Supplementary Figures from Geometric constraint of mechanosensing by modification of hydrogel thickness prevents stiffness-induced differentiation in bone marrow stromal cells
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journal contribution
posted on 2024-10-28, 07:28 authored by Maria L Hernandez-Miranda, Dichu Xu, Aya A Ben Issa, David A Johnston, Martin Browne, Richard B Cook, Bram G Sengers, Nicholas D EvansThis file provides 3 Supplementary Figures with captions shown in the file. They are:Supplementary Figure 1.Fluorescent microbeads are distributed evenly in stiff hydrogels but show alignment with gel wrinkles in soft hydrogels (A). Microbead incorporation did not affect the thickness of either thin or thick hydrogels, except for in 50 uL soft hydrogels. Three hydrogels per condition were imaged using a confocal microscope (B). Microbead incorporation had little effect on corrected elastic modulus on either soft (C) or thick (D) hydrogels. Statistical differences were calculated by the 2-way ANOVA method (****=p<0.0001). Three soft or stiff hydrogels per PAAM volume were used to measure hydrogel thickness from top to bottom using a confocal microscope.Supplementary Figure 2Addition of osteogenic supplements leads to a significant increase in Ca2+ deposition at 14 days, as measured by alizarin red S staining and image quantification.Supplementary Figure 3Stiff matrices exhibit small deformations by BMSCs regardless of hydrogel thickness. Small coloured arrows show small deformations on stiff, thick hydrogels compared to their thin counterparts. Phase contrast (BF) and fluorescent (Cy3) images were obtained at 10X magnification under a Nikon Eclipse Ti inverted microscope. One microscopic field per stiff/thin, stiff/thick hydrogel (n=3) was imaged. Scale bar = 100 um.
