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： 每年进行数百万次骨移植手术以修复口腔中的骨缺损。 虽然自体移植物和同种异体移植物在临床上被用于这些目的，但它们的局限性促使人们对移植物进行了更多的研究。 合成移植材料的研究尽管已经开发了许多生物相容性材料，但缺乏生物相容性材料。 生物活性的降低往往会导致次优结果。生物活性肽在细胞表面的应用 材料可以帮助在材料和活组织之间形成界面。利用噬菌体展示技术， 一个实验室已经开发出一种双功能肽DPI-VTK，它既含有间充质干细胞， (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 骨髓间充质干细胞与羟基磷灰石的粘附，但也可以促进宿主骨髓间充质干细胞的特异性迁移，绕过 细胞移植的必要性。此外，该肽可以与BMP肽组合，以 创建双功能骨传导和骨诱导界面，促进MSC归巢 沿着它们随后的分化。这一建议的总体假设是， 具有细胞特异性肽的材料的骨传导性和骨诱导性将导致增加的骨传导性和骨诱导性。 不依赖外源性细胞的骨再生。 本项目提出了以下目标：1）确定DPI-VTK是否可以增强再生 2）确定DPI-VTK是否可以与BMP协同作用 3）确定DPI-VTK的MSC结合机制。体内研究将 使用小鼠颅骨缺损模型完成，并使用Micro-CT沿着分析数据， 组织学染色。肽结合研究将使用基于ELISA的蛋白质测定完成， 亲和下拉测定法和质谱法。这些实验的结果将提高我们的 在组织工程的背景下理解细胞-材料界面，并确定潜在的 我们实验室的矿物质结合肽的临床应用。由于广泛使用羟基磷灰石基 结论DPI-VTK是一种理想的颅面骨移植生物材料，在临床上可用于促进颅面骨的整合 移植到原生骨组织中开发既能募集宿主细胞又能诱导 分化可能是再生大体积缺陷的关键策略，而不需要外源性 细胞或自体移植物。