In this work, bacterial cellulose/electrospun membrane three-dimensional (BC/ENM 3D) hybrid structures were fabricated using in-situ self-assembly and characterized by SEM and FTIR. SEM showed as the bacterium extrudes cellulose nanofibrils; they gradually self-assemble to form a unified network on membrane surface and also penetrate into its structure, hence securely binding electrospun nanofiber and interlocking multiple membrane layers to form a stable 3D hybrid scaffold. FTIR results confirmed the presence of BC in the resulting nanocomposites. The permeability and porosity of the scaffold decreased with prolonged fermentation and with increased oxygen ratio as the cellulose grew more quickly. [GRAPHICS]