The purpose of this study was to determine whether low-power pulsed laser irradiation could affect the regeneration of a 10-mm gap of rat sciatic nerve created between the proximal and distal nerve stumps, which were sutured into silicone rubber tubes. After 8 weeks of recovery, pulsed laser-irradiated groups at frequencies of 5 kHz and 20 kHz both had significantly lower success percentages of regeneration (50% and 44%, respectively) compared to sham-irradiated controls (100%). In addition, qualitative and quantitative histology of the regenerated nerves revealed a less mature ultrastructural organization with a smaller cross-sectional area and a lower number of myelinated axons in both pulsed laser-irradiated groups than in controls. These results suggest that pulsed laser irradiation could elicit suppressing effects on regenerating nerves.