PI3K inhibitor However, we still need a breakthrough in the form of a novel vector that will transform cells at high efficiency and with low risk of adverse effects. This is especially true in cardiovascular medicine, where malignant cellular transformation is rare [17]. One of the promising candidates for safe and efficacious gene transfection is a naked plasmid vector that has been modified to have high affinity for cardiovascular tissues but which has no built-in viral components [17, Inhibitors,research,lifescience,medical 18]. We have

developed a method for electroporation of a cytokine gene for treatment of cardiomyopathy [13]. However, using electric shock for transfection is not clinically practical. For this reason, we are pursuing the present sonoporation method as a protocol for gene transfection. The HGF protein used in the present study is found in a wide variety of cell types and has multiple biological properties, including mitogenic, motogenic, morphogenic and antiapoptotic activities [19]. Several lines of evidence indicate that this molecule has potential

for therapeutic use for treatment Inhibitors,research,lifescience,medical of heart failure, myocardial infarction, angina, and hypertension [20–22]. HGF may also have enormous therapeutic Inhibitors,research,lifescience,medical potential for hepatic and renal disorders, in addition to cardiovascular diseases [23–26]. In the present study, we showed variations in amount of HGF plasmid DNA, liposome concentration, the duration of insonification, and incubation time of the cardiomyocytes with liposome and DNA, and their dose relationships with the final amount of HGF protein released from the cultured neonatal

Inhibitors,research,lifescience,medical cardiomyocytes. We found that specific amounts of liposome and repetitions of insonification were needed for effective protein production from cardiomyocytes. However, high concentrations of bubble liposome and large numbers of repeat insonifications resulted in decreased cell viability. Plasma membrane sonoporation induced by ultrasound and subsequent self-sealing has been reported in previous investigations [27–29]. However, the exact mechanism by which membrane sonoporation causes substance incorporation into the cell is not yet understood. Some investigators speculate Inhibitors,research,lifescience,medical that the membrane poration results in both transfection efficiency and cellular damage. In the present study, scanning microscopy images revealed some microdimples or pores on the cell surface after sonoporation, which did not exist on the surface of others control cardiomyocytes. The numbers of dimples or pores tended to increase with higher concentrations of liposome. Thus, we speculate that these dimples or pores on the cell surface might be related to transfection efficiency and might be evidence of cellular injury by sonoporation. Previous studies of sonoporation of vascular walls revealed that microbubble destruction would cause rupture of microvessels and extravasation [30–33], which would cancel out some benefits of sonoporation. Thus, the poration and self-sealing mechanism needs to be fully investigated and optimized.