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.

项目摘要/摘要 牙髓由含有血管、神经、基质的支撑性网状基质组成 成纤维细胞和很少的特定干细胞。再生医学将牙髓干细胞作为一种 多潜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 这是牙髓功能正常的信号。其中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，采用创新和替代的方法来实现再生科学 牙髓成纤维细胞，可能用于神经再生治疗。在这里，我们探索加强这一点的方法 牙髓成纤维细胞通过一条新的C5L2通路发挥神经营养功能，该通路涉及p38丝裂原活化蛋白激酶(P38)信号。我们 最近对补体C5a受体(C5aR)的研究表明，补体C5a受体是控制脑源性 神经营养因子(BDNF)分泌。由此产生的BDNF分泌诱导轴突向外生长 受伤地点。虽然已知C5a与其G偶联蛋白受体C5aR相互作用，但另一个有争议的 C5a受体已被克隆。C5L2被认为是C5a信号的非功能性诱骗受体， 因此，与C5aR相比，它受到的关注要少得多。初步数据表明，siRNA对C5L2的抑制作用 显著增加牙髓成纤维细胞介导的脑源性神经营养因子的分泌和轴突生长。我们提供进一步的 有证据表明，p38在牙髓成纤维细胞介导的脑源性神经营养因子的调控中起关键作用。根据我们的观察， 我们假设牙髓成纤维细胞通过分泌的旁分泌功能促进神经再生。 神经营养因子通过C5L2和p38途径。在这项提案中，我们将描述 C5L2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(轴)，以确定与C5L2和p38的相互作用，以及C5L2功能是依赖还是 独立于C5a/C5aR。我们将在体内进一步确定C5L2是否作用于牙髓成纤维细胞功能以 C5L2沉默牙髓移植小鼠Ian再生模型控制Ian再生 成纤维细胞。在我们的后续R01中，我们将研究牙髓成纤维细胞如何介导神经再生和C5L2 信号影响功能可塑性：(A)基因敲除方法是否显示神经再生 潜在利用体内Ian再生模型，(B)是否移植工程化牙髓成纤维细胞 在Ian失神经模型中结构性表达和BDNF的牙髓干细胞可以增强 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(即疼痛)效应的神经再生的后果。从这个过程中得到的结果 该项目将为协调神经再生的最初步骤的细胞和分子事件提供新的线索 通过C5L2和p38通路将突起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。