Project Summary/Abstract The coordinated development of craniofacial jaw bones and teeth are the result of a complex interplay between a surprisingly large and growing number of tissues - including blood and vasculature, and nerves – combined with the immune system, and a multitude of growth factor signaling pathways. Although knowledge of signaling pathways resulting in human mineralized tissue development has improved over the past few decades, detailed knowledge and understanding of how to regenerate human mineralized tissues in a functional and timely manner remains elusive – particularly with respect to the craniofacial complex and teeth. Here we propose novel strategies to effectively and coordinately regenerate alveolar jaw bone and tooth tissues, using three dimensional tissue engineering strategies. The objectives of the proposed studies, which build on our prior published reports, are to characterize two potential models for biomimetic three dimensional alveolar bone-tooth constructs: 1) natural decellularized tooth bud extracellular matrix scaffolds; and 2) Gelatin Methacrylate (GelMA) hydrogel scaffolds. Our published expertise in in the field of regenerative dentistry, mineralized tissue development and disease, strong preliminary data, and team of developmental biologists, clinicians and bioengineers supports our ability to accomplish the proposed Aims. We anticipate that the completion of the proposed studies will result in significantly improved knowledge and understanding of new methods to repair craniofacial defects caused by a variety of insults. The significance of the proposed studies and relevance to public health includes the facts that skeletal and craniofacial defects occur in as many as 1 in 700 live births in the United States alone, and that over a quarter of a million Maxillofacial surgeries were performed in 2014, including oral cancer resections. In addition, injuries to the craniofacial complex that include teeth are increasingly common due to accidents and sports injuries, and particularly in the battlefield where they represent up to 20% of injuries. As such, the proposed studies will make a significant contribution towards improved quality of life of both civilian and military populations.