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

• Published in: International journal of molecular sciences 2015-05, Vol.16 (5), p.9936-9948
• Main Authors: El-Khatib, Mustafa, Tepe, Carolin, Senger, Brigitte, Dibue-Adjei, Maxine, Riemenschneider, Markus Johannes, Stummer, Walter, Steiger, Hans Jakob, Cornelius, Jan Frederick
• Format: Article
• Language: English
• ISSN: 1422-0067
• DOI: 10.3390/ijms16059936

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 anin 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 mu L of 0, 25, 50 and 100 mu g/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/cm super(2)). 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 mu g/mL was 96.5% + or - 7.6%, 67.9% + or - 29.9%, 24.0% + or - 16.7% and 13.8% + or - 7.5%, respectively. For the negative controls (no 5-ALA/PDT and ALA/no PDT), the viability rates were 101.72% + or - 3.5% and 100.17% + or - 3.6%, respectively. The LD sub(50) for 5-ALA was estimated between 25 and 50 mu g/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-PDTin vivo.