Research Papers: General

Real-time analysis of endogenous protoporphyrin IX fluorescence from δ-aminolevulinic acid and its derivatives reveals distinct time- and dose-dependent characteristics in vitro

[+] Author Affiliations
Tobias Kiesslich

Paracelsus Medical University, Institute of Physiology and Pathophysiology, Strubergasse 21, Salzburg A-5020, Austria

Paracelsus Medical University/Salzburger Landeskliniken, Department of Internal Medicine I, Muellner Hauptstrasse 48, Salzburg A-5020, Austria

Linda Helander

Norwegian University of Science and Technology, Department of Cancer Research and Molecular Medicine, Erling Skjalgssons gate 1, N-7491 Trondheim, Norway

Romana Illig

Paracelsus Medical University/Salzburger Landeskliniken, Institute of Pathology, Muellner Hauptstrasse 48, Salzburg A-5020, Austria

Christian Oberdanner

Tecan Austria, Untersbergstrasse 1, Grödig A-5082, Austria

Andrej Wagner

Paracelsus Medical University/Salzburger Landeskliniken, Department of Internal Medicine I, Muellner Hauptstrasse 48, Salzburg A-5020, Austria

Herbert Lettner

University of Salzburg, Department of Materials Science and Physics, Division of Physics and Biophysics, Hellbrunnerstraße 34, Salzburg A-5020, Austria

Martin Jakab

Paracelsus Medical University, Institute of Physiology and Pathophysiology, Strubergasse 21, Salzburg A-5020, Austria

Kristjan Plaetzer

University of Salzburg, Department of Materials Science and Physics, Laboratory of Photodynamic Inactivation of Microorganisms, Hellbrunnerstraße 34, Salzburg A-5020, Austria

J. Biomed. Opt. 19(8), 085007 (Aug 12, 2014). doi:10.1117/1.JBO.19.8.085007
History: Received April 25, 2014; Revised June 17, 2014; Accepted July 14, 2014
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Abstract.  Photodynamic therapy (PDT) and photodiagnosis based on the intracellular production of the photosensitizer protoporphyrin IX (PPIX) by administration of its metabolic precursor δ-aminolevulinic acid (ALA) achieved their breakthrough upon the clinical approval of MAL (ALA methyl ester) and HAL (ALA hexyl ester). For newly developed ALA derivatives or application in new tumor types, in vitro determination of PPIX formation involves multiparametric experiments covering variable pro-drug concentrations, medium composition, time points of analysis, and cell type(s). This study uses a fluorescence microplate reader with a built-in temperature and atmosphere control to investigate the high-resolution long-term kinetics (72 h) of cellular PPIX fueled by administration of either ALA, MAL, or HAL for each 10 different concentrations. For simultaneous proliferation correction, A431 cells were stably transfected with green fluorescent protein. The results indicate that the peak PPIX level is a function of both, incubation concentration and period: maximal PPIX is generated with 1 to 2-mM ALA/MAL or 0.125-mM HAL; also, the PPIX peak shifts to longer incubation periods with increasing pro-drug concentrations. The results underline the need for detailed temporal analysis of PPIX formation to optimize ALA (derivative)-based PDT or photodiagnosis and highlight the value of environment-controlled microplate readers for automated in vitro analysis.

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© 2014 Society of Photo-Optical Instrumentation Engineers

Citation

Tobias Kiesslich ; Linda Helander ; Romana Illig ; Christian Oberdanner ; Andrej Wagner, et al.
"Real-time analysis of endogenous protoporphyrin IX fluorescence from δ-aminolevulinic acid and its derivatives reveals distinct time- and dose-dependent characteristics in vitro", J. Biomed. Opt. 19(8), 085007 (Aug 12, 2014). ; http://dx.doi.org/10.1117/1.JBO.19.8.085007


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