Huang Yufang, Obana Akira, Gohto Yuko, Nakajima Susumu
Department of Ophthalmology and Visual Sciences, Osaka City University Graduate School of Medicine, Osaka, Japan.
Lasers Surg Med. 2004;34(3):216-26. doi: 10.1002/lsm.10251.
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).
STUDY DESIGN/MATERIALS AND 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.
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.
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.
比较ATX-S10(Na)和维替泊芬在光动力疗法（PDT）中对人微血管内皮细胞（HMVEC）、猴脉络膜和视网膜血管内皮细胞（CRVEC）以及人视网膜色素上皮细胞（HRPE）的光毒性。
研究设计/材料与方法：PDT以两种不同方式进行。在短时间染料暴露PDT中，将HMVEC和CRVEC暴露于每种光敏剂5分钟，然后分别用波长670nm（针对ATX-S10(Na)）或689nm（针对维替泊芬）的激光照射，培养基中的光敏剂不冲洗掉。在长时间染料暴露PDT中，细胞暴露于光敏剂的时间为5分钟至2小时，冲洗掉光敏剂，然后在新鲜培养基中进行激光照射。对HRPE进行PDT时采用导致90%的HMVEC受损的PDT剂量（ED(90)）。在PDT后1天通过MTS法测定光毒性。
光毒性程度取决于染料浓度、激光剂量和染料暴露时间。在激光剂量为50J/cm²的短时间染料暴露PDT中，HMVEC的ED(90)对于ATX-S10(Na)为6.3μg/ml，对于维替泊芬为0.04μg/ml；而在长时间染料暴露PDT中，当培养基添加5%胎牛血清时，ATX-S10(Na)的ED(90)为50.0μg/ml，维替泊芬为0.04μg/ml。与含有10%血清的培养基相比，当培养基含有5%血清时，HMVEC的光毒性率更高。在短时间染料暴露PDT中，当培养基添加10%血清时，CRVEC的ED(90)对于ATX-S10(Na)为100μg/ml且辐照度为100J/cm²，对于维替泊芬为0.08μg/ml且辐照度为100J/cm²。在某些短时间染料暴露PDT剂量下，ATX-S10(Na)对HMVEC和CRVEC的光毒性率高于对HRPE的光毒性率。然而，ATX-S10(Na)的长时间染料暴露PDT以及维替泊芬的短时间和长时间染料暴露PDT在HMVEC和CRVEC上未能获得比对HRPE更高的光毒性率。
在HMVEC和CRVEC上，维替泊芬的光毒性高于ATX-S1(Na)。两种光敏剂进行PDT后，CRVEC比HMVEC更具耐受性。在短时间染料暴露PDT中，ATX-S10(Na)对HMVEC和CRVEC的光毒性比对HRPE更具选择性。
