Electrospinning is a method to fabricate nanofiber scaffold for tissue engineering. One of the parameters that affects the fiber size and morphology and the other physical characteristics is relative humidity of the room in which the electrospinning process is undertaken. The synthesis of electrospun Poly Vinyl Alcohol (PVA)-Aloe vera was performed with a variation of relative humidity (52%, 55%, 58%, and 64%). The physical characteristics of the fiber was tested by using functional group test, morphology test, thermal analysis test, and degradation test. The result of functional group test by using Fourier Transform Infrared (FTIR) showed that there was a crosslinking bond when the Aloe vera was introduced to the PVA. The morphology test result by using Scanning Electron Microscope (SEM) indicated that the addition of Aloe vera could increase the average of fiber diameter and the increase of relative humidity reduced the fiber diameter. The thermal analysis test result by using Differential Scanning Calorimetry (DSC) showed that the increase in the relative humidity could increase the temperature of glass transition (Tg) and decrease the temperature of melting (Tm). The degradation test implied that all the fibers could be degraded in 12 minutes.