BMC Oral Health. 2025 Nov 22;25(1):1822. doi: 10.1186/s12903-025-07145-1.
ABSTRACT
BACKGROUND: In periodontitis, the classical inflammatory NF-κB signaling pathway is activated, which promotes alveolar bone resorption and inhibits osteogenic differentiation of periodontal ligament stem cells. Low-energy laser therapy can reduce inflammation and promote healing. This in vitro study aimed to investigate the effects of different low-energy lasers on inflammation and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) under inflammatory conditions, and to determine whether these effects are associated with the NF-κB signaling pathway at both the protein and gene levels.
METHODS: Healthy premolars or third molars were collected during oral and maxillofacial surgery, and periodontal ligament tissues were obtained. hPDLSCs were cultured using the tissue block method and identified by osteogenic differentiation potential and flow cytometry. Based on treatment conditions, the cells were divided into five groups (using abbreviations from key elements of the grouping): control group (C, control), lipopolysaccharide group (L, LPS), LPS + Nd:YAG laser group (N, Nd:YAG), LPS + Er:YAG laser group (E, Er:YAG), and LPS + semiconductor laser group (D, semiconductor). Enzyme-linked immunosorbent assay (ELISA), quantitative real-time polymerase chain reaction (qRT-PCR), alizarin red staining, and immunofluorescence (IF) were performed to detect inflammatory factors, osteogenic ability, and NF-κB signaling pathway activity. Fluorescence intensity of P65 was quantified using ImageJ software. Statistical analysis was conducted using SPSS 21.0.
RESULTS: The cultured cells were confirmed as periodontal stem cells. Statistical significance was set at P < 0.05. The CCK-8 assay showed that on day 7, the optical density (OD) values of all three laser groups were higher than those of group L (P < 0.05). Compared with group C, IL-6 and TNF-α levels in the supernatant were increased in group L (P < 0.05). After laser treatment, IL-6 and TNF-α levels in groups N, E, and D were lower than in group L (P < 0.05), with the lowest levels observed in group N. Expression of Runx-2 and OSX mRNA was higher in groups N, E, and D than in group L, with group N showing the highest expression of osteogenic genes. The fluorescence intensity of P65 in group L was significantly higher than in group C (P < 0.05), indicating increased nuclear translocation of P65. Among the laser-treated groups, group N showed the lowest nuclear P65 fluorescence intensity (P < 0.05).
CONCLUSIONS: Low-energy lasers promote anti-inflammatory activity and osteogenic differentiation of hPDLSCs in LPS-induced inflammatory environments by regulating the NF-κB signaling pathway, with Nd:YAG lasers showing the strongest effect.
PMID:41275224 | DOI:10.1186/s12903-025-07145-1
