Every year millions of adults and children in the United States and in the world suffer from caries and its treatment related problems. Current clinical practices are not adequate to enable regeneration of the lost dental pulp tissue. The National Institute of Dental and Craniofacial Research (NIDCR), a part of the National Institute of Health (NIH) has devoted much of its resources towards research that targets regeneration of these tissues. In accordance with the mission and the focus of the NIDCR, this project focuses on the development of a biomimetic scaffold for regeneration of complete and functional dental pulp tissue. Specifically, we intend to use the extracellular matrix (ECM) of pulp cells incorporated into 3D scaffolds to drive the differentiation of somatic stem cells in vivo in a mini pig pulpectomy model. This study consists of two specific aims directed at regeneration of dental pulp in a large animal pre-clinical model and understanding the molecular mechanism of regeneration and the involvement of unique multifunctional proteins in this process. In the in vivo experiments, we plan to compare the ECM scaffolds side by side with the current clinical standards to analyze if they are able to perform better and justify the potential that they have shown in our preliminary in vitro and in vivo experiments. Results from this project have the potential to replace current root canal therapy with patient- specific regenerative therapy. The ECM incorporated scaffolds can be mass-produced and their use will remove the need growth factor delivery systems from dental pulp tissue engineering applications. Additionally, the study will also lay the platform for a totally new system to analyze extracellular roles of multifunctionl proteins in stem cell differentiation and tissue regeneration.