Bioengineered Dental Tissues from Human Tooth Bud Cells

描述(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#R01 DE016132-01补助金的延伸。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体和方法。识别和操作成年牙祖细胞或牙齿干细胞(DSCs)的能力将极大地促进这一过程。我们之前已经在已发表的报告中证明，成功地使用猪和大鼠的牙蕾细胞来生成生物工程牙齿组织。这些结果表明，我们有能力在培养中分离和保持出生后的上皮性和间叶性DSCs，并使用这些细胞来生物工程包含牙本质、釉质、牙髓、牙骨质和牙周韧带组织的准确形成的小牙冠。基于这些有希望的结果，我们假设人的牙蕾细胞可以用来产生生物工程牙齿组织。在这里，我们提议成功完成的研究将确定生物工程人类牙齿组织的方法。具体地说，我们将：1)优化人类出生后牙蕾细胞的采集/培养条件；2)对人类出生后牙齿干细胞进行分子/细胞鉴定；以及3)产生和表征生物工程人类牙齿组织。这些目标的成功完成将大大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，并将为最终在人类体内进行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.