We investigated the correlation between the therapeutic effect by early irradiation Photodynamic Therapy (PDT) and vascular response. The early irradiation PDT has been proposed by our group. This PDT protocol is that pulse laser irradiates to tumors 1 h after intravenous injection of water-soluble photosensitizer. The intact layer appeared over the well treated layer, when the early irradiation PDT was performed at rat prostate subcutaneous tumors with high intensity pulse laser (over 1 MW/cm2 in peak intensity) and Talaporfin sodium. In order to clarify the phenomenon mechanism, we monitored blood volume, surface temperature, photosensitizer amount, and oxygen saturation during the PDT. The rat prostate subcutaneous tumor was irradiated with excimer dye laser light at 1 h after the intravenous injection. The photosensitizer dose wa 2.0 mg/kg, and the pulse energy density was 2.5 mJ/cm2 (low intensity) or 10 mJ/cm2 (high intensity). Under the low intensity pulsed PDT, the fluorescence amount was decreasing gently during the irradiation, and the blood volume and oxygen saturation started decreasing just after the irradiation. Under the hgh intensity pulsed PDT, the fluorescence amount was decreaased rapidly for 20 s after the irradiation started. The blood volume and oxygen saturation were temporally decreased during the irradiation, and recovered at 48 hrs after the irradiation. According to these results, under the low intensity pulsed PDT, the blood vessel located near the surface started closing just after the irradiation. On the other hand, under the high intensity pulsed PDT the blood vessel was closing for 20 s after the irradiation started, moreover, the blood flow recovered at 48 hrs after the irradiation. We concluded that the vascular response depended on the pulse energy density, and then the therapeutic effect was attributed to the difference of the vascular response. In other words, the surface intact layer could be considered to be induced the temporal drug and oxygen depletion effect associated with the temporal vascular shutdown.
We propose the application of early state photodynamic therapy (PDT) to treatment of atrial fibrillation, which is a kind
of arrhythmia characterized by irregular rapid beating of heart. We had demonstrated that our PDT can block the propagation of electrical excitation in cardiac myocytes. However, the mechanism of the PDT-induced electrical blockade was not clear. In order to clarify this mechanism, changes in intracellular Ca2+ concentration during the PDT with Talaporfin sodium (water soluble photosensitizer) were measured by fluorescence Ca2+ indicator, fluo-4 AM. The PDT led to the rapid increase of intracellular Ca2+ concentration and the changes in cell shapes. These results indicated that extracellular Ca2+ flowed into the cells mediated by cell membrane. Moreover, we found bubble generation in the cells after the PDT. In conclusion, the PDT-induced electrical blockade in myocytes can be caused by cell death following the bubble generation, which is accompanied by the increase in intracellular Ca2+ concentration due to the cell membrane malfunction with the PDT.
In order to investigate the interaction between the triplet state T1 and ground state oxygen 3O2 during pulsed
excitation photodynamic therapy (PDT), we measured the phosphorescence and singlet oxygen 1O2 fluorescence time-resolved
waveform. The phosphorescence time-resolved waveform from clinical photosensitizers has not been
obtained because this signal was buried in the photosensitizer fluorescence. We constructed the experimental setup
with a spectral and temporal filter to select the phosphorescence signals from the Photofrin(II)(R) and Talaporfin sodium
solution. The lifetimes and spectrums of the measured luminescence coincided with the phosphorescence
characteristics, respectively. We obtained the phosphorescence time-resolved waveforms from the clinical
photosensitizer solutions successfully. The 1O2 fluorescence time-resolved waveforms from these photosensitizers
were measured with an IR-PMT with a photon counter. The fluorescence time-resolved waveforms of each
photosensitizer were also obtained by the authors. We could consequently describe sequential generation of three
time-resolved waveforms throughout the photosensitive reaction in the clinical photosensitizers. We think we may
evaluate the photoseisitizer characteristics by these waveforms.
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