Dual Delivery of Engineered EVs and Growth Factor for Bone Regeneration

工程电动汽车和生长因子的双重输送用于骨再生

• 批准号: 10718684
• 负责人: CHUN-CHIEH HUANG
• 金额: $ 42.04万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-16 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10718684
• 关键词: Acute; Adopted; Affect; Alginates; Anti-Inflammatory Agents; Antiinflammatory Effect; BMP2 gene; Biological Assay; Bone Marrow; Bone Regeneration; Calcium; Calvaria; Characteristics; Chronic; Clinical; Complex; Cues; Data; Defect; Diabetes Mellitus; Diabetic mouse; Diameter; Disease; Encapsulated; Engineering; Epidemic; Failure; Foundations; Gel; Generations; Growth Factor; Histology; Hydrogels; Immune response; Immune system; Immunohistochemistry; Impaired wound healing; Individual; Inflammation; Inflammatory; Kinetics; Knowledge; Leucine Zippers; Link; Macrophage; Mediating; Mesenchymal Stem Cells; Methods; MicroRNAs; Modeling; Molecular; Morbidity - disease rate; Mus; Non-Insulin-Dependent Diabetes Mellitus; Outcome; Paracrine Communication; Peptides; Phase; Physiological; Physiology; Play; Process; Property; Protocols documentation; Publishing; Quantitative Reverse Transcriptase PCR; RGD (sequence); Rattus; Role; Signal Transduction; Standardization; System; Systemic disease; Testing; Therapeutic; Tissue Engineering; Training; Treatment outcome; Vesicle; Weight; bone; bone growth factor; bone healing; bone quality; bone repair; combinatorial; comparison control; controlled release; cost; crosslink; cytokine; design; diabetic; differential expression; engineered stem cells; extracellular vesicles; healing; immunoregulation; in vivo regeneration; innovation; novel; overexpression; physical property; recombinant human bone morphogenetic protein-2; regeneration function; regenerative; scaffold; self assembly; tool; wound; wound healing

Project Summary/Abstract The inflammatory conditions and the complex physiological changes associated with chronic immunological responses to wound healing in type 2 diabetes (T2DM) challenge the outcomes of the treatments for bone regeneration. The related morbidity and costs associated with bone healing failures are considerable and increasing. Recent discoveries have demonstrated that the immune system is tightly linked to bone physiology and that MSC derived extracellular vesicles (EVs) play a prominent role in immunomodulation of bone repair. While recombinant human bone morphogenetic protein 2 (rhBMP2) mediated bone regeneration is well established, the challenges of controlling inflammation arising from rhBMP2 usage is exacerbated in the presence of immunomodulating systemic diseases such as T2DM present a significant knowledge gap that limit clinical outcomes. Based on preliminary studies of inflammation informed MSC EV microRNA (miRNA) composition and effects on immunomodulation and bone regeneration, we hypothesize that a dual delivery of engineered MSC EVs with anti-inflammatory properties and rhBMP2 using a combinatorial delivery system with self-assembling leucine zipper (LZ) peptide-based hydrogels and alginate beads can enhance bone healing in T2DM. We propose three specific aims to test this hypothesis. In aim 1, we will test our preliminary data driven hypothesis that a subset of altered miRNAs in preconditioned MSC EVs contributes to the enhanced immunomodulatory function of the inflammatory preconditioned MSC derived EVs. We will also generate engineered MSC EVs that overexpress candidate miRNA with anti-inflammatory properties to further study the mechanistic aspects by which individual EV related miRNAs control the immunomodulation functionality. In aim 2, we will create a novel dual delivery system with LZ-hydrogels and alginate beads that contain engineered EVs and rhBMP2 respectively. We will test our hypothesis that 1) EVs with anti-inflammatory properties (generated from aim 1) can be released during the inflammatory phase of healing (up to 7 days) from a self-assembling LZ- hydrogel and 2) rhBMP2 that is encapsulated in alginate beads with an extended-release profile (4-6 weeks) can provide a long term osteoinductive signal to promote/enhance bone regeneration. In aim 3, using a diabetic mouse calvarial defect model, we will test the hypothesis that engineered MSC EVs containing anti-inflammatory miRNA will suppress inflammation upon wounding while sustained released rhBMP2 promote bone healing. Overall, the results of these first studies of the combinatorial usage of engineered and MSC EVs rhBMP2 will provide a foundation for continued definition of MSC EV-mediated immunomodulation, and the regenerative properties and applicability of the dual delivery system to bone healing in T2DM. Further, these studies will also provide a framework for the use of growth factors and engineered EVs in combination for tissue engineering applications.

项目摘要/摘要 炎症条件和与慢性免疫学相关的复杂生理变化 对2型糖尿病（T2DM）中伤口愈合的反应挑战骨骼治疗的结果 再生。与骨骼愈合失败相关的相关发病率和成本是相当大的，并且 增加。最近的发现表明，免疫系统与骨骼生理密切相关 MSC衍生的细胞外囊泡（EV）在骨修复的免疫调节中起着重要作用。 重组人骨形态发生蛋白2（RHBMP2）介导的骨再生很好 已建立，控制RHBMP2使用引起的炎症的挑战在此中加剧了 T2DM等免疫调节的系统性疾病的存在列出了一个显着的知识差距 限制临床结果。基于炎症知情MSC EV microRNA（miRNA）的初步研究 组成和对免疫调节和骨再生的影响，我们假设双重递送 具有抗炎特性的工程MSC EV和使用组合输送系统的RHBMP2 自组装亮氨酸拉链（LZ）基于肽的水凝胶和藻酸盐珠可以增强骨骼愈合 T2DM。我们提出了三个特定的目的来检验这一假设。在AIM 1中，我们将测试我们的初步数据驱动 假设在预处理MSC EV中改变的miRNA的子集有助于增强 炎症预处理MSC衍生的EV的免疫调节功能。我们还将生成 设计了具有抗炎特性的过表达候选miRNA，以进一步研究 各个EV相关的miRNA控制免疫调节功能的机械方面。目标 2，我们将使用LZ-Hydregels和含有工程EVS的藻酸盐珠创建一个新颖的双递送系统 和RHBMP2。我们将测试我们的假设，即1）具有抗炎特性的EV（生成 从AIM 1）可以在愈合的炎症阶段释放（最多7天），从自组装LZ-中释放 水凝胶和2）包裹在具有延长释放曲线（4-6周）的藻酸盐珠中的RHBMP2可以 提供长期的骨诱导信号，以促进/增强骨骼再生。在AIM 3中，使用糖尿病患者 小鼠颅缺陷模型，我们将测试以下假设：含有抗炎的工程MSC EV miRNA会在受伤时抑制炎症，而持续释放的RHBMP2会促进骨骼愈合。 总体而言，这些对工程和MSC EVS RHBMP2组合使用的首次研究的结果将 为继续定义MSC EV介导的免疫调节和再生提供了基础 双递送系统对T2DM中骨骼愈合的特性和适用性。此外，这些研究也将 提供了使用增长因子和工程电动汽车组合进行组织工程的框架 申请。