Neurobiological control of periodontal homeostasis through microRNA, TGF-beta, and Wnt signaling

通过 microRNA、TGF-β 和 Wnt 信号传导对牙周稳态的神经生物学控制

• 批准号: 9756189
• 负责人: Tom Diekwisch
• 金额: $ 15.39万
• 依托单位: TEXAS A&M UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-04 至 2020-02-18
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9756189
• 关键词: Affect; Animal Model; Animals; Ankylosis; Bioinformatics; Biological Assay; Bone Regeneration; Bone remodeling; Cicatrix; Collagen; Cytoskeletal Modeling; Cytoskeletal Proteins; Data; Data Set; Defect; Dentition; Diagnostic radiologic examination; Down-Regulation; Emotional; Engineering; Epithelial; Epithelial Cells; G-Protein-Coupled Receptors; Galanin; Ganglia; Gene Expression; Gene Expression Profiling; Goals; Health; Height; Homeostasis; Hybrids; In Vitro; Interruption; Jaw; Laboratories; Messenger RNA; MicroRNAs; Modeling; Nanosphere; Nerve; Nerve Fibers; Neurobiology; Neuropeptides; Operative Surgical Procedures; Osteoblasts; Osteoclasts; Osteocytes; Osteogenesis; Outcome; Pathway interactions; Patients; Peptides; Periodontal Diseases; Periodontal Ligament; Periodontal Pocket; Periodontitis; Periodontium; Phenotype; Prevention; Proteins; Rattus; Research; Rest; Reverse Transcriptase Polymerase Chain Reaction; Roentgen Rays; Role; Sensory; Signal Transduction; Signaling Molecule; Testing; Tissue Engineering; Tissues; Tooth Ankylosis; Tooth structure; Transforming Growth Factor beta; Trigeminal nerve structure; Up-Regulation; WNT Signaling Pathway; Walkers; alveolar bone; base; bone; clinically significant; craniofacial bone; design; glycogen synthase kinase 3 beta; inferior alveolar nerve; inhibitor/antagonist; innovation; mandibular nerve; mineralization; nerve supply; nerve transection; osteoclastogenesis; osteogenic; prevent; progenitor; response; sample fixation; skeletogenesis; soft tissue; spatial relationship; trend

The periodontium is a richly innervated tissue that undergoes continuous modelling and remodeling by alveolar bone osteoblasts, osteocytes, and osteoclasts. Studies from our laboratory have demonstrated a close spatial relationship between trigeminal nerve ganglia and the Epithelial Rests of Malassez (ERM), an epithelial cell network residing within the non-mineralized periodontal ligament. Inferior alveolar nerve (IAN) transection studies resulting in dento-alveolar ankylosis and a reduction in ERM have confirmed the essential role of sensory innervation for periodontal homeostasis. For the present application, we have established the IAN transection model in our laboratory and provided radiographic evidence for enhanced mineralization and ankylosis in the periodontal region of rat molars. Gene expression profiling comparing IAN transected and control tissues demonstrated an unexpected 28-fold significant increase in galanin (GAL) and a more than two-fold decrease in the TGF-β signaling molecules Smad2, Smad3 and Tgf-β1, and the Wnt inhibitors Dkk1, Dkk2 and Gsk-3β. In the same IAN transection group, microRNA miR-92b expression was more than two-fold upregulated, as verified via miRNA profiling and RT-PCR. MiR-92b upregulation after IAN transection in conjunction with bioinformatics data implicating Wnt and Tgf-β as possible miR-92b targets prompted us to speculate that GAL affects its skeletogenic downstream effects through miR-92b. In vitro studies revealed that GAL treatment promoted osteogenic differentiation of PDL progenitors and increased mineralization, while reducing osteoclastogenesis of BMMCs. Block of RhoA or application of GAL antagonist affected PDL cytoskeletal organization and gene expression, indicating that GAL functions through G protein coupled receptors. IWhen applied to periodontal pockets of animals suffering from periodontitis, GAL tissue engineering constructs accomplished a 20% increase in alveolar bone levels compared to controls, resulting in a clinically significant increase in alveolar bone height. Based on this promising new set of data we have designed a research plan to define the role of GAL in response to IAN transection, determine the mechanism underlying its effect on skeletogenesis, and exploit its applicability for bone regeneration and the prevention of periodontal ankylosis. The overall goal of our research plan is to test the hypothesis that periodontal nerves affect alveolar bone homeostasis through a GAL–GPCR–miR-92b–TGF-β/Wnt regulatory loop and that application of GAL neurosecretory peptides will stimulate Wnt signaling and new bone formation. We anticipate that the outcomes of our study will lead to innovative neuropeptide based/engineering hybrid approaches that will restore periodontal health in millions of patients and prevent the emotional and functional scars associated with lost teeth and incomplete dentitions.

牙周是一种富裕的组织，通过 牙槽骨成骨细胞，骨细胞和破骨细胞。我们实验室的研究表明 三叉神经神经节和上皮静止之间的紧密空间关系（erm） 位于非矿物化牙周韧带内的上皮细胞网络。下肺泡神经（IAN） 导致牙道肺内甲性甲性疾病和ERM减少的翻译研究证实了这一点 牙周稳态的感觉神经的重要作用。对于本申请，我们有 在我们的实验室中建立了IAN交易模型，并提供了放射线证据以增强 大鼠磨牙的牙周区域中的矿化和强直性。基因表达分析比较 伊恩（Ian）转移和控制组织表现出意外的28倍galanin显着增加 （GAL）和TGF-β信号分子SMAD2，SMAD3和TGF-β1的降低超过两倍。 WNT抑制剂DKK1，DKK2和GSK-3β。在同一IAN转导组中，microRNA mir-92b 通过miRNA分析和RT-PCR进行了验证，表达更新了两倍以上。 mir-92b IAN翻译后的上调与生物信息学数据含义Wnt和TGF-β一起 可能的miR-92b目标促使我们推测GAL会影响其骨骼发育的下游效应 通过mir-92b。体外研究表明，GAL处理促进了PDL的成骨分化 祖细胞和矿化增加，同时减少BMMC的破骨细胞生成。 Rhoa或 GAL拮抗剂的应用影响PDL细胞骨架组织和基因表达，表明 通过G蛋白偶联受体的gal功能。当我应用于牙周口袋时 患有牙周炎的gal组织工程构建体的肺泡增加了20％ 与对照相比，骨水平导致牙槽骨高度的临床显着增加。基于 在这一承诺新的数据集中，我们设计了一个研究计划，以定义GAL的作用 要伊恩翻译，确定其对骨骼生成的影响的机制，并利用它 适用于骨骼再生和预防牙周钙化病。我们的总体目标 研究计划是测试牙周神经影响牙槽骨稳态的假设 通过GAL – GPCR-MIR-92B – TGF-β/Wnt调节环和GAL的应用 神经分泌肽会刺激Wnt信号传导和新的骨形成。我们预计 我们研究的结果将导致创新的基于神经肽/工程的混合方法 恢复数百万患者的牙周健康，并防止与情绪和功能疤痕相关 牙齿失去和不完整的牙列。