Chitosan (Cs) and polyvinyl alcohol (PVA) are the natural and synthetic polymers, respectively, that have biocompatible, biodegradable, and non-toxic properties. Thus, they have been widely developed for wound dressing material. Studies on the preparation of electrospun PVA/Cs have been extensively carried out, but for in vivo testing is rare. This research aims to study the effect of the ratio of chitosan to PVA on the morphology and mechanical properties of PVA/Cs nanofiber membranes and the effectiveness of the membrane for ulcus diabetic wound healing. The PVA/Cs nanofiber membranes were fabricated by mixing 3% chitosan and 10% PVA with varying ratio of Cs to PVA 90:10, 85:15, and 80:20 (w/w) using electrospinning method at applied voltage 18kV, tip to collector distance (TCD) 16.5 cm, a diameter of syringe needle 0.6 mm and flow rate 0.33 μl/min. The viscosity and conductivity of PVA/Cs solutions were measured with viscometer and conductometer, respectively. A scanning electron microscope (SEM) characterized the morphology of the nanofiber membranes, and tensile properties were tested with a universal testing machine (UTM) according to ASTM D882. In vivo testing was carried out on the wound of mice which have diabetic for 12 days by dividing in three groups namely positive group (Sofra-tulle), negative group (Pure PVA) and PVA/Cs group (PVA/Cs membrane at the ratio 80:20). The results indicated that an increase in the ratio of CS into PVA increased tensile properties of the membranes and the average diameter of the fiber. The effectiveness of ulcus diabetic wound healing on mice showed that the PVA/Cs nanofiber membrane at the ratio of 80:20 is almost similar to the commercial Sofra-tulle which is a wound dressing with commercialized antibiotics. Therefore, the PVA/Cs nanofiber membrane resulted from this research is recommended as a wound dressing material candidate.