Raman Spectroscopy of Live Cell Invasion of 3D nano-fabricated scaffolds

点击翻译按钮获取中文摘要

In recent years there has been a vast development in the engineering of artificial tissues to repair or replace damaged tissue. Scientists can now grow cartilage, produce artificial skin and even print 3D tracheas and are advancing towards the ideal goal of growing replacement organs from a patient's own cells. Whilst present advancements have had some patient benefit this has been limited and there is still a need for a greater understanding of how to control and determine the growth of artificial tissues if we are to achieve extensive improvements to health. The difficulty is that cells do not form tissue in isolation but also require a cell matrix or scaffold. The most promising analytical techniques for tissue engineering at present focus on the growth of cells on artificially printed 3D scaffolds. However, the majority of these techniques either require the labelling of cells with specific markers to detect their presence or the removal of the cells and scaffolds from cell culture conditions thus ending the process and only providing limited information. Our aim is to develop the use of Raman spectroscopy, an alternative non-destructive and label free approach, to investigate in situ cell growth on 3D scaffolds of differing topology.One process that happens when light is shone at a substance is that it is scattered and sometimes the light scatters at a different wavelength; an effect named Raman scattering. The resultant Raman scattering from a molecule will depend on the chemical structure and for biomolecules it will also depend on the biophysical structure. Recent advancements in Raman spectrometers has enabled Raman maps of live cells to be rapidly collected from which images of cells can be produced identifying biochemical and biophysical changes that occur as cells grow. We will therefore develop Raman spectroscopy as a novel method for the direct in situ analysis of live cells growing on 3D scaffolds of different shapes. Various methods exist to produce artificial 3D scaffolds made from a variety of polymers. One of the most successful is direct laser writing which enables the printing of scaffolds on a microscale using a range of shapes, sizes and materials. By understanding how different scaffold topography affect cells grown in a 3D cell culture environment we will be a step nearer to controlling and determining cell and ultimately tissue growth necessary to advance further the field of tissue engineering.