Nanoparticles of zinc oxide are widely used in commercial applications and the harmful effects of nanoparticles on the development of oocytes have been studied in previous investigations. In this paper, we aimed to grow an eco-friendly procedure for the green synthesis of zinc oxide nanoparticles (G/ZnO-NPs) utilizing aqueous extract of Ferula gummosa gum resin (Galbanum) and evaluated G/ZnO-NPs effects on in vitro pre-antral follicle (PF) maturation compared to Galbanum extract alone and commercial zinc oxide nanoparticles (C/ZnO-NPs). Synthesized, G/ZnO-NPs have an absorbance band at 320 nm and spherical shapes, the mean particle size of 36 nm and are reasonably stable with a zeta potential value of -20 mV. FTIR results indicated that Galbanum extract covered the surface of ZnO-NPs. The results showed that Galbanum extract inhibiting free radicals as well as increasing the production of estradiol and testosterone improves PF growth. Also, the Galbanum extract increased expression of GDF-9 and BMP-15 versus the expression of Foxo1 and VNN1 that associated with atresia of PF were reduced. However, the evidence suggests G/ZnO-NPs and C/ZnO-NPs, have negative effects on PF through increasing the production of free radicals, reducing the expression of estradiol and testosterone, the reduced expression of GDF-9 and BMP-15, as well as the increased expression of Foxo1 and VNN1. Overall, the findings of this study showed that Galbanum extract due to antioxidant effects, supports PF development while C/ZnO-NPs and G/ZnO-NPs inhibit PF growth, and the Galbanum extract covered the surface of ZnO-NPs only slightly offsets the disadvantages of these nanoparticles.