Pumpkin seed oil (PSO) provides many health benefits including antioxidant, cardiovascular health boost, treatment of benign prostatic hyperplasia (BPH), and reduction of hair loss. The main objective of this study was to design a suitable formulation of niosomes encapsulated PSO for topical delivery. Formulation of PSO-loaded niosomes was optimised by altering the types of surfactant and the amount of PSO:surfactant:cholesterol. The developed PSO-loaded niosomes were spherical shape with the size range of 138-366 nm. The niosomes formulated with Tween 20 provided the smallest particle size. An increase in the ratio of PSO:surfactant:cholesterol from 2:2:1 to 2:4:1 led to reduction of the particle size of the niosomes. The PSO-loaded niosomes formulation F1 (PSO:Tween 20:cholesterol = 2:2:1) provided the highest percent entrapment efficacy at 75.99 ± 14.65%. The in vitro release study suggested that the release mechanism was followed Korsmeyer-Peppas. The physical stability study indicated good stability over 3 months of storage at 30 °C.