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. 2024 Mar 29;15:2766. doi: 10.1038/s41467-024-46774-y

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — a Schematic of study design in which MFC spheroids are seeded on a gelatin substrate and additional material is placed atop to form an interface. b 3D reconstructions of the representative spheroid (magenta) from no material condition prior to (left panel) and after (right panel) spreading along the interface (gelatin: green). Scale bar = 200 μm. c, d Representative top view images of spheroids spreading along interfaces after 1 day (c, Scale bar = 200 μm) and corresponding quantification (d). n = 4 (GH), 5 (0.25 wt% Agarose), 6 (gelatin), or 7 (No Material) spheroids from 2-3 biologically independent experiments. No Material vs. Gelatin ****p ≤ 0.0001; No-Material vs. 0.25 wt% Agarose ***p = 0.0009; Gelatin vs. Guest-Host ****p ≤ 0.0001; Gelatin vs. 0.25 wt% Agarose ***p = 0.0002; Guest-Host vs. 0.25 wt% Agarose ***p = 0.0006; one-way ANOVA with Tukey post hoc. e Schematic of interface materials in which agarose particles undergo low, medium, or high densification within a continuous gelatin material. f Quantification of connectivity of gelatin (left panel), connectivity of particles (middle panel), and interfacial surface area normalized to sample volume across hydrogels (right panel). n = 9 (None) or 10 (Low, Medium, High) regions across 3 hydrogels per condition. Left panel: n.s. indicates no statistical significance. Middle panel: None vs. Low, None vs. Medium, None vs. High, Low vs. Medium, Low vs. High, ****p ≤ 0.0001. Right panel: None vs. Low, None vs. Medium, None vs. High, ****p ≤ 0.0001; Low vs. High, ***p = 0.0001; one-way ANOVA with Tukey post hoc. g, h Representative Z-sections of MFC spheroids (actin: magenta) spread throughout interface materials (gelatin: green) at day 3 (g, Scale bar = 200 μm), and corresponding quantification (h). n = 6 (None), 8 (Low, High), or 13 (Medium) spheroids per condition from 2 biologically independent experiments. None vs. Medium **p = 0.0042; None vs. High, Low vs. High, Medium vs. High, ****p ≤ 0.0001; one-way ANOVA with Tukey post hoc. Data are mean ± s.d. Source data for (d, f, h) provided as a source data file.