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

Fig. 3. In vitro macroscopic, biochemical, and histological evaluation of engineered hMSC condensate tube maturation.

Fig. 3

a Representative gross macroscopic images of hMSC tubes containing TGF-β1-loaded, BMP-2-loaded, or TGF-β1 + BMP-2-loaded microparticles at week 5. Scale bar, 1 mm. b, c Quantification of hMSC tube wall width and height (n = 3 biologically independent samples per group; black circles = TGF-β1; red squares = BMP-2; blue triangles = TGF-β1 + BMP-2). d, e Representative histological Hematoxylin & Eosin (H&E), Safranin-O/Fast-green (Saf-O), and Alizarin Red S (ARS) staining of transaxial and sagittal hMSC tube sections. Scale bars, 100 μm (dotted squares in insets show region of interest in high magnification image). Quantification of f GAG/DNA content, g Ca2+/DNA content, h ALP activity/DNA, i DNA content, j GAG content, k Ca2+ content; dashed line represents the amount of Ca2+ initially contributed by mineral-coated hydroxyapatite microparticles assuming 100% incorporation, and l ALP activity (n = 3; derived from three hMSC donors (average of averages) each with 3 (TGF-β1; BMP-2) or 4 (TGF-β1 + BMP-2) biologically independent samples; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; black circles = TGF-β1; red squares = BMP-2; blue triangles = TGF-β1 + BMP-2). Individual data points are shown with mean ± SD. Analyzed by one-way or two-way ANOVA with Tukey’s post hoc test (p < 0.05 or lower considered significant).