Artikel
5-ALA fluorescence in native pituitary adenoma cell suspensions: basis for resection control and photodynamic therapy (PDT)?
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Autoren
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Thomas Fortmann
- Klinik für Neurochirurgie, Westfälische Wilhelms Universität, Münster, Deutschland
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Stefan Poeschke
- Klinik für Neurochirurgie, Westfälische Wilhelms Universität, Münster, Deutschland
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Ralf Reeker
- Klinik für Neurochirurgie, Westfälische Wilhelms Universität, Münster, Deutschland
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Hilko Ardon
- Klinik für Neurochirurgie, Westfälische Wilhelms Universität, Münster, Deutschland; Katholieke Universiteit Leuven, Leuven, Belgium; Sint Elisabeth Ziekenhuis, Tilburg, Netherlands
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Hans-Jakob Steiger
- Klinik für Neurochirurgie, Heinrich Heine Universität, Düsseldorf, Deutschland
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Daniel Hänggi
- Klinik für Neurochirurgie, Heinrich Heine Universität, Düsseldorf, Deutschland
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Daniel Prevedello
- Department of Neurological Surgery, Ohio State University, Columbus, USA
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Bernhard Fischer
- Klinik für Neurochirurgie, Westfälische Wilhelms Universität, Münster, Deutschland
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Walter Stummer
- Klinik für Neurochirurgie, Westfälische Wilhelms Universität, Münster, Deutschland
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Christian Ewelt
- Klinik für Neurochirurgie, Westfälische Wilhelms Universität, Münster, Deutschland
| Veröffentlicht: | 21. Mai 2013 |
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Gliederung
Text
Objective: Aminolevulinic acid (5-ALA) may represent a diagnostic intra-operative tool and treatment option as photosensitizer for recurrent or subtotally operated pituitary adenomas. We used cell suspensions of native pituitary adenomas and immortalized pituitary adenoma cell line GH3 in order to provide evidence for 5-ALA induced intracellular porphyrin accumulation and for acute effects of the photodynamic therapy (PDT) in these cells.
Method: Cell suspensions of pituitary adenomas in primary cell culture, immortalized pituitary adenoma cell line GH3 and glioma cell line U373 were incubated for 4 h in 50 and 100 µg/ml ALA in 5% CO2 in room air. 5-ALA fluorescence accumulation was measured by fluorescence activated cell sorting (FACS) after 4/6/8/10/12/18/33/40 hours. Further, subsequent irradiation using a diode laser (lambda = 635 nm, 40 mW/cm2, total fluence 25 J/cm2) was performed for 5-ALA-incubated cell suspensions of GH3 cell line and U373 cell line and three control groups ("laser only", "ALA only", "no drug no light"). Cell death was analyzed by colorimetric assay (WST-1 based) for the nonradioactive quantification of cell proliferation, cell viability and cytotoxicity. Furthermore, clinical and histopathological features were correlated to our experimental findings.
Results: Incubation with 5-ALA in 50 and 100 µg/ml concentrations revealed the strongest fluorescence accumulation in ten pituitary adenoma cell suspensions of 80% after 8–12 hours vs. 96% in the immortalized pituitary adenoma cell line GH3 after 33–40 hours and 91% in the control glioma cell line U373 after 4–6 hours. In first trials, ALA incubation in 5% CO2 with subsequent PDT resulted in significant cell death of more than 70% and growth arrest in the immortalized adenoma cell line and the glioma cell line as well. The rate of cell death in all control groups was negligible
Conclusions: Pituitary adenoma cell suspensions show porphyrin fluorescence after incubations with ALA. Furthermore, PDT of experimental pituitary adenoma cell suspensions results in significant cell death after irradiation. Therefore, 5-ALA may offer a diagnostic intra-operative tool for resection control and further as treatment option for recurrent and not completely resectable pituitary adenomas in-vivo.
