Targeting the AMPK pathway to enhance dentin repair with novel metformin-releasing dental cements

靶向 AMPK 通路，利用新型二甲双胍释放牙科水泥增强牙本质修复

• 批准号: 10657804
• 负责人: Abraham Schneider
• 金额: $ 23.18万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10657804
• 关键词: 5' -AMP-activated protein kinase; Accidents; Adult; Affect; Antidiabetic Drugs; Biguanides; Biocompatible Materials; Bioenergetics; Biological; Biology; Biopolymers; Calcium; Cell Survival; Cells; Cementation; Chemicals; Chitosan; Clinical; Complex; Data; Dental; Dental Care; Dental Cements; Dental Materials; Dental Pulp; Dental Pulp Capping; Dental Pulp Exposure; Dental caries; Dentin; Dentin Formation; Dentinogenesis; Dentistry; Development; Differentiation Antigens; Drug usage; Elasticity; Excision; FDA approved; Formulation; Gene Expression; Gene Family; Glucose; Goals; Hardness; Human; Hydroxyapatites; Hypoglycemic Agents; Impairment; In Vitro; Inflammatory Response; Injury; Ions; Legal patent; Mechanics; Metformin; Minerals; Modeling; Modulus; Molecular Target; Natural regeneration; Odontoblasts; Oral; Oral health; Organic Cation Transporter; Organic Cation Transporter 1; Outcome; Oxides; Pathway interactions; Persons; Pharmaceutical Preparations; Plant Resins; Positioning Attribute; Procedures; Property; Rattus; Reporting; Signal Pathway; Signal Transduction; Silicates; Source; Stress; Structure; Technology Transfer; Testing; Therapeutic; Thick; Time; Tissue Engineering; Tissues; Tooth structure; Translating; Traumatic injury; United States; Up-Regulation; Work; biomaterial compatibility; calcium phosphate; cost; cost effective; diabetic patient; genetic approach; hydrophilicity; improved; in vivo; innovation; insight; mechanical properties; mineralization; nanoparticle; novel; permanent tooth; preservation; prevent; procedure cost; public health relevance; regenerative; repaired; response; restorative material; solute; stem cells; therapy outcome; uptake

PROJECT SUMMARY The normal structure and function of the dentin-pulp complex in adult permanent teeth can be affected by the exposure of a vital pulp following deep caries removal, traumatic injuries, or accidental restorative procedures. To stimulate dentin repair, preserve pulp vitality and avoid more invasive and costly procedures, vital pulp therapy relies on direct pulp-capping agents. These are mainly composed of inorganic hydraulic calcium-silicate cements, where mineral trioxide aggregate (MTA) is often considered the gold standard. Despite its well-accepted therapeutic value, it remains unclear which specific underlying signaling mechanisms orchestrate reparative dentinogenesis through the differentiation of dental pulp stem cells (DPSCs) into odontoblast-like cells. Also, common drawbacks associated with MTA include long setting times and high cost. Thus, enhancing dentin repair through novel, substantially more affordable bioactive formulations with improved physico-mechanical properties that molecularly target the pulp cells responsible for its synthesis could translate into truly beneficial and highly cost-effective therapeutic outcomes. We provide the first evidence supporting the development of a novel biomaterial formulated with calcium phosphate cement/chitosan (CPCC) and metformin (Met), that triggered a significant upregulation in the expression of odontoblastic differentiation markers and mineral synthesis in DPSCs. Met is a widely used, safe and low-cost oral anti-diabetic biguanide drug, and potent activator of the AMP-activated protein kinase (AMPK) signaling pathway, a master sensing mechanism of cellular bioenergetics. These promising preliminary data imply that Met could be safely repurposed within locally delivered formulations to enhance reparative dentin by molecularly targeting AMPK. In the proposed studies, we seek to maximize dentin repair by developing a new Met-CPCC pulp-capping agent with similar mechanical and flowability properties like MTA but with a substantial, several folds of reduction in setting time and cost. This innovative formulation relies on Met to induce AMPK activation and odontoblastic differentiation in DPSCs, and CPCC to provide the alkaline, ionic building blocks for hydroxyapatite formation. To that end, we will test the central hypothesis that dentin repair following vital pulp exposure is significantly potentiated by a Met-releasing CPCC bioactive pulp-capping agent through AMPK activation and delivery of mineralized tissue-building ions. In vitro and in vivo studies will expand our initial findings through two specific aims. Aim 1 will test the hypothesis that in DPSCs, a novel Met-CPCC pulp-capping agent induces odontogenic responses in an AMPK-dependent manner. In Aim 2, we will test the hypothesis that Met-CPCC pulp-capping agent significantly enhances dentin repair and increases the hardness and elastic modulus of new dentin in a rat dentin injury model with pulp exposure in vivo. The long-term goal of this proposal is to potentiate reparative dentinogenesis with novel biologically active Met-containing dental materials targeting AMPK activation, yielding new mechanisms and improved treatments that are widely applicable to restorative and regenerative dentistry.

项目摘要 成人恒牙中牙本质 - 羽状络合物的正常结构和功能可能会受到暴露的影响 深层龋齿去除，创伤性伤害或意外修复程序后，重要的果肉。刺激牙本质 维修，保留果肉活力并避免更具侵入性和昂贵的手术，重要的纸浆治疗依赖于直接纸浆粘贴 代理商。这些主要由无机液压钙硅酸盐水泥组成 （MTA）通常被认为是黄金标准。尽管具有良好的治疗价值，但仍不清楚哪种特定 潜在的信号传导机制通过分化牙髓干细胞来协调修复性牙本质生成 （DPSC）进入牙糖细胞样细胞。此外，与MTA相关的常见缺点包括长时间的时间和高 成本。因此，通过小说增强牙本质修复，并有所改善，可以实现更便宜的生物活性配方 分子靶向负责其合成的纸浆细胞的物理机械特性可以转化为真正的 有益且具有成本效益的治疗结果。我们提供了支持发展的第一个证据 用磷酸钙水泥/壳聚糖（CPCC）和二甲双胍（MET）制成的新型生物材料，该材料触发了A 在DPSC中，牙粒细胞分化标记和矿物合成的表达显着上调。 Met是 一种广泛使用的，安全和低成本的口服抗糖尿病型Biguanide药物，以及AMP激活的蛋白激酶的有效激活剂 （AMPK）信号通路，这是细胞生物能学的主传感机理。这些有希望的初步数据暗示 可以在本地传递的制剂中安全地重新利用该MET，以通过分子来增强牙本质 定位AMPK。在拟议的研究中，我们试图通过开发新的MET-CPCC纸浆上限来最大化牙本质修复 具有类似的机械和流动性能的代理，例如MTA，但具有很大的折叠率降低了 设定时间和成本。这种创新的配方依赖于MET诱导AMPK激活和ODOntoblastic分化 在DPSC和CPCC中，为碱性构建块提供了羟基磷灰石形成的离子构建块。为此，我们将测试 中心假设是，通过释放MET的CPCC，牙本质的维修在重要的果肉暴露后显着增强 通过AMPK激活和矿化组织建造离子的递送，生物活性纸浆封顶剂。体外和 体内研究将通过两个具体目标扩大我们的初始发现。 AIM 1将检验以下假设：在DPSC中，一种新颖 MET-CPCC纸浆封面剂以AMPK依赖性方式诱导牙源性反应。在AIM 2中，我们将测试 MET-CPCC纸浆贴剂的假设显着增强了牙本质的修复并增加了硬度和弹性 在大鼠牙本质损伤模型中，新牙本质的模量在体内暴露于果肉。该提议的长期目标是 用新型的生物活性含有元的牙科材料靶向AMPK的新型生物活性牙科的增强修复性牙本质发生 激活，产生新的机制和改进的治疗方法，可广泛适用于恢复和再生 牙科。