Fracture healing assessment by real-time and noninvasive Raman spectorscopy

描述（由申请人提供）：骨折破坏骨骼的基本代谢和机械功能，从而威胁健康和生活质量。骨折愈合试图恢复骨的形状、功能和材料特性。生物矿化被认为在达到稳定形式之前经历了一系列短暂的阶段，这一过程已经在无脊椎动物和牙本质的体外观察到，最近莫里斯小组在体内发现了胎儿小鼠组织的生物矿化。然而，骨组织矿化背后的生化机制，特别是在骨折愈合过程中，仍然没有得到很好的理解。虽然骨折愈伤组织矿化被认为与生长板矿化相似，但在骨折修复过程中，暂态矿物相的发生尚未得到实时检测。本研究的假设是：(1)骨折愈合包括矿物从无序阶段到短暂阶段到最终稳定阶段的转变，无论骨骼年龄和发育阶段如何，这一机制都是相似的；(2)骨折愈伤组织的生化成分可以在整个愈合过程中进行无创监测，这些测量结果可以预测愈伤组织的材料质量。

DESCRIPTION (provided by applicant): Bone fractures disrupt essential metabolic and mechanical functions of the skeleton and thus threaten health and quality of life. Fracture healing attempts to restore the shape, function, and material properties of bone. Biomineralization is thought to proceed through a sequence of transient phases before reaching a stable form, a progression that has been observed in vitro for invertebrates and dentin and recently was discovered in vivo for fetal murine tissue by the Morris group. However, the biochemical mechanisms behind bone tissue mineralization, particularly during fracture healing, are still not well understood. While fracture callus mineralization is thought to be similar to growth plate mineralization, the occurrence of transient mineral phases during fracture repair have not been examined in real time. The hypotheses for this study are that (1) fracture healing includes a mineral transformation from a disordered phase through a transient phase to a final stable phase, and this mechanism is similar regardless of skeletal age and stage of development; and (2) biochemical composition of the fracture callus can be monitored noninvasively throughout the healing process, and these measurements are predictive of callus material quality. To test the hypotheses, mineralization kinetics will be followed for the fracture callus at the femoral middiaphysis of male wild type mice. Raman spectroscopy will examine the kinetics over a 24-hour period in the early stages of fracture repair for mice of different ages. Next, bone matrix and mineral properties will be studied in the fracture callus throughout healing in skeletally mature male wild type mice. The biochemical properties observed by noninvasive Raman spectroscopy will be correlated with geometric and mechanical properties to assess the ability of this technique to characterize the fracture callus integrity. The proposed study will provide unique information about the progression of tissue mineralization during fracture healing and will assess the ability of a noninvasive Raman spectroscopic system to monitor the healing process. Overall, the goals of these experiments are to gain insights into the mechanisms of fracture healing mineralization and to provide a clinical imaging and analysis tool to assess the fracture site noninvasively, regardless of age. This type of information could be invaluable in the detection and correction of abnormal healing and thus may be useful for developing strategies to promote successful bone repair.