Journal: Journal of Biological Engineering
Article Title: Rapid osteoinduction of human adipose-derived stem cells grown on bioactive surfaces and stimulated by chemically modified media flow
doi: 10.1186/s13036-025-00491-2
Figure Lengend Snippet: Fluid shear stress strengthens the osteogenic effects of rhBMP-2, PD98059 and Phenamil in ASCs cultured on SBG-PLGA composites. mRNA levels of osteoblastic markers after 7-day ASC culture on SBG-PLGA composites in ( A ) standard osteogenic medium under either static conditions or with fluid shear stress; and ( C ) osteogenic medium supplemented with 100 ng/ml rhBMP-2, 50 µM PD98059 and 20 µM Phenamil under either static conditions or with fluid shear stress. Results are presented as relative mRNA expression levels compared to mRNA levels in a control, static culture on PLGA (marked as a black line at 1). ( B ) The method of fluid shear stress application in ASC cultures using a standard laboratory see-saw rocker (7° tilt angle, 6 RPM frequency). ( D ) F-actin distribution in ASCs (Phalloidin-Atto488, magenta colored) at day 3 of culture in osteogenic medium supplemented with rhBMP-2, PD98059 and Phenamil after continuous static or dynamic culture conditions applied for 3 days. Scale bar represents 100 μm. ( E ) Western blot (WB) analysis of p-ERK1/2 and p-SMAD1/5/8 in ASCs after 1-h treatment with rhBMP-2 or rhBMP-2, PD98059 and Phenamil in static or dynamic conditions (upper panel) along with densitometric quantifications of WB results normalized to GAPDH levels. Averages ± SD are indicated. Two-way ANOVA test, * p < 0.05, ** p < 0.001, *** p < 0.0001 relative to the static PLGA control or between marked groups
Article Snippet: ASC52telo cells (ASC; ATCC, SCRC-4000) and normal human ASCs (ATCC, PCS-500-011) were expanded in the dedicated medium (ATCC, Mesenchymal Stem Cell Basal Medium with Mesenchymal Stem Cell Growth Kit and G418).
Techniques: Shear, Cell Culture, Expressing, Control, Western Blot