The Nd:YAG laser has been proposed to apply in minor soft tissue surgery, including various periodontal procedures. However, little information is available regarding the direct effect of Nd:YAG laser on gingival fibroblasts, which play an important role in the early healing processes of periodontal repair. Nd:YAG laser irradiation was performed in pulsed mode on human gingival fibroblasts, which was derived from healthy human gingiva by an explant method. The size of laser diode was 400 microm in diameter. The parameters in laser delivery were pulse energy (50-150 mJ), power output (1.0-3.0 W), pulse rate (10-30 pps), and fixed duration of irradiation (10 seconds). The cell cultures were analysed by cytomorphologic examination under phase-contrast and scanning electron microscope. The vitality was also examined with the help of MTT staining. The area of laser damage on cell culture was circular in shape, with diameter beyond the size of laser diode. By scanning electron microscopy, we observed the cellular damage of cultured gingival fibroblasts induced by Nd:YAG laser irradiation, comparable with the progressive increased power settings. The cytomorphologic changes ranged from disappearance of cellular boundary, loss of identifiable cellular nucleus, and finally cell contraction and vacuolization. Significant decrease in cellular vitality (14% approximately 44%) after laser treatment with irradiation distance of nearly contact was noted. However, 2 mm defocusing irradiation with the same power settings did not significantly decrease cellular vitality. Our study demonstrated the cell damaging effects of Nd:YAG laser, ranging from degeneratively cytomorphologic change to cell death, on the cultured human gingival fibroblasts. It provided the dentist a chance to understand the potential hazard of laser application in periodontal treatment. If the energy output is enough for the clinical purposes, Nd:YAG laser with lower pulse energy and corresponding pulse rate should be selected to minimize the damage on adjacent soft tissue.