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

基本信息

批准号：
10450657
负责人：
Dong Rim Seol
金额：
$ 15.26万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-14 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10450657
关键词：
Alkaline Phosphatase Anti-Bacterial Agents Applications Grants Attention Behavior Biological Assay Blood Body Temperature Cell Differentiation process Cell Lineage Cells Chemotactic Factors Chemotaxis Communicable Diseases Conditioned Culture Media 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 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

项目摘要

[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] 目前针对龋齿的牙髓治疗，龋齿是世界上最流行的传染病之一世界上，是一种用合成盖髓材料代替活髓的程序。无髓牙可能会脱落它们具有感知环境变化和维持牙本质再生的功能，以及合成材料存在细菌渗入牙髓、粘结强度差、变色等缺点齿数多，凝固时间长。作为替代方案，活体牙髓治疗 (VPT) 被定义为一种修复性牙齿治疗方法。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。由于以下原因，成功 VPT 和纸浆再生的潜力正在增加了解可分化为特化细胞的间充质干细胞 (MSC)。然而，间充质干细胞移植会产生与离体细胞扩增相关的高成本和风险。因此，一个招募内源性牙髓干细胞（DPSC）的细胞归巢策略是治疗牙髓病的有效方法。牙髓病学。近年来，外泌体因其促进细胞间质转化的巨大潜力而受到关注。通讯导致增强细胞募集、分化为特定细胞谱系和组织再生。特别是，在谱系特异性分化下培养的条件培养基或外泌体具有血管生成和牙髓再生的巨大潜力。长期目标是开发一种用于人类牙髓活髓治疗的盖髓材料系统。该应用的总体目标是（1）阐明特征外泌体的治疗潜力作为化学引诱剂刺激 DPSC 迁移和牙髓样分化，以及 (2) 确定它们的体内兔部分活髓切断术模型中牙髓再生的效果。我们的中心假设是外泌体会通过促进 DPSC 趋化性和谱系特异性分化来刺激牙髓再生。外泌体将从在生长或谱系特异性分化条件下培养的兔 DPSC 中分离出来（牙发育或血管生成）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行为。这项研究的具体目标是 (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牙髓再生的体内影响。在拟议的 R03 项目完成后，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调节细胞归巢和牙髓组织的 miRNA 形成。这些结果将为我们的未来提供初步数据，产生非常重要的积极影响 R01 资助申请，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。