2. Academic Validation
  3. ILK inhibition reduces osteophyte formation through suppression of osteogenesis in BMSCs via Akt/GSK-3β/β-catenin pathway

ILK inhibition reduces osteophyte formation through suppression of osteogenesis in BMSCs via Akt/GSK-3β/β-catenin pathway

  • Mol Biol Rep. 2024 Mar 14;51(1):421. doi: 10.1007/s11033-024-09336-5.
Zhixiang Huang # 1 2,  Lixin Huang # 1 2 3,  Jiali Ding 2 4,  Yukai Huang 1 2,  Xuechan Huang 1 2,  Tianwang Li 5 6 7 8
Affiliations

Affiliations

  • 1 The Second School of Clinical Medicine, Southern Medical University, Guangzhou, China.
  • 2 Department of Rheumatology and Immunology, Guangdong Second Provincial General Hospital, No. 466, Xingangzhong Road, Guangzhou, 510317, China.
  • 3 Department of Rheumatology, Fujian Medical University Union Hospital, Fuzhou, China.
  • 4 Guangdong Medical University, Zhanjiang, China.
  • 5 The Second School of Clinical Medicine, Southern Medical University, Guangzhou, China. litian-wang@163.com.
  • 6 Department of Rheumatology and Immunology, Guangdong Second Provincial General Hospital, No. 466, Xingangzhong Road, Guangzhou, 510317, China. litian-wang@163.com.
  • 7 Guangdong Medical University, Zhanjiang, China. litian-wang@163.com.
  • 8 Department of Rheumatology and Immunology, Zhaoqing Central People's Hospital, Zhaoqing, China. litian-wang@163.com.
  • # Contributed equally.
PMID: 38483756 DOI: 10.1007/s11033-024-09336-5
Abstract

Background: Osteophyte development is a common characteristic of inflammatory skeletal diseases. Elevated osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) participates in pathological osteogenesis. Integrin-linked kinase (ILK) positively regulates the osteoblastic differentiation of osteoprogenitors, but whether the ILK blockage prevents osteophytes and its potential mechanism is still unknown. Furthermore, the low-dose tumor necrosis factor-α (TNF-α) promotes osteogenic differentiation, but a lack of study reports on the relationship between this cytokine and ILK. OSU-T315 is a small ILK inhibitor, which was used to determine the effect of ILK inhibition on osteogenesis and osteophyte formation.

Methods and results: The osteogenesis of BMSCs was evaluated using Alizarin red S staining, Alkaline Phosphatase, collagen type I alpha 2 chain, and bone gamma-carboxyglutamate protein. The expression and phosphorylation of protein were assessed through western blot. Immunofluorescence was employed to display the distribution of β-catenin. microCT, hematoxylin-eosin, and safranin O/fast green staining were utilized to observe the osteophyte formation in collagen antibody-induced arthritis mice. We found that ILK blockage significantly declined calcium deposition and osteoblastic markers in a dose- and time-dependent manner. Furthermore, it lowered osteogenesis in the TNF-α-induced inflammatory microenvironment by diminishing the effect of ILK and inactivating the Akt/ GSK-3β/ β-catenin pathway. Nuclear β-catenin was descended by OSU-T315 as well. Finally, the ILK suppression restrained osteophyte formation but not inflammation in vivo.

Conclusions: ILK inhibition lowered osteogenesis in TNF-α-related inflammatory conditions by deactivating the Akt/ GSK-3β/ β-catenin pathway. This may be a potential strategy to alleviate osteophyte development in addition to anti-inflammatory treatment.

Keywords

Bone marrow mesenchymal stem cells; Collagen antibody-induced arthritis; Integrin-linked kinase; Osteophyte; Tumor necrosis factor-α.

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