Effects of photodynamic therapy on glioma spheroids

Abstract

The present study describes the sensitivity of glioma cells to a haematoporphyrin derivative (Photosan-3TM) under laser activation (argon-pumped dye laser). The effects of photodynamic therapy (PDT) on cell growth, directional migration and cell invasion were investigated on two human glioma cell lines (GaMg and U-251Mg). The directional cell migration and spheroid growth was determined for both cell lines exposed to increasing laser energy output (15-35J/cm2) with concentrations of 5 and 7 mg/ml of Photosan-3TM. Both cell lines showed a dose-dependent migratory response to increasing laser irradiation, that was more prominent in the 7 mg/ml treatment group. This effect occurred during the first 4 days after drug exposure. Also, spheroids from both cell lines showed a drug and laser output energy dose-dependent inhibition of growth which became apparent after a lag period of 6 days. The lag period was characterized by a decreased growth rate as compared with the control group. During this period the outer cell layers of the spheroids fell apart. The remaining spheroid tissue was not able to migrate and to regrow when exposed to the highest laser energy outputs (30-35J/cm2, 5 and 7 mg/ml Photosan-3TM). These spheroids showed, however, the ability for invasion when confronted with normal brain cell aggregated in vitro. Light microscopic observations of co-cultures between tumour tissue and brain cell aggregates revealed a normal tumour morphology. This indicates that the remaining tumour cells were not dead and could be stimulated to invade the normal tissue when exposed to a normal brain microenvironment.

• Key Words: Glioma, Haematoporphyrin Derivative, Invasiveness, Photodynamic Therapy, Three-dimensional Culture.