Enhanced wound healing and osteogenic potential of photodynamic therapy on human gingival fibroblasts

Photodynamic therapy (PDT) has shown ideal antibacterial effects in clinical treatment of periodontal diseases. However, little is known about the specific potential of PDT on human gingival fibroblasts (HGFs) especially cells in the inflamed state, which may contribute to the repairi of periodontal tissue.

The effect of PDT with different concentrations of methylene blue (5 μM, 10 μM, 20 μM) on cell vitality of healthy and inflamed human gingival fibroblasts was evaluated by CCK-8, and cell migration was assessed by cell scratching assay. The gene expression of interleukin-6 (IL-6), interleukin-8 (IL-8), type I collagen (Col I), fibronectin (FN) and basic fibroblast growth factor (bFGF) were measured with real-time fluorescent quantitative polymerase chain reaction. The alkaline phosphatase (ALP) production and alizarin red staining of mineralized nodules in healthy and inflamed human gingival fibroblasts was evaluated to explore the effect on osteogenic differentiation.

PDT with relatively low concentration of methylene blue (5 μM) inhibited the cell vitality of inflamed human gingival fibroblasts (I-HGFs) slightly (P < 0.05), but had no adverse effect on healthy human gingival fibroblasts (H-HGFs) (P > 0.05). As the concentration increased, PDT with 20 μM methylene blue had significantly negative effect on both healthy and inflamed cells. Further, PDT with 5 μM methylene blue was observed to be able to promote the migration of HGFs especially the healthy state, and increases the expression of wound healing related genes including IL-6, COL1, FN, bFGF in healthy and inflamed HGFs (P < 0.05). PDT with 5 μM methylene blue was also capable of increasing the production of ALP and mineralized nodules (P < 0.05), although the better effect was observed in the laser treatment group.

The relatively low concentration of methylene blue mediated PDT is conducive to the growth of H-HGFs while inhibiting the I-HGFs, and it also has the potential to promote the wound healing and osteogenic related functions of both healthy and inflamed HGFs.