228 Chapter 9 Figure 4: Investigation of collagen fiber curvature 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 the differences in collagen fiber curvature and compared with their corresponding empty hydrogel samples. A) Schematic representation of low and high curvature windows with respect to periodicity of peaks and peak height, B) Day 7 response analysis for fiber curvature with respect to number of peaks and peak height, C) Day 14 response analysis for fiber curvature with respect to number of peaks and peak height. 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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