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. 2021 Jan 19;4:89. doi: 10.1038/s42003-020-01576-y

Fig. 2. In vitro macroscopic, biochemical, molecular, and histological evaluation of engineered hMSC condensate tube early chondrogenic priming.

Fig. 2

a Representative gross macroscopic images of hMSC tubes containing TGF-β1-loaded, BMP-2-loaded, or TGF-β1 + BMP-2-loaded microparticles at day 8. Scale bar, 4 mm. b Normalized mRNA fold-change over control of key chondrogenic and osteogenic markers by qRT-PCR (n = 3 biologically independent samples per group; *p < 0.05 vs. control; orange circles = chondrogenic markers; purple diamonds = osteogenic markers). c Immunoblots and d relative fold-change over control of p-SMAD3/SMAD3, and e p-SMAD5/SMAD5. β-Actin served as a loading control (n = 3 biologically independent samples per group; **p < 0.01, ***p < 0.001; black circles = TGF-β1; red squares = BMP-2; blue triangles = TGF-β1 + BMP-2). f Representative histological Hematoxylin & Eosin (H&E), Safranin-O/Fast-green (Saf-O), and Alizarin Red S (ARS) staining of transaxial hMSC tube sections. Scale bars, 100 μm (dotted squares in insets show region of interest in high magnification image). Individual data points are shown with mean ± SD. Groups with shared letters have no significant differences. Analyzed by one-way ANOVA with Tukey’s post hoc test (p < 0.05 or lower considered significant).