Interleukin-17A Promotes Proliferation and Osteogenic Differentiation of Human Ligamentum Flavum Cells Through Regulation of β-Catenin Signaling

• Published in: Spine (Philadelphia, Pa. 1976) Pa. 1976), 2023-11, Vol.48 (21), p.E362-E371
• Main Authors: Lin, Jialiang, Jiang, Shuai, Xiang, Qian, Zhao, Yongzhao, Wang, Longjie, Fan, Dongwei, Zhong, Woquan, Sun, Chuiguo, Chen, Zhongqiang, Li, Weishi
• Format: Article
• Language: English
• ISSN: 0362-2436; 1528-1159
• DOI: 10.1097/BRS.0000000000004789

Basic experimental study. To elucidate the role and mechanism of IL-17A in thoracic ossification of the ligamentum flavum (TOLF). TOLF is characterized by the replacement of the thoracic ligamentum flavum with ossified tissue and is one of the leading causes of thoracic spinal stenosis. IL-17A is an important member of the IL-17 family that has received widespread attention for its key contributions to the regulation of bone metabolism and heterotopic ossification. However, it is unclear whether IL-17A is involved in TOLF. Cell counting kit-8 assay and 5-Ethynyl-2'-deoxyuridine staining were performed to assess the proliferation of ligamentum flavum cells (LFCs). Alkaline phosphatase activity assay, Alizarin red staining, and protein level expression of osteogenic-related genes were used to evaluate the osteogenic differentiation potential of LFCs. The effect of IL-17A on the proliferation and osteogenic differentiation of LFCs was further assessed after silencing β-catenin by transfection with siRNA. In addition, the possible source of IL-17A was further demonstrated by co-culture assays of T helper 17 (Th17) cells with LFCs. Student's t test (t-test) was used for comparisons between groups and the one-way analysis of variance (ANOVA) followed by Tukey's post hoc test was used for comparison of more than two groups. IL-17A was elevated in TOLF tissue compared to normal ligamentum flavum. IL-17A stimulation promoted the proliferation and osteogenic differentiation of LFCs derived from TOLF patients. We found that IL-17A promoted the proliferation and osteogenic differentiation of LFCs by regulating the β-catenin signaling. Co-culture of Th17 cells with LFCs enhanced β-catenin signaling-mediated proliferation and osteogenic differentiation of LFCs. However, these effects were markedly attenuated after the neutralization of IL-17A. This is the first work we are aware of to highlight the importance of IL-17A in TOLF. IL-17A secreted by Th17 cells in the ligamentum flavum may be involved in ossification of the microenvironment by regulating β-catenin signaling to promote proliferation and osteogenic differentiation of LFCs.