Microengineering the Dental Pulp Vascular Microenvironment

牙髓血管微环境的微工程

基本信息

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
项目类别：
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

项目摘要

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：龋齿是一种传染病，影响全世界约 90% 的成年人。龋齿晚期影响牙髓，导致组织坏死，最终需要根管治疗。通常，根恒牙根管的治疗方法是去除坏死组织并用人造材料替换。再生牙髓学已被提议作为这些病症的改进治疗选择。然而，如果没有可控的策略来设计牙髓脉管系统，有效的牙髓再生实际上是不可能的不可能的。最近已经证明，功能性脉管系统可以通过培养来设计不同来源的内皮细胞和干细胞处于正确的微环境条件下。然而，对于牙髓脉管系统再生的具体要求仍然知之甚少。这个项目将系统地调查我们提出的三个重叠方面，它们是再生的关键决定因素牙髓脉管系统：(1) 基质物理和机械特性，(2) 成分，(3) 微结构。在目标 1 中，我们将研究不同物理和机械特性对能力的贡献人内皮集落形成细胞（ECFC）和牙髓干细胞（DPSC）形成微血管嵌入水凝胶中的网络可以通过光交联系统地获得其特性调整。然后，我们将设计用预制内皮细胞预血管化的牙髓组织结构微通道可增强体内全长根管的牙髓再生。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水凝胶，并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，测试它们的体外和体内再生潜力。在目标 3 中，我们将制造架构控制的梯度使用微流体技术检测 ECFC 和 DPSC 旁分泌因子以测试组织的贡献微结构影响牙髓脉管系统的形成。然后我们将模仿微架构通过 3D 生物打印组织结构来复制天然牙髓的组织，从而形成血管化牙髓。在这个项目结束时，我们期望通过微工程设计出一个 3D 血管化牙髓微环境，该环境将改进用于恒牙再生牙髓治疗的转化方法。