Fig. 5

A short ASC preculture on ZnO- or SrO-modified SBG-PLGA growth surfaces, combined with BMP-based chemical treatment and fluid shear stress, leads to robust mineral deposition in further cultures on typical tissue culture plates. (A) The mRNA levels of OPG and COL1A1 in 18-day ASC cultures on PLGA-based composites containing either unmodified or SrO- or ZnO-modified SBGs. Cells were treated with 100 ng/ml rhBMP-2, 50 µM PD98059 and 20 µM Phenamil in either static conditions or with fluid shear stress. The upper panel shows the scheme of ASC treatment. Results are presented as relative mRNA expression vs. mRNA levels in a control, static culture on PLGA (marked as a black line at 1). (B) Mineral levels (Alizarin Red S staining) in ASC cultures after 7-day preculture on the indicated composites followed by 12-day culture in standard tissue culture plates (left panel). The upper panel shows the scheme of ASC treatment. Colorimetric quantification of Alizarin Red S staining normalized to the number of live cells (right panel). Averages ± SD are indicated. Two-way ANOVA test, *p < 0.05, **p < 0.001, ***p < 0.0001 relative to the PLGA static control