In vitro effects of pulsed near-ultraviolet laser exposure on human larynx carcinoma cells.

Effects of pulsed near-ultraviolet laser beam on structural characteristics and macromolecular synthesis of carcinoma HEp2 cells were investigated. Laser irradiation damage induced in these eukaryotic cells could be characterized by two development stages: a) a reversible stage with minor morphological damages (1.5 kJ/m2) and 2) an irreversible one, at higher fluences, characterized by cellular membrane damage, necrobiosis and cells detachment from the substrate (4.5 kJ/m2). A. Studies performed referring to macromolecular syntheses of low laser fluences (1.5 kJ/m2)--irradiated HEp2 cells showed the following aspects: a) syntheses inhibiton phase in the first cycles of cellular replication and b) syntheses stimulation phases in the following cycle with total repair of laser-induced molecular lesions. B. At high laser fluences (3-4.5 kJ/m2), metabolic lesions repair was partially or totally blocked after prolonged culturing at 37 degrees C. Ths paper suggests some mechanisms of laser action on macromolecular synthesis and correlates them with morphological changes induced by laser exposure of carcinoma cells.

Laser effects on cellular adhesion and influence on the interrelation between HeLa S3 and human diploid cells.

The interactions between HeLa S3 tumoral cells and human fibroblasts after nitrogen-laser irradiation (337.1 nm) have been studied by using an in vitro cell invasion model. For the quantitative and morphological evaluation of nitrogen-laser radiation action upon tumoral adhesion to the fibroblast monostrate, we used: a) 3H-thymidine labelling of HeLa S3 tumoral cells; b) morphological modifications studies by phase contrast and scanning electron microscopy. The results emphasized the following aspects: 1. In non-irradiated cell cultures we noticed three interaction stages: adhesion, tumoral spreading and displacement with fibroblasts destruction; on the other side, we found a reduced adhesion to non-irradiated human fibroblasts of laser irradiated tumoral cells. 2. Significant percent increasing of non-irradiated tumoral cells adhesion to fibroblast monostrate, irradiated with various laser fluences (e.g. 0.2 kJ/m2--48.1%; 0.8 kJ/m2--63.8% and for 1.6 kJ/m2--79.5%). This phenomenon evidenced the close interrelation between irradiation fluences and tumoral adhesion rates. 3. The importance of numerical ratio between tumoral cells and fibroblasts in tumoral adhesion and invasion processes (e.g. ratio 1:10 tumoral adhesion reached 8.1%; in 1:5--25.9%; in 1:1--59.4% and for 2:1--83.9%). 4. Marked cytotoxic effects for both cell types after exposure to high and very high laser fluences (1.6--6.4 kJ/m2). Our results emphasize near UV-laser irradiation effects upon some of tumoral adhesion and invasion mechanisms and demonstrate the interrelations between cell populations manifesting a different vital potential.

Studies of the effects of associated photodynamic therapy and drugs on macromolecular synthesis of tumoral cells grown in vitro.

HeLa S3 tumoral cells were used as an experimental model for studying the association of photodynamic therapy (PDT) and antitumoral agents. Tumoral monolayer cultures were incubated 18 hours at 37 degrees C with Photofrin II, trypsinized and suspended in Eagle medium supplemented with 10% FCS and then treated with antitumoral agents 90 minutes before He-Ne laser exposure. The tumoral cells were exposed to antitumoral agents in the following concentrations (equivalent to ED70): adriamycin (0.0297 micrograms); mitomycin C (0.0199 micrograms); 5-FU (0.4937 micrograms) and vinblastine (0.0109 micrograms) per 10(5) cells. Macromolecular syntheses (DNA, RNA and proteins) were investigated by use of radioactive precursors: 3H-thymidine, 3H-uridine and 3H-leucine, as expressed in percent referring to Photofrin II-pretreated controls; they were exposed to He-Ne laser but not treated with antitumoral agents. All experiments were followed for 72 hours incubation at 37 degrees C. The conclusions of the results of PDT associated with antitumoral agents sustain the following aspects: a) the antitumoral agents activity (adriamycin, mitomycin C, 5-FU, vinblastine) was more noticeable when applied 90 minutes before He-Ne laser irradiation; b) inhibition of radioactive precursors uptake in DNA, RNA and proteins was accompanied by suppression of in vitro tumoral cells development and c) PDT association with antitumoral agents could manifest at least three positive effects upon animals; 1) PDT potentiating effects with antitumoral agents; 2) suppressing effects on tumoral macromolecular synthesis; 3) antitumoral agents cytotoxic elimination (due to the low doses used).