Dimensional Metrology of Cell-matrix Interactions in 3D Microscale Fibrous Substrates

Abstract

The significant potential of engineered tissue models is bounded by the current lack of robust scalable additive biomanufacturing processes that reliably capture the cell's structural microenvironments. To address this bottleneck, a melt electrospinning writing system is designed to fabricate 3D fibrous substrates within a tight cellular dimensional scale window. The biological relevance of the produced 3D experimental substrates over its 2D monolayer controls is demonstrated with respect to cell morphology. Cell confinement states are quantitatively characterized using an automated single-cell bioimage data analysis workflow. A multidimensional data set composed of size, shape and distribution related metrics of cellular and sub-cellular focal adhesions is extracted to build a classifier that can, with 91-93% classification accuracy, distinguish cell shape phenotypes in 3D confined versus 2D unconfined cell states.