Cellular effects of hematoporphyrin derivative photodynamic therapy on normal and neoplastic rat bladder cells

Abstract

HPD is known to localize in neoplastic cells and when exposed to the appropriate wavelength of light causes cytotoxicity. The authors have established a rat urothelial cell model for use in comparing and contrasting the effects of HPD photodynamic therapy (PDT) in normal (RBL-01) and transitional cell carcinoma (AY27) bladder cell lines. Uptake, toxicity, and morphologic damage following exposure to HPD PDT were evaluated. Trypan blue exclusion was used for determination of the toxicity of several HPD concentrations (1, 10, 25, and 50 micrograms/ml) with increasing duration of incubation with HPD (0, 1, 2, 4, 12, 24, and 48 hours). Both cell lines displayed increased toxicity with higher concentrations of HPD; however, the AY27 cells were more susceptible to the toxic effects of HPD PDT than the RBL-01 cells at the higher HPD doses studied (25 and 50 micrograms/ml). Viability decreased with increased duration of HPD incubation in RBL-01 cells up until 4 hours, after which it showed a steady increase. Viability decreased in the AY27 cells with increased duration of HPD incubation. An increase in serum concentration in the medium resulted in an increase in viability for both cell lines. Both cell lines demonstrated fast initial uptake of HPD followed by slower uptake over the time studied. By 24 and 48 hours the AY27 cells contained twice the amount of methanol-extractable porphyrins as the RBL-01 cells. The initial morphologic change following HPD PDT was damage to mitochondria. Mitochondrial damage occurred immediately after PDT in the AY27 cells and 30 minutes after PDT in the RBL-01 cells. Both cell lines exhibited a similar progression of cell injury; however, morphologic damage was observed earlier after PDT and appeared more extensive in the AY27 cells.