Fig. 4 Effects of morphogen priming of engineered mesenchymal condensations on in vitro chondrogenic lineage specification at the time of implantation.

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Fig. 4 Effects of morphogen priming of engineered mesenchymal condensations on in vitro chondrogenic lineage specification at the time of implantation. Effects of morphogen priming of engineered mesenchymal condensations on in vitro chondrogenic lineage specification at the time of implantation. Histological Safranin-O/Fast green staining of representative microparticle-containing hMSC sheets at the time of implantation (2 days; n = 3 per group). Scale bars, 100 μm (top, x10; bottom, x40 magnification of dotted squares). (B) Normalized mRNA fold change over control of key chondrogenic or osteogenic markers by quantitative reverse transcription polymerase chain reaction (qRT-PCR; n = 3 per group; *P < 0.05, **P < 0.01, ***P < 0.001 versus empty/control; $P < 0.05 versus BMP-2–containing hMSC sheets). (C) Representative immunoblots and (D) relative quantification of phosphorylated SMAD5 (p-SMAD5)/SMAD5 and (E) p-SMAD3/SMAD3 in lysates of day 2 hMSC sheets (n = 3 per group). β-Actin served as the loading control. Individual data points are shown as means ± SD. Comparisons between groups were evaluated by one-way ANOVA with Tukey’s post hoc tests. Repeated significance indicator letters (a, b, and c) signify P > 0.05, while groups with distinct indicators signify P < 0.05. S. Herberg et al. Sci Adv 2019;5:eaax2476 Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).