3D morphometry of endothelial cells angiogenesis in an extracellular matrix composite hydrogel.

Human umbilical vein endothelial cells (HUVECs) play a fundamental role in angiogenesis. Herein, we introduce digital holographic microscopy (DHM) for the 3D quantitative morphological analysis of HUVECs in extracellular matrix (ECM)-based biomaterials as an angiogenesis model. The combination of volumetric information from DHM and the physicochemical and cytobiocompatibility data provided by fluorescence microscopy and cytology offers a comprehensive understanding of the angiogenesis-related parameters of HUVECs within the ECM. DHM enables label-free, non-contact, and non-invasive 3D monitoring of living samples in real time, in a quantitative manner. In this study, the human amniotic membrane (HAM) is decellularized, pulverized, and combined with sodium alginate hydrogel to provide an in vitro substrate for modeling HUVEC angiogenesis. Our results demonstrate that modifying alginate hydrogel with HAM enhances its biofunctionality due to the presence of ECM components. Moreover, the DHM results reveal an increase in its porous properties, which, in turn, aids in interpreting the tubulation results.