Su Guan-Chin, Wei Yau-Huei, Wang Hsing-Wen
Institute of Biophotonics, National Yang-Ming University, 155 Li-Nong St., Sec. 2, Taipei 112, Taiwan.
Opt Express. 2011 Oct 24;19(22):21145-54. doi: 10.1364/OE.19.021145.
Photodynamic therapy (PDT) dosimetry is complex as many factors are involved and varied interdependently. Monitoring the biological consequence of PDT such as cell death is the most direct approach to assess treatment efficacy. In this study, we performed 5-aminolevlinic acid (ALA)-PDT in vitro to induce different cell death modes (i.e., slight cell cytotoxicity, apoptosis, and necrosis) by a fixed fluence rate of 10 mW/cm(2) and varied fluences (1, 2, and 6 J/cm(2)). Time course measurements of cell viability, caspase-3 activity, and DNA fragmentation were conducted to determine the mode of cell death. We demonstrated that NADH fluorescence lifetime together with NADH fluorescence intensity permit us to detect apoptosis and differentiate it from necrosis. This feature will be unique in the use of optimizing apoptosis-favored treatments such as metronomic PDT.
光动力疗法（PDT）剂量测定很复杂，因为涉及许多因素且相互依存各不相同。监测PDT的生物学后果，如细胞死亡，是评估治疗效果的最直接方法。在本研究中，我们在体外进行5-氨基乙酰丙酸（ALA）-PDT，通过固定光通量率10 mW/cm²和不同光通量（1、2和6 J/cm²）诱导不同的细胞死亡模式（即轻微细胞毒性、凋亡和坏死）。进行细胞活力、半胱天冬酶-3活性和DNA片段化的时间进程测量以确定细胞死亡模式。我们证明，NADH荧光寿命与NADH荧光强度一起使我们能够检测凋亡并将其与坏死区分开来。这一特性在优化如节拍式PDT等有利于凋亡的治疗方法的应用中将是独一无二的。
