226 Chapter 9 Figure 3: Individual collagen fiber structure in empty and fibroblast-seeded control and IPF lung ECM-derived hydrogels. Control and IPF primary lung fibroblasts were seeded in control or IPF lung ECM-derived hydrogels and cultured for 7 or 14 days. Fluorescence images of PicroSirius Red (PSR) stained sections of paraffin-embedded hydrogels were analyzed for individual fiber characteristics and compared with their corresponding empty hydrogel samples. Day 7 responses with respect to A) average fiber length (AFL) (μm), B) number of endpoints per 1000 μm fiber total length, C) number of branchpoints per 1000 μm fiber total length. Day 14 responses with respect to D) average fiber length (AFL) (μm), E) number of endpoints per 1000 μm fiber total length, F) number of branchpoints per 1000 μm fiber total length. The dotted line shows the intrinsic difference between empty IPF and control hydrogels. The estimate (± 95% confidence interval) shows the difference between the IPF and control hydrogels seeded with control (light blue, triangle) or IPF (dark blue, square) fibroblasts. P values below each fibroblast group represent the differences induced by fibroblasts in IPF versus control hydrogels compared to the intrinsic difference between IPF and control empty hydrogels. P-values above the estimates indicate the differences between the responses of IPF and control fibroblasts in the different hydrogels. Applied statistical test: mixed-model analysis. ns: not significant. IPF: Idiopathic pulmonary fibrosis. n=6 for fibroblast donors, 6 images per sample were captured and analyzed.
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