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 这项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， 联合收割机二甲双胍与干细胞再生大的，临界尺寸的口腔和颅面骨骼缺损。虽然组织 使用干细胞、支架和生长因子的工程学为自体骨提供了一种有吸引力的、侵入性较小的替代方法 移植后，其成功高度依赖于细胞对局部缺氧微环境的适当适应， 功能性微血管系统血管内皮生长因子（VEGF）是血管内皮细胞生长的关键介质， 骨再生中的成骨/血管生成偶联。因此，开发新的和负担得起的基于干细胞的组织工程 增强VEGF介导的血管生成的策略可以显著增强骨骼再生。我们集团和其他 最近报道，二甲双胍，一种全球数百万糖尿病患者使用的低成本药物， 干细胞的分化来源于各种组织来源。这表明二甲双胍可以在当地的 递送制剂以增强基于干细胞的骨再生。我们已经通过制定一种钙， 含有二甲双胍的磷酸盐骨水泥（CPC），当在培养物中释放时， 标记物和增加的矿化细胞外沉积物的牙髓干细胞（DPSC），一个容易获得， 取之不尽的产后干细胞来源。有趣的是，我们发现二甲双胍还诱导了 VEGF分泌在暴露于缺氧条件的DPSC中进一步扩增。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，AMPK是一种主要的感知信号通路。 细胞生物能量学机制。而在肝细胞中，二甲双胍通过激活 AMPK通过上游激酶肝激酶B1（LKB 1），一个机制，诊断相关的问题，仍然存在 目前尚不清楚DPSC是否主要依赖于LKB 1来增强骨形成和新生血管形成，以响应局部 给药二甲双胍。我们将测试基于DPSC的颅面骨再生和 响应于局部递送的二甲双胍的新血管形成通过AMPK活化而增强， 催化活性LKB 1。体外和体内研究将通过两个具体目标扩展我们的结果。目标1将 确定在DPSC中，从CPC支架释放的二甲双胍是否诱导成骨和促血管生成反应， LKB 1/AMPK依赖性。目的2将检验在DPSC中，功能性LKB 1/AMPK细胞表达的假设。 对于增强颅面骨再生和新生血管形成， 从CPC支架释放的二甲双胍。我们预计我们的结果将产生新的，有价值的基础和翻译 信息，将导致成本效益的组织工程平台，其中二甲双胍加载的支架组合 将增强颅面和矫形骨再生。