Development of Diagnosis Method for Early Stage of Osteoarthritis using Fluorescence Correlation Spectroscopy

为了建立骨关节炎（OA）早期诊断方法，我们研制了一种基于共聚焦显微镜的荧光相关光谱（FCS）仪器。在这种新的方法中，我们研究的细胞外基质（ECMs），形成不均匀的空间与网格结构的异常扩散发生的分子运输。在反常扩散中，与正常扩散不同，扩散系数随扩散距离和扩散时间而变化。因此，我们开发了一种新的方法称为“采样体积控制（SVC）FCS”，使我们能够记录扩散系数的距离依赖性（DDDC）在一个直接的方式。在本研究期间，我们建立了仪器和理论治疗和异常扩散来被认为是一个重要的过程，不仅在ECM，而且在细胞内空间。我们发展了一种诊断关节软骨的方法， ...更多信息 f 20μm厚放入染料溶液中。观察到蛋白酶处理时扩散系数的变化，表明关节软骨中胶原网状结构的破坏，这模拟了OA的症状，并证实了我们新诊断方法的原理。在本研究的早期，我们使用透明质酸的水溶液作为滑液的模型系统。然而，由于流体含有另一个重要的组成部分，粘蛋白型蛋白聚糖，我们寻找一个合适的模型系统的基础研究。结果发现了一种新的粘蛋白，命名为qniumucin。我们对这种新材料进行了纯化方法的改进和结构分析，发现它是一种非常纯和均一的粘蛋白，适合于药用物质。我们使用qniumucin和透明质酸构建了一种用于滑液的新的实现，并使用新的FCS进行了研究。我们还发现，这种新的液体在受损关节软骨的再生方面表现出显着的改善。现在，我们计划扩展我们的方法，以发展和改进这种固化方法。少

To establish a diagnosis method for the early stage of Osteoarthritis (OA), we developed specially designed instruments of fluorescence correlation spectroscopy (FCS) using a confocal microscope. In this new method, we investigate the molecular transports in extracellular matrices (ECMs) which form inhomogeneous space with a mesh structure where anomalous diffusion occurs. In anomalous diffusion, different from normal diffusion, diffusion coefficient changes depending on the diffusion distance and the diffusion times. Therefore we developed a new method called "Sampling Volume Controlled (SVC) FCS" which enable us to record the distance dependence of diffusion coefficient (DDDC) in a direct manner. Within this research term we established the instrumentation and the theoretical treatment and anomalous diffusion comes to be recognized as an important process not only in ECM but also in intracellular spaces. We developed a method for diagnosis for an articular cartilage using its slice o … More f 20μm thickness put in a dye solution. The change in diffusion coefficients on protease treatment was observed showing that the destruction of the collagen mesh structure in the articular cartilage which mimics the symptom of OA and the principle of our new diagnosis method was confirmed. In the early term of this research we used the aqueous solution of hyaluronan as the model system of the synovial fluid. However, since the fluid contains another important component, mucin type proteoglycans, we searched an appropriate model system for basic research. As the result, we discovered a novel mucin in jellyfishes and named qniumucin. We performed the improvement of purification and the structural analysis to find that this new material is an exceptionally pure and homogenous mucin which is suitable to medical substances. We constructed a novel fulfillment for synovial fluid using qniumucin and hyaluronan and investigated using new FCS. We also found that this new fluid showed significant improvement in regeneration of damaged articular cartilage. Now we plan to extend our method for the development and improvement of this curing method. Less