Osteoinductive effects of glyceollins on adult mesenchymal stromal/stem cells from adipose tissue and bone marrow.
BACKGROUND
While current therapies for osteoporosis focus on reducing bone resorption, the development of therapies to regenerate bone may also be beneficial. Promising anabolic therapy candidates include phytoestrogens, such as daidzein, which effectively induce osteogenesis of adipose-derived stromal cells (ASCs) and bone marrow stromal cells (BMSCs).


OBJECTIVE
To investigate the effects of glyceollins, structural derivatives of daidzein, on osteogenesis of ASCs and BMSCs.


METHODS
Herein, the osteoinductive effects of glyceollin I and glyceollin II were assessed and compared to estradiol in ASCs and BMSCs. The mechanism by which glyceollin II induces osteogenesis was further examined.


METHODS
The ability of glyceollins to promote osteogenesis of ASCs and BMSCs was evaluated in adherent and scaffold cultures. Relative deposition of calcium was analyzed using Alizarin Red staining, Bichinchoninic acid Protein Assay, and Alamar Blue Assay. To further explore the mechanism by which glyceollin II exerts its osteoinductive effects, docking studies of glyceollin II, RNA isolation, cDNA synthesis, and quantitative RT-PCR (qPCR) were performed.


RESULTS
In adherent cultures, ASCs and BMSCs treated with estradiol, glyceollin I, or glyceollin II demonstrated increased calcium deposition relative to vehicle-treated cells. During evaluation on PLGA scaffolds seeded with ASCs and BMSCs, glyceollin II was the most efficacious in inducing ASC and BMSC osteogenesis compared to estradiol and glyceollin I. Dose-response analysis in ASCs and BMSCs revealed that glyceollin II has the highest potency at 10nM in adherent cultures and 1µM in tissue scaffold cultures. At all doses, osteoinductive effects were attenuated by fulvestrant, suggesting that glyceollin II acts at least in part through estrogen receptor-mediated pathways to induce osteogenesis. Analysis of gene expression demonstrated that, similar to estradiol, glyceollin II induces upregulation of genes involved in osteogenic differentiation.


CONCLUSIONS
The ability of glyceollin II to induce osteogenic differentiation in ASCs and BMSCs indicates that glyceollins hold the potential for the development of pharmacological interventions to improve clinical outcomes of patients with osteoporosis.
Center for Stem Cell Research and Regenerative Medicine, Tulane University School of Medicine, New Orleans, LA, USA.
Marjorie E
ME
Bateman
Center for Stem Cell Research and Regenerative Medicine, Tulane University School of Medicine, New Orleans, LA, USA.
Amy L
AL
Strong
Center for Stem Cell Research and Regenerative Medicine, Tulane University School of Medicine, New Orleans, LA, USA.
Ryan S
RS
Hunter
Cell and Molecular Biology Core Facility, Xavier University of Louisiana, New Orleans, LA, USA.
Melyssa R
MR
Bratton
Department of Chemistry, Xavier University of Louisiana, New Orleans, LA, USA.
Rajesh
R
Komati
Department of Chemistry, Xavier University of Louisiana, New Orleans, LA, USA.
Jayalakshmi
J
Sridhar
Department of Chemistry, Xavier University of Louisiana, New Orleans, LA, USA.
Kevin E
KE
Riley
Department of Chemistry, Xavier University of Louisiana, New Orleans, LA, USA.
Guangdi
G
Wang
Department of Biological and Agricultural Engineering, Louisiana State University and Agricultural Center, Baton Rouge, LA, USA.
Daniel J
DJ
Hayes
Southern Regional Research Center, US Department of Agriculture, 1100 Robert E. Lee Blvd, New Orleans, LA, USA.
Stephen M
SM
Boue
Department of Medicine, Tulane University School of Medicine, New Orleans, LA, USA.
Matthew E
ME
Burow
Center for Stem Cell Research and Regenerative Medicine, Tulane University School of Medicine, New Orleans, LA, USA; Department of Pharmacology, Tulane University School of Medicine, New Orleans, LA, USA; Division of Regenerative Medicine, Tulane National Primate Research Center, Tulane University, Covington, LA USA. Electronic address: bbunnell@tulane.edu.
Bruce A
BA
Bunnell
D000273:Adipose Tissue / Q000187:drug effects*; D000328:Adult; D001854:Bone Marrow Cells / Q000187:drug effects*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured / Q000187:drug effects; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000187:drug effects*; D008875:Middle Aged; D010012:Osteogenesis / Q000187:drug effects*; D010024:Osteoporosis / Q000188:drug therapy*; D048789:Phytoestrogens / Q000494:pharmacology; D036343:Pterocarpans / Q000494:pharmacology*; D013025:Glycine max / Q000737:chemistry; D013234:Stem Cells / Q000187:drug effects*; D014481:United States
D048789:Phytoestrogens; D036343:Pterocarpans; D004958:Estradiol
Adipose tissue; Bone marrow; Glyceollin; Mesenchymal stem cells; Osteogenesis; Tissue scaffolds