Rania S Abdel-Rashid,1 Doaa A Helal,2 Mahmoud M Omar,3,4 Amani M El Sisi51Pharmaceutics and Industrial Pharmacy Department, Faculty of Pharmacy, Helwan University, Cairo, Egypt; 2Pharmaceutics and Industrial Pharmacy Department, Faculty of Pharmacy, Fayoum University, Fayoum, Egypt; 3Pharmaceutics and Industrial Pharmacy Department, Faculty of Pharmacy, Deraya University, El-Minia, Egypt; 4Pharmaceutics and Industrial Pharmacy Department, Faculty of Pharmacy, Sohag University, Sohag, Egypt; 5Pharmaceutics and Industrial Pharmacy Department, Faculty of Pharmacy, Beni-Suef University, Banī Suwayf, EgyptObjective: Intraocular pressure has always been a great challenge for topical ophthalmic drugs. The study aimed to develop ocular surfactant based nanovesicles (NVs) carried in mucoadhesive nanogel providing efficient topical delivery of acetazolamide (ACZ).Methods: For the sake of optimizing formulation parameters, the effect of the type of edge activator and its ratio to sorbitan monostearate (Span 60) on the mean particle size, entrapment efficiency (%EE), and zeta potential (ZP) of produced NVs was investigated.Results: The selected formulation composed of Span 60:sodium deoxycholate with ratio 80:20 showed an average diameter of 202.90 nm, %EE of 90.2%, and ZP of −38.1 mV with a spherical and smooth surface. The ACZ loaded nanovesicles (ACZ-NVs) were embedded in different concentrations of Chitosan–sodium tripolyphosphate (CS-TPP) nanogels. The nanogel prepared using 1.5% CS showed the most promising viscosity, adhesion time, and rheological behavior (118,246 cP, 290 min, and thixotropic behavior, respectively). The in vitro release of ACZ showed a controlled release profile after incorporation in nanogels. The in vivo irritation test showed minimal irritation for the nanogel formulation compared to ACZ topical suspension. The effect of intraocular pressure lowering was significantly prolonged using ACZ-NV nanogels compared to ACZ oral tablets. Histopathological examination emphasized the healing power of CS on retinal atrophy.Conclusion: The research work indicated a promising potential for successful topical delivery of ACZ.Keywords: nanogels, acetazoleamide, intraocular pressure, nanovesicles, edge activators, ocular delivery