Microengineering the Dental Pulp Vascular Microenvironment

牙髓血管微环境的微工程

• 批准号: 9158576
• 负责人: Luiz Eduardo Bertassoni
• 金额: $ 38.5万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9158576
• 关键词: Address; Adult; Affect; Angiogenic Factor; Architecture; Biocompatible; Biocompatible Materials; Biological; Blood Vessels; Blood capillaries; Cell Differentiation process; Cells; Coculture Techniques; Communicable Diseases; Cues; Dental Materials; Dental Pulp; Dental caries; Dentin; Dentistry; Development; Endodontics; Endothelial Cells; Engineering; Environment; Evaluation; Excision; Extracellular Matrix; Gelatin; Goals; Heart; Human; Hydrogels; Implant; In Vitro; Individual; Injectable; Learning; Length; Light; Mechanics; Mediating; Methacrylates; Microfluidics; Mus; Natural regeneration; Necrosis; Nutrient; Odontoblasts; Pepsin A; Phenotype; Plant Roots; Process; Property; Pulp Canals; Role; Root Canal Therapy; Side; Signaling Molecule; Site; Skin; Source; Staging; Stem cells; Swelling; Techniques; Testing; Tissue Engineering; Tissue Model; Tissue Survival; Tissues; Tooth Apex; Tooth structure; Vascular blood supply; Vascularization; Waste Products; base; bioprinting; blood vessel development; capillary; cell motility; cellular engineering; crosslink; dental structure; improved; in vivo; monolayer; novel; paracrine; permanent tooth; physical property; regenerative; response; scaffold; success; tissue regeneration; translational approach; vasculogenesis

Project Summary: Dental caries is an infectious disease affecting approximately 90% of adults worldwide. Late stages of caries affect the dental pulp, leading to tissue necrosis and ultimately requiring root canal therapy. Typically, root canals in permanent teeth are treated by removing the necrotic tissue and replacing it with an artificial material. Regenerative endodontics has been proposed as an improved treatment option for these conditions. However, without controllable strategies to engineer the pulp vasculature, effective pulp regeneration is virtually impossible. It has been recently demonstrated that a functional vasculature can be engineered by culturing endothelial cells and stem cells from various sources in the correct microenvironmental conditions. However, the precise requirements specific to regenerating the pulp vasculature remain poorly understood. This project will systematically investigate three overlapping aspects that we propose are key determinants to regenerate the pulp vasculature: (1) matrix physical and mechanical properties, (2) composition, and (3) microarchitecture. In aim 1 we will investigate the contributions of different physical and mechanical properties to the ability of human endothelial colony forming cells (ECFCs) and dental pulp stem cells (DPSCs) to form microvascular networks when embedded in hydrogels that can be photo-crosslinked to have their properties systematically adjusted. We will then engineer pulp tissue-constructs that are pre-vascularized with pre-fabricated endothelial microchannels to enhance pulp regeneration in full-length root canals in-vivo. In aim 2 we will develop injectable and photo-curable hydrogels synthesized from the natural matrix of dentin and modified with methacrylates to test the contribution of matrix composition to the regeneration of the pulp vasculature. Further, we will combine these hydrogels with angiogenic components extracted from the dentin matrix and test their regenerative potential in vitro and in vivo. In aim 3 we will fabricate architecturally controlled gradients of ECFC and DPSC paracrine factors using microfluidics techniques to test the contribution of tissue microarchitecture to the formation of the pulp vasculature. We will then mimic the microarchitectures of vascularized dental pulp by 3D bioprinting tissue constructs that reproduce the organization of the native pulp. In the end of this project we expect to have microengineered a 3D vascularized pulp microenvironment that will improve translational approaches for use in regenerative endodontics in adult teeth.

项目概要： 龋齿是一种感染性疾病，影响全球约90%的成年人。晚期龋齿 影响牙髓，导致组织坏死，最终需要根管治疗。通常，根 恒牙的根管是通过去除坏死组织并用人造材料代替来治疗的。 再生牙髓学已被提出作为这些条件的一种改进的治疗选择。然而，在这方面， 如果没有可控的策略来设计牙髓脉管系统， 不可能的最近已经证明，功能性脉管系统可以通过培养来工程化。 内皮细胞和干细胞从各种来源在正确的微环境条件。然而，在这方面， 对于再生牙髓脉管系统的具体要求仍然知之甚少。这个项目 我将系统地研究我们认为是再生的关键决定因素的三个重叠方面 牙髓脉管系统：（1）基质物理和机械性质，（2）组成，和（3）微结构。 在目标1中，我们将研究不同的物理和机械性能对聚合物聚合能力的贡献。 人内皮集落形成细胞（ECFC）和牙髓干细胞（DPSC）以形成微血管 当嵌入水凝胶中时，网络可以光交联以系统地具有其特性 调整然后，我们将设计牙髓组织结构，用预制的内皮细胞预血管化， 微通道，以增强牙髓再生全长根管体内。在目标2中，我们将开发 由天然牙本质基质合成的可注射和光固化水凝胶， 使用甲基丙烯酸酯来测试基质组合物对牙髓脉管系统再生的贡献。 此外，我们将联合收割机这些水凝胶与从牙本质基质中提取的血管生成成分结合， 在体外和体内测试它们的再生潜力。在目标3中，我们将制造建筑控制梯度 ECFC和DPSC旁分泌因子使用微流体技术测试组织的贡献 微结构对牙髓脉管系统形成的影响。然后，我们将模仿 通过3D生物打印组织构建体来复制天然牙髓的组织，从而实现血管化牙髓。 在这个项目的最后，我们希望有一个3D血管化牙髓微环境， 用于成人牙齿再生牙髓学改进的平移方法。