Comparative study of the phototoxicity of two chrolin type photosensitizers, ATX-S10(Na) and verteporfin, on vascular endothelial and retinal pigment epithelial cells.
OBJECTIVE
To compare the phototoxicity in photodynamic therapy (PDT) of ATX-S10(Na) and Verteporfin on human microvascular endothelial cells (HMVEC), vascular endothelial cells of monkey choroid and retina (CRVEC), and human retinal pigment epithelial cells (HRPE).


METHODS
PDT was performed in two different ways. In short dye-exposure PDT, HMVEC and CRVEC were exposed to each photosensitizer for 5 minutes followed by laser irradiation of 670 nm wavelength for ATX-S10(Na) or 689 nm for Verteporfin without washing out the photosensitizer in the medium. In long dye-exposure PDT, the cells were exposed to photosensitizers for times ranging from 5 minutes to 2 hours, washed out the photosensitizers, followed by laser irradiation in a fresh medium. PDT was performed on HRPE with PDT doses that resulted in damaging 90% of the HMVEC (ED(90)). Phototoxicity was determined by MTS Assay 1 day after PDT.


RESULTS
The degree of phototoxicity depended on the dye concentration, laser dose, and dye exposure time. In short dye-exposure PDT on HMVEC with a laser dose of 50 J/cm(2), the ED(90) was 6.3 microg/ml of ATX-S10(Na) and 0.04 microg/ml of Verteporfin, while in long dye-exposure PDT the ED(90) was 50.0 microg/ml of ATX-S10(Na) and 0.04 microg/ml of Verteporfin when the medium was supplemented with 5% fetal calf serum. The phototoxic rate on HMVEC was higher when the medium contained 5% as contrasted with 10% of serum. In short dye-exposure PDT, the ED(90) of CRVEC was 100 microg/ml of ATX-S10(Na) and an irradiance of 100 J/cm(2), and 0.08 microg/ml of Verteporfin and an irradiance of 100 J/cm(2) when the medium was supplemented with 10% serum. With some doses of short dye-exposure PDT, the ATX-S10(Na) achieved higher phototoxic rates on HMVEC and CRVEC than on the HRPE. However, long dye-exposure PDT with ATX-S10(Na) and short and long dye-exposure PDT with Vereteporfin failed to obtain higher phototoxic rates on HMVEC and CRVEC than on HRPE.


CONCLUSIONS
Verteporfin had a higher phototoxicity than ATX-S10(Na) on HMVEC and CRVEC. The CRVEC resisted more than HMVEC following PDT with both photosensitizers. In short dye-exposure PDT, ATX-S10(Na) had a more selective phototoxicity on HMVEC and CRVEC than on HRPE.
Huang
Yufang
Y
Department of Ophthalmology and Visual Sciences, Osaka City University Graduate School of Medicine, Osaka, Japan.
Obana
Akira
A
Gohto
Yuko
Y
Nakajima
Susumu
S
D000818:Animals; D004730:Endothelium, Vascular / Q000187:drug effects*; D006801:Humans; D007834:Lasers; D008253:Macaca mulatta; D008268:Macular Degeneration; D010778:Photochemotherapy; D017319:Photosensitizing Agents / Q000633:toxicity*; D010857:Pigment Epithelium of Eye / Q000187:drug effects*; D011166:Porphyrins / Q000633:toxicity*; D000077362:Verteporfin
C115550:ATX-S10; D017319:Photosensitizing Agents; D011166:Porphyrins; D000077362:Verteporfin