Dental Stem Cells and Tooth Tissue Engineering

描述（由申请人提供）：本研究项目的最终目标是开发新的基于生物学的人类牙齿和颅面骨骼修复和再生疗法。我们假设，对牙科干细胞（DSC）特性和特征的进一步了解，结合对组织工程策略的进一步了解，可以可靠地生成可预测大小和形状的矿化牙齿和颅面组织，将导致人类开发出新颖有效的临床相关治疗方法。为了解决这一假设，提出了四个具体目标，所有这些目标都将专门使用从人类牙齿中提取的人类dsc。首先，我们将描述个体采集的人类牙齿组织的特性，建立DSC特性的正常范围，将DSC特性与供体的年龄和整体健康状况联系起来，并将这些特性与成功使用采集的牙齿干细胞进行牙齿组织再生治疗的能力联系起来。接下来，我们将测试四种不同的支架材料，这些材料是根据其明确的物理特性精心挑选的，因为它们有能力制造出可预测大小和形状的生物工程人类牙冠。第三，我们将扩展我们的研究结果，证明丝支架可以用来产生预测大小和形状的骨牙素，生物工程的全尺寸，功能的人类牙根等同物，可以支持合成或生物工程的牙冠。丝绸支架的制造方法，孔隙率，降解率和添加的生长因子肽，将被系统地评估。根据先前综述的建议，Aims 2和3将在皮下和微型猪下颌种植体模型中进行。最后，在Aim 4中，我们将结合我们的牙冠、牙根和颅面骨骼再生策略，再次使用尤卡坦迷你猪下颌种植体模型，连续生成第一代、第二代和第三代替代牙。对于每一个提出的目标，详细的发展分析生物工程牙种植组织将进行，以便更好地了解，并制定策略，以提高牙组织再生。我们的牙齿修复和再生的新方法，使用人类dsc，和最先进的支架

DESCRIPTION (provided by applicant): The ultimate goal of this research project is to develop novel biologically based, dental and craniofacial skeletal repair and regeneration therapies in humans. We hypothesize that improved knowledge of dental stem cell (DSC) properties, and characteristics, combined with improved knowledge of tissue engineering strategies to reliably generate mineralized dental and craniofacial tissues of predictable size and shape, will result in the development of novel and effective, clinically relevant therapies for humans. To address this hypothesis, four specific aims are proposed, all of which will exclusively use human DSCs harvested from extracted human teeth. First, we will characterize the properties of individually harvested human dental tissues, to establish the normal range of DSC properties, to correlate DSC properties with the age and overall health of the donor, and to correlate these properties with the ability to successfully use harvested dental stem cells for dental tissue regeneration therapies. Next, we will test four different scaffold materials, carefully chosen for their defined physical properties, for their ability to generate bioengineered human tooth crowns of predictable size and shape. Third, we will expand upon our results demonstrating that silk scaffolds can be used to generate osteodentin of predicted size and shape, to bioengineer full sized, functional human tooth root equivalents that can support a synthetic or bioengineered tooth crown. Silk scaffold fabrication methods, porosity, degradation rates, and added growth factor peptides, will be systematically evaluated. As recommended in prior review, Aims 2 and 3 will be performed in both subcutaneous and minipig mandibular implant models. Finally, in Aim 4, we will combine our tooth crown, root, and craniofacial skeletal regenerative strategies, again using the Yucatan mini pig mandibular implant model, to generate successive 1st, 2nd and 3rd generation replacement teeth. For each of the proposed aims, detailed developmental analyses of bioengineered dental implant tissues will be performed in order to better understand, and devise strategies to improve dental tissue regeneration. Our novel approach to tooth repair and regeneration, using human DSCs, and state of the art scaffold fabrication methods, combined with our extensive expertise in Developmental Biology and Tissue Engineering, have the potential to provide new and improved, biologically based repair and regeneration strategies, using autologous tissues. The successful accomplishment of the proposed studies would dramatically alter the field of dentistry as it currently exists, extending clinically relevant dental repair therapies to include biologically based dental materials with properties closely matching those of naturally formed dental tissues.