Targeting Collagen Mechanical Damage using Collagen Hybridizing Peptides

摘要 使用胶原蛋白杂交肽检测胶原蛋白机械损伤。 承重组织的机械损伤会导致许多具有临床意义的病症（例如肌腱变性、旋转肌变性） 袖口病），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获得对负载生物力学的新理解 将胶原蛋白杂交肽（CHP）技术发展成一种新的承载胶原组织的方法 机械损伤检测方法。据报道，CHP 与源自的变性胶原蛋白链结合 蛋白酶活性或机械损伤，其方式类似于引物与熔化的 DNA 结合 PCR。我们建议通过开发新的热电联产来大幅扩展热电联产损坏检测的能力 更小，可以更快地扩散到致密的肌肉骨骼组织中，并表现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，它仅在与胶原蛋白结合时才会发出荧光， 无需染色和清洗组织，使 CHP 能够充当超载情况下的损伤测量仪 组织。然后，我们将为现有和新热电联产的使用制定优化协议，确定 肌肉骨骼软组织中胶原纤维应变与 CHP 结合之间的关系，并定量 将 CHP 定位与其他技术进行比较。最后，我们将应用CHP目标来阐明这种关系 组织水平机械负荷和分子水平胶原蛋白机械损伤之间的关系。我们会 重点关注具有相当临床相关性的两种重要的肌肉骨骼组织：肌腱和关节 软骨。考虑到胶原蛋白损伤对肌肉骨骼损伤和疾病的广泛影响， 深入理解分子水平胶原蛋白损伤与机械过载之间的关系 将为承重组织的生物力学提供新的见解，并帮助开发新的诊断方法 以及治疗肌肉骨骼疾病的疗法。

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.