Fig. 3

Effect of biomaterials on the osteogenic differentiation of MSCs under FSS culture. MSCs were grown in conjunction with MC, B30, and B30Str biomaterials under standard culture condition for 48 h to establish adhesion (N = 8 per experimental group). The cells/biomaterial complex was induced to the osteogenic differentiation (OD) fate under mechanical FSS in comparison to static (ST) culture up to 21 days. In parallel, MSCs which were grown in basal medium BM were served as negative control. Cells which were cultivated in osteogenic medium without adding biomaterials were used as negative control (NC). a, b Average cell viability was measured in triplicates at day 14 and day 21 using MTT assay. The data show enhanced cell viability in the presence of OD together with MC compared to B30, B30Str, and BM condition. c, d Spectrophotometric measurement of ALP activity in triplicates at 405 nm following 10 min and 30 min incubation. e Runx2 protein western blot following combined osteogenic differentiation (OD) induction with biomaterials (NC/OD, OD/MC, OD/B30, and OD/B30Str) under static (ST) and fluid shear stress (FSS) conditions at day 14. Approximately 50 μg of cell lysates were loaded in 7.5% SDS-PAGE in a reduced condition. Protein bands were transferred into a nitrocellulose membrane and were probed with anti Runx2 and β actin antibodies. Cells which were grown in basal medium (BM/NC) were used as negative controls. f Semi-quantification of the solubilized bound alizarin red S (ARS) in triplicates using 10% CPC at day 21 post osteogenic induction. Analysis of CPC (N = 8 per experimental group) revealed improved matrix mineralization as shown with MC biomaterial and control osteogenic condition without biomaterial (NC). All data presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001