantly decreased, and the expression level of collagen type II was significantly increased with metformin concentration increasing. (4) In summary, metformin can alleviate the degradation of cartilage and cartilage matrix of early osteoarthritis and inhibit the sclerosis and remodeling of subchondral bone, thereby playing a role in delaying the progress of early osteoarthritis.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: subchondaral bone, metformin, osteoarthritis, articular cartilage, osteoarthritic cartilage, cartlilage matrix

CLC Number:

R446
R318

Feng Xiaofeng, Zhang Rongkai, Qi Weizhong, Pan Jianying, Li Junyan, Bai Xiaochun, Cai Daozhang . Metformin affects changes of cartilage and subchondral bone in mouse models of early osteoarthritis[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(19): 3031-3036.

Figures/Tables 4

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