Bioengineered Dental Tissues from Human Tooth Bud Cells

描述（由申请人提供）： 这项研究将主要在巴西圣保罗大学进行，与Silvio和Monica Duailibi合作，作为NIH资助#R 01 DE 016132 -01的扩展。我们的长期目标是确定可用于人类自体牙齿组织工程的细胞群和方法。识别和操纵成年牙齿祖细胞或牙齿干细胞（DSCs）的能力将显著促进这一过程。我们以前已经证明，在已发表的报告中，成功地使用猪和大鼠牙芽细胞来产生生物工程牙齿组织。这些结果表明，我们有能力分离和维持培养，上皮和间充质出生后的DSCs，并使用这些细胞的生物工程小，准确形成的牙冠含有牙本质，釉质，牙髓，牙骨质和牙周韧带组织。基于这些有希望的结果，我们假设人牙芽细胞可以用于产生生物工程牙齿组织。在这里，我们提出的研究，其成功完成将定义方法，生物工程人类牙齿组织。具体而言，我们将：1）优化人类出生后牙芽细胞的收获/培养条件; 2）进行人类出生后牙齿干细胞的分子/细胞表征;以及3）生成和表征生物工程化人类牙齿组织。所提出的目标的成功完成将显著提高我们对人类出生后DSCs及其在牙齿组织工程中的潜在用途的认识和理解，并将为人类最终的自体牙齿组织工程提供基础。

DESCRIPTION (provided by applicant): This research will be done primarily in Brazil at University Sao Paolo, in collaboration with Silvio and Monica Duailibi, as an extension of NIH grant # R01 DE016132-01. Our long-term goal is to define cell populations and methods that can be used for autologous tooth tissue engineering in humans. The ability to identify and manipulate adult tooth progenitor cells, or dental stem cells (DSCs), would significantly facilitate this process. We have previously demonstrated, in published reports, the successful use of both pig and rat tooth bud cells to generate bioengineered dental tissues. These results demonstrate our ability to isolate and maintain in culture, both epithelial and mesenchymal post-natal DSCs, and to use these cells to bioengineer small, accurately formed tooth crowns containing dentin, enamel, pulp, cementum, and periodontal ligament tissues. Based on these promising results, we hypothesize that human tooth bud cells can be used to generate bioengineered dental tissues. Here we propose studies whose successful completion will define methods to bioengineer human dental tissues. Specifically, we will: 1) Optimize harvesting/culturing conditions for human post-natal tooth bud cells; 2) Perform molecular/cellular characterization of human post-natal dental stem cells; and 3) Generate and characterize bioengineered human tooth tissues. The successful completion of the proposed aims will significantly improve our knowledge and understanding of human postnatal DSCs and their potential utility in tooth tissue engineering, and will provide the foundation for eventual autologous tooth tissue engineering in humans.