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: 10505282
• 负责人: Abraham Schneider
• 金额: $ 19.31万
• 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/10505282
• 关键词: 5' -AMP-activated protein kinase; Adult; Affect; Antidiabetic Drugs; Biguanides; Biocompatible Materials; Bioenergetics; Biological; Biology; Biopolymers; Calcium; Cations; Cell Survival; Cells; 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; Excision; FDA approved; Formulation; Gene Expression; Gene Family; Glucose; Goals; Gold; Hardness; Human; Hydroxyapatites; Hypoglycemic Agents; Impairment; In Vitro; Inflammatory Response; Injury; Ions; Lead; Legal patent; Mechanics; Metformin; Minerals; Modeling; Modulus; Molecular Target; Natural regeneration; Odontoblasts; Oral; Oral health; Organic Cation Transporter; Organic Cation Transporter 1; Outcome; Pathway interactions; Persons; Pharmaceutical Preparations; Plant Resins; Positioning Attribute; Procedures; Property; Protein Kinase; 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; alkalinity; base; biomaterial compatibility; calcium phosphate; cost; cost effective; diabetic patient; genetic approach; hydrophilicity; improved; in vivo; innovation; insight; mechanical properties; 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.

项目概要 成人恒牙中牙本质-牙髓复合体的正常结构和功能可能会受到暴露的影响。 深度龋齿去除、外伤或意外修复手术后的活髓。刺激牙本质 修复、保持牙髓活力并避免更具侵入性和昂贵的手术，活牙髓治疗依赖于直接盖髓 代理。这些水泥主要由无机水硬硅酸钙水泥组成，其中三氧化二矿物骨料 (MTA) 通常被认为是黄金标准。尽管其治疗价值已被广泛接受，但仍不清楚其具体作用是什么 潜在的信号机制通过牙髓干细胞的分化协调修复性牙本质发生 (DPSC) 转化为成牙本质细胞样细胞。此外，与 MTA 相关的常见缺点包括凝固时间长和温度高 成本。因此，通过新颖的、更实惠的生物活性配方来增强牙本质修复 分子靶向负责其合成的牙髓细胞的物理机械特性可以转化为真正的 有益且极具成本效益的治疗结果。我们提供了支持开发的第一个证据 由磷酸钙水泥/壳聚糖（CPCC）和二甲双胍（Met）配制而成的新型生物材料，引发了 DPSC 中成牙本质细胞分化标记物的表达和矿物质合成显着上调。梅特是 一种广泛使用、安全且低成本的口服抗糖尿病双胍药物，也是 AMP 激活蛋白激酶的有效激活剂 (AMPK) 信号通路，细胞生物能量学的主要传感机制。这些有希望的初步数据意味着 Met 可以安全地在局部交付的配方中重新利用，以通过分子方式增强修复性牙本质 靶向 AMPK。在拟议的研究中，我们寻求通过开发一种新的 Met-CPCC 盖髓来最大限度地修复牙本质 具有与 MTA 类似的机械和流动性特性的试剂，但显着降低数倍 设定时间和成本。这种创新配方依靠 Met 诱导 AMPK 激活和成牙本质细胞分化 在 DPSC 和 CPCC 中为羟基磷灰石的形成提供碱性离子结构单元。为此，我们将测试 中心假设是，释放 Met 的 CPCC 显着增强了活髓暴露后的牙本质修复 通过 AMPK 激活和输送矿化组织构建离子的生物活性盖髓剂。体外和体内 体内研究将通过两个具体目标扩展我们的初步发现。目标 1 将检验以下假设：在 DPSC 中，一种新颖的 Met-CPCC 盖髓剂以 AMPK 依赖性方式诱导成牙反应。在目标 2 中，我们将测试 假设Met-CPCC盖髓剂显着增强牙本质修复并增加硬度和弹性 体内牙髓暴露的大鼠牙本质损伤模型中新牙本质的模量。该提案的长期目标是 利用靶向 AMPK 的新型生物活性含 Met 牙科材料增强修复性牙本质生成 激活，产生新的机制和改进的治疗方法，广泛适用于恢复和再生 牙科。