School of Electrical Engineering and Computer Science, Seoul National University, Seoul 151-742, Korea.
Biotechnol Bioeng. 2011 Jun;108(6):1473-8. doi: 10.1002/bit.23049. Epub 2011 Jan 17.
Matrix mineralization is a terminal process in osteoblast differentiation, and several approaches have been introduced to characterize the process in tissues or cultured cells. However, an analytical technique that quantitates in vitro matrix mineralization of live cells without any labeling or complex treatments is still lacking. In this study, we investigate a simple and enhanced optical method based on surface plasmon resonance (SPR) detection that can monitor the surface-limited refractive index change in real-time. During monitoring MC3T3-E1 cells in vitro culture every 2 days for over 4 weeks, the SPR angle is shifted with a greater resonance change in cells cultured with osteogenic reagents than those without the reagents. In addition, the SPR results obtained have a close relevance with the tendency of conventional mineralization staining and an inductively coupled plasma-based calcium content measure. These results suggest a new approach of a real-time SPR monitoring in vitro matrix mineralization of cultured cells.
基质矿化是成骨细胞分化的终末过程，已经提出了几种方法来对组织或培养细胞中的该过程进行特征化描述。然而，仍然缺乏一种无需任何标记或复杂处理即可定量检测活细胞体外基质矿化的分析技术。在本研究中，我们研究了一种基于表面等离子体共振（SPR）检测的简单增强光学方法，该方法可以实时监测表面受限的折射率变化。在监测体外培养的 MC3T3-E1 细胞的过程中，每隔 2 天进行一次，持续 4 周以上，结果表明，用成骨试剂培养的细胞的 SPR 角度发生了更大的共振变化，而没有用成骨试剂培养的细胞则没有这种变化。此外，SPR 结果与传统矿化染色和电感耦合等离子体基钙含量测量的趋势密切相关。这些结果表明了一种实时 SPR 监测培养细胞体外基质矿化的新方法。
