Calculation of Singlet Oxygen Dose from Photosensitizer Fluorescence and Photobleaching During mTHPC Photodynamic Therapy of MLL Cells ¶

Predicting the therapeutic outcome of photodynamic therapy (PDT) requires knowledge of the amount of cytoxic species generated. An implicit approach to assessing PDT efficacy has been proposed where changes in photosensitizer (PS) fluorescence during treatment are used to predict treatment outcome. To investigate this, in vitro experiments were performed in which Mat‐LyLu cells were incubated in meta‐tetra(hydroxyphenyl)chlorin (mTHPC) and then irradiated with 652 nm light. PS concentration, fluence rate and oxygenation were independently controlled and monitored during the treatment. Fluorescence of mTHPC was monitored during treatment and, at selected fluence levels, cell viability was determined using a colony‐formation assay. Singlet oxygen dose was calculated using four different models and was compared with cell survival. For the dose metric based on singlet oxygen–mediated PS photobleaching, a universal relationship between cell survival and singlet oxygen dose was found for all treatment parameters. Analysis of the concentration dependence of bleaching suggests that the lifetime of singlet oxygen within the cell is 0.05–0.25 μs. Generation of about 9 × 108 molecules of singlet oxygen per cell reduces the surviving fraction by 1/e.

[1]  P. Monnier,et al.  Optimizing light dosimetry in photodynamic therapy of early stage carcinomas of the esophagus using fluorescence spectroscopy , 1996, Lasers in surgery and medicine.

[2]  R. Redmond,et al.  A Compilation of Singlet Oxygen Yields from Biologically Relevant Molecules , 1999, Photochemistry and photobiology.

[3]  Michael S Patterson,et al.  Direct Near-infrared Luminescence Detection of Singlet Oxygen Generated by Photodynamic Therapy in Cells In Vitro and Tissues In Vivo¶ , 2002, Photochemistry and photobiology.

[4]  H. S. de Bruijn,et al.  Monitoring In Situ Dosimetry and Protoporphyrin IX Fluorescence Photobleaching in the Normal Rat Esophagus During 5-Aminolevulinic Acid Photodynamic Therapy¶ , 2003 .

[5]  J. Moan,et al.  Photodynamic effect on DNA and cell survival of human cells sensitized by hematoporphyrin , 1981 .

[6]  T. Hasan,et al.  In vivo fluence rate and fractionation effects on tumor response and photobleaching: photodynamic therapy with two photosensitizers in an orthotopic rat tumor model. , 1999, Cancer research.

[7]  C. Foote.  DEFINITION OF TYPE I and TYPE II PHOTOSENSITIZED OXIDATION , 1991, Photochemistry and photobiology.

[8]  J. Kanofsky,et al.  QUENCHING OF SINGLET OXYGEN BY BIOMOLECULES FROM L1210 LEUKEMIA CELLS , 1992, Photochemistry and photobiology.

[9]  H. Bergh,et al.  Rapid Communication: Stability of the Fluorescence Measurement of Foscan® in the Normal Human Oral Cavity as an Indicator of its Content in Early Cancers of the Esophagus and the Bronchi , 1999, Photochemistry and photobiology.

[10]  T J Dougherty,et al.  Energetics and efficiency of photoinactivation of murine tumor cells containing hematoporphyrin. , 1976, Cancer research.

[11]  Arjun G Yodh,et al.  Correlation of in vivo photosensitizer fluorescence and photodynamic-therapy-induced depth of necrosis in a murine tumor model. , 2003, Journal of biomedical optics.

[12]  H. Oppelaar,et al.  Oxygen Depletion during and after mTHPC-Mediated Photodynamic Therapy in RIF1 and H-MESO1 Tumors , 2003, Radiation research.

[13]  E. Hull,et al.  Porphyrin Bleaching and PDT-induced Spectral Changes are Irradiance Dependent in ALA-sensitized Normal Rat Skin In Vivo¶ , 2001, Photochemistry and photobiology.

[14]  J F Fowler,et al.  DIFFERENCES IN SURVIVAL CURVE SHAPES FOR FORMAL MULTI-TARGET AND MULTI-HIT MODELS , 1964 .

[15]  Michael S Patterson,et al.  Relationship Between mTHPC Fluorescence Photobleaching and Cell Viability During In Vitro Photodynamic Treatment of DP16 Cells¶ , 2002, Photochemistry and photobiology.

[16]  V. Melnikova,et al.  Photodynamic properties of meta-tetra(hydroxyphenyl)chlorin in human tumor cells. , 1999, Radiation research.

[17]  T. Foster,et al.  Singlet Oxygen‐Versus Nonsinglet Oxygen‐Mediated Mechanisms of Sensitizer Photobleaching and Their Effects on Photodynamic Dosimetry , 1998, Photochemistry and photobiology.

[18]  N. Mak,et al.  Cellular uptake, subcellular localization and photodamaging effect of temoporfin (mTHPC) in nasopharyngeal carcinoma cells: comparison with hematoporphyrin derivative. , 2000, Cancer letters.

[19]  L. Kunz,et al.  Intracellular Photobleaching of 5,10,15,20-Tetrakis(m-hydroxyphenyl) chlorin (Foscan®) Exhibits a Complex Dependence on Oxygen Level and Fluence Rate¶ , 2002, Photochemistry and photobiology.

[20]  L. Lilge,et al.  Implicit and explicit dosimetry in photodynamic therapy: a New paradigm , 1997, Lasers in Medical Science.

[21]  J Moan,et al.  Evaluation of a new photosensitizer, meso‐tetra‐hydroxyphenyl‐chlorin, for use in photodynamic therapy: A comparison of its photobiological properties with those of two other photosensitizers , 1994, International journal of cancer.

[22]  Thomas H. Foster,et al.  In Vivo mTHPC Photobleaching in Normal Rat Skin Exhibits Unique Irradiance-dependent Features¶ , 2002, Photochemistry and photobiology.

[23]  H. Oppelaar,et al.  Influence of fractionation and fluence rate in photodynamic therapy with Photofrin or mTHPC. , 1996, Radiation research.

[24]  F. Guillemin,et al.  Subcellular localization of meta-tetra (hydroxyphenyl) chlorin in human tumor cells subjected to photodynamic treatment. , 1999, Journal of photochemistry and photobiology. B, Biology.

[25]  Jarod C Finlay,et al.  Photobleaching kinetics of Photofrin in vivo and in multicell tumour spheroids indicate two simultaneous bleaching mechanisms. , 2004, Physics in medicine and biology.

[26]  T. G. Truscott,et al.  Photophysical properties of 5,10,15,20-tetrakis(m-hydroxyphenyl)porphyrin (m-THPP), 5,10,15,20-tetrakis(m-hydroxyphenyl)chlorin (m-THPC) and 5,10,15,20-tetrakis(m-hydroxyphenyl)bacteriochlorin (m-THPBC): a comparative study , 1999 .

[27]  A. Moor,et al.  In vitro Fluence Rate Effects in Photodynamic Reactions with AIPcS4 as Sensitizer , 1997, Photochemistry and photobiology.

[28]  T. Foster,et al.  Effect of Irradiation Fluence Rate on the Efficacy of Photodynamic Therapy and Tumor Oxygenation in Meta-Tetra (Hydroxyphenyl) Chlorin (mTHPC)-Sensitized HT29 Xenografts in Nude Mice1 , 2002, Radiation research.

[29]  M R Arnfield,et al.  Oxygen dependency of tumor cell killing in vitro by light-activated Photofrin II. , 1991, Radiation research.

[30]  William R. Potter,et al.  The Theory Of Photodynamic Therapy Dosimetry: Consequences Of Photodestruction Of Sensitizer , 1987, Other Conferences.

[31]  Michael S Patterson,et al.  In vitro tests of the validity of singlet oxygen luminescence measurements as a dose metric in photodynamic therapy. , 2003, Cancer research.

[32]  M S Patterson,et al.  Modeling of photosensitizer fluorescence emission and photobleaching for photodynamic therapy dosimetry. , 1998, Applied optics.

[33]  B. Henderson,et al.  Reduction of tumour oxygenation during and after photodynamic therapy in vivo: effects of fluence rate. , 1998, British Journal of Cancer.

[34]  Hubert van den Bergh,et al.  In-vivo measurement of fluorescence bleaching of meso-tetra hydroxy phenyl chlorin (mTHPC) in the esophagus and the oral cavity , 1995, European Conference on Biomedical Optics.

[35]  J Moan,et al.  The Biophysical Foundations of Photodynamic Therapy , 1998, Endoscopy.

[36]  S. Mitra,et al.  Photodynamic therapy: Biophysical mechanisms and molecular responses , 2004 .

[37]  Michele T. Cooper,et al.  Photofrin photodynamic therapy can significantly deplete or preserve oxygenation in human basal cell carcinomas during treatment, depending on fluence rate. , 2000, Cancer research.

[38]  L. Gossner,et al.  Photodynamic therapy of human Barrett's cancer using 5-aminolaevulinic acid-induced protoporphyrin IX: an in-vivo dosimetry study in athymic nude mice , 2002, European journal of gastroenterology & hepatology.

[39]  A. Rauth,et al.  An experimental and analytical study of oxygen depletion in stirred cell suspensions. , 1980, Radiation research.

[40]  L. Lilge,et al.  Light Dosimetry for Intraperitoneal Photodynamic Therapy in a Murine Xenograft Model of Human Epithelial Ovarian Carcinoma , 1998, Photochemistry and photobiology.

[41]  Stanley B. Brown,et al.  Fluorescence Photobleaching of ALA‐induced Protoporphyrin IX during Photodynamic Therapy of Normal Hairless Mouse Skin: The Effect of Light Dose and Irradiance and the Resulting Biological Effect , 1998, Photochemistry and photobiology.

[42]  J. Blais,et al.  A comparative study of the cellular uptake, localization and phototoxicity of meta-tetra(hydroxyphenyl) chlorin encapsulated in surface-modified submicronic oil/water carriers in HT29 tumor cells. , 2000, Journal of photochemistry and photobiology. B, Biology.

[43]  K. Berg,et al.  THE PHOTODEGRADATION OF PORPHYRINS IN CELLS CAN BE USED TO ESTIMATE THE LIFETIME OF SINGLET OXYGEN , 1991, Photochemistry and photobiology.

[44]  Qian Peng,et al.  An outline of the hundred-year history of PDT. , 2003, Anticancer research.

[45]  M G Nichols,et al.  The Mechanism of Photofrin Photobleaching and Its Consequences for Photodynamic Dosimetry , 1997, Photochemistry and photobiology.

[46]  T. Foster,et al.  Effects of Fluence Rate on Cell Survival and Photobleaching in Meta‐Tetra‐(hydroxyphenyl)chlorin–photosensitized Colo 26 Multicell Tumor Spheroids ¶ , 2001 .

[47]  J. Spikes.  QUANTUM YIELDS AND KINETICS OF THE PHOTOBLEACHING OF HEMATOPORPHYRIN, PHOTOFRIN II, TETRA(4‐SULFONATOPHENYL)‐PORPHINE AND UROPORPHYRIN , 1984 .