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Characterization of Photofrin photobleaching for singlet oxygen dose estimation during photodynamic therapy of MLL cells in vitro

Jonathan S Dysart et al 2005 Phys. Med. Biol. 50 2597-2616   doi: 10.1088/0031-9155/50/11/011  Help

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Jonathan S Dysart1,2 and Michael S Patterson1,2
1 Juravinski Cancer Centre, Physics Research, 699 Concession St., Hamilton, Ontario L8V 5C2, Canada
2 Department of Medical Physics and Applied Radiation Sciences, McMaster University, 1260 Main St. West, Hamilton, Ontario L8S 4L8, Canada
E-mail: jon.dysart@hrcc.on.ca and mike.patterson@hrcc.on.ca

Abstract. A singlet oxygen dose model is developed for PDT with Photofrin. The model is based on photosensitizer photobleaching kinetics, and incorporates both singlet oxygen and non-singlet oxygen mediated bleaching mechanisms. To test our model, in vitro experiments were performed in which MatLyLu (MLL) cells were incubated in Photofrin and then irradiated with 532 nm light. Photofrin fluorescence was monitored during treatment and, at selected fluence levels, cell viability was determined using a colony formation assay. Cell survival correlated well to calculated singlet oxygen dose, independent of initial Photofrin concentration or oxygenation. About 2 × 108 molecules of singlet oxygen per cell were required to reduce the surviving fraction by 1/e. Analysis of the photobleaching kinetics suggests that the lifetime of singlet oxygen in cells is 0.048 ± 0.005 µs. The generation of fluorescent photoproducts was not a result of singlet oxygen reactions exclusively, and therefore did not yield additional information to aid in quantifying singlet oxygen dose.

Print publication: Issue 11 (7 June 2005)
Received 1 February 2005, in final form 7 April 2005
Published 18 May 2005

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