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Fig. 2 | Stem Cell Research & Therapy

Fig. 2

From: Combined macromolecule biomaterials together with fluid shear stress promote the osteogenic differentiation capacity of equine adipose-derived mesenchymal stem cells

Fig. 2

Effect of biomaterials on the osteogenic differentiation of MSCs under static culture. MSCs were cultivated in the presence of MC, B30, and B30Str biomaterials under standard culture condition for 48 h (N = 8 per experimental group). Cells were induced to osteogenic differentiation (OD) using 5% FBS in DMEM, 0.05 mM ascorbic acid-2-phosphate, 10 mM β-glycerolphosphate, and 0.1 μM dexamethasone up to 21 days. ac Spectrophotometric quantification of alkaline phosphatase (ALP) activity at days 7, 14, and 21 shows promoted ALP activity post osteogenic induction. ALP activity was detected by measuring the metabolism of p-Nitrophenylphosphate into p-Nitrophenol (PNP) in the presence of ALP. The values of PNP were measured in triplicates at 405 nm absorbance, and the average was calculated. MSCs were grown in basal medium (BM) composed of 5% FBS in DMEM served as a negative control. Cells which were cultivated in osteogenic medium without adding biomaterials were used as negative control (NC). d, e Semi-quantification of the solubilized bound alizarin red S (ARS) using 10% cetylpyridinium chloride (CPC, Roth Germany) at days 14 and 21 post osteogenic induction (N = 8 per experimental group). The absorbance was measured in triplicates at 562 nm. Analysis of CPC revealed improved matrix mineralization as shown with MC biomaterial scaffold. fo Representative ARS stained images (red) for MSCs cultivated in basal medium (BM, f, g), osteogenic differentiation medium (OD, h, i), OD + MC (j, k), OD + B30 (l, m), and OD + B30Str (n, o) under static conditions at day 14 and day 21. Osteogenic nodules could be shown in the presence of OD medium compared to BM. Diffused ARS staining was observed in the presence of OD + MC and scattered mineralization was observed in the condition of OD + B30 and OD + B30/Str. All data presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001. Scale bar in fo = 100 μm

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