The effect of femtosecond laser irradiation in air and in O 2 and CF 4 gas flows on the wettability of electrospun poly(ɛ-caprolactone) fiber tissue scaffolds was studied. Laser power, focus spot size, raster scan spacing and gas atmosphere were varied in experiments. SEM imaging showed the average fiber diameter and surface porosity sizes were both altered by ablation. The micro-scale surface roughness measured by scanning laser profilometry was found to have a non-monotonic relationship to the surface wettability measured by the contact angle of sessile water droplets. In contrast, surface water contact angle continuously decreased with increased oxygen atomic percentage and oxygen-containing group fraction as measured by XPS. Further, the oxygen content was larger for more extensively ablated fiber surfaces, regardless of whether the increased ablation was caused by high laser power, smaller scanning space or smaller defocusing distance. Of the three gas atmospheres, O 2 gas flow was the most favorable environment for increasing surface oxidization, resulting in the largest water contact angle decrease for given laser power. For CF 4 gas flow, the least oxidization occurred, and the magnitude of water contact angle decrease was smallest for treatment at a given laser power.