Atomic level structure of Extracellular Matrix (ECM): spectroscopic approaches to the systems biology of intact tissue

细胞外基质（ECM）是一种固体蛋白质和其他物质的网络，支持构成我们各种组织的细胞。健康组织在老年的维持对经济和我们的生活质量有着巨大的影响。此外，当组织磨损或受损时，组织替代物的设计至关重要，并且ECM对于将这些替代物忠实地整合到宿主组织中至关重要。该项目将开发在实验室中从细胞中生长ECM的方法，这些细胞在分子和纳米尺度上看起来尽可能像骨骼中发现的ECM。然后，这些ECM可用于在实验室中生长骨组织，用于移植目的，并用于进一步的基础科学研究，例如骨骼的详细结构及其对药物和其他化学品的反应。为了做到这一点，我们将首先开发使用光谱学形式，固态核磁共振（SSNMR）检查ECM分子结构的方法。为了测试这些方法，我们将使用类似于骨ECM中发现的分子，但可以通过实验室中的正常化学反应合成。然后，我们将使用各种细胞培养方法从几种不同的细胞类型中生长ECM，这些细胞类型能够产生骨样材料，在不同的材料上和各种不同的条件下支持细胞，以研究产生最接近骨的ECM所必需的特征，正如我们在项目的第一部分开发的SSNMR方法和电子显微镜检查材料的纳米结构所判断的那样。最后，我们将应用我们已经开发的SSNMR方法来研究骨，其中我们引入NMR活性同位素作为分子结构的“报告者”，以表明我们使用细胞培养产生的材料代表了真实的材料的有效模型。

The extracellular matrix (ECM) is a network of solid protein and other substances which supports the cells that make up our various tissues. The maintenance of healthy tissues into older age has huge implications for the economy and our quality of life. Moreover, the design of tissue replacements is of vital importance when tissues wear out or become damaged, and the ECM is critical to the faithful integration of these replacements into the host tissue. This project will develop methods for growing ECMs from cells in the laboratory that on molecular and nano-lengthscales look as far as possible like the ECM found in bone. These ECMs may then be used to grow bone tissue in the laboratory for grafting purposes and for further basic science research into the detailed structure of bone and its reaction to drugs and other chemicals, for instance. In order to do this, we will first develop ways of examining the molecular structure of the ECM using a form of spectroscopy, solid-state nuclear magnetic resonance (SSNMR). To test these methods, we will use molecules which resemble those found in the ECM of bone, but which can be synthesised by normal chemical reactions in the laboratory. We will then use various cell culturing methods to grow ECMs from a several different cell types that are capable of producing bone-like material, supporting the cells on different materials and under a variety of different conditions in order to investigate what features are necessary to produce an ECM which most closely resembles that in bone, as judged by the SSNMR methods we have developed in the first part of the project and electron microscopy to examine the nanostructure of the material. Finally we will apply the SSNMR methods we have developed to study bone into which we have introduced NMR-active isotopes as 'reporters' of molecular structure in order to show that the material we have produced using cell culture represents a valid model of the real material.