Synchrotron Radiation Infrared Microscopy Analysis of Mouse Trabecular Meshwork Cells and Myofibroblasts
HE Ming-xia1, TIAN Tian2, LIU Li-yuan*, BU Shao-chong, DONG Li-jie, ZHANG Xin-xin, ZHANG Hong-zhen
1. State Key Laboratory of Precision Measuring Technology and Instruments, Tianjin University, Tianjin 300072, China
2. School of Precision Instrument and Optoelectronics Engineering, Tianjin University, Tianjin 300072, China
3. State Key Laboratory of Chemical Engineering, Tianjin University, Tianjin 300072, China
4. Key Laboratory of Opto-electronics Information and Technical Science, Ministry of Education, Tianjin University, Tianjin 300072, China
5. Tianjin Medical University Eye Hospital, Tianjin 300384, China
Abstract：Primary open angle glaucoma is a common blind eye disease. Elevated intraocular pressure is the most important risk factor for the occurrence and development of primary open-angle glaucoma. It is caused by the lesion of aqueous humor outflow system and the increase of aqueous humor outflow resistance in trabecular meshwork pathway. It has been shown that TGF- β in aqueous humor can make trabecular cells fibrosis and induce excessive proliferation of trabecular cells, thus hindering the outflow of aqueous humor, leading to the occurrence of primary open-angle glaucoma. POAG is a covert disease with slow progression and no symptoms in the early stage. It is often found only when the visual field is significantly impaired in the late stage. Therefore, the early diagnosis of it is particularly important. Synchrotron radiation infrared microimaging combined with high luminance and high resolution synchrotron radiation source with Fourier transform infrared spectrometer and infrared microscope can realize cell detection. It is very important to obtain cellular change information from molecular level and to understand the pathogenesis of disease and the early diagnosis of disease. Although there are many research reports on infrared spectroscopy in biomedical field, the application of infrared spectroscopy microscopic imaging technology to the study of biomedical systems such as cells is still an area in urgent need of development. At present, no infrared spectroscopy has been found for the detection of trabecular meshwork cells. In this paper, TGF-β was used to induce rat trabecular meshwork cells into myofibroblasts in vitro, simulating the process of trabecular meshwork cells fibrosis. The meshwork cells and the myofibroblasts induced by TGF-β were studied by synchrotron radiation infrared microscopy and spectral analysis, and the feasibility of using synchrotron radiation in the early diagnosis of primary open-angle glaucoma was discussed. Study have shown that the elastin in myofibroblasts was significantly higher than that in trabecular meshwork cells, and 95% of elastins were nonpolar amino acids. Comparing the IR spectra of the two kinds of cells, it was found that the stretching vibration of CH3, CH2 and CH of myofibroblasts at 2 934，2 900 and 2 845 cm-1 was stronger than that of trabecular meshwork cells, which might be due to the increase of intracellular elastin induced by TGF-β. In this paper, we detected the excessive proliferation of trabecular meshwork cells at the cellular level, which laid a foundation for obtaining the infrared spectrum of cells directly and detecting the proliferation of trabecular meshwork cells in the future, and then for detecting diseases such as primary open angle glaucoma. It is concluded that synchrotron radiation infrared spectroscopy and microscopic imaging are expected to be new methods for detecting POAG and provide a basis for real-time clinical detection of glaucoma by portable infrared microspectrometer.
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