Construction and evaluation of charge-reversal nanocarrier loaded with hematoporphyrin monomethyl ether / 药学学报

Charge-reversal nanocarrier was constructed to enhance lysosomal escape and improve an-titumor effect. We synthesized the cholesterol-polyethyleneimine-hexahydrophthalic anhydride (Chol-PEI-HHPA) polymer and characterized by 1H NMR. The charge-reversal liposomes (Lipo-HHPA) were synthesized and the hematoporphyrin monomethyl ether (HMME) was loaded. pH-triggered charge conversion was determined at different pH values. The lysosomal escape and cytotoxicity of the Lipo-HHPA were evaluated in MCF-7 cells. The Lipo-HHPA was uniform with an average particle size of 102 nm. Upon the irradiation of ultrasound, burst release of HMME could be observed. The zeta potential of Lipo-HHPA changed sharply from negative (-23.5 mV) to positive (+21.2 mV) over the pH range of 7.4-4.5. In the cellular uptake experiment, the lysosomal escape of Lipo-HHPA was observed. HMME loaded Lipo-HHPA displayed obviously enhanced cytotoxicity towards MCF-7 cells. These results indicate that the charge-reversal liposomes hold a great potential in improving the cytotoxicity and antitumor effect.

Hematoporphyrin monomethyl ether combined with He-Ne laser irradiation-induced apoptosis in canine breast cancer cells through the mitochondrial pathway

Hematoporphyrin monomethyl ether (HMME) combined with He-Ne laser irradiation is a novel and promising photodynamic therapy (PDT)-induced apoptosis that can be applied in vitro on canine breast cancer cells. However, the exact pathway responsible for HMME-PDT in canine breast cancer cells remains unknown. CHMm cells morphology and apoptosis were analyzed using optical microscope, terminal deoxynucleotidyl transferase dUTP nick end labeling fluorescein staining and DNA ladder assays. Apoptotic pathway was further confirmed by Real-time-polymerase chain reaction and Western blotting assays. Our results showed that HMME-PDT induced significant changes in cell morphology, such as formation of cytoplasmic vacuoles and the gradual rounding of cells coupled with decreased size and detachment. DNA fragmentation and cell death was shown to occur in a time-dependent manner. Furthermore, HMME-PDT increased the activities of caspase-9 and caspase-3, and released cytochrome c from mitochondria into the cytoplasm. HMME-PDT also significantly increased both mRNA and protein levels of Bax and decreased P53 gene expression in a time-dependent manner, while the mRNA and protein expression of Bcl-2 were repressed. These alterations suggest that HMME-PDT induced CHMm cell apoptosis via the mitochondrial apoptosis pathway and had anti-canine breast cancer effects in vitro.

Dose effect of laser irradiation in photodynamic therapy for laryngocarcinoma in BALB/c mouse / 国际生物医学工程杂志

Objective The dose and type of light and photosensitizer could seriously affect the curative effect of photodynamic therapy (PDT).The purpose of this study was to observe whether or not PDT with hematoporphyrin monomethyl ether (HMME) can cure laryngocarcinoma in the solid tumor model,and to define the proper laser amount for killing the cancer cells.Methods Forty eight BALB/c mouse models with subcutaneous Hep-2 laryngeal carcinomas were prepared.Mice were divided into six groups depending on the amount of laser received from 30 J/cm2 to 480 J/cm2 including a control group,tumor size in each group was between 8 mm and 10 mm.Tail vein injection were given with HMME prior to applying the laser light,and then illumination was carried out on the tumor at 3 h after HMME administration.Tumor volume,animal weight and histopathologic changes were observed after PDT.Results All mice apparently showed positive results via PDT,and the cancer had been cured in 120 J/cm2 and 480 J/cm2 groups.The laryngeal cancer lesions began to form scab 1 d after PDT and the scab became hard and black after 5 d.The tumor regression began simultaneously and completed around 30 d after PDT.Conclusions PDT may treat laryngeal cancers which sized less than 10 mm in mouse models.The optimum energy to destruct the laryngeal cancer cells may be 120 J/cm2.

Impact of HMME-PDT on Pg of the surface of implants and cementum tablets / 国际生物医学工程杂志

Objective To evaluate the bactericidal effect of diode laser on Porphyromonas gingivalis (Pg),and to explore an optimized protocol for a safe dose of photodynamic therapy (PDT) to eliminate periodontal pathogens as well as the impact on the implant surfaces,so as to provide theoretical and experimental basis for PDT in periimplantitis therapy.Methods Artificial in vitro models were formatted by culturing Pg standard strain and ITI (International Team for Implantology) implants together in CDC broth.Then artificial in vitro models were treated by different doses of hematoporphyrin monomethyl ether (HMME) and different energy density of laser (EDL) for 60 s.The cultures were counted by colony form unit (CFU),and SPSS 17.0 statistical software was used for data statistical analysis to select the best EDL and HMME dose.Finally,ITI implants were observed by scanning electron microscope (SEM) to evaluate the impact of HMME-PDT on Pg of implant surfaces.Results When EDL was 12 J/cm2 and mass concentration of HMME was 25 μg/ml,SEM observations showed that PDT could effectively kill Pg ((13.00±5.00) CFU)without damaging the implant surfaces.Conclusions PDT therapy combining 630 nm diode laser with photosensitizer HMME have good bactericidal effect on Pg,and the EDL and HMME dose is as small as the clinical applicable safe dose.

Investigating the killing effect of photodynamic therapy induced by different photosensitizers on human leukemia cell in vitro / 国际生物医学工程杂志

Objective To investigate and compare the killing effect of photodynamic therapy (PDT)induced by hematoporphyrin derivative (HpD),hematoporphyrin monomethyl ether (HMME) and photocarcinorin (PsD007) on human leukemia cells K562 in vitro.Methods Human leukemia cells were cultured with serial concentrations of photosensitizers followed by irradiation of different dosage of laser light,then MTT colorimetric assay was applied to measure the relative survival rate of PDT for the cells.Results Significant difference in the inhibitory between the PDT group and control group was observed (P＜0.05).The survival rate of PDT for the cells elevated along with the increase in the concentration of sensitizer and dose of laser light.When the photosensitizer concentration was bigger (25 μg/ml) or the energy density was bigger (7.2 J/cm2),the effect of PsD007 was better than HMME,and they were significantly better than HpD (P＜0.05).Conclusion PDT has significant killing effect on human leukemia cells K562,and its relative inhibitory rate appears to be correlated with the dose of sensitizer and laser light irritation.The effect of PDT is related to the photosensitizers.The effect of HpD-PDT is not as effective as PsD007 and HMME.On the conditions of higher energy density and larger photosensitizer concentration,the effect of PsD007-PDT is better than HMME-PDT.

The inhibitory effect of photodynamic therapy and of an anti-VCAM-1 monoclonal antibody on the in vivo growth of C6 glioma xenografts

We investigated the effect of photodynamic therapy (PDT) and of an anti-vascular cell adhesion molecule-1 (VCAM-1) monoclonal antibody on the in vivo growth of C6 glioma. Seven days after inoculation with C6 cells, adult male Wistar rats weighing 280-300 g with MRI-confirmed glioma were randomly assigned to 4 groups (N = 15 per group): PDT + VCAM-1 antibody group; PDT group; VCAM-1 antibody group; control group. Eight days after inoculation, hematoporphyrin monomethyl ether (HMME) was administered as a photosensitizer and PDT was performed at 630 nm (illumination intensity: 360 J/cm²) for 10 min. VCAM-1 antibody (50 µg/mL) was then administered (0.5 mL) through the tail vein every other day from day 8 to day 16. At day 21, 5 rats in each group were sacrificed and cancers were harvested for immunohistochemistry and Western blot assay for the detection of VCAM-1, and TUNEL assay was used to detect apoptosis. Survival and tumor volume were recorded in the remaining 10 rats in each group. In the PDT group, tumor growth was significantly suppressed (67.2 percent) and survival prolonged (89.3 percent), accompanied by an increase in apoptosis (369.5 percent), when compared to control. Furthermore, these changes were more pronounced in the PDT + VCAM-1 antibody group. After PDT, VCAM-1 expression was markedly increased (121.8 percent) and after VCAM-1 monoclonal antibody treatment, VCAM-1 expression was significantly reduced (58.2 percent). PDT in combination with VCAM-1 antibody can significantly inhibit the growth of C6 glioma and prolong survival. This approach may represent a promising strategy in the treatment of glioma.

Apoptosis of hypertrophic scar fibroblasts induced by HMME-PDT / 中华医学美学美容杂志

Objective To investigate the apoptotic effects of hypertrophic scar fibroblast (HSF) induced by HMME-PDT.Methods Fibroblasts were cultured from nontreated hypertrophic scars,and cells at passages 4-6 were used for the experiments (photosensitizer dose 4 μg/ml,λ630 nm,pow er density 10 mw/cm2,energy fluence 2.5 J/cm2).Morphological and biochemical changes in fibroblasts were assessed by Hoechst 33258 staining and fluorescence microscopy.The rate of apoptotic or necrotic cells was detected by flow cytometry (FCM) through double staining of Annexin V -FITC and popodium iodide (PI),respectively.Results Marked morphological features of cell apoptosis were viewed under the fluorescent microscope through Hoechst 33258 staining.The analysis of FCM indica ted that the apoptotic rate was significantly increased after HMME PDT [(34.82 ± I.42) ％ vs (3.12±0.28) ％,P＜0.05],and apoptotic rate was higher than necrosis rate [(14.65±1.02) ％ vs (34.82±1.42) ％,P＜0.05].Conclusions Low level exposure to 630 nm PDT mediated by HMME appears to induce fibroblast apoptosis.

Distribution of Hematoporphyrin Monomethyl Ether in Tumor-bearing Mice / 第二军医大学学报

spleen(?)heart(?)muscle.The pharmacokinetics of PsD-044 in mice was fitted by an open three compartment model. The pharmacokinetic parameters calculated were shown to be essentially consistent with the determination of tissue distribution. It was also shown that PsD-044 can't pass through blood-brain barrier.