Emiyu Ogawa

Nonthermal Cardiac Catheter Ablation Using Photodynamic Therapy

Background— Radiofrequency ablation has limitations, largely related to creation of lesions by heating. Here, we report the first nonthermal ablation by applying photodynamic therapy (PDT) to cardiac tissues using a custom-made deflectable laser catheter. The present study investigated the feasibility of PDT for cavotricuspid isthmus ablation in a canine model. Methods and Results— We evaluated the pharmacokinetic profiles of 17 canines after administration of a photosensitizer (talaporfin sodium) by various protocols. We succeeded in maintaining the photosensitizer concentration at a level in excess of the clinically effective dose for humans. Using a 4-polar 7-French deflectable laser catheter, we performed PDT-mediated cavotricuspid isthmus ablation in 8 canines. PDT caused oxidative injury only to the irradiated area and successfully produced a persistent electric conduction block. No acute, gross changes such as edematous degeneration, thrombus formation, steam pops, or traumatic injury were observed after irradiation. Hematoxylin and eosin staining of tissues samples also showed well-preserved endothelial layers. Testing of the blood samples taken before and after the procedure revealed no remarkable changes. Lesion size at 2 weeks after the procedure and the temperature data collected during irradiation were compared between the PDT and irrigated radiofrequency ablation procedures. A ventricular cross-section revealed a solid PDT lesion, which was as deep as a radiofrequency lesion. In addition, endocardial, surficial, and intramural temperature monitoring during the PDT irradiation clearly demonstrated the nonthermal nature of the ablation technique. Conclusions— Nonthermal PDT-mediated catheter ablation is a potentially novel treatment for cardiac arrhythmias.

Detailed in vitro study of the photosensitization reaction of extracellular talaporfin sodium in rat myocardial cells

We proposed a new non‐thermal treatment for tachyarrhythmia that employs an extracellular photosensitization reaction. Oxygen depletion may easily occur in in vitro studies of this reaction because the photosensitizing agent is often highly concentrated in such studies. The aim of the current study was to examine the progress of the extracellular photosensitization reaction and the photocytotoxicity of extracellular Talaporfin sodium on myocardial cells for application in tachyarrhythmia therapy.

Optimal conditions for cardiac catheter ablation using photodynamic therapy

AIMS Photodynamic therapy (PDT) is based on non-thermal injury mediated by singlet oxygen species and is used clinically in cancer therapy. In our continuing efforts to apply this technology to cardiac catheter ablation, we clarified the optimal condition for creating PDT-mediated lesions using a laser catheter. METHODS AND RESULTS In a total of 35 canines, we applied a laser directly to the epicardium of the beating heart during open-chest surgery at 15 min after administration of a photosensitizer, talaporfin sodium. We evaluated the lesion size (depth and width) using hematoxylin-eosin staining under varying conditions as follows: laser output (5, 10, 20 W/cm(2)), irradiation time (0-60 s), photosensitizer concentration (0, 2.5, 5 mg/kg), blood oxygen concentration (103.5 ± 2.1 vs. 548.0 ± 18.4 torr), and contact force applied during irradiations (low: <20 g, high: >20 g). A laser irradiation at 20 W/cm(2) for 60 s under 5 mg/kg (29 µg/mL) of photosensitizer induced a lesion 8.7 ± 0.8 mm deep and 5.2 ± 0.2 mm wide. The lesion size was thus positively correlated to the laser power, irradiation time, and photosensitizer concentration, and was independent of the applied contact force and oxygen concentration. In addition, the concentration of the photosensitizer strongly correlated with the changes in the pulse oximetry data and fluorescence of the backscattering laser, suggesting that a clinically appropriate condition could be estimated in real time. CONCLUSION Photodynamic therapy-mediated cardiac lesions might be controllable by regulating the photosensitizer concentration, laser output, and irradiation time.

Effects of albumin binding on photocytotoxicity of extracellular photosensitization reaction using talaporfin sodium to rat myocardial cells

BACKGROUND We previously proposed a new treatment for tachyarrhythmia using an extracellular photosensitization reaction occurring in the interstitial space of myocardia shortly after the injection of talaporfin sodium. Using myocardial cells, we studied the photocytotoxicity of this extracellular photosensitization reaction between talaporfin sodium and albumin. METHODS The albumin concentrations tested spanned the physiological range found in the interstitial space (0-15 mg/ml) while the talaporfin sodium concentration were varied from 0 to 40 μg/ml. The reactions were conducted in 96-well plates. To obtain the binding ratio and the amount of energy deposited into the photosensitizer, we measured the change in the absorbance spectra of talaporfin sodium solutions containing different concentrations of albumin. RESULTS Photocytotoxicity to myocardial cell due to the reaction decreased when physiological concentrations of albumin were added to the reaction mix, and decreased sharply when the molar concentration ratio of albumin to talaporfin sodium was between 0.3 and 1.2. A monotonic binding ratio was obtained, ranging from 10 to 80%, at albumin concentrations of 0.1-1.0 mg/ml. We found that the lethality of the extracellular photosensitization reaction towards myocardial cells had a threshold albumin concentration, even though the energy deposited into the talaporfin sodium solution was calculated to be almost constant (4.23 ± 0.19 J/well) in the presence of 0-15 mg/ml albumin. CONCLUSIONS Based on the likely concentration of albumin in the interstitial space, we conclude that the photodynamic efficacy of talaporfin, under conditions used here, will markedly decrease if the albumin level exceeds 0.65 mg/ml.

Comparison of myocardial cell survival 2 h and 24 h after extracellular talaporfin sodium-induced photodynamic reaction.

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.

Electrical superior vena cava isolation using photodynamic therapy in a canine model

AIMS With the new era of multi-tip radiofrequency or balloon ablation catheters replacing the point-to-point ablation strategy, we aimed to determine the feasibility of a ring-laser catheter ablation technology to electrically isolate the superior vena cava (SVC) by exploring the advantages of the limitless catheter tip size possibly with the photodynamic therapy (PDT)-mediated ablation. METHODS AND RESULTS We developed a first-generation prototype of a circular-laser-mapping catheter by fitting a 7 cm plastic optical fibre onto a circular variable-loop Lasso™ mapping catheter. Following SVC venography, both the laser catheter and another ring catheter for monitoring the SVC potentials were placed at the SVC. After the systemic infusion of a photosensitizer (talaporfin sodium), we initiated the irradiation with an output of 1 W in three canines and 0.3 W in four. The creation of electrical isolation as well as occurrence of phrenic nerve injury, sinus node injury, and SVC stenosis were evaluated before, immediately after, and 1 month after the procedure. A PDT-mediated SVC isolation was successfully performed in all seven canines. The isolation was completed with a laser irradiation of 70.4 ± 71.4 J/cm under 30.9 ± 5.0 µg/mL of a photosensitizer without any sinus node injury, phrenic nerve palsy, or SVC stenosis in both the acute and chronic evaluations. The minimum isolation time of 270 s was not correlated with the laser input power or the photosensitizer concentration. CONCLUSION The electrical SVC isolation was successfully and instantly achieved using the PDT laser-ring catheter without any complications.

Dependence of damage within 10 min to myocardial cells by a photodynamic reaction with a high concentration of talaporfin sodium outside cells in vitro on parameters of laser irradiation

BACKGROUND To investigate the immediate occurrence of irreparable severe damage to myocardial cells up to 10min after a photodynamic reaction with a high concentration of photosensitizer outside cells, we measured the damage response time and the parameters that govern the response time via rat myocardial Ca(2+) concentration. In our proposed method for catheter ablation of tachyarrhythmia by photodynamic reaction, there are two components to the electrical conduction block: an immediate electrical conduction block of several tens of seconds to several minutes, and a permanent electrical conduction block. METHODS Rat myocardial intracellular Ca(2+) concentration changes before, during and after the photodynamic reaction with a high concentration of photosensitizer outside myocardial cells were continuously observed using a Fluo-4 AM Ca(2+) probe. Talaporfin sodium with 663-nm excitation was used to induce the photodynamic reaction. Talaporfin concentration was 10-30μg/ml, radiant exposure was 10-40J/cm(2), and irradiance was 30-290mW/cm(2). We evaluated the response time of irreparable severe damage to myocardial cells, according to Ca(2+) concentration. RESULTS The response time of the defined severe damage occurrence to myocardial cells ranged from 200 to 500s. The response time decreased with increasing irradiance and photosensitizer concentration, but exhibited no significant change with total radiant exposure. CONCLUSIONS We found that severe myocardial cell damage caused by a photodynamic reaction with a high concentration of photosensitizer outside cells occurred within a few minutes, which might be useful for catheter ablation for tachyarrhythmia that needs immediate response during the ablation procedure.

Evaluation of human and bovine serum albumin on oxidation characteristics by a photosensitization reaction under complete binding of talaporfin sodium

BACKGROUND In order to investigate the therapeutic interaction of an extra-cellular photosensitization reaction, we evaluated the oxidation characteristics of human and bovine serum albumin by this reaction with talaporfin sodium under complete binding with albumin by spectroscopic analysis in a cell-free solution. METHODS The solution was composed of 20μg/ml talaporfin sodium and 2.1mg/ml human or bovine serum albumin. A 662nm laser light was used to irradiate the solution. Visible absorbance spectra of solutions were measured to obtain the oxidized and non-oxidized relative densities of albumin and talaporfin sodium before and after the photosensitization reaction. The defined oxidation path ratio of talaporfin sodium to albumin reflected the oxidation of the solution. Absorbance wavelengths at approximately 240 and 660nm were used to calculate normalized molecular densities of oxidized albumin and talaporfin sodium, respectively. RESULTS AND CONCLUSIONS The oxidation path ratio of talaporfin sodium to albumin when binding human serum albumin was approximately 1.8 times larger than that of bovine serum albumin during the photosensitization reaction from 1 to 50J/cm(2). We hypothesized that the oxidation path ratio results might have been caused by talaporfin sodium binding affinity or binding location difference between the two albumins, because the fluorescence lifetimes of talaporfin sodium bound to human and bovine serum albumin were 7.0 and 4.9ns, respectively. Therefore, the photodynamic therapeutic interaction might be stronger with human serum albumin than with bovine serum albumin in the case of extracellular photosensitization reaction.

Extracellular talaporfin sodium-induced photosensitization reaction with various albumin animal species on myocardial cells in vitro

It is reported that the albumin has different structure among animal species. We have proposed a new methodology of cardiac ablation using talaporfin sodium-induced photosensitization reaction with short drug-light interval to realize immediate and permanent therapeutic effect by singlet oxygen production mainly in the interstitial space. The photosensitization reaction efficacy with different animal species should be investing to consider the optimal animal therapeutic model to evaluate the therapeutic effect of new cardiac ablation methodology. We studied the cell-killing efficacy of extracellular talaporfin sodium-induced photosensitization reaction using talaporfin sodium on myocardial cells in vitro with different albumin animal species: human, canine, bovine, and porcine serum albumin. We obtained that the albumin concentration tendency on the binding ratio and cell lethality was different among the animal species but there was no correlation between binding ratio and cell lethality. We found that the cell lethality dependence on albumin concentration showed 2 different groups, human-canine and bovine-porcine. We think that the canine might be useful as a therapeutic animal model since the cytotoxicity tendency on albumin concentration was similar with that of human albumin. These cell lethality tendency difference would be suggested to explain by the existence of the diazepam site that talaporfin sodium binds mainly.

Temperature Influence on Myocardial Cell Cytotoxicity of the Extracellular Photosensitization Reaction with Talaporfin Sodium and Serum Proteins at 17–37°C

BACKGROUND We investigated the binding of talaporfin sodium with albumin and its photocytotoxicity during temperature changes by measuring absorbance spectra. The targeted tissue temperature differs according to the procedure. The photocytotoxicity efficiency should be investigated quantitatively because efficiency changes arising from temperature changes are expected. MATERIALS AND METHODS The temperature dependence of talaporfin sodium binding with human serum albumin (0-20 mg/mL), high-density lipoprotein (0-0.04 mg/mL), and low-density lipoprotein (0-0.14 mg/mL) was investigated at 17°C, 27°C, and 37°C by measurement of absorbance spectra. Cell lethality was measured using a water-soluble tetrazolium-8 assay at 2 h after the photosensitization reaction at 17°C and 37°C. RESULTS The binding ratios of talaporfin sodium with high-density lipoprotein decreased by 6.3% and those with low-density lipoprotein decreased by 12.8% when the temperature increased from 17°C to 37°C. Cell lethality increased significantly with a temperature rise from 17°C to 37°C at irradiation exposure of 20 and 40 J/cm2 and talaporfin sodium concentration of 20 μg/mL. CONCLUSIONS From our in vitro data, we can predict that the change in photocytotoxicity efficiency would be negligible with a temperature decrease of <5°C from the body temperature in the case of photodynamic ablation with a short drug-light interval.