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The aim of the current work is to develop a thermo-sensitive hydrogel system of moxifloxacin hydrochloride (MOX) for improved ocular delivery. Fifteen formulations were prepared at different concentrations of β-glycerophosphate disodium salt (β-GP) 12–20% (w/v) and chitosan (CS) 1.7–1.9% (w/v). The optimized MOX loaded thermo-sensitive hydrogel system (F8), consisting of CS (1.8%, w/v) and β-GP (16%, w/v), showed optimum gelation temperature (35 °C) and gelation time (2 min), thus was selected for further investigations. It showed a significant decrease (p < 0.05) in the zeta potential value compared to CS solution with a favorable pH value (7.1) and confirmed thermoreversible behavior. MOX loaded F8 displayed a porous structure under scanning electron microscopy. Rheological investigation of MOX loaded F8 revealed the presence of a strong hydrogel network with high elasticity along with a small …