To explore the bacterial blocking effect of oriented multilayer MXene/polyvinyl alcohol (PVA) nanocomposite hydrogels and their effect on the repair of intestinal defects.MXene/PVA nanocomposite hydrogels were prepared using the traditional freezing method and the bidirectional freezing ice template method. The structures of the different hydrogels were observed using scanning electron microscopy (SEM) and micro-CT reconstruction. The rheological properties of the hydrogels were measured using a dynamic rheometer, and their mechanical properties were assessed using a universal testing machine. The burst pressure of the hydrogels was determined through burst experiments, and bacterial colony growth was observed by the osmosis method to assess the bacteria blocking ability of the hydrogels . A rat model of cecal perforation was established, and the hydrogels were used for intestinal repair. Gram staining was performed to observe the bacterial blocking ability of the hydrogels, HE staining was performed to observe the intestinal inflammation, and CD31 and CD68 immunofluorescence staining and proliferating cell nuclear antigen (PCNA) staining were performed to observe the repair effect of the hydrogels on intestinal defects.SEM and micro-CT reconstruction revealed that the hydrogel prepared by the traditional freezing method exhibited a random porous structure, while the hydrogel prepared by the bidirectional freezing method showed an oriented multilayer structure. Rheological and tensile tests indicated that the oriented hydrogel had superior mechanical properties, and the burst pressure of the oriented multilayer hydrogel was as high as 27 kPa, significantly higher than that of the non-oriented hydrogel (<0.001). Bacterial colony growth was observed by the osmosis method and it was found that, compared with the non-oriented hydrogel, the oriented multilayer hydrogel could effectively prevent the infiltration of and . Gram staining results showed that the oriented multilayer hydrogel could effectively block intestinal bacteria from entering the abdominal cavity . HE staining results showed that the oriented multilayer hydrogel could effectively reduce intestinal inflammation . CD31 and CD68 immunofluorescence staining and PCNA staining results showed that the oriented multilayer hydrogel had a repairing effect on intestinal defects .The oriented multilayer hydrogel prepared by bidirectional freezing effectively prevents bacterial infiltration and reduces intestinal inflammation.