Functionalization of biomimetic apatite with mineral binding peptides for bone tissue engineering

用于骨组织工程的矿物结合肽仿生磷灰石的功能化

• 批准号: 10080724
• 负责人: Eric J Madsen
• 金额: $ 5.18万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10080724
• 关键词: Adhesions; Affinity; Allografting; Apatites; Autologous Transplantation; BMP2 gene; Binding; Biocompatible Materials; Biological Assay; Biomimetics; Bone Regeneration; Bone Tissue; Bone Transplantation; Bypass; Calvaria; Cell Transplantation; Cells; Clinic; Clinical; Collagen; Cues; Data; Data Analyses; Defect; Enzyme-Linked Immunosorbent Assay; Fibronectins; Genes; Goals; Hematopoietic; Hematoxylin and Eosin Staining Method; Histology; Homing; Human; Hydroxyapatites; Implant; Incubated; Integrin Binding; Integrins; Laboratories; Left; Mass Spectrum Analysis; Measures; Mechanics; Mediating; Mesenchymal Differentiation; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Minerals; Modeling; Morbidity - disease rate; Mus; Natural regeneration; Oral cavity; Organ Transplantation; Osteoblasts; Osteogenesis; Outcome; Peptide Library; Peptides; Phage Display; Population; Problem Solving; Procedures; Proteins; Research; Research Personnel; Sampling; Signal Transduction; Site; Specificity; Stains; Surface; Techniques; Technology; Time; Tissue Engineering; Tissues; Transplantation; Up-Regulation; Work; base; bone; cell motility; cost; craniofacial bone; experimental study; histological stains; improved; in vivo; insight; microCT; migration; nanoindentation; neurotensin mimic 2; novel; osteogenic; protein aminoacid sequence; receptor; recruit; regenerative therapy; scaffold; side effect; stem cell migration

Project Summary Abstract: Millions of bone grafting procedures are performed annually to repair bone defects in the oral cavity. While autografts and allografts are used clinically for such purposes, their limitations have motivated much research into synthetic graft materials. Although numerous biocompatible materials have been developed, lack of bioactivity often times results in suboptimal results. The use of bioactive peptides on the surface of a material can help create an interface between materials and living tissue. Using phage-display technology, our laboratory has developed a dual-functional peptide DPI-VTK that contains both a mesenchymal stem cell (MSC) binding domain and a hydroxyapatite binding domain. Such a peptide could not only enhance the adhesion of MSCs to hydroxyapatite, but could also promote the specific migration of host MSCs, bypassing the need for cell transplantation. Additionally, this peptide could be combined with BMP peptides in order to create dual-functional osteoconductive and osteoinductive interfaces that encourage the homing of MSCs along with their subsequent differentiation. The overall hypothesis of this proposal is that enhancing the osteoconductivity and osteoinductivity of materials with cell-specific peptides will result in increased bone regeneration without reliance on exogenous cells. This project proposes the following aims: 1) Determine if DPI-VTK can enhance the regeneration of bone defects in acellular scaffold models, 2) Determine if DPI-VTK can behave synergistically with BMP derived peptides, and 3) Determine the MSC binding mechanism of DPI-VTK. The in-vivo studies will be accomplished using a mouse calvarial defect model and the data analyzed using Micro-CT along with histological staining. The peptide binding studies will be accomplished using ELISA based protein assays, affinity pull-down assays, and mass spectrometry. The outcomes of these experiments will enhance our understanding of cell-material interfaces in the context of tissue engineering as well as determine the potential clinical utility of our laboratory's mineral binding peptides. Due to the widespread use of hydroxyapatite-based biomaterials in craniofacial bone grafting, DPI-VTK could be very useful clinically for enhancing their integration into native bone tissue. Developing dual-functional material that both recruit host cells and induce differentiation could be a key strategy for regenerating large-volume defects without the need for exogenous cells or autografts.

项目摘要摘要： 每年进行数百万个骨移植手术以修复口腔中的骨缺损。 虽然自体移植和同种异体移植用于临床目的，但它们的局限性却有很大的动机 研究合成移植物材料。尽管已经开发了许多生物相容性材料，但缺乏 生物活性通常会导致次优结果。在A表面使用生物活性肽 材料可以帮助建立材料和活组织之间的接口。使用噬菌体播放技术，我们的 实验室已经开发了一种双功能肽DPI-VTK，其中包含间充质干细胞 （MSC）结合结构域和羟基磷灰石结合结构域。这样的肽不仅可以增强 MSC粘附于羟基磷灰石，但也可以促进宿主MSC的特定迁移，绕过 细胞移植的需求。此外，该肽可以与BMP肽合并 创建双功能的骨导导和骨诱导界面，以鼓励MSC的归宿 以及随后的分化。该提议的总体假设是增强 具有细胞特异性肽的材料的骨导导性和骨诱导性将导致增加 骨再生，不依赖外源细胞。 该项目提出以下目的：1）确定DPI-VTK是否可以增强 细胞支架模型中的骨缺损，2）确定DPI-VTK是否可以与BMP协同作用 衍生的肽，3）确定DPI-VTK的MSC结合机理。体内研究将是 使用鼠标钙钙缺损模型和使用Micro-CT分析的数据以及 组织学染色。肽结合研究将使用基于ELISA的蛋白质分析来完成 亲和力下拉测定法和质谱法。这些实验的结果将增强我们的 了解组织工程背景下的细胞材料界面，并确定潜力 我们实验室矿物结合肽的临床实用性。由于基于羟基磷灰石的广泛使用 颅面骨移植中的生物材料，DPI-VTK在临床上可能非常有用，可用于增强其整合 进入天然骨组织。开发既募集宿主细胞又诱导的双功能材料 分化可能是再生大批量缺陷而无需外源的关键策略 细胞或自体移植。