Emiyu Ogawa1, Naoki Machida2, Arisa Ito3, Tsunenori Arai4. 1. School of Fundamental Science and Technology, Graduate School of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522, Japan. Electronic address: e.ogawa@arai.appi.keio.ac.jp. 2. School of Fundamental Science and Technology, Graduate School of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522, Japan. Electronic address: n.machida@arai.appi.keio.ac.jp. 3. School of Fundamental Science and Technology, Graduate School of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522, Japan. Electronic address: a.ito@arai.appi.keio.ac.jp. 4. School of Fundamental Science and Technology, Graduate School of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522, Japan. Electronic address: tsunearai@appi.keio.ac.jp.
Abstract
BACKGROUND: We have proposed an application of photodynamic reaction for less-heated myocardial ablation which employs talaporfin sodium. Intracellular photodynamic reactions with ongoing uptake have the ability to induce apoptosis over time, raising the possibility of extending the lesion depth. The objective of this study was to understand how, in myocardial cells, the late cell survival levels change by incubation time with talaporfin sodium, and what dependence talaporfin sodium uptake has on the duration of incubation with talaporfin sodium in vitro. METHODS: Rat myocardial cells were incubated with talaporfin sodium for 5-360 min and intracellular concentrations measured using a fluorescence micro-plate reader after wash. Cell survival was measured using a water-soluble tetrazolium assay at 2 and 24 h after a photodynamic reaction using a red diode laser of 660 nm, following 15-180 min of incubation with talaporfin sodium. Cells were stained with Hoechst 33342 to observe nuclear changes. RESULTS: Intracellular talaporfin sodium concentration increased with incubation time, with a marked increase between 0 and 60 min. Cell survival at 24 h decreased by 20% when the duration of incubation with talaporfin sodium was extended from 15 to 30 min. Following incubation time of 30-180 min with talaporfin sodium, cell survival was decreased by approximately 30% between measurements at 2 and 24 h. The intracellular talaporfin sodium concentration that induced higher levels of late cell death with cell nuclei fragmentation in these cells was approximately 0.2 μg/mL. CONCLUSION: We obtained the characteristics of late cell death occurrence and talaporfin sodium uptake to myocardial cell with various incubation times with talaporfin sodium.
BACKGROUND: We have proposed an application of photodynamic reaction for less-heated myocardial ablation which employs talaporfin sodium. Intracellular photodynamic reactions with ongoing uptake have the ability to induce apoptosis over time, raising the possibility of extending the lesion depth. The objective of this study was to understand how, in myocardial cells, the late cell survival levels change by incubation time with talaporfin sodium, and what dependence talaporfin sodium uptake has on the duration of incubation with talaporfin sodium in vitro. METHODS:Rat myocardial cells were incubated with talaporfin sodium for 5-360 min and intracellular concentrations measured using a fluorescence micro-plate reader after wash. Cell survival was measured using a water-soluble tetrazolium assay at 2 and 24 h after a photodynamic reaction using a red diode laser of 660 nm, following 15-180 min of incubation with talaporfin sodium. Cells were stained with Hoechst 33342 to observe nuclear changes. RESULTS: Intracellular talaporfin sodium concentration increased with incubation time, with a marked increase between 0 and 60 min. Cell survival at 24 h decreased by 20% when the duration of incubation with talaporfin sodium was extended from 15 to 30 min. Following incubation time of 30-180 min with talaporfin sodium, cell survival was decreased by approximately 30% between measurements at 2 and 24 h. The intracellular talaporfin sodium concentration that induced higher levels of late cell death with cell nuclei fragmentation in these cells was approximately 0.2 μg/mL. CONCLUSION: We obtained the characteristics of late cell death occurrence and talaporfin sodium uptake to myocardial cell with various incubation times with talaporfin sodium.