This study aims to develop scaffold for transdermal drug delivery method (TDDM) using electrospinning technique from polyvinyl alcohol (PVA) and hydroxyethylcellulose (HEC). The fluorescein isothiocyanate (FITC) loaded on ethosomes (FITC@Eth) was used as a drug model. The prepared PVA/HEC/FITC@Eth scaffold was characterized via scanning electron microscope (SEM) that show morphology change by adding FITC@Eth. Also, Fourier transform infrared spectroscopy (FTIR), mechanical properties, X-ray diffraction (XRD), thermal gravimetric (TGA) analysis show that the addition of FITC@Eth to PVA/HEC does not change the scaffold properties. Franz diffusion cells were used for in vitro skin permeation experiments using rat dorsal skins. The FITC@Eth penetration was better than that of free FITC due to the presence of ethosome which enhance the potential skin targeting. In conclusion, the prepared PVA/HEC/FITC@Eth scaffold can serve as a promising transdermal scaffold for sustained FITC release.