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

细胞外基质（ECM）是一个由固体蛋白质和其他物质组成的网络，它支持构成我们各种组织的细胞。在老年时期保持健康的组织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，ECM对于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培养ECM的方法，使细胞在分子和纳米尺度上尽可能与骨骼中的ECM相似。然后，这些ecm可用于在实验室中培养骨组织，用于移植目的，并用于进一步的基础科学研究，例如，研究骨骼的详细结构及其对药物和其他化学物质的反应。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外基质中发现的分子，但这些分子可以在实验室中通过正常的化学反应合成。然后我们将使用各种细胞培养方法增加ECM的几种不同的细胞,能够产生骨突材料,支持细胞在不同的材料和不同的条件下为了调查哪些特性是必要的生产一个ECM最接近,在骨,根据SSNMR方法我们已经开发出的第一部分项目和电子显微镜检查材料的纳米结构。最后，我们将应用我们开发的ssmr方法来研究骨骼，我们引入了核磁共振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生产的材料代表了真实材料的有效模型。

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.