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: 10327307
• 负责人: Eric J Madsen
• 金额: $ 5.26万
• 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 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10327307
• 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; bioscaffold; bone; bone repair; 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.

项目概要摘要： 每年进行数百万例骨移植手术来修复口腔骨缺损。 虽然自体移植物和同种异体移植物在临床上用于此类目的，但它们的局限性引起了人们的极大关注。 合成移植材料的研究。尽管已开发出多种生物相容性材料，但仍缺乏 生物活性的降低常常导致次优结果。生物活性肽在生物体表面的应用 材料可以帮助在材料和活组织之间建立界面。利用噬菌体展示技术，我们的 实验室开发出双功能肽DPI-VTK，既含有间充质干细胞 (MSC) 结合域和羟基磷灰石结合域。这种肽不仅可以增强 MSCs 与羟基磷灰石的粘附，但也可以促进宿主 MSCs 的特异性迁移，绕过 细胞移植的需要。此外，该肽可以与 BMP 肽结合，以便 创建双功能骨传导和骨诱导界面，促进 MSC 归巢 以及它们随后的分化。该提案的总体假设是，增强 具有细胞特异性肽的材料的骨传导性和骨诱导性将导致骨传导性和骨诱导性增加 骨再生不依赖外源细胞。 该项目提出以下目标：1）确定DPI-VTK是否可以增强再生 非细胞支架模型中的骨缺陷，2) 确定 DPI-VTK 是否可以与 BMP 发挥协同作用 衍生肽，3) 确定 DPI-VTK 的 MSC 结合机制。体内研究将是 使用小鼠颅骨缺损模型完成，并使用 Micro-CT 分析数据 组织学染色。肽结合研究将使用基于 ELISA 的蛋白质测定来完成， 亲和力下拉测定和质谱分析。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 我们实验室的矿物质结合肽的临床应用。由于羟基磷灰石基材料的广泛使用 颅面骨移植中的生物材料，DPI-VTK 在临床上对于增强其整合非常有用 进入天然骨组织。开发既招募宿主细胞又诱导宿主细胞的双功能材料 分化可能是无需外源性再生大体积缺陷的关键策略 细胞或自体移植物。