Exosome-based Cell Homing and Lineage-Specific Differentiation Strategies for Dental Pulp Regeneration

基于外泌体的牙髓再生细胞归巢和谱系特异性分化策略

• 批准号: 10189073
• 负责人: Dong Rim Seol
• 金额: $ 15.09万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-14 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10189073
• 关键词: Alkaline Phosphatase; Anti-Bacterial Agents; Applications Grants; Attention; Behavior; Biological Assay; Blood; Body Temperature; Cell Culture Techniques; Cell Differentiation process; Cell Lineage; Cells; Chemotactic Factors; Chemotaxis; Communicable Diseases; Conditioned Culture Media; Cultured Cells; Data; Dental; Dental Pulp; Dental Pulp Capping; Dental caries; Dentin; Disadvantaged; Encapsulated; Endodontics; Endothelium; Extravasation; Fibrin; Future; Goals; Growth; Histologic; Homing; Human; Hyaluronic Acid; Hydrogels; In Vitro; Incisor; Inflammatory; Injectable; Knowledge; Long-Term Effects; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; MicroRNAs; Minerals; Modeling; Morphology; Natural regeneration; Necrosis; Nonvital Tooth; Odontoblasts; Odontogenesis; Operative Surgical Procedures; Oral cavity; Oryctolagus cuniculus; Outcome; Paste substance; Patients; Play; Procedures; Production; Property; Prosthesis; Pulpitis; Pulpotomy; Risk; Role; Structure; System; Temperature; Testing; Therapeutic; Time; Tissues; Tooth Discoloration; Tooth structure; Toxic effect; Tube; Undifferentiated; angiogenesis; base; biomaterial compatibility; blood vessel development; calcium hydroxide; cell motility; cohesion; cost; engineered exosomes; environmental change; exosome; healing; in vivo; intercellular communication; miRNA expression profiling; next generation sequencing; novel; preservation; progenitor; protein biomarkers; pulp capping material; recruit; regenerative; response; scaffold; seal; stem cell migration; stem cells; tissue regeneration; tissue repair; uptake

[PROJECT SUMMARY/ABSTRACT] Current endodontic therapy for dental caries, which are one of the most prevalent infectious diseases in the world, is a procedure for replacing the vital pulp with synthetic pulp-capping materials. Pulpless teeth can lose their functions to sense environmental changes and maintain dentin regeneration, and the synthetic materials have several disadvantages such as bacterial leakage into the dental pulp, poor cohesive strength, discoloration of tooth, and long setting time. As an alternative, vital pulp therapy (VPT), which is defined as a restorative dental treatment that aims to preserve and maintain pulp tissue, is beneficial for young patients who have high healing capacity for pulp regeneration. Potential for successful VPT and pulp regeneration is increasing due to the knowledge of mesenchymal stem cells (MSCs) that can differentiate into specialized cells. However, the transplantation of MSCs incurs high costs and risks associated with the ex vivo cell expansion. Consequently, a cell homing strategy which recruits endogenous dental pulp stem cells (DPSCs) is the effective approach in endodontics. Recently, exosomes have attracted attention due to their great potential to promote intercellular communication leading to enhanced cell recruitment, differentiation to specific cell lineage, and tissue regeneration. In particular, conditioned medium or exosomes cultured under lineage-specific differentiation have a great potential for angiogenesis and odontogenesis for pulp regeneration. The long-term goal is to develop a pulp capping material system for vital pulp therapy of human dental pulp. The overall objectives for this application are (1) to elucidate the therapeutic potential of characterized exosomes as a chemoattractant to stimulate DPSC migration and pulp-like differentiation and (2) to determine their in vivo effect of pulp regeneration in a rabbit partial pulpotomy model. Our central hypothesis is that exosomes will stimulate dental pulp regeneration by promoting DPSC chemotaxis and lineage-specific differentiation. Exosomes will be isolated from rabbit DPSCs cultured under growth or lineage-specific differentiation conditions (odontogenesis or angiogenesis) and will be encapsulated in injectable hydrogel (F-127/hyaluronic acid) which has temperature-sensitive gelation behavior at body temperature. Specific aims in this proposed study are (1) characterize DPSC-Exos and determine the effect of DPSC-Exos on in vitro cell homing and lineage-specific differentiation and (2) evaluate the in vivo effects of DPSC-Exos on pulp regeneration in a rabbit partial pulpotomy. At the completion of the proposed R03 project, our expected outcomes are to define the therapeutic potential of DPSC-Exos for dental pulp regeneration and to identify miRNAs that may regulate cell homing and pulp tissue formation. These results will have a very important positive impact by providing preliminary data for our future R01 grant application in which we plan to synthesize miRNAs loaded in engineered exosomes, thereby replacing DPSC culture as a potential means of exosome production.

[项目概要/摘要] 龋病是世界上最常见的感染性疾病之一， 是一种用合成盖髓材料替代活髓的方法。无髓牙会失去 它们具有感知环境变化和维持牙本质再生的功能， 具有几个缺点，例如细菌渗漏到牙髓中、内聚强度差、变色 牙齿，和长的设置时间。作为一种替代方法，活髓疗法（VPT），被定义为一种修复性牙科治疗。 旨在保护和维持牙髓组织的治疗，对于愈合率高的年轻患者有益 纸浆再生能力。成功的VPT和纸浆再生的潜力正在增加， 间充质干细胞（MSC）可以分化成专门的细胞的知识。但 MSC的移植引起与离体细胞扩增相关的高成本和风险。因此， 细胞归巢策略招募内源性牙髓干细胞（DPSCs）是治疗牙髓病的有效方法。 牙髓病学近年来，外泌体由于其促进细胞间相互作用的巨大潜力而引起人们的关注。 细胞间通讯导致增强的细胞募集、向特定细胞谱系的分化和组织分化。 再生特别地，在谱系特异性分化下培养的条件培养基或外来体具有 牙髓再生的血管生成和牙发生的巨大潜力。 长期目标是开发一种用于人牙髓活髓治疗的盖髓材料体系。 本申请的总体目标是（1）阐明表征的外泌体的治疗潜力 作为化学引诱物刺激DPSC迁移和髓样分化，以及（2）测定它们的体内 兔部分活髓切断模型中牙髓再生效果。我们的中心假设是外泌体 通过促进DPSC趋化性和谱系特异性分化来刺激牙髓再生。 将从在生长或谱系特异性分化条件下培养的兔DPSC中分离外泌体 （牙生成或血管生成）并将被封装在可注射水凝胶（F-127/透明质酸）中， 在体温下具有温度敏感的凝胶化行为。本研究的具体目标是：（1） 表征DPSC-Exos并确定DPSC-Exos对体外细胞归巢和谱系特异性的影响 （2）评估DPSC-Exos对兔部分活髓切断术中的牙髓再生的体内作用。 在拟议的R 03项目完成时，我们的预期结果是确定治疗潜力 DPSC-Exos用于牙髓再生，并鉴定可能调节细胞归巢和牙髓组织的miRNA 阵这些结果将通过为我们的未来提供初步数据而产生非常重要的积极影响 R 01赠款申请，我们计划合成装载在工程外泌体中的miRNA，从而取代 DPSC培养作为外泌体生产的潜在手段。