Multidrug-resistant organisms (MDRO), including MDR Enterobacter cloacae, may emerge due to the extensive usage of antibiotics and threaten the lives of millions of people around the world. Developing new antibiotic-free strategies to combat E. cloacae infections and curb the spread of drug-resistant genes is crucial. Bacteriophage therapy has garnered widespread attention as a promising approach to tackle bacterial infections. Herein, we isolated a specific bacteriophage (vB_Ent31) targeting E. cloacae from sewage using E. cloacae Ent31 as the host bacterium. vB_Ent31 is a tadpole-like phage with double-stranded DNA belonging to the Siphoviridae family. It exhibits narrow-spectrum activity against Enterobacter spp. and remains stable across a temperature range of 4-50 °C and pH 4 to 11. Significantly, vB_Ent31 prevents proliferation of Ent31 and inhibits inflammation, which further accelerate wound healing. Our findings suggest that bacteriophage therapy could offer an alternative to combating drug-resistant bacteria.