A bilayered polylactic-co-glycolic acid (PLGA)/multiwall carbon nanotubes (MWNTs)/bacterial cellulose (BC) composite membrane was prepared by vacuum freeze-drying and electrospinning for use as a guided tissue regeneration (GTR) membrane to repair maxillary canine periodontal defects in beagle dogs. Morphological and histological observations and analyses were performed to characterize the membrane. The bilayered membrane comprised a lower fibrous BC film layer and an upper electrospun PLGA/MWNTs film layer with a three-dimensional network structure. The histological results showed that the bilayered membrane promoted periodontal tissue regeneration. The BC/PLGA/MWNTs membrane may be an ideal GTR membrane candidate and thus has potential therapeutic applications in periodontal disease. (C) 2017 Elsevier B.V. All rights reserved.