A novel metformin-nanomineral scaffold as enhancer of craniofacial bone regeneration and angiogenesis via dental pulp stem cells

一种新型二甲双胍纳米矿物质支架通过牙髓干细胞增强颅面骨再生和血管生成

• 批准号: 10256799
• 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: 2020
• 资助国家: 美国
• 起止时间: 2020-09-08 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10256799
• 关键词: 5' -AMP-activated protein kinase; Antidiabetic Drugs; Area; Autologous; Biguanides; Bioenergetics; Biological; Biopolymers; Blood Glucose; Bone Growth; Bone Regeneration; Bone Tissue; Bone Transplantation; Calvaria; Cell Survival; Cells; Chemosensitization; Chitosan; Clinical; Conditioned Culture Media; Coupling; Data; Defect; Dental Models; Dental Pulp; Deposition; Development; Drug Costs; Drug usage; Endothelial Cells; Enhancers; Exposure to; FDA approved; Formulation; Goals; Growth Factor; Hepatocyte; Hypoglycemic Agents; Hypoxia; Implant; In Vitro; Infection; Lead; Malignant Neoplasms; Mediating; Mediator of activation protein; Mesenchymal Differentiation; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Metformin; Minerals; Modeling; Nanostructures; Natural regeneration; Oral; Orthopedic Surgery; Orthopedics; Osteogenesis; Patients; Pharmaceutical Preparations; Pharmacology; Phosphotransferases; Play; Protein Kinase; Rattus; Reporting; Research; STK11 gene; Signal Pathway; Signal Transduction; Source; Surgeon; Testing; Therapeutic; Time; Tissue Engineering; Tissues; Trauma; Vascular Endothelial Growth Factors; Work; angiogenesis; base; bone; calcium phosphate; cost; cost effective; craniofacial; craniofacial bone; dental transplantation; diabetic; diabetic patient; extracellular; glucose production; improved; in vivo; insight; maxillofacial; multidisciplinary; neovascularization; novel; oral surgery specialty; osteoblast differentiation; osteogenic; postnatal; public health relevance; response; scaffold; skeletal; skeletal regeneration; stem cell differentiation; stem cells; success; upstream kinase

PROJECT SUMMARY The long-term goal of this proposal is to develop widely applicable, cost-effective bone tissue engineering platforms that combine metformin with stem cells to regenerate large, critical-sized oral and craniofacial skeletal defects. Although tissue engineering using stem cells, scaffolds and growth factors offers an attractive, less invasive alternative to autologous bone grafts, its success highly depends on the proper adaptation of cells to a local hypoxic microenvironment, and reestablishment of a functional microvasculature. It is well established that vascular endothelial growth factor (VEGF) is a key mediator of osteogenic/angiogenic coupling in bone regeneration. Thus, developing novel and affordable stem cell-based tissue engineering strategies that potentiate VEGF-mediated angiogenesis may significantly enhance skeletal regeneration. Our group and others recently reported that metformin, a low-cost drug used by millions of diabetics worldwide induces the osteoblastic differentiation of stem cells derived from various tissue sources. This suggests that metformin could be repurposed in a local delivery formulation to potentiate stem cell-based bone regeneration. We have advanced this concept by formulating a calcium phosphate cement (CPC) containing metformin that when released in culture upregulated the expression of osteogenic markers and increased mineralized extracellular deposits in dental pulp stem cells (DPSCs), an easily accessible and inexhaustible source of postnatal stem cells. Intriguingly, we have found that metformin also induces a significant increase in VEGF secretion that is further amplified in DPSCs exposed to hypoxic conditions. The osteogenic action of metformin has been associated with the activation of the AMP-activated protein kinase (AMPK) signaling pathway, a master sensing mechanism of cellular bioenergetics. While in hepatocytes, metformin reduces high blood glucose production by activating AMPK via the upstream kinase liver kinase B1 (LKB1), a mechanistic, translationally relevant question that still remains elusive is whether DPSCs rely mainly on LKB1 to enhance bone formation and neovascularization in response to locally delivered metformin. We will test the central hypothesis that DPSC-based craniofacial bone regeneration and neovascularization in response to locally delivered metformin is enhanced by AMPK activation through a functional, catalytically active LKB1. In vitro and in vivo studies will expand our results through two specific aims. Aim 1 will determine whether in DPSCs, metformin released from CPC scaffolds induces osteogenic and pro-angiogenic responses in an LKB1/AMPK-dependent manner. Aim 2 will test the hypothesis that in DPSCs, a functional LKB1/AMPK cellular response is necessary to enhance craniofacial bone regeneration and neovascularization in response to locally delivered metformin released from CPC scaffolds. We anticipate our results will yield new, valuable basic and translational information that will lead to cost-effective tissue engineering platforms where metformin-loaded scaffolds in combination with stem cells will enhance craniofacial and orthopedic bone regeneration.

项目摘要 该提案的长期目标是开发广泛适用的，具有成本效益的骨组织工程平台 将二甲双胍与干细胞结合起来，以再生大型，临界大小的口服和颅面骨骼缺陷。虽然组织 使用干细胞，脚手架和生长因子的工程提供了一种自体骨的吸引人，侵入性较低的替代品 移植物，其成功很大程度上取决于细胞对局部低氧微环境的适当适应和重建 功能性微脉管系统。众所周知，血管内皮生长因子（VEGF）是 骨再生中的成骨/血管生成耦合。因此，开发新颖且负担得起的干细胞组织工程 增强VEGF介导的血管生成的策略可能会显着增强骨骼再生。我们的小组和其他人 最近报道，二甲双胍是一种低成本药物，全世界数百万糖尿病患者诱导了成骨细胞 来自各种组织来源的干细胞的分化。这表明二甲双胍可以在局部重新使用 递送配方以增强基于干细胞的骨再生。我们通过制定钙提出了这个概念 含有二甲双胍的磷酸盐水泥（CPC）在培养物中释放时上调成骨的表达 标记物并增加了牙髓干细胞（DPSC）中的矿化细胞外沉积物，易于访问和 产后干细胞的无尽来源。有趣的是，我们发现二甲双胍也诱导了显着增加 在暴露于低氧条件下的DPSC中进一步扩增的VEGF分泌。二甲双胍的成骨作用 与AMP激活蛋白激酶（AMPK）信号通路的激活有关 细胞生物能力的机理。在肝细胞中，二甲双胍通过激活减少高血糖的产生 AMPK通过上游激酶肝激酶B1（LKB1），这是一个机械，翻译相关的问题，仍然存在 难以捉摸的是DPSC是否主要依赖LKB1来增强骨骼形成和新血管形成。 递送二甲双胍。我们将测试以DPSC为基于DPSC的颅面骨再生和 通过功能，通过功能性，AMPK激活来增强对局部递送的二甲双胍的响应的新血管化 催化活性LKB1。体外和体内研究将通过两个具体目标扩大我们的结果。目标1意志 确定在DPSC中，从CPC支架释放的二甲双胍是否在诱导成骨和亲血管生成反应中 LKB1/AMPK依赖性方式。 AIM 2将检验以下假设：在DPSC中，功能性LKB1/AMPK细胞 为了增强颅面骨再生和新血管形成，需要反应 从CPC脚手架释放的二甲双胍。我们预计我们的结果将产生新的，有价值的基本和翻译 将导致具有成本效益的组织工程平台的信息，其中结合了二甲双胍的脚手架 使用干细胞可以增强颅面和骨科骨再生。