Fabrication of human myocardium using multidimensional modelling of engineered tissues
Pilar Montero-Calle 1 , María Flandes-Iparraguirre 1 , Konstantinos Mountris 2 , Ana S de la Nava 3 , Nicolás Laita 4 , Ricardo M Rosales 4 , Olalla Iglesias-García 1 , Elena M De-Juan-Pardo 5 , Felipe Atienza 6 , María Eugenia Fernández-Santos 7 , Estefanía Peña 4 , Manuel Doblaré 8 , Juan J Gavira 9 , Francisco Fernández-Avilés 3 , Felipe Prosper 10 , Esther Pueyo 11 , Manuel M Mazo Vega 12
Biofabrication of human tissues has seen a meteoric growth triggered by recent technical advancements such as human induced pluripotent stem cells (hiPSCs) and additive manufacturing.
However, generation of cardiac tissue is still hampered by lack of addequate mechanical properties and crucially by the often unpredictable post-fabrication evolution of biological components. In this study we employ melt electrowriting (MEW) and hiPSC-derived cardiac cells to generate fibre-reinforced human cardiac minitissues. These are thoroughly characterized in order to build computational models and simulations able to predict their post-fabrication evolution.
Our results show that MEW-based human minitissues display advanced maturation 28 post-generation, with a significant increase in the expression of cardiac genes such as MYL2, GJA5, SCN5A and the MYH7/MYH6 and MYL2/MYL7 ratios. Human iPSC-cardiomyocytes are significantly more aligned within the MEW-based 3D tissues, as compared to conventional 2D controls, and also display greater expression of CX43. These are also correlated with a more mature functionality in the form of faster conduction velocity.
We used these data to develop simulations capable of accurately reproducing the experimental performance. In-depth gauging of the structural disposition (cellular alignment) and intercellular connectivity (CX43) allowed us to develop an improved computational model able to predict the relationship between cardiac cell alignment and functional performance.
This study lays down the path for advancing in the development of in silico tools to predict cardiac biofabricated tissue evolution after generation, and maps the route towards more accurate and biomimetic tissue manufacture.