Cardiac Extracellular Matrix Scaffold Generated Using Sarcomeric Disassembly and Antigen Removal

Xenogeneic cardiac extracellular matrix (cECM) scaffolds for reconstructive cardiac surgery applications have potential to overcome the limitations of current clinically utilized patch materials. A potentially ideal cECM scaffold would be immunologically acceptable while preserving the native cECM niche. Production of such a scaffold necessitates removal of cellular and antigenic components from cardiac tissue while preserving cECM structure/function properties. Existing decellularization methodologies predominantly utilize denaturing detergents which might irreversibly alter cECM material properties. To overcome potential deficiencies of current approaches, the effect of sarcomere relaxation and disassembly on resultant cECM scaffold cellularity was investigated. Additionally, the ability of sequential differential protein solubilization (antigen removal-AR) to reduce cECM scaffold antigenicity was examined. Sarcomeric relaxation and disassembly were necessary to achieve scaffold acellularity. All groups in which AR was employed displayed statistically significant decreases in residual antigenicity regardless of their degree of acellularity. AR combined with sarcomeric disassembly preserved structural, biochemical, mechanical and recellularization properties of the cECM scaffold. However, sodium dodecyl sulfate significantly altered cECM properties. This study demonstrates the importance of solubilizing cellular elements and antigenic components in a stepwise manner for production of a potentially ideal cECM scaffold and may have implications for future tissue engineering and regenerative medicine applications.