Extending the vitamin D pathway to vitamin D₃ and CYP27A1 in periodontal ligament cells

Background: In periodontal connective tissue cells, the vitamin D pathway has been elucidated, and vitamin D₃ in the main storage form, 25-hydroxy vitamin D₃ (25[OH]D₃ ), and the functional form, 1,25-dihydroxy vitamin D₃ (1,25[OH]₂ D₃ ), have been found to induce the expression of human cationic antimicrobial protein (hCAP-18)/LL-37. Moreover, synergistic effects between Toll-like receptor agonists and 25(OH)D₃ have been reported. This research aimed at extending the vitamin D pathway to vitamin D₃ and CYP27A1 in human periodontal ligament cells (hPDLCs) to further explore its function in periodontal inflammatory reaction.

Methods: Vitamin D₃ was used to stimulate hPDLCs in the presence or absence of Porphyromonas gingivalis lipopolysaccharide (Pg-LPS). Conversely, CYP27A1 RNA interference was performed to further validate the findings. The mRNA expression of hCAP-18 was determined with real-time polymerase chain reaction. Monocyte chemotactic protein-1 (MCP-1) and interleukin-8 (IL-8) were also detected. The cell supernatant levels of LL-37 were detected with enzyme-linked immunosorbent assay.

Results: Vitamin D₃ significantly enhanced the generation of hCAP-18/LL-37. A combination of Pg-LPS and vitamin D₃ significantly promoted hCAP-18/LL-37 expression. When the expression of CYP27A1 was knocked down with RNA interference, the induction of hCAP-18/LL-37 expression was significantly inhibited. Therefore, the mRNA levels of MCP-1 and IL-8 in hPDLCs were significantly decreased through the vitamin D pathway.

Conclusion: The vitamin D pathway from vitamin D₃ to hCAP-18/LL-37 exists in hPDLCs, and CYP27A1 might be involved in periodontal immune defense.