Real-time analysis of endogenous protoporphyrin IX fluorescence from δ-aminolevulinic acid and its derivatives reveals distinct time- and dose-dependent characteristics in vitro
| Autor: | Romana Illig, Andrej Wagner, Tobias Kiesslich, Martin Jakab, Kristjan Plaetzer, Linda Helander, Herbert Lettner, Christian Benno Oberdanner |
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| Rok vydání: | 2014 |
| Předmět: |
Radiation-Sensitizing Agents
Cell Survival medicine.medical_treatment Biomedical Engineering Protoporphyrins Photodynamic therapy Endogeny Apoptosis Green fluorescent protein Biomaterials chemistry.chemical_compound Computer Systems Cell Line Tumor medicine Humans Protoporphyrin IX Dose-Response Relationship Drug Aminolevulinic Acid Neoplasms Experimental Fluorescence Atomic and Molecular Physics and Optics In vitro Electronic Optical and Magnetic Materials Dose–response relationship Spectrometry Fluorescence chemistry Photochemotherapy Biophysics A431 cells |
| Zdroj: | 19:085007 Journal of Biomedical Optics |
| ISSN: | 1560-2281 |
| Popis: | Photodynamic therapy (PDT) and photodiagnosis based on the intracellular production of the photo- sensitizer 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 com- position, 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 pro- liferation 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. © 2014 Society of Photo-Optical Instrumentation Engineers (SPIE) (DOI: 10.1117/1.JBO.19.8 .085007) |
| Databáze: | OpenAIRE |
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