Quantitative analysis of fibrillar collagen organization in the immediate proximity of embedded fibroblasts in 3D collagen hydrogels
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Date
2024-06
Journal Title
Journal ISSN
Volume Title
Publisher
The Indian Academy of Sciences
Abstract
Fibroblasts embedded in a 3D matrix microenvironment can remodel the matrix to regulate cell adhesion and function. Collagen hydrogels are a useful in vitro system to study cell–matrix interactions in a 3D microenvironment. While major matrix reorganizations are easily recognizable, subtle changes in response to environmental or biochemical cues are challenging to discern in 3D hydrogels. Three-dimensional collagen gels at 1.0 mg/ml vs 1.5 mg/ml were labelled with DQ-collagen and imaged by confocal reflectance microscopy to evaluate these small changes. An image analysis pipeline was developed, hydrogel area and number of cross-sections analysed were optimized, and fibrillar collagen properties (number of branches, number of junctions, and average branch length) were quantified. While no significant changes were seen in fibrillar collagen organization between 1.0 mg/ml and 1.5 mg/ml collagen hydrogels, embedded mouse fibroblasts caused a significant increase in collagen branching and organization. Using the phalloidin-labelled cells, this change was quantitated in immediate proximity of the cell. A distinct increase in branch and junction numbers was observed, significantly altered by small changes in collagen concentration (1.0 mg/ml vs 1.5 mg/ml). Together, this analysis gives a quantitative evaluation of how cells respond to and modify their immediate microenvironment in a 3D collagen hydrogel.
Description
Keywords
Collagen organization, extracellular matrices, fibroblast, reflectance imaging, 3D hydrogel
Citation
KANADE SHAUNAK, DESAI MILIE, BHATAVADEKAR NEEL, BALASUBRAMANIAN NAGARAJ. Quantitative analysis of fibrillar collagen organization in the immediate proximity of embedded fibroblasts in 3D collagen hydrogels. Journal of Biosciences. 2024 Jun; 49: 1-11