25‐Hydroxycholesterol induces odontoclastic differentiation through RANK–RANKL upregulation and NF‐κB activation in odontoblast‐like MDPC‐23 cells: An in vitro study. (12th December 2022)

• Record Type: Journal Article
• Authors: Lim, HyangI; Oh, Ji‐Su; Kang, Kyeong‐Rok; Seo, Jeong‐Yeon; Kim, Do Kyung; Yu, Sun‐Kyoung; Kim, Heung‐Joong; Park, Joo‐Cheol; Kim, Jae‐Sung
• Is Part Of: International endontic journal. Volume 56:Number 4(2023)
• Page Start: 432
• Page End: 446
• Publication Date: 2022-12-12
• Subjects: 25‐hydroxycholesterol -- odontoclast -- odontoclastogenesis -- oxysterol -- RANK‐RANKL signalling pathway; Endodontics -- Periodicals; 617.6342
• DOI: 10.1111/iej.13878
• Languages: English
• ISSNs: 0143-2885

Abstract

Abstract: Aim: The physiological effects and cellular mechanism of 25‐hydroxycholesterol (25‐HC), which is an oxysterol synthesized from cholesterol by cholesterol‐25‐hydroxylase (CH25H) expressed under inflammatory conditions, are still largely unknown during odontoclastogenesis. This study aimed to evaluate 25‐HC‐induced odontoclastogenesis and its cellular mechanisms in odontoblast‐like MDPC‐23 cells. Methodology: To investigate 25‐HC‐induced odontoclastogenesis of MDPC‐23 cells and its cellular mechanism, haemotoxylin and eosin staining, tartrate‐resistant acid phosphatase (TRAP) staining, dentine resorption assay, zymography, reactive oxygen species (ROS) detection, immunocytochemistry, and nuclear translocation were performed. The experimental values are presented as mean ± standard deviation and were compared using analysis of variance, followed by post hoc multiple comparisons (Tukey's test) using SPSS software version 22 (IBM Corp.). A p ‐value <.05 was considered statistically significant. Results: Lipopolysaccharide or receptor activator of nuclear factor‐κB ligand (RANKL) induced the synthesis of 25‐HC via the expression of CH25H in MDPC‐23 cells ( p < .01). Multinucleated giant cells with morphological characteristics and TRAP activity of the odontoclast were increased by 25‐HC in MDPC‐23 cells ( p < .01). Moreover, 25‐HC increased dentine resorption through the expression and activity of matrix metalloproteinases in MDPC‐23 cells. It not only increased the expression of odontoclastogenic biomarkers but also translocated cytosolic nuclear factor‐κB (NF‐κB) to the nucleus in MDPC‐23 cells. Additionally, 25‐HC not only increased the production of ROS ( p < .01), expression of inflammatory mediators ( p < .01), pro‐inflammatory cytokines, receptor activator of NF‐κB (RANK), and RANKL but also suppressed the expression of osteoprotegerin (OPG) in MDPC‐23 cells. In contrast, CDDO‐Me, a chemical NF‐κB inhibitor, decreased TRAP activity ( p < .01) and downregulated the expression of the odontoclastogenic biomarkers, including RANK and RANKL, in MDPC‐23 cells. Conclusion: 25‐HC induced odontoclastogenesis by modulating the RANK–RANKL–OPG axis via NF‐κB activation in MDPC‐23 cells. Therefore, these findings provide that 25‐HC derived from cholesterol metabolism may be involved in the pathophysiological etiological factors of internal tooth resorption.