In situ heart valve tissue engineering aims at developing next-generation heart valve replacements, named tissue engineered heart valves (TEHVs), to substitute diseased (pulmonary/aortic) heart valves. Compared to currently available clinical-grade valvular prostheses, TEHVs have the potential to regenerate and remodel upon implantation. In our project, we manufacture TEHV by using an in vitro grown human tissue engineered matrix (hTEM) obtained by culturing human dermal fibroblasts onto a bioresorbable scaffold for up to 6 weeks. This results in an extracellular matrix- (ECM) rich environment capable of providing crucial mechanical and biochemical cues to ensure in-vivo functionality and native-like remodelling upon implantation.
However, there is still a limited understanding on what is the complete hTEM composition, and on how it develops over tissue culture time. Therefore, the aim of this project is to perform an in-depth longitudinal evaluation of the hTEM by 1) multiscale analysis of hTEM composition during culture time, using (immuno)histology, biochemical assays, and mass spectrometry (LC-MS/MS); 2) analysis of protein pathways involved in ECM development using gene set enrichment analysis (GSEA); 3) assessment of hTEM mechanical properties using uniaxial tensile testing; 4) in vitro assessment of hTEM immunocompatibility by using hiPSC-derived macrophages (IPSCDMs) as a model for resident tissue macrophages; and 5) manufacturing of hTEM-based TEHV for in vitro testing at both pulmonary and aortic pressure conditions in a pulse duplicator.