Irradiance dependence of the conduction block of an in vitro cardiomyocyte wire.

BACKGROUND: To obtain therapeutic condition precisely by in vitro experiment, we studied the irradiance dependence of the electrical conduction blockage caused by a photodynamic reaction using a high extracellular concentration of talaporfin sodium on a novel in vitro cardiomyocyte electrical conduction wire. METHODS: The cardiomyocyte wires were constructed on patterned cultivation cover glass, which had cultivation areas 60µm in width, and a maximum length of 10mm. The talaporfin sodium concentration was set to 20µg/mL. The photodynamic reaction with a high extracellular photosensitizer concentration was performed with a short time interval (approximately 15min) between photosensitizer exposure and irradiation. A 663-nm laser was applied to the cardiomyocyte wire, and the irradiance was varied between 3 and 120mW/cm2. The cardiomyocyte electrical conduction was evaluated using the cross-correlation function of intracellular Ca2+ probe fluorescence brightness from an upper and lower section outside the laser irradiation area of a wire every 10s, which lasted up to 600s. RESULTS: The onset of electrical conduction blockage was defined by an 85% decrease in the cross-correlation function, compared with its initial value. The time for the electrical conduction blockage decreased from 600 to 300s as the irradiance was increased. Also, the probability of electrical conduction blockage was found to increase with increasing irradiance. CONCLUSIONS: We found a strong dependence on the irradiance for the time and probability of electrical conduction blockage.

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.

Conduction block in novel cardiomyocyte electrical conduction line by photosensitization reaction.

We developed a novel cardiomyocyte electrical conduction line. We studied electrical conduction block by extra-cellular photosensitization reaction with this conduction line to study electrical blockade by the photosensitization reaction in vitro.