Greater cardiomyocyte density on aligned compared with random carbon nanofibers in polymer composites

Carbon nanofibers (CNFs) randomly embedded in poly (lactic-co-glycolic-acid) (PLGA) composites have recently been shown to promote cardiomyocyte growth when compared with conventional PLGA without CNFs. It was shown then that PLGA:CNF composites were conductive and that conductivity increased as greater amounts of CNFs were added to pure PLGA. Moreover, tensile tests showed that addition of CNFs increased the tensile strength of the PLGA composite to mimic that of natural heart tissue. Most importantly, throughout all cytocompatibility experiments, cardiomyocytes were viable and expressed important biomarkers that were greatest on 50:50 wt% CNF:PLGA composites. The increased selective adsorption of fibronectin and vitronectin (critical proteins that mediate cardiomyocyte function) onto such composites proved to be the mechanism of action. However, the natural myocardium is anisotropic in terms of mechanical and electrical properties, which was not emulated in these prior PLGA:CNF composites. Thus, the aim of this in vitro study was to create and characterize CNFs aligned in PLGA composites (at 50:50 wt%, including their mechanical and electrical properties and cardiomyocyte density), comparing such results with randomly oriented CNFs in PLGA. Specifically, CNFs were added to soluble biodegradable PLGA (50:50 PGA:PLA weight ratio) and aligned by applying a voltage and then allowing the polymer to cure. CNF surface micron patterns (20 μm wide) on PLGA were then fabricated through a mold method to further mimic myocardium anisotropy. The results demonstrated anisotropic mechanical and electrical properties and significantly improved cardiomyocyte density for up to 5 days on CNFs aligned in PLGA compared with being randomly oriented in PLGA. These results indicate that CNFs aligned in PLGA should be further explored for improving cardiomyocyte density, which is necessary in numerous cardiovascular applications.