Aminolevulinic Acid-Mediated Photodynamic Therapy of Human Meningioma: An in Vitro Study on Primary Cell Lines

International Journal of Molecular Sciences

Mustafa El-Khatib¹  Carolin Tepe¹  Brigitte Senger¹  Maxine Dibué-Adjei¹  Markus Johannes Riemenschneider³  Walter Stummer²  Hans Jakob Steiger¹  Jan Frຝérick Cornelius¹  Michael R. Hamblin⁴ 

[1] Department of Neurosurgery, Universitätsklinikum Düsseldorf, Heinrich Heine Universität, 40225 Düsseldorf, Germany; E-Mails:;Department of Neurosurgery, Universitätsklinikum Münster, Westfälische Wilhelms-Universität, 48149 Münster, Germany; E-Mail:;Department of Neuropathology, Regensburg University Hospital, 93042 Regensburg, Germany; E-Mail:Department of Neurosurgery, Universitätsklinikum Düsseldorf, Heinrich Heine Universität, 40225 Düsseldorf, Germany;

关键词: 5-ALA;    aminolevulinic acid;    PDT;    photodynamic therapy;    meningioma;    in vitro;    primary cell culture;   

DOI  :  10.3390/ijms16059936

来源: mdpi

【 摘 要 】

Objective: Five-aminolevulinic acid (5-ALA)-induced porphyrins in malignant gliomas are potent photosensitizers. Promising results of ALA-PDT (photodynamic therapy) in recurrent glioblastomas have been published. Recently, 5-ALA-induced fluorescence was studied in meningioma surgery. Here, we present an experimental study of ALA-PDT in an in vitro model of primary meningioma cell lines. Methods: We processed native tumor material obtained intra-operatively within 24 h for cell culture. Epithelial membrane antigen (EMA) immunohistochemistry was performed after the first passage to confirm that cells were meningioma cells. For 5-ALA-PDT treatment, about 5000 cells per well were seeded in 20 wells of a blank 96-well plate. Each block of 4 wells was inoculated with 150 µL of 0, 25, 50 and 100 µg/mL 5-ALA solutions; one block was used as negative control without 5-ALA and without PDT. Following incubation for 3 h PDT was performed using a laser (635 nm, 18.75 J/cm²). The therapeutic response was analyzed by the water soluble tetrazolium salt (WST-1) cell viability assay 90 min after PDT. Results: 5-ALA-PDT was performed in 14 primary meningioma cell lines. EMA expression was verified in 10 primary cell cultures. The remaining 4 were EMA negative and PDT was without any effect in these cultures. All 10 EMA-positive cell lines showed a significant and dose-dependent decrease in viability rate (p < 0.001). Cell survival at 5-ALA concentrations of 12.5, 25, 50 and 100 μg/mL was 96.5% ± 7.6%, 67.9% ± 29.9%, 24.0% ± 16.7% and 13.8% ± 7.5%, respectively. For the negative controls (no 5-ALA/PDT and ALA/no PDT), the viability rates were 101.72% ± 3.5% and 100.17% ± 3.6%, respectively. The LD₅₀ for 5-ALA was estimated between 25 and 50 µg/mL. Conclusion: This study reveals dose-dependent cytotoxic effects of 5-ALA-PDT on primary cell lines of meningiomas. Either 5-ALA or PDT alone did not affect cell survival. Further efforts are necessary to study the potential therapeutic effects of 5-ALA-PDT in vivo.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.