Microengineering the Dental Pulp Vascular Microenvironment

牙髓血管微环境的微工程

• 批准号: 9981727
• 负责人: Luiz Eduardo Bertassoni
• 金额: $ 50.31万
• 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 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9981727
• 关键词: 3-Dimensional; Address; Adult; Affect; Angiogenic Factor; Architecture; 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; Diffuse; Endodontics; Endothelial Cells; Endothelium; Engineering; Environment; Evaluation; Excision; Extracellular Matrix; Gelatin; Goals; Heart; Human; Hydrogels; Implant; In Vitro; Individual; Injectable; Length; Light; Mediating; Methacrylates; Microfluidics; Mus; Natural regeneration; Necrosis; Nutrient; Odontoblasts; Pepsin A; Phenotype; Plant Roots; Polymers; Process; Property; Pulp Canals; Role; Root Canal Therapy; Side; Signaling Molecule; Site; Skin; Source; Swelling; Techniques; Testing; Tissue Engineering; Tissue Model; Tissue Survival; Tissues; Tooth Apex; Tooth structure; Vascular blood supply; Vascularization; Waste Products; base; biomaterial compatibility; bioprinting; blood vessel development; cell motility; crosslink; dental structure; improved; in vivo; mechanical properties; monolayer; novel; paracrine; permanent tooth; physical property; regenerative; response; scaffold; stem cells; success; tissue regeneration; translational approach; vasculogenesis; virtual

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.

项目总结： 龋齿是一种感染性疾病，全世界约90%的成年人受到影响。龋齿晚期 影响牙髓，导致组织坏死，最终需要进行根管治疗。通常，根用户 恒牙的根管是通过移除坏死组织并用人造材料取代它来治疗的。 再生性牙髓治疗已被提议作为这些疾病的一种改进的治疗选择。然而， 如果没有可控的策略来设计牙髓血管系统，有效的牙髓再生几乎是 不可能。最近已经证明，通过培养可以设计出具有功能的血管系统。 来自各种来源的内皮细胞和干细胞在正确的微环境条件下。然而， 再生牙髓血管系统的确切要求仍然知之甚少。这个项目 我将系统地研究三个重叠的方面，我们认为这三个方面是再生的关键决定因素 牙髓血管系统：(1)基质物理和机械特性，(2)组成，和(3)微结构。 在目标1中，我们将调查不同的物理和机械性能对以下能力的贡献 人内皮细胞集落形成细胞和牙髓干细胞形成微血管 当网络嵌入到水凝胶中时，可以通过光交联使其具有系统的性质 调整过了。然后，我们将设计牙髓组织--用预制的内皮细胞预血运的牙髓组织 促进全长根管内牙髓再生的微通道。在目标2中，我们将开发 由牙本质天然基质合成的可注射和光固化的水凝胶 测试基质成分对牙髓血管再生的贡献。 此外，我们将把这些水凝胶与从牙本质基质中提取的血管生成成分结合起来，并 在体外和体内测试它们的再生能力。在目标3中，我们将制作建筑控制的渐变 用微流控技术检测ECFC和DPSC旁分泌因子对组织的贡献 微结构对牙髓血管系统的形成。然后我们将模拟的微体系结构 通过3D生物打印组织构建的血管化牙髓，复制了天然牙髓的组织。 在这个项目的最后，我们希望已经完成了一个3D血管化的牙髓微环境的微工程，它将 改进用于成人牙齿再生牙髓治疗的翻译方法。