Various synthetic and natural biomaterials have been used for regeneration of tissues and hollow organs. However, clinical outcome of reconstructive procedures remained challenging due to the lack of appropriate scaffold materials, supporting the needs of various cell types and providing a barrier function required in hollow organs. To address these problems, we have developed a bilayered hybrid scaffold comprising unique traits of polymeric microfibers and naturally derived acellular matrices and tested its potential for hollow organ regeneration in a rat bladder model. Hybrid scaffolds were fabricated by electrospinning of PLGA microfibers directly onto the abluminal surface of a bladder acellular matrix. Stability of this bilayered construct was established using modified spinning technique. The resulting 3-dimensional framework provided good support for growth, attachment and proliferation of primary bladder smooth muscle cells. Histological analysis in vivo at 4 and 8 weeks post implantation, revealed regeneration of bladder tissue structures consisting of urothelium, smooth muscle and collagen rich layers infiltrated with host cells and micro vessels. Furthermore, hybrid scaffolds maintained normal bladder capacity, whereas BAM recipients showed a significant distension of the bladder. These results demonstrate that this adaptable hybrid scaffold supports bladder regeneration and holds potential for engineering of bladder and other hollow organs.