Dental Stem Cells and Tooth Tissue Engineering

描述(申请人提供)：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。鉴定和操作成人牙齿干细胞(DSCs)的能力将大大促进这一过程。尽管有相当多的文献支持上皮和间充质干细胞的存在，但相对于其他干细胞，包括神经干细胞、骨髓基质干细胞、造血干细胞、精原干细胞和角质形成干细胞，DSC本身的特性仍然很差。特别是，DSC表面标志物的抗体可用于产生丰富的DSC群体，识别成人DSC群体，并检查非牙齿组织中成牙潜能的诱导，目前尚缺乏工具。我们推测，结合最先进的组织工程技术，出生后牙齿干细胞(DSCs)的鉴定和特征将促进全牙组织工程的努力。我们最近的牙齿组织工程结果表明，我们有能力在培养中分离和保持出生后的上皮和间充质DSC，并使用这些细胞来生物工程包含牙本质和釉质的小牙冠。基于这些有希望的初步结果，我们在这里提出了一些研究，这些研究的成功完成将定义预定大小和形状的生物工程牙齿的方法。我们建议：1)使用特征完善的干细胞分选技术建立和鉴定丰富的上皮和间充质DSC群体，并使用我们的功能性牙齿组织工程实验来确认它们的身份；2)确定支架材料和设计，以优化特定大小和形状的牙齿组织的生物工程。这些目标的成功完成将大大提高我们对DSCs及其在牙齿组织工程中的潜在用途的认识和理解，并将为最终实现人类自体牙齿组织工程奠定基础。

DESCRIPTION (provided by applicant): 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 dental stem cells (DSCs) would significantly facilitate this process. Although a considerable body of literature supports the existence of epithelial and mesenchymal DSCs, DSCs themselves remain poorly characterized relative to other stem cells including neural, bone marrow stromal, hematopoetic, spermatogonial and keratinocyte stem cells. In particular, tools such as antibodies to DSC surface markers that could be used to generate enriched DSC populations, to identify adult DSC populations, and to examine the induction of odontogenic potential in non-dental tissues, are currently lacking. We hypothesize that the identification and characterization of post-natal dental stem cell (DSCs), combined with state-of the art tissue engineering techniques, will facilitate whole tooth tissue engineering efforts. Our recent tooth tissue engineering results demonstrate our ability to isolate, and maintain in culture, epithelial and mesenchymal post-natal DSCs, and to use these cells to bioengineer small tooth crowns containing dentin and enamel. Based on these promising initial results, here we propose studies whose successful completion will define methods to bioengineer teeth of predetermined size and shape. We propose: 1) to establish and characterize enriched epithelial and mesenchymal DSC populations using well-characterized stem cell sorting techniques, and to confirm their identity using our functional tooth tissue-engineering assay; and 2) to define scaffold materials and design for optimized bioengineering of tooth tissues of specified size and shape. The successful completion of the proposed aims will significantly improve our knowledge and understanding of DSCs and their potential utility in tooth tissue engineering, and will provide the foundation for eventual autologous tooth tissue engineering in humans.