miR-23b mediates TNF-α-Inhibited Osteogenic Differentiation of Human Periodontal Ligament Stem Cells by Targeting Runx2

Abstract

Periodontitis is the most prevalent oral infection disease, which causes the destruction of periodontal supporting tissues and eventual tooth loss. This study aimed to investigate the molecular mechanism of miRNA-23b (miR-23b) in regulating the osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) in an inflammatory environment. Results revealed that tumor necrosis factor-α (TNF-α), a notoriously inflammatory cytokine, remarkably attenuated the osteogenic differentiation of hPDLSCs, which were partially rescued by SKL2001 (Wnt/β-catenin agonist). We further explored the underlying roles of miRNAs involved in TNF-α-inhibited osteogenesis of hPDLSCs. The miR-23b significantly increased with TNF-α stimulation, which was abolished by SKL2001. Similar to the effect of TNF-α, miR-23b agonist (agomir-23b) dramatically reduced the expression of runt-related transcription factor 2 (Runx2) and suppressed the osteogenic differentiation of hPDLSCs. The inhibition of miR-23b significantly increased Runx2, which is the major transcription factor during osteogenesis, thereby indicating that miR-23b was an endogenous regulator of Runx2 in hPDLSCs. Bioinformatic analysis and dual luciferase reporter assays confirmed that Runx2 was a target gene of miR-23b. Furthermore, the gain function assay of Runx2 revealed that the Runx2 overexpression efficiently reversed the suppression of the osteogenic differentiation of hPDLSCs with miR-23b agonist, suggesting that the suppressing effect of miR-23b on osteogenesis was mediated by Runx2 inhibition. Our study clarified that miR-23b mediated the TNF-α-inhibited osteogenic differentiation of hPDLSCs by targeting Runx2. Therefore, the expanded function of miR-23b in the osteogenesis of hPDLSCs under inflammatory conditions. This study might provide new insights and a novel therapeutic target for periodontitis.

The role of TNF-α in the fate regulation and functional reprogramming of mesenchymal stem cells in an inflammatory microenvironment

“…High concentrations of TNF-α were shown to induce the expression of miR-23b and inhibit RUNX2 expression via the NF-κB in human bone marrow MSCs ( 65 ) or the Wnt/β-catenin signaling pathways in human dental MSCs ( 66 ), thus suppressing the osteogenic differentiation of human MSCs. 10 ng/ml TNF-α also inhibited the osteogenic differentiation ability of human periodontal stem cells by decreasing miR-21 expression, leading to a rise in recombinant Sprouty Homolog 1 (SPRY1) levels, thereby suppressing ALP and RUNX2 production ( 67 ).…”

Tetrahedral Framework Nucleic Acid Loaded miR‐23b Inhibits Synovial Inflammation and Cartilage Matrix Degradation in the Treatment of Rheumatoid Arthritis

“…These functions may be achieved by targeting key regulatory factors and signaling pathways, such as inhibiting the expression of matrix metalloproteinase (MMP) family members or modulating the activity of the Wnt/𝛽-catenin signaling pathway. [13,14] However, the single-stranded structure of pure miRNAs is unstable and can be easily degraded, which makes it difficult to exert their function alone. As a result, plasmids are often employed to deliver miRNAs.…”

“…These functions may be achieved by targeting key regulatory factors and signaling pathways, such as inhibiting the expression of matrix metalloproteinase (MMP) family members or modulating the activity of the Wnt/β‐catenin signaling pathway. [ 13,14 ]…”

Identifying genes for regulating osteogenic differentiation of human periodontal ligament stem cells in inflammatory environments by bioinformatics analysis

“…23,24 However, hPDLSCs from periodontitis patients show poor osteogenic ability, including decreased expression of osteoblast genes and decreases in calcified nodular deposition and alkaline phosphatase (ALP) activity. 25 Therefore, methods to improve the osteogenic differentiation ability of hPDLSCs in the inflammatory condition is promising for periodontitis treatment. Previous studies have claimed that the osteogenic differentiation of hPDLSCs is mainly regulated with the following signaling pathways: MAPK, Wnt/β-catenin, and PI3K/AKT.…”