Bioengineered Dental Tissues from Human Tooth Bud Cells

描述（由申请人提供）：本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# R01 DE016132-01的延伸。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。鉴定和操作成牙祖细胞或牙干细胞（DSCs）的能力将大大促进这一过程。我们之前已经在发表的报告中证明，成功地使用猪和大鼠的牙芽细胞来产生生物工程牙齿组织。这些结果证明了我们在培养中分离和维持上皮细胞和间充质干细胞的能力，并使用这些细胞来生物工程小的、准确形成的牙冠，包括牙本质、牙釉质、牙髓、牙骨质和牙周韧带组织。基于这些有希望的结果，我们假设人类牙芽细胞可以用来产生生物工程牙齿组织。在这里，我们提出的研究，其成功完成将定义生物工程人类牙齿组织的方法。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)对人出生后牙干细胞进行分子/细胞表征；3)生成和表征生物工程人类牙齿组织。本研究的成功完成将大大提高我们对人类出生后dsc及其在牙齿组织工程中的潜在应用的认识和理解，并将为最终的人类自体牙齿组织工程奠定基础。

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.