Pulp fibroblast-mediated inferior alveolar nerve regeneration

牙髓成纤维细胞介导的下牙槽神经再生

• 批准号: 9979112
• 负责人: Seung Chung
• 金额: $ 15.99万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9979112
• 关键词: Action Potentials; Address; Adopted; Advocate; Allogenic; Area; Attention; Axon; Behavioral; Binding Proteins; Biological; Biological Assay; Blood Vessels; Brain; Brain-Derived Neurotrophic Factor; C5a anaphylatoxin receptor; CCAAT-Enhancer-Binding Proteins; Calcium; Cell Therapy; Cell Transplantation; Collagen Type I; Complement; Complement 5a; Complement Activation; Cues; Data; Denervation; Dental; Dental Care; Dental Implants; Dental Pulp; Dentin; Devices; Digastric muscle; Distal; Electric Stimulation; Electron Microscopy; Engineering; Esthesia; Event; Excision; Fibroblasts; Foundations; Future; G-Protein-Coupled Receptors; GTP-Binding Proteins; Gene Expression; Goals; Growth Factor; Human; Immunohistochemistry; In Vitro; Individual; Infection; Injury; Investigation; Knowledge; Light; Link; Measures; Mediating; Mesenchymal; Modeling; Molecular; Motor; Mus; Natural regeneration; Nerve; Nerve Regeneration; Neurites; Outcome; Pain; Pathway interactions; Patients; Pharmacology; Phosphotransferases; Play; Quality of life; Receptor Signaling; Recovery; Regenerative Medicine; Regulatory Pathway; Repression; Role; Science; Signal Transduction; Site; Small Interfering RNA; Source; Structure; System; Testing; Therapeutic; Therapeutic Intervention; Tissues; Tooth structure; Transactivation; Transplantation; Trauma; Work; adult stem cell; angiogenesis; base; cell growth; experimental study; functional outcomes; functional plasticity; healing; in vivo; in vivo regeneration; inferior alveolar nerve; inhibitor/antagonist; injured; injury and repair; innovation; knockout gene; maxillofacial; mind control; negative affect; nerve agent; nerve injury; neurogenesis; neurotrophic factor; neurotropic; novel; p38 Mitogen Activated Protein Kinase; paracrine; peripheral nerve regeneration; preservation; receptor; regenerative; regenerative therapy; repaired; stem cells; tool; transcription factor; transplant model; wisdom tooth

Project Summary/Abstract The dental pulp is composed of a supporting reticular matrix containing blood vessels, nerves, stromal fibroblasts and few specific stem cells. Regenerative medicine has focused on the dental pulp stem cell as a ready source of multipotential adult stem cells. However, the dental pulp fibroblast also may provide important cues for the functioning dental pulp. Included is the dental pulp fibroblast’s support of nerve repair. This project adopts an innovative and alternative approach to regenerative sciences by leveraging the neurotrophic features of dental pulp fibroblasts, potentially for nerve regeneration therapies. Here we explore ways of enhancing this neurotropic function of pulp fibroblasts via a novel C5L2 pathway involving p38 map kinase (p38) signaling. We recently studied the complement C5a receptor (C5aR) as one of the initial events that control the brain-derived neurotrophic factor (BDNF) secretion. This resultant BDNF secretion induces neurite outgrowth towards the injury site. Whereas C5a is known to interact with its G-coupled protein receptor C5aR, another controversial C5a receptor has been cloned. The C5L2 was considered as a non-functional decoy receptor of C5a signaling, thus has received much less attention than C5aR. Preliminary data demonstrate that C5L2 inhibition by siRNA significantly increases the pulp fibroblast-mediated BDNF secretion and neurite outgrowth. We provide further evidence that p38 plays a key role in the pulp fibroblast-mediated BDNF modulation. Based on our observations, we hypothesize that pulp fibroblasts enhance nerve regeneration by a paracrine function of secreted neurotrophic factors via the C5L2 and p38 pathways. In this proposal, we will characterize the mechanisms of C5L2 action in the pulp fibroblast-mediated nerve outgrowth in vitro. We will use an axon investigation system device (AXIS) to identify the interaction with C5L2 and p38, and whether C5L2 function is dependent or independent of C5a/C5aR. We will further determine, in vivo, whether C5L2 acts on pulp fibroblast functions to control IAN regeneration using our mouse IAN regeneration model by transplantation of the C5L2-silenced pulp fibroblasts. In our follow-on R01, we will investigate how pulp fibroblast-mediated nerve regeneration and C5L2 signaling impact functional plasticity: (a) whether the gene knockout approaches demonstrate nerve regenerative potential using in vivo IAN regeneration model, (b) whether the transplantation of engineered pulp fibroblasts and dental pulp stem cells that constitutively express and BDNF into the IAN denervation model can enhance nerve regeneration, (c) the consequence of nerve regeneration regarding pulp and dentin healing, and (d) the consequence of nerve regeneration regarding a behavioral (i.e., pain) effect. The results obtained from this project will shed new light onto cellular and molecular events which orchestrate initial steps of nerve regeneration by linking the neurite outgrowth to pulp fibroblast function through C5L2 and p38 pathways. These studies will provide the basis for future potential therapeutic interventions of nerve repair and vital tooth preservation.

项目总结/摘要 牙髓是由含有血管、神经、基质和纤维的网状基质构成的 成纤维细胞和很少的特异性干细胞。再生医学已经将牙髓干细胞作为一种 多能成体干细胞的现成来源。然而，牙髓成纤维细胞也可以提供重要的 牙髓功能的线索包括牙髓成纤维细胞对神经修复的支持。这个项目 通过利用神经营养功能，采用创新和替代方法进行再生科学 牙髓成纤维细胞，潜在的神经再生疗法。在这里，我们探讨如何提高这一点 牙髓成纤维细胞的神经营养功能通过一种新的C5 L2途径，涉及p38映射激酶（p38）信号。我们 最近研究了补体C5 a受体（C5 aR）作为控制脑源性 神经营养因子（BDNF）分泌。这种合成的BDNF分泌诱导神经突向轴突生长。 受伤部位。尽管已知C5 a与其G偶联蛋白受体C5 aR相互作用，另一个有争议的 C5 a受体已被克隆。C5 L2被认为是C5 a信号传导的非功能性诱饵受体， 因此，它受到的关注远低于C5 aR。初步数据表明，通过siRNA抑制C5 L2 显着增加牙髓成纤维细胞介导的脑源性神经营养因子分泌和神经突生长。我们进一步提供 p38在牙髓成纤维细胞介导的BDNF调节中起关键作用的证据。根据我们的观察， 我们假设牙髓成纤维细胞通过分泌的神经元的旁分泌功能促进神经再生， 神经营养因子通过C5 L2和p38途径。在本提案中，我们将描述 C5 L2在牙髓成纤维细胞介导的神经生长中的作用我们将使用轴突调查系统 设备（AXIS），以确定与C5 L2和p38的相互作用，以及C5 L2功能是否依赖或 与C5 a/C5 aR无关。我们将进一步确定，在体内，C5 L2是否作用于牙髓成纤维细胞功能， 通过移植C5 L2沉默的牙髓，使用我们的小鼠IAN再生模型控制IAN再生 成纤维细胞在我们后续的R 01中，我们将研究牙髓成纤维细胞介导的神经再生和C5 L2 信号传导影响功能可塑性：（a）基因敲除方法是否证明神经再生 可能使用体内IAN再生模型，（B）是否移植工程化牙髓成纤维细胞 而组成性表达BDNF的牙髓干细胞进入IAN去神经模型， 神经再生，（c）神经再生对牙髓和牙本质愈合的影响，以及（d） 关于行为的神经再生的结果（即，疼痛）效果。由此获得的结果 该项目将为细胞和分子事件提供新的线索，这些事件协调了神经再生的初始步骤 通过C5 L2和p38通路将神经突生长与牙髓成纤维细胞功能联系起来。这些研究将 为未来神经修复和活齿保存的潜在治疗干预提供基础。