Human amnion cells are very important type of stem cells, which are able to differentiate into cell types of all three germ layers, so this study was investigated the impact of He-Ne laser on viability, proliferation and differentiation of cultured human amnion cells grown over a synthetic nanoscaffold. This nanoscaffold was prepared from 13% polycaprolactone (PCL) dissolved in 1:1 chloroform: dimethyle formamide (DMF) as a solvent by using electrospinning technique. Human amnion cells were grown over the scaffold and were exposed to He:Ne laser at doses 1, 2.5 and 5 J/cm² every 48 hours for a duration of 12 days. It was concluded that scanning electron microscope (SEM) photographs showed good colony spreading of seeded amnion cells grown over the electrospun scaffold, and He:Ne laser induced cell proliferation as detected by MTT, nitric oxide and lipid peroxidation production.