Modeling ECM fiber formation: structure information extracted by analysis of 2D and 3D image sets

Recent evidence supports the notion that biological functions of extracellular matrix (ECM) are highly correlated to its structure. Understanding this fibrous structure is very crucial in tissue engineering to develop the next generation of biomaterials for restoration of tissues and organs. In this paper, we integrate confocal microscopy imaging and image-processing techniques to analyze the structural properties of ECM. We describe a 2D fiber middle-line tracing algorithm and apply it via Euclidean distance maps (EDM) to extract accurate fibrous structure information, such as fiber diameter, length, orientation, and density, from single slices. Based on a 2D tracing algorithm, we extend our analysis to 3D tracing via Euclidean distance maps to extract 3D fibrous structure information. We use computer simulation to construct the 3D fibrous structure which is subsequently used to test our tracing algorithms. After further image processing, these models are then applied to a variety of ECM constructions from which results of 2D and 3D traces are statistically analyzed.