Adenoviral Mediated Expression of BMP2 by Bone Marrow Stromal Cells Cultured in 3D Copolymer Scaffolds Enhances Bone Formation
Fig 2
BMP2 over-expression led to up-regulation of osteogenic and angiogenic molecules in vitro.
5 × 104 ad-BMP2 or ad-GFP BMSC were seeded in each Poly(LLA-co-CL) scaffolds and harvested after 3 and 14 days for custom PCR array and TaqMan based qRT-PCR. (A) With SAM analysis, ALPL and FLT1 mRNA levels were found to be significantly (FDR = 0) up-regulated at 3-days in ad-BMP2 BMSC. (B) More osteogenic and angiogenic markers were significantly induced at 14-days in ad-BMP2 BMSC as compared to the ad-GFP BMSC group. (C and D) Array results for ALPL, RUNX2, BGLAP and VEGFA genes were independently validated in ad-BMP2 BMSC by using TaqMan qRT- PCR. Error bars represent SEM of 3 repeated experiments (n = 3) done in 3 technical replicates. ANOVA test with Bonferroni post hoc analysis was performed for statistical analysis in C and D. ***, p<0.001; *, P<0.01; ns, non-significant. (E) Representative images demonstrating higher alkaline phosphatase activity in ad-BMP2 BMSC in monolayer as compared to the ad-GFP BMSC and untransduced BMSC. Experiments were repeated at least three times.