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In vitro photodynamic therapy of MG-63 osteosarcoma cells mediated by aminolevulinic acid

机译:氨基乙酰丙酸介导的MG-63骨肉瘤细胞的体外光动力治疗

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This is an in vitro study of photodynamic therapy (PDT) in the MG-63 line of human osteosarcoma cells, as mediated by aminolevulinic acid (ALA). The primary goal of this work is to determine the feasibility and effectiveness of treating osteosarcoma through PDT. In addition, this work is aimed at determining whether the resulting cell death occurs through apoptosis or cellular necrosis. The MG-63 cells are treated with increasing concentrations of ALA from 0.1-10 mM ALA, leading to the accumulation of the photosensitizer protoporphyrin IX (PpIX) within the cells. After incubation periods of 4 and 24 hours in ALA, the cells are illuminated by 0-10 J/cm2 of 636 nm light in order to activate the PpIX and induce oxidative damage to the cells. Light is administered by an 8x12 array of LED's, which are controlled by an Arduino Duemilanove microcontroller board in order to assure ease of use along with accurate levels of exposure. Controls for this experiment include 0 J/cm2 of light exposure for all experimental concentrations of ALA, as well as illuminating cells that have not been incubated in ALA at all experimental levels of illumination. MG-63 cells are analyzed through fluorimetry and MTT assays in order to determine the effectiveness of ALA mediated PDT of osteosarcoma.
机译:这是由氨基乙酰丙酸(ALA)介导的人骨肉瘤MG-63细胞中光动力疗法(PDT)的体外研究。这项工作的主要目的是确定通过PDT治疗骨肉瘤的可行性和有效性。此外,这项工作旨在确定导致的细胞死亡是通过凋亡还是细胞坏死发生的。使用浓度从0.1-10 mM ALA递增的ALA处理MG-63细胞,导致光敏剂原卟啉IX(PpIX)在细胞内积聚。在ALA中孵育4和24小时后,以0-10 J / cm2的636 nm光照射细胞,以激活PpIX并诱导对细胞的氧化损伤。光线由8x12 LED阵列管理,该阵列由Arduino Duemilanove微控制器板控制,以确保易于使用以及准确的曝光水平。该实验的对照包括所有实验浓度的ALA的0 J / cm2曝光量,以及在所有实验照明水平下未在ALA中孵育的照明细胞。为了确定ALA介导的骨肉瘤PDT的有效性,通过荧光法和MTT法对MG-63细胞进行了分析。

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