Figure 3
Rab27a knockdown in PDLSC-attenuated co-culture-enhanced bone marrow mesenchymal stromal cell (BMMSC) osteogenesis. (A) Schematic diagram of a non-contact co-culture system in which BMMSCs were cultured separately with PDLSCsiControl or PDLSCsiRab27a through a porous membrane (pore size 0.4 µm). (B) Osteogenic gene expression of BMMSCs in the Transwell system after 3 d of co-culture. *p < 0.05, **p < 0.01, as compared with the control group; #p < 0.05, ##p < 0.01, as compared with the PDLSCsiControl co-culture group (n = 3). (C) Representative Alizarin Red-S (ARS)-stained images of BMMSCs in the Transwell system after 3 weeks of co-culture. Scale bar = 200 µm. ARS intensity was evaluated using densitometry at 562 nm. **p < 0.01, as compared with the control group; #p < 0.05, as compared with the PDLSCsiControl co-culture group (n = 3). (D) Western blotting analysis revealed that co-culture with PDLSCsiControl enhanced RUNX2 expression by 1.41-fold and ALP expression by 1.38-fold. Rab27a knockdown in PDLSCs attenuated the co-culture-enhanced osteogenesis of BMMSCs. *p < 0.05 and **p < 0.01, as compared to the control group; #p < 0.05, as compared to the PDLSCsiControl co-culture group. Representative results from triplicate experiments are expressed as mean ± standard deviation.
