Photodynamic therapy in HeLa and mice fibrosarcoma cells using m-tetrahydroxyphenyl chlorin and 650 nm Light

ETCHEVERRY, MARÍA EUGENIA , GALARZA, CELESTE , PASQUALE, MIGUEL ANGEL, BIBE, SOLANGE , GUTIERREZ ANABELA , POINZINIBBIO, CARLOS , GARAVAGLIA, MARIO

Córdoba

Congreso; ELAFOT XI; 2012

Universiodad Nacional de Rio Cuarto, CONICET, Agencia Nacional de Promoción Cietífica y Tecnológica, Asociación Argentina de Investigación Ficoquimica

The photodynamic therapy (PDT) consists in the administration of a non toxic photosensitizer (PS) drug and after a certain period of time in which the drug is accumulated in the tumor it is irradiated with visible light, usually a long wavelength red light [1]. In this contribution we present results of PDT employing a 0.8 W LED lamp and a Laser (Medligth FD1, with 0.9 W) both emitting at 650 nm, and (meta-tetra(hydroxyfenyl)) chlorine ? Foscan as photosensitizer (PS). HeLa, cells passage 44-60, and fibrosarcoma cells (TMC) obtained from an induced tumor on the flank of BalbC mices were employed. Cell cultured were obtained seeding 2 ml of RPMI medium (Gibco, Invitrogen Corp.) supplemented with 10% FBS, 2g/l bicarbonate, and 100µm/ml streptomycin containing 50000?75000 cells in 3.6 cm Petri dishes. The cultures were incubated overnight in an oven at 37º C in a 5 % carbon dioxide and 97 % humidity atmosphere. Then the medium was replaced by new one supplemented with 2 % FBS with and without PS and incubated for different times (tF). Before PDT treatment cultures were washed and fresh complete medium was added. Cell cultures containing PS were manipulated in black boxes to avoid influence of natural light. PS concentration (cF) in the range 0.05 ≤ cF ≤ 80 g/ml, tF in the range 90 ≤ tF ≤ 4320 min and the radiation time in the range 2 ≤ tR ≤ 18 min were employed. Cell viability before and after PDT treatment was evaluated with standard supra-vital dyes and optical phase contrast and fluorescence microscopy. The effectiveness of PDT treatment was evaluated by cell counting. For some conditions apoptosis and necrosis were investigated. The presented results suggest that the proposed irradiation source appears to be useful for in vitro photodynamic studies and in vivo animal models.