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Objective@#To evaluate the clinical efficacy of hematoporphyrin monomethyl ether-based photodynamic (HMME-PDT) therapy for the treatment of port-wine stain (PWS) and its sonographic changes.@*Methods@#A total of 45 patients with confirmed PWS were enrolled from the Department of Dermatology, Third Affiliated Hospital of Soochow University from March 2017 to June 2018, including 5 with pink PWS, 39 with purplish red PWS and 1 with thickened PWS. All the patients received 3 sessions of HMME-PDT therapy. The skin thickness and density were compared before and after the treatment by using high-frequency ultrasound. Ranked data were analyzed by using nonparametric test. Measurement data were expressed as mean ± standard deviation, and analyzed using ony-way analysis of variance. Multiple comparisons were performed using Student-Newman-Keuls-q (SNK-q) test. The results were considered to be statistically significant if P < 0.05.@*Results@#Among the 45 patients with PWS who completed the treatment and follow-up, 10 were cured, 21 received marked improvement, 12 received improvement, and 1 showed no response. The total response rate was 97.78%, and the response rate in the patients with pink PWS was higher than that in the patients with purplish red PWS (U = 12.50, P < 0.001) . The difference value of the skin thickness or skin density before and after the treatment significantly differed among the cured patients, patients receiving marked improvement and those receiving improvement (skin thickness:0.65 ± 0.21, 0.56 ± 0.88, 0.37 ± 0.12 mm respectively; skin density: -8.65 ± 2.19, -6.86 ± 2.79, -4.92 ± 2.91 g/cm3 respectively; F = 14.528, 5.428 respectively, both P < 0.001) , and the difference values of the skin thickness and density were significantly higher in the cured patients than in those receiving improvement (q = 5.82, 4.63, both P < 0.05) . Erythematous swelling to different extents occurred at the laser-exposed sites in the zygomatic and cheek region in 23 patients with PWS and in the frontal-zygomatic region in 6 with PWS after the HMME-PDT therapy, but gradually regressed about 1 week later. Pale brown crusts were observed at the laser-exposed sites in 35 patients, and shed spontaneously about 3 weeks later. Post-inflammatory hyperpigmentation at the laser-exposed sites was observed in 4 patients, and gradually regressed after 2-month follow-up.@*Conclusions@#HMME-PDT therapy is effective for the treatment of PWS, with high safety and few adverse reactions. High-frequency ultrasound can be used for objectively evaluating the clinical efficacy of HMME-PDT therapy.
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Objective To evaluate the clinical efficacy of hematoporphyrin monomethyl etherbased photodynamic (HMME-PDT) therapy for the treatment of port-wine stain (PWS) and its sonographic changes.Methods A total of 45 patients with confirmed PWS were enrolled from the Department of Dermatology,Third Affiliated Hospital of Soochow University from March 2017 to June 2018,including 5 with pink PWS,39 with purplish red PWS and 1 with thickened PWS.All the patients received 3 sessions of HMME-PDT therapy.The skin thickness and density were compared before and after the treatment by using high-frequency ultrasound.Ranked data were analyzed by using nonparametric test.Measurement data were expressed as mean ± standard deviation,and analyzed using ony-way analysis of variance.Multiple comparisons were performed using Student-Newman-Keuls-q (SNK-q) test.The results were considered to be statistically significant if P ＜ 0.05.Results Among the 45 patients with PWS who completed the treatment and follow-up,10 were cured,21 received marked improvement,12 received improvement,and 1 showed no response.The total response rate was 97.78％,and the response rate in the patients with pink PWS was higher than that in the patients with purplish red PWS (U =12.50,P ＜ 0.001).The difference value of the skin thickness or skin density before and after the treatment significantly differed among the cured patients,patients receiving marked improvement and those receiving improvement (skin thickness:0.65 ± 0.21,0.56 ± 0.88,0.37 ± 0.12 mm respectively;skin density:-8.65 ± 2.19,-6.86 ± 2.79,-4.92 ± 2.91 g/cm3 respectively;F =14.528,5.428 respectively,both P ＜ 0.001),and the difference values of the skin thickness and density were significantly higher in the cured patients than in those receiving improvement (q =5.82,4.63,both P ＜ 0.05).Erythematous swelling to different extents occurred at the laser-exposed sites in the zygomatic and cheek region in 23 patients with PWS and in the frontal-zygomatic region in 6 with PWS after the HMME-PDT therapy,but gradually regressed about 1 week later.Pale brown crusts were observed at the laser-exposed sites in 35 patients,and shed spontaneously about 3 weeks later.Post-inflammatory hyperpigmentation at the laser-exposed sites was observed in 4 patients,and gradually regressed after 2-month follow-up.Conclusions HMME-PDT therapy is effective for the treatment of PWS,with high safety and few adverse reactions.High-frequency ultrasound can be used for objectively evaluating the clinical efficacy of HMME-PDT therapy.
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Objective To compare the clinical efficacy and adverse effects of photodynamic therapy (PDT) versus pulsed dye laser(PDL)for the treatment of port wine stains(PWS). Methods Forty?five patients with PWS were enrolled in this study. The PWS lesions in each patient were randomly divided into PDT and PDL areas. Hematoporphyrin monomethyl ether of 5 mg/kg was injected intravenously into the PDT area protected from light, followed by 20?minute irradiation with a 532?nm, solid?state, continuous?wave laser(power density:80-100 mw/cm2;spot diameter: 7 cm)10 minutes later. The PDL area was treated with a single session of 595?nm pulsed dye laser radiation(spot diameter:7 mm;pulse width:10 ms;energy density:10-12 J/cm2). The interval between PDT and PDL treatment was no shorter than two months. Follow up visits were scheduled on day 4 and week 8 after each treatment. Adverse reactions were recorded, and photographs were taken before and 8 weeks after the treatment for evaluation of lesion regression. Results In the case of PDT area, 10 cases(22.22%)were nearly cured, 22(48.89%)achieved marked improvement, 9(20.00%)improvement, 4(8.89%)no improvement. As far as the PDL area is concerned, 6 cases(13.33%)were nearly cured, 16(35.56%)achieved marked improvement, 18(40.00%)improvement, and 5 (11.11%)no improvement. The response rate was significantly higher in the PDT area than in the PDL area(Z=2.48, P0.05). Conclusion For the treatment of PWS, both PDT and PDL are effective and safe, and single?session PDT appears to be superior to single?session PDL.
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Objective To observe the phosphorylation of Smad3 in hyperplastic scar fibroblasts (HSFs) induced by hematoporphyrin monomerthyl ether (HMME) followed by photodynamic therapy (PDT).Methods Fibroblasts were isolated from the hypertrophic scar tissues of 10 patients and subjected to culture in vitro.After 3-5 passages,the HSFs were divided into 4 groups:control group receiving no treatment,PDT group pretreated with HMME of 4 μg/ml followed by PDT,HMME group induced by HMME alone,and laser group irradiated with laser alone.Fluorescence microscopy was used to observe the expression of Smad3 after immunofluorescent staining with anti-Smad3 antibody,and Western blot to detect the expression of Smad3 and phosphorylated Smad3 in these HSFs.Paired t test was conducted to compare the difference in Smad3 and phosphorylated Smad3 expression between these groups.Results The total fluorescence intensity of Smad3 was similar between these groups,but the intranuclear fluorescence signal was significantly weaker in the PDT group than in the control group.The level of phosphorylated Smad3 was statistically decreased in the PDT group compared with the control group (0.20 ± 0.02 vs.0.92 ± 0.15,P ＜ 0.05),but no significant difference was observed between the HMME group and laser group (P ＞ 0.05).Conclusion PDT may inhibit the proliferation of HSFs via attenuating the phosphorylation of Smad3.
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Objective To compare the absorption characteristics of HMME by a human umbilical vein endothelial cell line ECV 304 versus a human keratinocyte cell line HaCaT.Methods Exponentially growing ECV304 and HaCaT cells were incubated with various concentrations (50,100,150,200 and 250 mg/L) of HMME for 16 h or HMME of 150 mg/L for various durations (15 min,30 min,1 h,3 h,8 h,12 h and 24 h).The quantity of HMME absorbed by the cells were determined by laser scanning confocal microscopy (LSCM).Results The fluorescence intensity was 74.00,125.57,135.24,141.99 and 132.09 for ECV304 cells,93.88,102.45,112.59,108.23 and 104.70 for HaCaT cells,after incubation with HMME of 50,100,150,200 and 250 mg/L,respectively.After treated with HMME of 150 mg/L for 15 min,30 min,1 h,3 h,8 h,12 h and 24 h,ECV304 cells showed a fluorescence intensity of 95.07,103.97,105.96,108.99,112.93,115.36 and 122.91,respectively,and HaCaT cells displayed a fluorescence intensity of 104.25,106.60,108.72,113.75,117.66,114.90 and 118.14,respectively.Conculsions Within a defined range of concentration and duration,the absorption of HMME by both ECV304 and HaCaT cells is,to some extent,concentration- and time-dependent.
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ObjectiveTo investigate the role of caspase 3 in HMME-induced apoptosis in hypertrophic scar fibroblasts (HSFs).MethodsFibroblasts were obtained from 10 patients with untreated hypertrophic scar,and subjected to a primary culture.After 4 to 6 passages of culture,the HSFs were divided into 3 groups to remain untreated(control group),be treated with HMME followed by photodynamic therapy (HMME-PDT group),or the combination of HMME and Z-DEVD-FMK followed by photodynamic therapy (caspase 3 inhibitor group).At 12 hours after the therapy,HSFs were collected and immunofluorescence microscopy was used to observe the fluorescence intensity of caspase 3 after staining with fluorescein isocyanate (FITC) and popodium iodide (PI),flow cytometry was performed to determine the percentage of caspase 3-positive HSFs and apoptosis rate in HSFs after single staining with FITC and PI respectively.Results The fluorescence intensity of caspase 3 was weak in the control group and caspase 3 inhibitor group,but was strong in the HMME-PDT group.An increased percentage of caspase 3-positive HSFs was noted in the HMMEPDT group compared with the control group and caspase 3 inhibitor group(30.86％ ± 1.21％ vs.3.12％ ±0.28％ and 2.46％ ± 0.18％,t =19.92,21.76,both P ＜ 0.05).The apoptosis rate in HSFs was significantly higher in the HMME-PDT group and caspase 3 inhibitor group than in the control group(30.54％ ± 3.78％ and 10.46％ ± 2.15％ vs.2.45％ ± 0.22％,t =35.90,27.97,both P＜ 0.05),and higher in the HMME-PDT group than in the caspase 3 inhibitor group.ConclusionsThe apoptosis in HSFs induced by HMME-PDT is closely related to the activation of caspase 3,while caspase 3 seems to be dispensable for the apoptosis.