224 Chapter 9 Figure 2: Changes in the global fiber organization 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 global fiber characteristics and compared with their corresponding empty hydrogel samples. A) Schematic representation and example fluorescence images of high and low percentages of high-density matrix (HDM). B) Day 7 response analysis for HDM (% area), C) Day 14 response analysis for HDM (% area), D) Schematic representation and example fluorescence images of high and low percentages of fiber alignment. E) Day 7 response analysis for fiber alignment (% fibers), F) Day 14 response analysis for fiber alignment (% fibers). The dotted line shows the intrinsic difference between the 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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