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Abstract:  The combined use of guided tissue/bone regeneration (GTR/GBR) membranes and bone filling grafts represents a classical therapy for guiding the regeneration and functional reconstruction of oral soft and hard tissues. Nevertheless, due to its displacement and poor mechanical support, bone meal is not suitable for implantation in the case of insufficient cortical bone support and large dimensional defects. The combination of GTR/GBR membrane with a three-dimensional (3D) porous scaffold may offer a resolution for the repair and functional reconstruction of large soft and hard tissue defects. In this study, a novel integrated gradient biodegradable porous scaffold was prepared by bonding a poly(lactic-co-glycolic acid) (PLGA)/fish collagen (FC) electrospun membrane (PFC) to a 3D-printed PLGA/nano-hydroxyapatite (HA) (PHA) scaffold. The consistency of the composition (PLGA) ensured strong interfacial bonding between the upper fibrous membrane and the lower 3D scaffold. In vitro cell experiments showed that the PFC membrane (upper layer) effectively prevented the unwanted migration of L929 cells. Further in vivo investigations with an oral soft and hard tissue defect model in beagles revealed that the integrated scaffold effectively guided the regeneration of defective oral tissues. These results suggest that the designed integrated scaffold has great potential for guiding the regeneration and reconstruction of large oral soft and hard tissues.