Juravinski Cancer Centre, Physics Research, Hamilton, Ontario, Canada.
Photochem Photobiol. 2011 Sep-Oct;87(5):1129-37. doi: 10.1111/j.1751-1097.2011.00942.x. Epub 2011 Jul 7.
Photodynamic therapy (PDT) oxygen consumption, clonogenic cell survival, fluorescence photobleaching and photoproduct formation were investigated during benzoporphyrin derivative monoacid (BPD-MA)-PDT of MAT-LyLu cells in vitro. Cells were incubated with BPD-MA concentrations of 0.1, 0.5 or 2.5 μg mL(-1) for 2 h and then treated with 405 nm light under oxygenated and hypoxic conditions. Fluorescence spectra were acquired during treatment, and photobleaching and photoproduct generation were quantified using singular value decomposition of the spectra. Cell survival was measured at set times during the treatment using a colony-forming assay. The amount of oxygen consumed by PDT per photon absorbed decreased with BPD-MA intracellular concentration. Survival was correlated with the total amount of oxygen consumed by PDT per unit volume, which is assumed to be equivalent to the amount of singlet oxygen that reacted. A photobleaching-based singlet oxygen dose metric was also found to predict survival independent of intracellular BPD-MA concentration. The BPD-MA photoproduct was bleached during the treatment. Two singlet oxygen dose metrics based on photoproduct kinetics could not be correlated with cell survival over the full range of intracellular BPD-MA concentrations used.
光动力疗法 (PDT) 的氧消耗、集落形成细胞存活、荧光光漂白和光产物形成在体外 MAT-LyLu 细胞的苯并卟啉衍生物单酸 (BPD-MA)-PDT 中进行了研究。细胞用 0.1、0.5 或 2.5μg mL(-1) 的 BPD-MA 孵育 2 小时,然后在有氧和缺氧条件下用 405nm 光处理。在处理过程中获取荧光光谱,并使用光谱的奇异值分解来量化光漂白和光产物的生成。在处理过程中的设定时间使用集落形成测定法测量细胞存活。每吸收一个光子消耗的 PDT 氧气量随 BPD-MA 细胞内浓度的降低而降低。存活率与每单位体积 PDT 消耗的氧气总量相关,这被认为与反应的单线态氧的量相当。还发现基于光漂白的单线态氧剂量指标与细胞存活独立于细胞内 BPD-MA 浓度相关。BPD-MA 光产物在治疗过程中被漂白。基于光产物动力学的两个单线态氧剂量指标不能与在使用的全范围内的细胞内 BPD-MA 浓度相关的细胞存活相关。