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Noninvasive oxygen and perfusion measurements in tumors after PDT
Poster Abstracts P003 Effect of high fluence light emitting diode mediated photodynamic therapy with Photofrin II on high-grade solid tumor model
185 Conclusion: Changes in pO2 and blood perfusion in the tumors treated with PDT depend on applied PDT protocol, i.e. time between photosensitizer and light application.
H.-J. Lim 1 , C.-H. Oh 1,2
doi:10.1016/j.pdpdt.2011.03.211
1
P005
Department of Medical Laser, Dankook University, South Korea Department of Oral Physiology, Dankook University, South Korea Keywords: Oral cancer; Photodynamic therapy; Light emitting diode; High fluence rate; High-grade solid tumor model 2
Backgrounds: Low fluence rates enhance tumor oxygenation sustenance during photodynamic therapy (PDT) but they are not acceptable for surgery due to the prolonged treatment time. This study examined whether high fluence rate PDT is efficient in treating solid tumors by reducing the intraoperative time. Methods: Oral squamous cell carcinoma (KB cells) was implanted subcutaneously into nude mice (Balb/c nu/nu). The tumor was irradiated 24 h after the administration of Photofrin II (10 mg/kg) and the fluence rate was ranged from 360 J/cm2 to 600 J/cm2 using a prototype light emitting diode (LED) lamp equipped with a cooling system. Results: The emission peak of the prototype LED lamp was located at approximately 630 nm and the temperature of the LED head module was 25 ◦ C. At a 600 J/cm2 light dose (500 mW/cm2 for 20min), 72% complete tumor remission was observed, whereas 34% and 58% tumor remission at 360 J/cm2 and 410 J/cm2 , respectively. Conclusion: High fluence PDT with a high concentration of photosensitizer can be applied to clinical treatments to reduce the intraoperative treatment time dramatically. doi:10.1016/j.pdpdt.2011.03.210 P004 Noninvasive oxygen and perfusion measurements in tumors after PDT M. Krzykawska 1 , K. Soczek 1 , B. Kalinowska 1 , J.M. Dabrowski 2 , G. nska 1 , M. Elas 1 Stochel 2 , L.G. Arnaut 3 , M.M. Pereira 3 , K. Urba˜ 1
Jagiellonian University, Department of Biophysics, Kraków, Poland 2 Jagiellonian University, Faculty of Chemistry, Kraków, Poland 3 Univerity of Coimbra, Chemistry Department, Coimbra, Portugal
Background: PDT induces extensive changes in the level of oxygenation and in perfusion of the treated tumors. Studying kinetics of these changes gives insight into the PDT physiological action and allows to optimize the PDT protocol. We have chosen difluorinated sulfamoyl bacteriochlorin (TDFPBSNHethyl) as a photosensitizer. It absorbs light in the NIR range of light where human tissues are the most transparent and generate singlet oxygen and other ROS with high yield. It is an effective photosensitizer in pilot in vitro and in vivo studies. Methods: S-91 Cloudman melanoma and Levis Lung Cancer tumors were grown in the legs of DBA/2 and C57 mice. When the tumor reached 3 mm of mean diameter, oxymetry probe (LiPc) was injected into the tumor tissue. The bacteriochlorin (2 mg/kg b.w.) was administered intravenously into mice and tumors were treated with 100 J/cm2 of light at = 750 nm. The pO2 was determined from the linewidth of the LiPc spectrum measured with electron paramagnetic resonance using surface coil at S-band (2.12 GHz). The pO2 and blood perfusion (Laser Doppler Periscan II) was measured before, and 15 min, 3 h after the treatment and then every 24 h for 6 following days. Results: Single dose TDFPBSNHethyl PDT eliminated tumors within 3—14 days. PDT-induced changes in pO2 in the tumor were paralleled by changes in superficial blood perfusion in the tumor.
PDT using ultrashort pulses—–Investigation of induced necrosis C. Grecco, S. Pratavieira, L.T. Moriyama, C. Kurachi, V.S. Bagnato University of São Paulo, Instituto de Física de São Carlos, São Calos, Brazil One limitation of PDT is the light penetration in biologic tissues. High intensity pulsed-laser may constitute a potential tool to overcome this limitation. In this study, we evaluated the photodegradation for two photosensitizers (PS), photodithazine (PDZ) and Photogem when irradiated with pulsed and CW laser at same fluence rate. Photodithazine (6 g/ml) and Photogem (8 g/ml) in distilled water were illuminated with pulsed or CW laser for 15 min. The data was collected at several investigation times using fluorescence spectroscopy and investigated wavelength was the maximum fluorescence intensity when excited with 532 nm. Three fluence rates were investigated for PDZ (15, 56 and 112 mW/cm2 ) and Photogem (280, 340 and 400 mW/cm2 ). The necrosis profile induced by PDT was evaluated in healthy livers of rats. The animals were sensitized and after the optimal uptake time for each PS, livers were exposed and illuminated with specific wavelength for each PS (660 nm for PDZ and 630 nm for the Photogem). After 30 h, the animals were killed and the livers removed for HE analysis. The results showed that photodegradation rate by CW laser is higher than pulsed laser for the PDZ sensitizer. Photogem presented an opposite behavior, i.e. a higher degradation rate for the pulsed irradiation, as well as a higher depth of necrosis. The results are not concluded and future experiments are planned to further investigate and explain the observed differences using these sensitizers. doi:10.1016/j.pdpdt.2011.03.212 P006 Heme carrier protein 1 involves a cancer specific porphyrin accumulation H. Matsui, T. Kaneko, Y.N. Nagano, K. Hiyama, M. Tamura, T. Takada, T. Yamamoto, I. Hyodo University of Tsukuba, Graduate School of Comprehensive Human Sciences, Japan Backgrounds and aims: Photodynamic therapy (PDT) is one of chemotherapies for neoplasm, which uses a cancer specific porphyrins accumulation. In this therapy, singlet oxygen is produced by the reaction between laser and a cancer specific accumulated porphyrins to involve cellular apoptosis. However, the reason why porphyrins were particularly accumulated to cancer cells has still remained unknown. We hypothesize that a cancer specific over-expression of heme carrier protein 1 (HCP-1) involve these porphyrins accumulation: HCP-1 was reported to transport porphyrin ring including substrates such as heme in the duodenum. To elucidate this possibility, we examined the effect of RNAi for HCP-1 on cellular porphyrins incorporation in gastric cancer cell-lines besides gastric normal cells. Moreover, we measured the porphyrin accumulations after HCP-1 gene transfection in these cells. Results: We found that HCP-1 was particularly expressed in cancer cell-lines, whereas not in the normal cell-line. Porphyrins accumulations occurred and obvious fluorescence was observed in these HCP-1 expressing cells, whereas no fluorescence observed in the normal cell-line. The treatment with RNAi
185 Conclusion: Changes in pO2 and blood perfusion in the tumors treated with PDT depend on applied PDT protocol, i.e. time between photosensitizer and light application.
H.-J. Lim 1 , C.-H. Oh 1,2
doi:10.1016/j.pdpdt.2011.03.211
1
P005
Department of Medical Laser, Dankook University, South Korea Department of Oral Physiology, Dankook University, South Korea Keywords: Oral cancer; Photodynamic therapy; Light emitting diode; High fluence rate; High-grade solid tumor model 2
Backgrounds: Low fluence rates enhance tumor oxygenation sustenance during photodynamic therapy (PDT) but they are not acceptable for surgery due to the prolonged treatment time. This study examined whether high fluence rate PDT is efficient in treating solid tumors by reducing the intraoperative time. Methods: Oral squamous cell carcinoma (KB cells) was implanted subcutaneously into nude mice (Balb/c nu/nu). The tumor was irradiated 24 h after the administration of Photofrin II (10 mg/kg) and the fluence rate was ranged from 360 J/cm2 to 600 J/cm2 using a prototype light emitting diode (LED) lamp equipped with a cooling system. Results: The emission peak of the prototype LED lamp was located at approximately 630 nm and the temperature of the LED head module was 25 ◦ C. At a 600 J/cm2 light dose (500 mW/cm2 for 20min), 72% complete tumor remission was observed, whereas 34% and 58% tumor remission at 360 J/cm2 and 410 J/cm2 , respectively. Conclusion: High fluence PDT with a high concentration of photosensitizer can be applied to clinical treatments to reduce the intraoperative treatment time dramatically. doi:10.1016/j.pdpdt.2011.03.210 P004 Noninvasive oxygen and perfusion measurements in tumors after PDT M. Krzykawska 1 , K. Soczek 1 , B. Kalinowska 1 , J.M. Dabrowski 2 , G. nska 1 , M. Elas 1 Stochel 2 , L.G. Arnaut 3 , M.M. Pereira 3 , K. Urba˜ 1
Jagiellonian University, Department of Biophysics, Kraków, Poland 2 Jagiellonian University, Faculty of Chemistry, Kraków, Poland 3 Univerity of Coimbra, Chemistry Department, Coimbra, Portugal
Background: PDT induces extensive changes in the level of oxygenation and in perfusion of the treated tumors. Studying kinetics of these changes gives insight into the PDT physiological action and allows to optimize the PDT protocol. We have chosen difluorinated sulfamoyl bacteriochlorin (TDFPBSNHethyl) as a photosensitizer. It absorbs light in the NIR range of light where human tissues are the most transparent and generate singlet oxygen and other ROS with high yield. It is an effective photosensitizer in pilot in vitro and in vivo studies. Methods: S-91 Cloudman melanoma and Levis Lung Cancer tumors were grown in the legs of DBA/2 and C57 mice. When the tumor reached 3 mm of mean diameter, oxymetry probe (LiPc) was injected into the tumor tissue. The bacteriochlorin (2 mg/kg b.w.) was administered intravenously into mice and tumors were treated with 100 J/cm2 of light at = 750 nm. The pO2 was determined from the linewidth of the LiPc spectrum measured with electron paramagnetic resonance using surface coil at S-band (2.12 GHz). The pO2 and blood perfusion (Laser Doppler Periscan II) was measured before, and 15 min, 3 h after the treatment and then every 24 h for 6 following days. Results: Single dose TDFPBSNHethyl PDT eliminated tumors within 3—14 days. PDT-induced changes in pO2 in the tumor were paralleled by changes in superficial blood perfusion in the tumor.
PDT using ultrashort pulses—–Investigation of induced necrosis C. Grecco, S. Pratavieira, L.T. Moriyama, C. Kurachi, V.S. Bagnato University of São Paulo, Instituto de Física de São Carlos, São Calos, Brazil One limitation of PDT is the light penetration in biologic tissues. High intensity pulsed-laser may constitute a potential tool to overcome this limitation. In this study, we evaluated the photodegradation for two photosensitizers (PS), photodithazine (PDZ) and Photogem when irradiated with pulsed and CW laser at same fluence rate. Photodithazine (6 g/ml) and Photogem (8 g/ml) in distilled water were illuminated with pulsed or CW laser for 15 min. The data was collected at several investigation times using fluorescence spectroscopy and investigated wavelength was the maximum fluorescence intensity when excited with 532 nm. Three fluence rates were investigated for PDZ (15, 56 and 112 mW/cm2 ) and Photogem (280, 340 and 400 mW/cm2 ). The necrosis profile induced by PDT was evaluated in healthy livers of rats. The animals were sensitized and after the optimal uptake time for each PS, livers were exposed and illuminated with specific wavelength for each PS (660 nm for PDZ and 630 nm for the Photogem). After 30 h, the animals were killed and the livers removed for HE analysis. The results showed that photodegradation rate by CW laser is higher than pulsed laser for the PDZ sensitizer. Photogem presented an opposite behavior, i.e. a higher degradation rate for the pulsed irradiation, as well as a higher depth of necrosis. The results are not concluded and future experiments are planned to further investigate and explain the observed differences using these sensitizers. doi:10.1016/j.pdpdt.2011.03.212 P006 Heme carrier protein 1 involves a cancer specific porphyrin accumulation H. Matsui, T. Kaneko, Y.N. Nagano, K. Hiyama, M. Tamura, T. Takada, T. Yamamoto, I. Hyodo University of Tsukuba, Graduate School of Comprehensive Human Sciences, Japan Backgrounds and aims: Photodynamic therapy (PDT) is one of chemotherapies for neoplasm, which uses a cancer specific porphyrins accumulation. In this therapy, singlet oxygen is produced by the reaction between laser and a cancer specific accumulated porphyrins to involve cellular apoptosis. However, the reason why porphyrins were particularly accumulated to cancer cells has still remained unknown. We hypothesize that a cancer specific over-expression of heme carrier protein 1 (HCP-1) involve these porphyrins accumulation: HCP-1 was reported to transport porphyrin ring including substrates such as heme in the duodenum. To elucidate this possibility, we examined the effect of RNAi for HCP-1 on cellular porphyrins incorporation in gastric cancer cell-lines besides gastric normal cells. Moreover, we measured the porphyrin accumulations after HCP-1 gene transfection in these cells. Results: We found that HCP-1 was particularly expressed in cancer cell-lines, whereas not in the normal cell-line. Porphyrins accumulations occurred and obvious fluorescence was observed in these HCP-1 expressing cells, whereas no fluorescence observed in the normal cell-line. The treatment with RNAi