In vitro model assesses the susceptibility of polymeric scaffolds for material-driven heart valve regeneration to calcification.

PURPOSE: Material driven in situ heart valve tissue engineering (HVTE) prospects an alternative to non-living replacements. HVTE exploits bioresorbable (synthetic) scaffolds that guide neo-tissue formation. Proper scaffold design assesses and mitigates potential material-related risks, such as calcific nodule formation. Herein, we establish an in vitro model to investigate the calcification risk of materials for HVTE. METHODS: Calcification was studied by culturing 3D scaffolds with porcine valvular interstitial cells in a phosphate-enhanced calcification medium (CM) for 3 weeks. The model was applied by testing three electrospun polymeric Tissue engineering (TE) scaffolds (PCL, PCL-BU, and PC-BU) against a bovine pericardial patch control. Additionally, the model included a 10% cyclic strain environment to evaluate hemodynamic effects. RESULTS: TE constructs showed significantly less calcification compared to the pericardial tissue control, mirroring in vivo animal model findings. No differences in calcification were observed among the TE constructs, and cyclic strain did not affect calcification. CONCLUSION: The 3D in vitro model established in this study effectively mimics calcification in TE material constructs, aiding in systematic testing and comparison of cardiovascular TE materials. It can help understand calcification principles and evaluate potential risk factors (e.g., strain). As such, the model will support the design of biomaterials for in situ HVTE in particular and implantable polymer grafts in general. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s44164-025-00090-x.