Pudlas Marieke, Koch Steffen, Bolwien Carsten, Walles Heike
Fraunhofer Institute for Interfacial Engineering and Biotechnology, Stuttgart - Germany.
Int J Artif Organs. 2010 Apr;33(4):228-37.
At present, the production of tissue engineered cartilage requires the concurrent production of two identical transplants. One transplant is used for destructive quality control and the second one is implanted into the patient. A non-invasive characterization of such tissue engineering samples would be a promising tool to achieve a production process of just one transplant that is both implanted and tested. Raman spectroscopy is a method that satisfies this requirement by analyzing cells without lysis, fixation or the use of any chemicals. This pure optical technique is based on inelastic scattering of laser photons by molecular vibrations of biopolymers. Characteristic peaks in Raman spectra of cells could be assigned to typical biochemical molecules present in biological samples. For the analysis of chondrocytes present in cartilage transplants, the determination of the cell vitality as well as the discrimination of another cell type have been studied by Raman spectroscopy. Another bottleneck in such biological processes under GMP conditions is sterility control, as most of the commonly used methods require long cultivation times. Raman spectroscopy provides a good alternative to conventional methods in terms of time saving. In this study, the potential of Raman spectroscopy as a quality and sterility control tool for tissue engineering applications was studied by analyzing and comparing the spectra of cell and bacteria cultures.
目前，组织工程软骨的生产需要同时生产两个相同的移植物。一个移植物用于破坏性质量控制，另一个则植入患者体内。对这类组织工程样本进行非侵入性表征将是实现仅生产一个既用于植入又用于测试的移植物的生产过程的一个有前景的工具。拉曼光谱法是一种通过在不进行细胞裂解、固定或使用任何化学物质的情况下分析细胞来满足这一要求的方法。这种纯光学技术基于生物聚合物分子振动对激光光子的非弹性散射。细胞拉曼光谱中的特征峰可归因于生物样本中存在的典型生化分子。对于软骨移植物中存在的软骨细胞分析，已经通过拉曼光谱法研究了细胞活力的测定以及对另一种细胞类型的区分。在GMP条件下，此类生物过程中的另一个瓶颈是无菌控制，因为大多数常用方法需要较长的培养时间。在节省时间方面，拉曼光谱法为传统方法提供了一个很好的替代方案。在本研究中，通过分析和比较细胞及细菌培养物的光谱，研究了拉曼光谱法作为组织工程应用质量和无菌控制工具的潜力。
