Retinoic acid increases the effect of bone morphogenetic protein type 2 on osteogenic differentiation of human adipose-derived stem cells

Abstract

Bone morphogenetic protein type 2 (BMP-2) and retinoic acid (RA) are osteoinductive factors that stimulate endogenous mechanisms of bone repair which can be applied on management of osseous defects in oral and maxillofacial fields. Objective Considering the different results of RA on osteogenesis and its possible use to substitute/potency BMP-2 effects, this study evaluated the outcomes of BMP-2, RA, and BMP-2+RA treatments on i n vitro osteogenic differentiation of human adipose-derived stem cells (ASCs) and the signaling pathway(s) involved. Material and Methods ASCs were treated every other day with basic osteogenic medium (OM) alone or supplemented with BMP-2, RA, or BMP-2+RA. Alkaline phosphatase (ALP) activity was determined using the r-nitrophenol method. Extracellular matrix mineralization was evaluated using von Kossa staining and calcium quantification. Expression of osteonectin and osteocalcin mRNA were determined using qPCR. Smad1, Smad4, phosphorylated Smad1/5/8, BMP-4, and BMP-7 proteins expressions were analyzed using western blotting. Signaling pathway was evaluated using the IPA ® software. Results RA promoted the highest ALP activity at days 7, 14, 21, and 28, in comparison to BMP-2 and BMP-2+RA. BMP-2+RA best stimulated phosphorylated Smad1/5/8 protein expression at day 7 and Smad4 expression at days 7, 14, 21, and 28. Osteocalcin and osteonectin mRNA expressions were best stimulated by BMP-2+RA at day 7. Matrix mineralization was most improved by BMP-2+RA at days 12 and 32. Additionally, BMP-2+RA promoted the highest BMP signaling pathway activation at days 7 and 14, and demonstrated more activation of differentiation of bone-forming cells than OM alone. Conclusions In summary, RA increased the effect of BMP-2 on osteogenic differentiation of human ASCs.

Effect of dexamethasone as osteogenic supplementation in in vitro osteogenic differentiation of stem cells from human exfoliated deciduous teeth

“…Tests were performed to determine DNA content and differentiation into an osteogenic-like lineage. Experimental control was performed using murine preosteoblasts MC3T3-E1 subclone 4 (American Type Culture Collection, CRL 2593) cultured with modified α-Minimum Eagles’ Medium (Gibco, Thermo Fisher Scientific), combining 10% FBS, 50-µM ASC, and 10-mM βGLY [ 19 ].…”

Bone morphogenetic protein 2‑enhanced osteogenic differentiation of stem cell spheres by regulation of Runx2 expression

“…In a previous study, researchers sequentially applied BMP-2 and basic fibroblast growth factor to achieve synergistic promotion of osteogenesis ( 32 ). Similarly, retinoic acid has been shown to increase the effect of BMP-2 on osteogenic differentiation of stem cells ( 33 ). BMP-2 and Forkhead c2 synergistically enhance osteogenic differentiation and mineralization of stem cells ( 34 ).…”

Retinoic Acid Signal Negatively Regulates Osteo/Odontogenic Differentiation of Dental Pulp Stem Cells

“…In contrast, some studies have suggested a positive role for the RA pathway in osteogenic differentiation of MSCs [ 54 – 56 ] and dental pulp cells [ 57 – 59 ]. However, their in vitro induction time was mostly shorter than 7 days while osteogenesis is a long and dynamic process.…”