Mechanisms regulating afferent innervation in the dental pulp

调节牙髓传入神经支配的机制

基本信息

批准号：
10599553
负责人：
Sarah Peters
金额：
$ 6.74万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10599553
关键词：
Adult Age Axon Biological Assay Cells Cessation of life Chronic Disease Coculture Techniques Complement Factor B Complex Data Defect Dental Dental Pulp Dental caries Dentin Dentin Formation Dentinogenesis Development Developmental Process Endodontics Enterobacteria phage P1 Cre recombinase Exhibits Exposure to Extracellular Matrix Proteins Extracellular Protein Future Ganglia Genetic Transcription Goals Golgi Apparatus In Vitro Injury Laboratories Mesenchymal Mesenchyme Messenger RNA Modeling Mus Natural regeneration Nerve Fibers Neurites Neurons Odontoblasts Organ Organism Pain Pain management Paracrine Communication Pattern Penetration Phosphoproteins Play Preventive Process Pulp Chambers Reaction Regulation Reporting Research Rest Role Sensory Sequence Analysis Series Signal Transduction Site Stimulus Structure of trigeminal ganglion Temperature Testing Therapeutic Thick Tissues Tooth Germ Tooth Injuries Tooth structure Transforming Growth Factor beta Receptors Transforming Growth Factors Trigeminal nerve structure Wild Type Mouse afferent nerve axon guidance base bone confocal imaging experimental study healing improved in vivo mineralization mouse model nerve stem cell nerve supply neurite growth neuroblast neuron development neuron regeneration neurotrophic factor new therapeutic target osteopontin postnatal postnatal development preservation pressure promoter regenerative approach repaired response skeletal therapeutic development

项目摘要

Project Summary/Abstract Since teeth are exposed to environmental stimuli, tooth innervation is crucial to their lifelong protection and us- age. The tooth is primarily innervated with sensory nerve fibers from the trigeminal ganglion (TG) that help pro- tect the tooth organ by relaying noxious stimuli. During postnatal development, the dental pulp (DP) secretes neurotrophic factors to guide axonal penetration and sprouting within the tooth in a highly regulated manner. The transforming growth factor β (TGFβ) superfamily regulates many developmental processes in teeth, in- cluding mineralization and innervation. A secreted phosphoprotein downstream of TGFβ signals, osteopontin (OPN), is a major extracellular protein that regulates tooth mineralization and also promotes neurite outgrowth. Following tooth injury, DP cells release neurotrophic factors that modulate neurite growth and dentin repair. Interestingly, OPN-/- mice do not form reparative dentin during pulpal healing, suggesting that OPN plays a cru- cial role in this process. The long-term goal of this project is to understand the cellular mechanisms that protect teeth via sensory innervation. The overall objective is to determine the role of signals from the dental mesen- chyme that regulate tooth innervation. Our central hypothesis is that Tgfbr2 in the dental mesenchyme governs paracrine signaling via OPN to guide tooth sensory innervation. Our laboratory has established a mouse model in which Tgfbr2 is conditionally deleted in odontoblast-producing mesenchyme using an Osterix promoter- driven Cre recombinase (Tgfbr2cko). These mice survive postnatally, but have significant defects in their bones and teeth. We performed a series of experiments to determine whether Tgfbr2 signaling in the DP mesen- chyme regulates their differentiation and ability to guide neurite outgrowth during tooth development. We found that OPN-/- mice do not demonstrate reduced innervation at P7 as originally hypothesized, but do exhibit dis- rupted axonal targeting. The role of neurite outgrowth in regulating reactionary and reparative dentine when the mineralization capacity of the DP cells is compromised remains unknown. Guided by these data, we will test our hypothesis with the following two specific aims: 1) To analyze the OPN-governed signaling cascades in the dental mesenchyme regulating tooth sensory innervation during development; 2) To establish the degree to which the neurite outgrowth associated with dentin repair is regulated by Tgfbr2 and/or OPN signaling in the DP mesenchyme. Under the first aim, we will perform an mRNA Sequence Analysis of control and OPN-/- DP tissue to determine what signals could be disrupting the afferent organization. In the second aim, we will use an in vivo dental injury model and investigate neuronal regeneration in Tgfbr2cko, OPN-/-, and WT mice. The proposed research is significant because it is expected to advance and expand the understanding of how DP cells protect the tooth organ via mechanisms involving axonal guidance. Such information will enhance our un- derstanding of the complex interplay of mesenchymal-neuronal interactions in the tooth that could serve as a basis for future preventive, therapeutic, and regenerative strategies in endodontics to improve the preservation of teeth.

项目概要/摘要由于牙齿暴露在环境刺激下，牙齿神经支配对于牙齿的终生保护和使用至关重要。牙齿主要由三叉神经节 (TG) 的感觉神经纤维支配，有助于促进年龄的增长。通过传递有害刺激来保护牙齿器官在出生后发育过程中，牙髓 (DP) 会分泌。神经营养因子以高度调节的方式引导轴突穿透并在牙齿内发芽。转化生长因子 β (TGFβ) 超家族调节牙齿的许多发育过程，包括骨桥蛋白下游的 TGFβ 信号的矿化和神经支配。 (OPN) 是一种主要的细胞外蛋白，可调节牙齿矿化并促进神经突生长。牙齿损伤后，DP 细胞释放调节神经突生长和牙本质修复的神经营养因子。提示，OPN-/- 小鼠在牙髓愈合过程中不会形成修复性牙本质，这表明 OPN 在牙髓愈合过程中发挥着关键作用。该项目的长期目标是了解保护细胞的机制。总体目标是确定来自牙间神经的信号的作用。我们的中心假设是牙齿间充质中的 Tgfbr2 负责控制。通过 OPN 的旁分泌信号来引导牙齿感觉神经支配我们实验室建立了小鼠模型。其中使用 Osterix 启动子在成牙本质细胞产生的间充质中有条件地删除 Tgfbr2 - 这些小鼠出生后能够存活，但其骨骼存在明显缺陷。我们进行了一系列实验来确定 DP 间质中的 Tgfbr2 信号传导是否存在。我们发现，食糜调节它们的分化和引导神经突生长的能力。 OPN-/- 小鼠并未表现出最初发育时 P7 神经支配的减少，但确实表现出异常破裂的轴突靶向。神经突生长在调节反应性和修复性牙本质中的作用。 DP 细胞的矿化能力是否受到损害仍然未知，我们将根据这些数据进行测试。我们的假设有以下两个具体目标：1）分析 OPN 控制的信号级联牙齿间质在发育过程中调节牙齿感觉神经支配 2) 确定牙齿感觉神经支配的程度；与牙本质修复相关的神经突生长受到 Tgfbr2 和/或 OPN 信号传导的调节在第一个目标下，我们将对对照和 OPN-/- DP 进行 mRNA 序列分析。确定哪些信号可能会干扰传入组织在第二个目标中，我们将使用。体内牙齿损伤模型并研究 Tgfbr2cko、OPN-/- 和 WT 小鼠的神经元再生。拟议的研究意义重大，因为它有望推进和扩大对 DP 如何进行的理解细胞通过涉及轴突引导的机制来保护牙齿器官，这些信息将增强我们的能力。了解牙齿中间充质-神经元相互作用的复杂相互作用，可以作为为未来牙髓学的预防、治疗和再生策略奠定基础，以改善保存的牙齿。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A Co-Culture Method to Study Neurite Outgrowth in Response to Dental Pulp Paracrine Signals.

DOI：
10.3791/60809
发表时间：
2020-02-14
期刊：
Journal of visualized experiments : JoVE
影响因子：
0
作者：
Barkley C;Serra R;Peters SB
通讯作者：
Peters SB

Co-culture methods to study neuronal function and disease.

DOI：
10.4103/1673-5374.297066
发表时间：
2021-05
期刊：
Neural regeneration research
影响因子：
6.1
作者：
Peters SB
通讯作者：
Peters SB

Nociceptors are needed to guide tooth development, function, repair, and regeneration.

DOI：
10.4103/1673-5374.360280
发表时间：
2023-07
期刊：
Neural regeneration research
影响因子：
6.1
作者：
Peters SB;Emrick JJ
通讯作者：
Emrick JJ

Tgfbr2 in Dental Pulp Cells Guides Neurite Outgrowth in Developing Teeth.

DOI：
10.3389/fcell.2022.834815
发表时间：
2022
期刊：
Frontiers in cell and developmental biology
影响因子：
5.5
作者：
Stanwick M;Barkley C;Serra R;Kruggel A;Webb A;Zhao Y;Pietrzak M;Ashman C;Staats A;Shahid S;Peters SB
通讯作者：
Peters SB