Abstract: A preparation method for a decellularized matrix biomaterial, which combines specific pretreatment, degreasing, disinfection, decellularization, ?-Gal antigen removal and sterilization, and reserves the integrity of bioactive components and structures in a material to the maximum extent while effectively removing an antigenic substance of an animal tissue, so that a decellularized matrix biomaterial having low immunogenicity and high biological activity is obtained. In addition, the decellularized matrix biomaterial prepared by the method effectively reduces immunogen components, has good biocompatibility and biological safety, is beneficial to cell adhesion, aggregation and proliferation on the surface, and can be used as a high-activity biological matrix material to be applied to skin soft tissue defect, gingival recession and periodontal soft tissue repair.