Chinese Journal of Tissue Engineering Research ›› 2024, Vol. 28 ›› Issue (13): 1976-1982.doi: 10.12307/2024.126

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Chondrogenic differentiation of rabbit bone marrow mesenchymal stem cells induced by growth differentiation factor 5

Li Feifei, Wang Buyu, Yang Zhihang, Dong Xiaoyu, Deng Jiang   

  1. Third Affiliated Hospital of Zunyi Medical University (First People’s Hospital of Zunyi), Zunyi 563000, Guizhou Province, China
  • Received:2023-02-28 Accepted:2023-04-04 Online:2024-05-08 Published:2023-08-28
  • Contact: Deng Jiang, Professor, Chief physician, Doctoral supervisor, Third Affiliated Hospital of Zunyi Medical University (First People’s Hospital of Zunyi), Zunyi 563000, Guizhou Province, China
  • About author:Li Feifei, Master candidate, Physician, Third Affiliated Hospital of Zunyi Medical University (First People’s Hospital of Zunyi), Zunyi 563000, Guizhou Province, China
  • Supported by:
    National Natural Science Foundation of China, No. 81660367 (to DJ); Guizhou Provincial Health Commission Science and Technology Fund, No. gzwkj 2021-236 (to DJ)

Abstract: BACKGROUND: Growth differentiation factor 5 is a member of the transforming growth factor superfamily and one of the earliest markers of joint development. Growth differentiation factor 5 has an important role in cartilage repair. 
OBJECTIVE: To explore the action mechanism of growth differentiation factor 5-induced chondrogenic differentiation of bone marrow mesenchymal stem cells. 
METHODS: Rabbit bone marrow mesenchymal stem cells were isolated and cultured. CCK-8 assay was used to detect the effect of different mass concentrations of growth differentiation factor 5 on the proliferation activity of bone marrow mesenchymal stem cells. RT-PCR was utilized to detect the expression of genes related to chondrogenic differentiation of bone marrow mesenchymal stem cells induced by different mass concentrations of growth differentiation factor 5. To further investigate the action mechanism of growth differentiation factor 5-induced chondrogenic differentiation of bone marrow mesenchymal stem cells, we added inhibitor XAV-939 and activator Laduviglusib of Wnt/β-catenin signaling pathways to induce cell culture for 14 days. RT-PCR and western blot assay were performed to detect the expression of cartilage-related genes and Wnt/β-catenin signaling pathway proteins.
RESULTS AND CONCLUSION: (1) CCK-8 results showed no significant effect of growth differentiation factor 5 on the proliferation of bone marrow mesenchymal stem cells. (2) Growth differentiation factor 5 promoted the expression of cartilage-related genes type II collagen, aggrecan and Sox9, among which growth differentiation factor 5 induced a significant upregulation of cartilage-related genes in the 50 ng/mL group. (3) Addition of Laduviglusib, an activator of Wnt/β-catenin signaling pathway, upregulated Sox9, β-catenin and type II collagen expression (P < 0.05). Addition of XAV939, an inhibitor of Wnt/β-catenin signaling pathway, down-regulated Sox9, β-catenin and type II collagen expression (P < 0.05). (4) Taken together, growth differentiation factor 5-induced chondrogenic differentiation of bone marrow mesenchymal stem cells may be associated with the activation of the Wnt/β-catenin signaling pathway. 

Key words: growth differentiation factor 5, bone marrow mesenchyml stem cell, cartilage differentiation, chondrocyte, Wnt, β-catenin, mechanism

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