Spatiotemporal analysis of pericyte-induced dynamics in nascent angiogenic morphogenesis and heterogeneity using a microvessel-on-a-chip platform

Abstract

The recent emergence of in vitro microvessel models has boosted research on angiogenesis and vascularization in tissue engineering, due to their superior control over in controlling culture conditions and enabling real-time observation compared to in vivo models. However, conventional two-dimensional (2D) observation and analysis fail to capture the heterogeneity of three-dimensional (3D) morphological dynamics. To overcome this issue, here, a novel morphological registration method for spatiotemporal quantification of angiogenic deformation dynamics by combining confocal microscopy of an engineered microvessel with computer vision techniques has been proposed in this paper. Using a coculture system of a microvessel and pericytes, the spatiotemporal measurements reveals: (i) distinct deformation patterns and growth/regression zonation on the parent vessel and angiogenic sprouts; (ii) spatiotemporal changes in pericyte localization and coverage; and (iii) the enhancing effect of pericyte-microvessel contact on local Notch signaling activation, the distribution of matrix metalloproteinase-1 (MMP-1), the heterogeneity of angiogenic dynamics, and morphological maturation. This pilot system offers a characterization of the comprehensive effects of cocultured cells during angiogenesis and enables the interactive fusion of multimodal data in future studies on vascular morphogenesis.