Abstract: OBJECTIVE To prepare mupirocin hydrogels with thermo-sensitive for controlling drug release. METHODS Gelatin was modified by maleic anhydride to obtain acylated gelatin, and then five kinds of hydrogels were prepared via free radical polymerization of acylated gelatin and N-isopropyl acrylamide in the presence of the initiator and promoting initiator. Finally, mupirocin were loaded into as-prepared thermo-sensitive hydrogels by dry soaking methord with mupirocin solution of 1 mg·mL^-1 and 0.5 mg·mL^-1 to obtain hydrogels. The surface morphology, thermo-sensitivity, swelling rate, mechanical compression properties, drug-loading rate, controlled release rate and antibacterial performance of mupirocin hydrogels were investigated in detail. RESULTS The mupirocin hydrogels prepared in this study had obvious thermo-sensitivity. When the molar ratio of acylated gelatin to N-isopropyl acrylamide was 1:1, the prepared hydrogels had the most uniform and dense pore structure, exhibited the highest swelling rate at 25℃, and had the best drug-loading and drug-releasing rate as well as antibacterial properties. In addition, the compression property of mupirocin hydrogel was enhanced with the increase of acylated gelatin. The drug loading rate and antibacterial properties of hydrogels prepared with 1 mg·mL^-1 mupirocin solution were higher than those prepared with 0.5 mg·mL^-1 mupirocin solution, while the drug release rate was opposite. The overall drug-releasing of the mupirocin hydrogels can be equilibrium in 2 h. CONCLUSION The prepared mupirocin hydrogels can control the drug release which has a long-term significance for reducing drug resistance. When the ratio of acylated gelatin to N-isopropyl acrylamide is 1:1, the performance of mupirocin hydrogels is the relatively best.