Abstract 396: 3d-Engineered Aligned Co-axial Nanofibrous Cardiac Patch for Drug Screening and Cardiac Repair Applications

Abstract

Introduction: Recent studies have demonstrated the great potential of human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) for testing the efficacy of various cardiac drugs. Additionally, studies have shown that the hiPSC-CMs grown in a 3D environment express better physiological characteristics than 2D cultures. The convergence of polymeric cardiac patch technology with hiPSC-CMs has opened up innovative ways for generating biomimetic 3D cardiac tissues. Hypothesis: The central hypothesis of this study was to develop a 3D cardiac tissue model for pharmacological testing of various cardiac drugs on a 3D nanofibrous aligned co-axial cardiac patch. Methods: A co-axial (Co-A) PCL-gelatin aligned nanofibrous patch was fabricated using the electrospinning technique and its mechanical properties were assessed using Universal Test Machine. Then, the hiPSC-CMs were cultured on this Co-A patch for 2 weeks and the LDH assay was performed to determine the cell viability. The functionality of the cardiac patch was determined by an assessment of calcium cycling in hiPSC-CMs. Further, particle image velocimetry (PIV) and microelectrode array (MEA) was used to evaluate the physiological functionality of the cardiac patch in response to various cardiac drugs. Results: Our studies showed that the mean diameter and thickness of aligned Co-A nanofibrous patch was 578±184 nm and 115±11 μm respectively, while its tensile strength was 0.780 ± 0.098 MPa. Further, confocal imaging confirmed the core-shell structure of the Co-A patches with a core diameter of 2.21 ± 0.50 μm. Additionally, The hiPSC-CMs cultured on these aligned Co-A patches showed an aligned morphology and expressed Troponin-T, GATA4, α-sarcomeric actinin, and connexin-43. The hiPSC-CMs seeded on a 3D scaffold showed efficient calcium cycling properties, which were similar to the hiPSC-CMs cultured in 2D scaffold. Furthermore, PIV and MEA analysis showed that hiPSC-CMs cultured in 2D and 3D showed a similar response to various cardiac drugs, isoproterenol, verapamil and E4031. Conclusions: Overall, this study demonstrated a successful fabrication of aligned Co-A nanofibrous cardiac patch and its evaluation as a 3D cardiac tissue model in-vitro, which could be applied towards drug screening, toxicity studies and cardiac repair applications for ischemic heart disease.