A Study Of HMME-PDT On Port Wine Stains

Part I Comparison of the efficacy of PDT, PDL and IPL for the treatment of port wine stainsObject To compare the efficacy of port wine stains treated by photodynamic therapy (PDT), pulsed dye laser (PDL) and intense pulsed light (IPL).Methods An retrospective analysis was carried out to compare the efficacy of continuous KTP (532nm) laser PDT with hematoporphyrin monomethyl ether (HMME) as a photosensitizer,595nm PDL and IPL with 560nm,590nm and 640nm filter on patients on port wine stains with different gender, lesion types and locations. Results One hundred and thirty cases were enrolled in our study, in which 35 cases were in PDT group,56 in PDL group and 39 in IPL group. There was no statistical significant difference in constituent ratio among different gender, lesion types and locations (p= 0.904,0.929,0.987). The total effecacy rate showed that PDT was the best, following with PDL and IPL in sequence. And the obvious effective rates were 54.3%,33.9% and 20.5%, respectively. In PDT group, the effect of pink flat lesions was better than purple flat (p=0.021) and proliferation lesions (p=0.026). There was no statistical significant difference between purple flat and proliferation lesions (p=0.068). In both PDL and IPL group, there was no statistical significant difference among different lesion types (PDL group:p= 0.226,0.400,0.946; IPL group:p=0.803,0.095,0.069). Lesions on cervical region had a better effect than on face in both PDT (p=0.002) and PDL group (p=0.004), while no statistical significant difference was showed in IPL group (p=0.097). There was no statistical significant difference between different genders in all three groups (p=0.225,0.821,0.145). Conclusion PDT is an effective and safe option in treating port wine stains. Its total effect was better than PDL and IPL. PartⅡComparision of the Absorption Characteristics of HMME by ECV304 and HaCaTObjective To investigate and compare the absorption characteristics of hematoporphyrin monomethyl ether (HMME) by vascular endothelial cell (ECV304) and keratinocyte (HaCaT). Methods Exponentially growing ECV304 and HaCaT were incubated with a serial dose of HMME for 16h. The concentrations were 50,100, 150,200,250μg/ml respectively. Then ECV304 and HaCaT were incubated with 150μg/ml HMME for different time as 15min,30min, 1h,3h,8h,12h and 24h. The absorption of HMME was measured by laser scanning confocal microscope (LSCM).Results The absorption of HMME was dose-dependent. The absorption for different concentration by ECV304 was 74.00,125.57,135.24,141.99 and 132.09, respectively; As for HaCaT, it was 93.88,102.45,112.59,108.23 and 104.70, respectively. Time-dependent absorption of HMME was also found in our study. The absorption for different time point by ECV304 was 95.07,103.97,105.96,108.99,112.93,115.36 and 122.91, respectively, and was 104.25,106.60,108.72,113.75,117.66,114.90 and 118.14 respectively by HaCaT. Conculsions The absorption of HMME by both ECV304 and HaCaT was dose-dependent and time-dependent within a certain concentration and time range.PartⅢStudy on HMME Subcellular Localization and Targets of PDT in ECV304 and HaCaTObject To investigate the subcellular localization of hematoporphyrin monomethyl ether (HMME) and the targets of HMME-photodynamic therapy(PDT) in ECV304 and HaCaT. Methods Four fluorescent organelle probes were selected to label mitochondria, nuclear membrane, cytomembrane and peroxisome. Fluorescence intensity was observed under laser scanning confocal microscope (LSCM). ECV304 and HaCaT were incubated with 150μg/ml HMME for 1h and 18h respectively, and analysis of HMME fluorescence intensity ratio on four organelles were conducted to study the subcellular localization of HMME. Both cell lines were exposed to 532nm laser after incubated with 150μg/ml HMME for 20h. Fluorescence intensity of four organelles before and after PDT was compared. Results The HMME fluorescence intensity ratio of mitochondria, nuclear membrane, cytomembrane and peroxisome in ECV304 1h group was 1.18,0.72,0.95 and 0.68, respectively, and was 1.35,0.83, 0.73, and 0.91 respectively in 18h group. In HaCaT 1h group, it was 1.09,0.66,0.92, and 0.77, respectively, and was 1.13,0.86,0.72, and 1.10, respectively in 18h group. The decreases of fluorescence intensity in ECV304 were 18.22%,10.77%,7.44%, 8.56%, respectively. It was 11.90%,5.02%,3.82%, and 8.90%, respectively in HaCaT.Conclusions In ECV304, HMME was mainly located in mitochondria, while in HaCaT was in mitochondria and peroxisome. The target organelle of PDT might probably be mitochondria in ECV304 and mitochondria and peroxisome in HaCaT.