Substrate-dependent variability in viability and angiogenicmarker expression among three endothelial cell subtypesVajda, Jernej (Avtor) Vihar, Boštjan (Avtor) Milojević, Marko (Avtor) Bjelić, Dragana (Avtor) Brečko, Amadeja (Avtor) Maver, Uroš (Avtor) biomacromoleculesendothelial cell typesmaterialsmicrovascularizationvascularizationissue engineering faces the challenge of achieving effective vascularization within tissue constructs for sustained viability andoptimal function. The success of tissue-engineered constructs depends on selecting an optimal angiogenesis-stimulating ECMsubstitute material. This study compares four substrates made from three different biomacromolecules—fibrin, fibronectin,non-crosslinked, and crosslinked gelatin, and their effect on endothelial cells. Acknowledging the diverse range of endothelialcells that play a role in (micro)vascularization, human endothelial primary cells, human umbilical vein endothelial cells, andhuman microvascular endothelial cells are subjected to these materials for evaluation. Biocompatibility is assessed by measuringcell viability (Live/Dead assay), metabolic activity (alamarBlue assay), morphology (actin staining), phenotype expression(immunocytochemistry), and the production of von Willebrand factor, which promotes angiogenesis by promoting cell adhesionand migration. The results show that the use of biomaterials as culturing substrates significantly impacts the viability andmorphology of the cells. While the expression of angiogenic markers is shown to rely more on the cell lineage, the use of differentsubstrates has an impact on the expression timeline. Thus, combining cells and biomaterials in a favorable manner can be usedas a powerful tool for controlled vascularization in vitro, which requires the systematic assembly of different stimuli.Wiley-VCH20252025-07-16 03:09:39Članek v reviji93711UDK: 61COBISS_ID: 242531843DOI: 10.1002/mabi.202500333ISSN pri članku: 1616-5187sl© 2025 The Author(s).