Time response of electrical conduction block in novel cardiomyocyte wire by extra-cellular photosensitization reaction at various irradiances

Abstract

We studied time response of electrical conduction (EC) block in a novel cardiomyocyte wire by extra-cellular photosensitization reaction (EPR) at various irradiances. This EC block using the EPR has been studied to develop a non-thermal arrhythmia therapeutic methodology. Despite the EC block in acute phase is needed to judge therapeutic endpoint in clinical arrhythmia therapy, time response of the EC block by the EPR in acute phase hasn’t been studied. We measured the time to EC block occurrence by the EPR with intra-cellular Ca²⁺ concentration change using Fluo-4 AM fluorescence measurement by a confocal laser microscope system. The pattern cultivation cover glass with 10 mm Φ which had 60 μm width cultivation areas with 300 μm separations was used to form the cardiomyocyte wires. Rat cardiomyocyte with 10.8×105 cells was disseminated to the cover glasses installed in a 35 mmΦ dish. After 3 days from the dissemination, the EPR was operated to the cardiomyocyte wires for 10 min varying 3-120 mW/cm² in 663 nm laser irradiances with 20 μg/ml talaporfin sodium. An irradiation area was approximately 60×340 μm² on each wires. Cross correlation functions (CCF) in measured fluorescence images in every 10 s were calculated across the irradiation area. The time to EC block occurrence was defined as the time of the max difference between adjacent CCFs. By decreasing irradiances in 30-6 mW/cm², the time to EC block occurrence became longer from 294 to 434 s. In 30-120 mW/cm², the time to EC block occurrence was nearly constant in 300 s.