Proangiogenic Hypoxia-Mimicking Agents Attenuate Osteogenic Potential of Adipose Stem/Stromal Cells.
Adipose tissue-derived mesenchymal stem/stromal cells
Baicalein
Dimethyloxalylglycine
Hypoxia-inducible factor-1alpha
Osteogenesis
Journal
Tissue engineering and regenerative medicine
ISSN: 2212-5469
Titre abrégé: Tissue Eng Regen Med
Pays: Korea (South)
ID NLM: 101699923
Informations de publication
Date de publication:
08 2020
08 2020
Historique:
received:
01
11
2019
accepted:
31
03
2020
revised:
27
03
2020
pubmed:
26
5
2020
medline:
18
9
2021
entrez:
26
5
2020
Statut:
ppublish
Résumé
Insufficient vascularization hampers bone tissue engineering strategies for reconstructing large bone defects. Delivery of prolyl-hydroxylase inhibitors (PHIs) is an interesting approach to upregulate vascular endothelial growth factor (VEGF) by mimicking hypoxic stabilization of hypoxia-inducible factor-1alpha (HIF-1α). This study assessed two PHIs: dimethyloxalylglycine (DMOG) and baicalein for their effects on human adipose tissue-derived mesenchymal stem/stromal cells (AT-MSCs). Isolated AT-MSCs were characterized and treated with PHIs to assess the cellular proliferation response. Immunostaining and western-blots served to verify the HIF-1α stabilization response. The optimized concentrations for long-term treatment were tested for their effects on the cell cycle, apoptosis, cytokine secretion, and osteogenic differentiation of AT-MSCs. Gene expression levels were evaluated for alkaline phosphatase (ALPL), bone morphogenetic protein 2 (BMP2), runt-related transcription factor 2 (RUNX2), vascular endothelial growth factor A (VEGFA), secreted phosphoprotein 1 (SPP1), and collagen type I alpha 1 (COL1A1). In addition, stemness-related genes Kruppel-like factor 4 (KLF4), Nanog homeobox (NANOG), and octamer-binding transcription factor 4 (OCT4) were assessed. PHIs stabilized HIF-1α in a dose-dependent manner and showed evident dose- and time dependent antiproliferative effects. With doses maintaining proliferation, DMOG and baicalein diminished the effect of osteogenic induction on the expression of RUNX2, ALPL, and COL1A1, and suppressed the formation of mineralized matrix. Suppressed osteogenic response of AT-MSCs was accompanied by an upregulation of stemness-related genes. PHIs significantly reduced the osteogenic differentiation of AT-MSCs and rather upregulated stemness-related genes. PHIs proangiogenic potential should be weighed against their longterm direct inhibitory effects on the osteogenic differentiation of AT-MSCs.
Sections du résumé
BACKGROUND
Insufficient vascularization hampers bone tissue engineering strategies for reconstructing large bone defects. Delivery of prolyl-hydroxylase inhibitors (PHIs) is an interesting approach to upregulate vascular endothelial growth factor (VEGF) by mimicking hypoxic stabilization of hypoxia-inducible factor-1alpha (HIF-1α). This study assessed two PHIs: dimethyloxalylglycine (DMOG) and baicalein for their effects on human adipose tissue-derived mesenchymal stem/stromal cells (AT-MSCs).
METHODS
Isolated AT-MSCs were characterized and treated with PHIs to assess the cellular proliferation response. Immunostaining and western-blots served to verify the HIF-1α stabilization response. The optimized concentrations for long-term treatment were tested for their effects on the cell cycle, apoptosis, cytokine secretion, and osteogenic differentiation of AT-MSCs. Gene expression levels were evaluated for alkaline phosphatase (ALPL), bone morphogenetic protein 2 (BMP2), runt-related transcription factor 2 (RUNX2), vascular endothelial growth factor A (VEGFA), secreted phosphoprotein 1 (SPP1), and collagen type I alpha 1 (COL1A1). In addition, stemness-related genes Kruppel-like factor 4 (KLF4), Nanog homeobox (NANOG), and octamer-binding transcription factor 4 (OCT4) were assessed.
RESULTS
PHIs stabilized HIF-1α in a dose-dependent manner and showed evident dose- and time dependent antiproliferative effects. With doses maintaining proliferation, DMOG and baicalein diminished the effect of osteogenic induction on the expression of RUNX2, ALPL, and COL1A1, and suppressed the formation of mineralized matrix. Suppressed osteogenic response of AT-MSCs was accompanied by an upregulation of stemness-related genes.
CONCLUSION
PHIs significantly reduced the osteogenic differentiation of AT-MSCs and rather upregulated stemness-related genes. PHIs proangiogenic potential should be weighed against their longterm direct inhibitory effects on the osteogenic differentiation of AT-MSCs.
Identifiants
pubmed: 32449039
doi: 10.1007/s13770-020-00259-3
pii: 10.1007/s13770-020-00259-3
pmc: PMC7392999
doi:
Substances chimiques
COL1A1 protein, human
0
Collagen Type I, alpha 1 Chain
0
KLF4 protein, human
0
Kruppel-Like Factor 4
0
Vascular Endothelial Growth Factor A
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
477-493Subventions
Organisme : Ministry of Higher Education, Egypt
ID : 290243/2014-2015
Pays : International
Organisme : Helsingin Yliopisto
ID : WBS490302
Pays : International
Organisme : Helsingin Yliopisto
ID : WBS73714112
Pays : International
Organisme : Helsingin ja Uudenmaan Sairaanhoitopiiri
ID : Y1014SUL05
Pays : International
Organisme : Helsingin ja Uudenmaan Sairaanhoitopiiri
ID : TYH2016130
Pays : International
Références
Plast Reconstr Surg. 2017 Mar;139(3):695e-706e
pubmed: 28234841
J Clin Invest. 2015 Sep;125(9):3606-18
pubmed: 26280574
Tissue Eng. 2007 Dec;13(12):2827-37
pubmed: 18052823
Cytotherapy. 2013 Jun;15(6):641-8
pubmed: 23570660
Cells Tissues Organs. 2017;204(2):59-83
pubmed: 28647733
Bone Marrow Transplant. 2012 Jul;47(7):997-1000
pubmed: 21986637
Bone. 2006 Sep;39(3):513-22
pubmed: 16616713
PLoS One. 2015 Jul 14;10(7):e0132572
pubmed: 26172833
Front Bioeng Biotechnol. 2017 Nov 03;5:68
pubmed: 29164110
J Plast Reconstr Aesthet Surg. 2013 Nov;66(11):1494-503
pubmed: 23845909
Mol Biol Cell. 2008 Jan;19(1):86-94
pubmed: 17942596
Stem Cells Int. 2018 Mar 11;2018:9468085
pubmed: 29713352
Trends Biotechnol. 2008 Aug;26(8):434-41
pubmed: 18585808
PLoS One. 2014 Sep 17;9(9):e107044
pubmed: 25229501
Tissue Eng Part B Rev. 2012 Aug;18(4):246-57
pubmed: 22029448
Oncol Lett. 2018 Jan;15(1):1119-1124
pubmed: 29391899
Cytotherapy. 2009;11(7):958-72
pubmed: 19903107
Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5510-4
pubmed: 7539918
Mol Med Rep. 2017 Nov;16(5):6459-6466
pubmed: 28901500
Acta Biomater. 2013 Nov;9(11):9159-68
pubmed: 23811216
Br J Oral Maxillofac Surg. 2011 Jun;49(4):314-8
pubmed: 20554359
Mol Pharmacol. 2008 Jul;74(1):70-81
pubmed: 18426858
Phytother Res. 2014 Sep;28(9):1342-8
pubmed: 24596136
Cell Physiol Biochem. 2016;39(4):1391-403
pubmed: 27606625
J Vis Exp. 2011 Aug 12;(54):
pubmed: 21860378
PLoS One. 2013 Jun 11;8(6):e65940
pubmed: 23776575
J Clin Invest. 2007 Jun;117(6):1616-26
pubmed: 17549257
J Cell Physiol. 2018 Dec;233(12):9777-9785
pubmed: 30078218
Bone. 2007 Apr;40(4):1078-87
pubmed: 17276151
Cell Physiol Biochem. 2018;49(5):2035-2046
pubmed: 30244250
Stem Cells Dev. 2014 May 1;23(9):990-1000
pubmed: 24328551
J Cell Physiol. 2001 Jun;187(3):345-55
pubmed: 11319758
J Cell Biochem. 2012 Apr;113(4):1407-15
pubmed: 22135004
Cytotherapy. 2006;8(4):315-7
pubmed: 16923606
Adv Drug Deliv Rev. 2011 Apr 30;63(4-5):300-11
pubmed: 21396416
Biochem Biophys Res Commun. 2014 May 30;448(2):218-24
pubmed: 24785372
Mol Med Rep. 2017 Oct;16(4):5515-5522
pubmed: 28849067
Methods. 2001 Dec;25(4):402-8
pubmed: 11846609
Theranostics. 2012;2(6):541-52
pubmed: 22737191
Connect Tissue Res. 2018 Jan;59(1):12-20
pubmed: 28165799
Lab Invest. 2011 Dec;91(12):1684-94
pubmed: 21876537
J Biomater Appl. 2011 Nov;26(4):383-417
pubmed: 21926148
Mol Cell Biol. 1992 Dec;12(12):5447-54
pubmed: 1448077
Proc Natl Acad Sci U S A. 2014 Nov 4;111(44):15768-73
pubmed: 25324523
Eur Cell Mater. 2008 May 02;15:100-14
pubmed: 18454418
Int J Oral Maxillofac Surg. 2009 Mar;38(3):201-9
pubmed: 19168327
J Biol Chem. 2004 Sep 17;279(38):40007-16
pubmed: 15263007
Nature. 2000 Sep 14;407(6801):249-57
pubmed: 11001068
PLoS One. 2012;7(9):e46483
pubmed: 23029528