Based on the photobiomodulation on wound healing of animal models, the stimulative effects of He-Ne laser irradiation on proliferation of human normal skin fibroblasts were used as the cell model to investigate the role of ketamine as an anesthetics in animal model experiments in this paper. The cell proliferation was measured using MTT-based colorimetric assay. The effects of 1.56 mW/cm2 He-Ne laser irradiation on human normal skin fibroblasts for 0, 10, 30, 100, 300 s, and on the cells preincubated with different concentrations of ketamine (0, 0.6, 1.2, 2.0 μg/mL) for 300 s were investigated, respectively. The results show that human normal skin fibroblast proliferation induced by He-Ne laser irradiation (468.7 mJ/cm2) was inhibited when the cells were preincubated with different concentrations of ketamine (0.6, 1.2, 2.0 μg/mL). These results suggest that anesthetics may affect photobiomodulation on wound healing.