Inhibitory Effect of Indocyanine Green Mediated Photodynamic Therapy on Restenosis of Rat Carotid Arteries and Smooth Muscle Cells

• Main Authors: Chia-Jung Wang, 王家蓉
• Other Authors: Wen-Tyng Li
• Format: Others
• Language: zh-TW
• Published: 2018
• Online Access: http://ndltd.ncl.edu.tw/handle/ypf7ur

碩士 === 中原大學 === 生物醫學工程研究所 === 106 === Photodynamic therapy (PDT) has been widely investigated in recent years in the treatment of cancer and non-cancer diseases. The basis of PDT is the accumulation of photosensitizers in the lesion site and activated by light of suitable wavelength which produced cytotoxic reactive oxygen species to achieve target toxic cytotoxicity. In this study, we first demonstrated that PDT combined with Indocyanine Green (ICG) using 780 nm light emitting diode irradiations could effectively inhibit carotid endothelial hyperplasia. Next, we studied the PDT effect on rat aortic smooth muscle cells (A-10). A-10 cells were incudated with different concentrations of ICG. The inverted microscope observation showed accumulation of green color increased when increasing the concentration of ICG. Highest accumulation was found of 8 h incubated at a concentration of 20 μM. Different concentrations of ICG were incubated with A-10 cells for 8 hours and then submitted to light irradiation (4 J / cm2). The results of cell viability assay showed that the half-inhibitory dose was 7.73 μM. Lactate dehydrogenase (LDH) release analysis revealed that A-10 cells incubated with 9 μM ICG without light irradiation resulted in less than 5% LDH release, which indicted no dark toxicity. While PDT resulted in more than 50% LDH release. LDH release result corresponded to cell viability result. Malondialdehyde test after PDT showed the degree of lipid peroxidation increased. Hochest 33258 fluorescence staining revealed PDT caused chromatin condensation 2 J/cm2 of light irradiation and was more pronounced with increasing light exposure. Forty-five minutes after PDT, Annexin V/Propidium iodide fluorescent staining found that both neciosis and apoptosis of A-10 cells occurred after PDT. Flow cytometry showed that most of the cells were late apoptotic cells. At 12 hours after PDT, a rise in the sub-diploid (Sub-G1) ratio in the cell cycle was observed. Western blotting studies of PARP and Caspase-3 performance revealed that PARP with a molecular weight of 25 kDa had a clear band after PDT; Caspase-3 expression decreased in the PDT group.The results of this study show that ICG mediated PDT can induce apoptosis of A-10 cells and further inhibit carotid artery stenosis in rats.