Mimicking the Dentin-Pulp Complex In-Vitro

体外模拟牙本质牙髓复合物

• 批准号: 8283896
• 负责人: Stefan Friedrich Habelitz
• 金额: $ 23.18万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8283896
• 关键词: Adult; Ameloblasts; Basal lamina; Biomedical Engineering; Caliber; California; Cell membrane; Cell physiology; Cells; Chemotactic Factors; Coculture Techniques; Collaborations; Complex; Dental; Dental Enamel; Dental Pulp; Dental crowns; Dentin; Dentinogenesis; Development; Engineering; Epithelial; Epithelial Cells; Epithelium; Film; Future; Gene Expression; Human; Immunocompromised Host; Immunohistochemistry; Implant; In Vitro; Laboratories; Life; Location; Membrane; Mesenchyme; Microfabrication; Molecular Biology; Morphology; Mus; NIH 3T3 Cells; Natural regeneration; Odontoblasts; Phenotype; Polymers; Positioning Attribute; Process; Rest; Reverse Transcription; Silicon; Source; Stem cells; Structure; Techniques; Technology; Testing; Tissue Engineering; Tissues; Tooth structure; Tubular formation; Universities; design; improved; interfacial; interstitial; millimeter; mimicry; mineralization; novel; polarized cell; polycaprolactone; reconstitution; scaffold

DESCRIPTION (provided by applicant): This projects attempts to create the dentin-pulp complex in-vitro using microfabrication and dental pulp stem cells (DPSC). The dentin pulp interface is characterized by a lining of specialized cells of unique morphology that produce the collagenous dentin matrix and control its mineralization. Highly polarized odontoblasts with diameters of 5 to 10 ¿m are tightly packed along the pulpal wall. Their unique feature is the odontoblastic process that extends up to 10 mm deep along tubules into the mineralized dentin. The cells are anchored in this position for the lifetime of the tooth. Hence these cells can live fr decades and only respond to external insult by producing additional dentin matrix (reparative dentin). The surrounding dentin pulp contains progenitor cells that can differentiate into odontoblast cells and produce dentin matrix or form a dentin crown when implanted with epithelial or ameloblast-like stem cells into an immunocompromised mouse. This application proposes to reconstitute this highly organized arrangement of cells in-vitro using tissue engineering approaches. In collaboration with the bioengineering laboratory of Dr. Tejal Desai and the microfabrication facilities at the University of California at Berkeley, micropatterned polymer scaffolds will be generated that permit the positioning of cells in precise locations. In our design these locations will be aligned with an open cylindrical pore allowing for one cell being positioned on top of each pore. Chemotactic approaches will be used to encourage protrusion of odontoblastic processes into the pores resulting in a cellular configuration of polarized cells that is comparable to the dentin-pulp interface. These studies will produce a unique construct to investigate gene expression and cellular interactions in-vitro and improve our understanding of tooth development. Furthermore such pre-assembled mimicries of dental tissue will advance current approaches towards tooth regeneration and pulp revitalization. PUBLIC HEALTH RELEVANCE: This project proposes to fabricate micropatterned membranes that will position dental pulp stem cells in-vitro in a configuration similar to the dentin-pulp interface in human teeth. The cell tissue engineered construct will facilitate future molecular biology studies on epithelium/mesenchyme interactions as well further enhance approaches towards engineering of dental tissues.

描述（由申请人提供）：本项目尝试使用微细加工和牙髓干细胞（DPSC）在体外创建牙本质-牙髓复合体。牙本质牙髓界面的特征在于具有独特形态的特化细胞衬里，其产生胶原牙本质基质并控制其矿化。高度极化的成牙本质细胞直径为5 ~ 10 μ m，沿牙髓壁沿着紧密。其独特的特征是成牙本质细胞的过程，延伸到沿着小管进入矿化牙本质深达10 mm。细胞在牙齿的整个生命周期中都被固定在这个位置。因此，这些细胞可以存活数十年，并且仅通过产生额外的牙本质基质（修复性牙本质）来应对外部损伤。周围的牙本质牙髓含有祖细胞，当将上皮或成釉细胞样干细胞植入免疫受损小鼠时，祖细胞可以分化成成牙本质细胞并产生牙本质基质或形成牙本质冠。本申请提出使用组织工程方法在体外重建这种高度组织化的细胞排列。在与Tejal Desai博士的生物工程实验室和加州大学伯克利分校的微加工设施的合作下，将产生微图案化的聚合物支架，允许将细胞定位在精确的位置。在我们的设计中，这些位置将与开放的圆柱形孔对齐，允许一个细胞位于每个孔的顶部。趋化性方法将用于促进成牙本质细胞突起进入孔隙，导致极化细胞的细胞构型与牙本质-牙髓界面相当。这些研究将产生一个独特的构建体，以研究基因表达和体外细胞相互作用，并提高我们对牙齿发育的理解。此外，这种预组装的牙齿组织模拟物将推进目前的牙齿再生和牙髓再生方法。 公共卫生关系：该项目提出制造微图案化膜，将牙髓干细胞定位在体外类似于人类牙齿中的牙本质-牙髓界面的配置中。细胞组织工程构建体将促进未来的上皮/间充质相互作用的分子生物学研究，以及进一步加强对牙齿组织工程的方法。