Targeting Collagen Mechanical Damage using Collagen Hybridizing Peptides

摘要 用胶原杂交肽检测胶原蛋白机械损伤 承重组织的机械性损伤会导致许多临床上有意义的情况(如肌腱病、旋转肌 袖带病)，但我们对机械损伤组织的损伤过程了解有限 压力。拟议研究的总体目标是对负荷的生物力学有新的理解。 胶原杂交肽(CHP)技术在胶原组织中的应用 机械损伤检测方法。据报道，CHP与变性胶原链结合，起源于 从蛋白酶活性或机械损伤的方式，类似于引物结合到融化的DNA在 聚合酶链式反应。我们建议通过开发新的CHP来大幅扩展CHP损坏检测的能力 它们较小，可更快地扩散到致密的肌肉骨骼组织中，并显示出加速的结合动力学 允许更快的损坏报告。我们还将开发一种新的CHP，它只在与胶原结合时才会发光， 免去沾染和清洗纸巾的需要，并使生防护中心在超载情况下可作为损害量度 纸巾。然后，我们将为现有和新的CHP的使用开发优化的协议，确定 肌肉骨骼软组织中胶原原纤维应变与CHP结合的关系及其定量研究 将CHP目标定位与其他技术进行比较。最后，我们将应用生防护计划目标来阐明两者之间的关系。 组织水平的机械负荷和分子水平上对胶原的机械损伤之间的关系。我们会 关注两个具有相当临床意义的重要肌肉骨骼组织：肌腱和关节 软骨。考虑到胶原损伤在肌肉骨骼损伤和疾病中的广泛影响，在... 深入了解分子水平的胶原损伤与机械超负荷的关系 将为承重组织的生物力学提供新的见解，并有助于开发新的诊断方法 以及管理肌肉骨骼疾病的治疗方法。

SUMMARY Detection of Collagen Mechanical Damage using Collagen Hybridizing Peptides. Mechanical injury to load-bearing tissues leads to many clinically significant conditions (e.g. tendinosis, rotator cuff disease) but we have limited understanding of the injury process of tissues that are damaged by mechanical stress. The overall goal of the proposed research is to gain new understanding of the biomechanics of load bearing collagenous tissues by developing the collagen hybridizing peptide (CHP) technology into a new mechanical damage detection method. CHP has been reported to bind to denatured collagen strands originating from protease activity or by mechanical damage in a manner similar to primer binding to melted DNA during PCR. We propose to substantially expand the capabilities of CHP damage detection by developing new CHPs that are smaller for faster diffusion into dense musculoskeletal tissues and exhibit accelerated binding kinetics to allow faster damage reporting. We will also develop a new CHP that only fluoresces upon binding with collagen, eliminating the need to stain and wash tissues and enabling the CHP to serve as a damage gauge in overloaded tissues. We will then develop optimized protocols for the use of the existing and new CHPs, determine the relationship between collagen fibril strain and CHP binding in musculoskeletal soft tissues, and quantitatively compare CHP targeting to other techniques. Finally, we will apply CHP targeting to elucidate the relationship between tissue level mechanical loading and mechanical damage to collagen at the molecular level. We will focus on two important musculoskeletal tissues of considerable clinical relevance: tendons and articular cartilage. Considering the wide-spread impact of collagen damage in musculoskeletal injuries and diseases, in- depth understanding of the relationships between molecular level collagen damage and mechanical overloading will provide new insights into the biomechanics of load-bearing tissues as well as help develop new diagnostics and therapies for managing musculoskeletal disorders.