Protective effect of low-level laser irradiation on lipopolysaccharide-mediated inflammatory injury of human periodontal ligament fibroblasts.

OBJECTIVES To study the effect and mechanism of low-level laser irradiation (LLLI) on lipopolysaccharide (LPS)-induced inflammatory injury of human periodontal ligament fibroblasts (hPDLFs). METHODS hPDLFs were inoculated into well plates and randomly divided into the normal group, LPS group, and LPS+LLLI group. The cells in the normal group were cultured in conventional medium. The hPDLFs in the LPS and LPS+LLLI groups were cultured in RPMI1640 medium containing 1 mg·L-1 LPS. The three subgroups of the LPS+LLLI group were exposed to different LLLI. After 4 days, the cell apoptosis, viability, and intracellular free Ca2+ concentration of each group were measured. The contents of tumor necrosis factor-α (TNF-α), interleukin (IL)-8, IL-1β, and IL-6 were measured by enzyme linked immunosorbent assay (ELISA). Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to detect the expression of matrix metalloproteinase (MMP)-2, MMP-3, and MMP-9 genes and proteins of hPDLFs in each group. RESULTS Compared with the normal group, the LPS group showed increased apoptosis rate of hPDLFs and intracellular free Ca2+concentration and decreased cell viability (P<0.05). The TNF-α, IL-8, IL-1β, and IL-6 levels were higher in the cell supernatant (P<0.05), and the expression of MMP-2, MMP-3, and MMP-9 genes and proteins of hPDLFs was significantly increased (P<0.05). Compared with the LPS group, the LPS+LLLI group showed significantly decreased apoptosis rate and intracellular free Ca2+ concentration and significantly increased cell viability (P<0.05). The TNF-α, IL-8, IL-1β, and IL-6 levels in the supernatant of cells and the expression of MMP-2, MMP-3, and MMP-9 genes and proteins of hPDLFs were significantly decreased (P<0.05). CONCLUSIONS LLLI has a protective effect on the inflammatory injury of hPDLFs induced by LPS, and the effect is most obvious when the irradiation intensity is 4 J·cm-2.