IL-1β-regulating angiogenic factors expression in perforated temporomandibular disk cells via NF-κB pathway.

Abstract

A high density of blood vessels is observed in the perforated disks of temporomandibular joint (TMJ), but the underlying mechanism is unknown. This study aimed to explore the regulation of disk angiogenesis in the perforated disks. Expressions of vascular endothelial growth factor (VEGF), angiogenin-1 (Ang-1), chondromodulin-1 (ChM-1), and thrombospondins-1 (TSP-1) were compared between healthy and perforated TMJ disk cells with or without interleukin-1β (IL-1β) incubation. The tube formation, cell migration, and expressions of matrix-metalloproteinases (MMPs) in human umbilical vein endothelial cell line (HUV-EC-C) were investigated in conditional media of disk cells. Western blot was performed to determine protein level of VEGF, Ang-1, ChM-1 and TSP-1 in IL-1β-induced disk cells cultured by NF-κB- or P38-specific pathway inhibitors, respectively. Conditional media from perforated disk cells induced more tube formation, cell migration, and MMPs' expression in HUV-EC-C. Expressions of VEGF and Ang-1 were significantly higher, and ChM-1 and TSP-1 were lower in perforated disks compared to healthy disks. The VEGFA concentration was 291.1 ± 36.09 pg/ml in perforated disk cell conditioned media, markedly larger than that in NDCCM (144.9 ± 33.69 pg/ml). IL-1β induced VEGF through NF-κB signaling pathway and Ang-1 through p38 MAPK pathway, while repressed expression of ChM-1 and TSP-1 was through NF-κB pathway. Blockade of each pathway markedly restrained inducing effect of cultural media on HUV-EC-C tube formation and migration. Perforated disk cells secreted more angiogenic factors which might induced via NF-κB pathway.