Indirect detection of photosensitizer ex vivo
Indirect detection of photosensitizer ex vivo
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DOI：
10.1016/s1011-1344(02)00279-8
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发表时间：
2002-05-01
影响因子：
5.4
通讯作者：
Patrice, T
中科院分区：
文献类型：
--
作者：
Bourré, L;Thibaut, S;Patrice, T
光动力疗法在施用敏化剂后在暴露于激光的组织内诱导活性氧（ROS）的产生。在具有致敏特性的化学品的持续临床和商业开发的背景下，需要微创测定来确定荧光或非荧光光反应性药物的组织动力学。使用2 '-7'二氯荧光素二乙酸酯（DCFH-DA）从1 mm（3）活组织检查样品离体测定ROS水平，所述DCFH-DA是一种荧光探针，其在ROS存在下转化为高荧光二氯荧光素（DCF）。对Meta（四羟基苯基）二氢卟酚（m-THPC，FOSCAN（R））（一种强大的荧光敏化剂）和细菌叶绿素衍生物WST 09（TOOKAD（R））（一种仅具有轻微荧光的近红外吸收敏化剂）进行了测试。结合ROS测定，使用光纤荧光分光光度计（OFS）在活检样品上测定m-THPC的组织累积。DCF荧光与辣根过氧化物酶（用作对照）诱导的氧化水平成正比，与在试管中与DCFH一起照射的两种选定光敏剂的浓度（范围：0-5 μ g/ml）成正比。无论研究的器官，一个很好的相关性之间的荧光测量OFS和ROS测定m-THPC。m-THPC（2 mg kg（-1）iv）在肿瘤组织中的蓄积在48 h后最好，在肝脏中获得最佳信号。使用非荧光WST 09（2 mg kg（-1）），ROS测定显示注射后48 h肿瘤摄取最佳，肿瘤/肌肉比为5.4。ROS测定似乎可用于确定常规夹持活检组织样本中的致敏剂浓度。(C)2002 Elsevier Science B. V.保留所有权利。
Photodynamic therapy induces the production of reactive oxygen species (ROS) within tissues exposed to laser light after administration of a sensitizer. In the context of continuing clinical and commercial development of chemicals with sensitizing properties, a minimally invasive assay is needed to determine the tissue kinetics of fluorescent or non-fluorescent photoreactive drugs. The level of ROS was determined ex vivo from 1 mm(3) biopsy samples using 2'-7' dichlorofluorescin diacetate (DCFH-DA), a fluorescent probe which was converted into highly fluorescent dichlorofluorescein (DCF) in the presence of ROS. This assay was tested on meta(tetrahydroxyphenyl)chlorin (m-THPC, FOSCAN(R)), a powerful and fluorescent sensitizer, and bacteriochlorophyll derivative WST09 (TOOKAD(R)), a near-infrared absorbing sensitizer that is only slightly fluorescent. In conjunction with the ROS assay, the tissue accumulation of m-THPC was determined on biopsy samples using an optic fibre spectrofluorometer (OFS). DCF fluorescence was proportional to the level of oxidation induced by horseradish peroxidase used as a control and to the concentration (range: 0-5 mug ml(-1)) of both selected photosensitizers irradiated in a tube together with DCFH. Regardless of the organ studied, an excellent correlation was found between fluorescence measurement by OFS and ROS determination for m-THPC. m-THPC (2 mg kg(-1) iv) accumulation in tumour tissues was best after 48 h, and the best signal was obtained in liver. With non-fluorescent WST09 (2 mg kg(-1)), ROS determination showed the best tumour uptake 48 h after injection, with a tumour/muscle ratio of 5.4. The ROS assay appears to be feasible for determining sensitizer concentration in regular grip biopsy tissue samples. (C) 2002 Elsevier Science B.V. All rights reserved.