Elucidating the effect of deferoxamine, a hypoxia mimetic agent, on angiogenesis restoration in endothelial progenitor cells (EPCs) from diabetic mice

Document Type : Original Article

Authors

1 Department of Pharmacology and Razi Drug Research Center, School of Medicine, Iran University of Medical Sciences, Tehran, Iran

2 Department of Clinical Pathology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

3 Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran

4 Tissue Engineering Group (NOCERAL), Department of Orthopedic Surgery, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia

5 Department of Pharmacology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran

6 Stem cell and Regenerative Medicine Research Center, Iran University of Medical Sciences, Tehran, Iran

10.22038/ijbms.2025.81969.17737

Abstract

Objective(s): Diabetes increases the risk of heart disease and stroke, primarily through endothelial cell dysfunction and vascular damage. These vascular complications are partly due to defects in endothelial progenitor cells (EPCs). This study explores the efficacy of pharmacological priming of bone marrow EPCs (BMEPCs) with Deferoxamine (DFO), a hypoxia mimetic agent, in restoring dysregulated angiogenic pathways in streptozotocin (STZ)-induced mice with type-1 diabetes (T1D).
Materials and Methods: BMEPCs were isolated from both normal and STZ-induced mice with T1D. The effects of an optimal concentration of DFO (80 µM) on the viability, proliferation, and tubulogenesis of EPCs were assessed. Furthermore, the probable beneficial effects of the conditioned medium from EPCs treated in the presence and absence of DFO were examined in mice (T1D) wound healing models.
Results: DFO (80 µM) increased cell viability, proliferation, and tubulogenesis. EPCs isolated from diabetic mice showed significant impairments in the expression of HIF-1α, VEGF, and SDF-1 proteins compared to controls. DFO-preconditioning significantly enhanced protein expression of these genes. The conditioned medium from diabetic EPCs treated with DFO had a substantially greater favorable effect on wound healing in diabetic mice, connected with elevated levels of HIF-1α, VEGF, phosphorylated Tie2/Tie2, and Ang1.
Conclusion: DFO reactivates proliferation and restores the impaired angiogenic properties of EPCs from diabetic mice by stabilizing HIF-1α and VEGF. Additionally, DFO enhanced the pro-angiogenic activity in the EPC-secretome, leading to improved wound healing. This improvement is attributed to the dual activation of HIF-1α /VEGF and Ang-1/Tie2 pathways, which are crucial for initiating and maturing new blood vessels.

Keywords

Main Subjects

References

1. Vulesevic B, McNeill B, Geoffrion M, Kuraitis D, McBane JE, Lochhead M, et al. Glyoxalase-1 overexpression in bone marrow cells reverses defective neovascularization in STZ-induced diabetic mice. Cardiovasc Res 2013; 101: 306-316
2. Folkman J. Angiogenesis: An organizing principle for drug discovery? Nat Rev Drug Discov 2007; 6: 273-286
3. Xu L, Kanasaki K, Kitada M, and Koya D. Diabetic angiopathy and angiogenic defects. Fibrogenesis Tissue Repair 2012; 5: 13-22.
4. Botusan IR, Sunkari VG, Savu O, Catrina AI, Grünler J, Lindberg S, et al. Stabilization of HIF-1α is critical to improve wound healing in diabetic mice. Proc Natl Acad Sci U S A 2008; 105: 19426-19431
5. Chen J-X, Stinnett A. Disruption of Ang-1/Tie-2 signaling contributes to the impaired myocardial vascular maturation and angiogenesis in type II diabetic mice. Arterioscler Thromb Vasc Biol 2008; 28: 1606-1613
6. Hernandez SL, Gong JH, Chen L, Wu I-H, Sun JK, Keenan HA, et al. Characterization of circulating and endothelial progenitor cells in patients with extreme-duration type 1 diabetes. Diabetes Care 2014; 37: 2193-2201.
7. Liu H, Yu S, Zhang H, Xu J. Angiogenesis impairment in diabetes: role of methylglyoxal-induced receptor for advanced glycation endproducts, autophagy and vascular endothelial growth factor receptor 2. PLoS One 2012; 7: e46720-46734.
8. Westerweel PE, Teraa M, Rafii S, Jaspers JE, White IA, Hooper, AT, et al. Impaired endothelial progenitor cell mobilization and dysfunctional bone marrow stroma in diabetes mellitus. PLoS One 2013; 8: e60357-60365.
9. Mehrabani M, Najafi M, Kamarul T, Mansouri K, Iranpour M, Nematollahi M, et al. Deferoxamine preconditioning to restore impaired HIF‐1α‐mediated angiogenic mechanisms in adipose‐derived stem cells from STZ‐induced type 1 diabetic rats. Cell prolif 2015; 48:532-549.
10. Siavashi V, Nassiri SM, Rahbarghazi R, Vafaei R, Sariri R. ECM‐dependence of endothelial progenitor cell features. J Cell Biochem 2016;117: 1934-1946.
11. Niederseer D, Steidle-Kloc E, Mayr M, Müller EE, Cadamuro J, Patsch W, et al.  Effects of a 12-week alpine skiing intervention on endothelial progenitor cells, peripheral arterial tone and endothelial biomarkers in the elderly. Int J Cardiol 20016; 214: 343-347.
12. Siavashi V, Asadian S, Sharifi A, Esmaeilivand M, Norouzinia R, Azadbakht M, Nassiri SM. Circulation enrichment of functional endothelial progenitor cells by infantile phototherapy. J Cell Biochem 2017; 118: 330-340.
13. Siavashi V, Asadian S, Taheri‐Asl M, Keshavarz S, Zamani‐Ahmadmahmudi M, Nassiri SM. Endothelial progenitor cell mobilization in preterm infants with sepsis is associated with improved survival. J Cell Biochem 2017; 118: 3299-3307. 
14.  Jabarpour M, Siavashi V, Asadian S, Babaei H, Jafari SM, Nassiri SM. Hyperbilirubinemia-induced pro-angiogenic activity of infantile endothelial progenitor cells. Microvasc Res 2018; 118: 49-56.
15. Thum T, Fraccarollo D, Schultheiss M, Froese S, Galuppo P, Widder JD, et al. Endothelial nitric oxide synthase uncoupling impairs endothelial progenitor cell mobilization and function in diabetes. Diabetes 2007; 56: 666-674.
16. Hu X, Yu SP, Fraser JL, Lu Z, Ogle ME, Wang J-A, et al. Transplantation of hypoxia-preconditioned mesenchymal stem cells improves infarcted heart function via enhanced survival of implanted cells and angiogenesis. J Thorac Cardiovasc Surg 2008; 135: 799-808.
17.  Doorn J, Fernandes HA, Le BQ, van de Peppel J, van Leeuwen JP, De Vries MR, et al. A small molecule approach to engineering vascularized tissue. Biomater 2013; 34: 3053-3063.
18.  Mirzamohammadi S, Aali E, Najafi R, Kamarul T, Mehrabani M, Aminzadeh A, et al. Effect of 17β-estradiol on mediators involved in mesenchymal stromal cell trafficking in cell therapy of diabetes. Cytotherapy 2015; 17: 46-57.
19. Tajali R, Eidi A, Ahmadi-Tafti H, Pazouki A, Sharifi AM. Restoring the angiogenic capacity of the human diabetic adipose-derived mesenchymal stem cells primed with deferoxamine as a hypoxia mimetic agent: Role of HIF-1α. Adv Pharm Bull 2013; 13: 350-360.
20.  Wang C, Cai Y, Zhang Y, Xiong Z, Li G, and Cui L. Local injection of deferoxamine improves neovascularization in ischemic diabetic random flap by increasing HIF-1α and VEGF expression. PloS one 2014; 9: e100818-100826.
21. Zhu Y, Chang B, Pang Y, Wang H, Zhou Y. Advances in HIF-1α stabilizer deferoxamine in tissue engineering. Tissue Eng Part B Rev 2022; 29: 347-357.
22. Chang EI, Loh SA, Ceradini DJ, Chang EI, Lin S-e, Bastidas N, et al. Age decreases endothelial progenitor cell recruitment through decreases in hypoxia-inducible factor 1α stabilization during ischemia. Circ 2007; 116: 2818-2829.
23. Short WD, Emily T, Steen H, Balaj S. IL‐10 promotes endothelial progenitor cell infiltration and wound healing via STAT3. FASEB J 2022; 36: e22298-22318.
24. Fagiani E, Christofori G. Angiopoietins in angiogenesis. Cancer Lett 2013; 328: 18-26.
25. Graham ML, Janecek JL, Kittredge JA, Hering BJ, Schuurman H-J. The streptozotocin-induced diabetic nude mouse model: Differences between animals from different sources. Comp Med 2011; 61: 356-360.
26. Keshavarz S, Nassiri SM, Siavashi V, Alimi NS. Regulation of plasticity and biological features of endothelial progenitor cells by MSC-derived SDF-1. Biochim Biophys Acta 2019; 1866: 296-304.
27. Vafaei R, Nassiri SM, Siavashi V. β3‐adrenergic regulation of EPC features through manipulation of the bone marrow MSC niche. J Cell Biochem 2017;118: 4753-4761.
28. Hassanpour M, Cheraghi O, Siavashi V, Rahbarghazi R, Nouri M. A reversal of age-dependent proliferative capacity of endothelial progenitor cells from different species origin in in vitro condition. J Thorac Cardiovasc Res 2016; 8: 102-106.
29. Mohammadi E, Nassiri SM, Rahbarghazi R, Siavashi V, Araghi A. Endothelial juxtaposition of distinct adult stem cells activates angiogenesis signaling molecules in endothelial cells. Cell Tissue Res 2015; 362: 597-609.
30. Rahbarghazi R, Nassiri SM, Khazraiinia P, Kajbafzadeh A-M, Ahmadi SH, Mohammadi E, et al. Juxtacrine and paracrine interactions of rat marrow-derived mesenchymal stem cells, muscle-derived satellite cells, and neonatal cardiomyocytes with endothelial cells in angiogenesis dynamics. Stem Cells Dev 2012; 22: 855-865.
31. Salter AB, Meadows SK, Muramoto GG, Himburg H, Doan P, Daher P, et al. Endothelial progenitor cell infusion induces hematopoietic stem cell reconstitution in vivo. Blood 2009;113: 2104-2107.
32. Siavashi V, Sariri R, Nassiri SM, Esmaeilivand M, Asadian S, Cheraghi H, et al. Angiogenic activity of endothelial progenitor cells through angiopoietin-1 and angiopoietin-2. Anim Cells Syst  2016; 20:118-129.
33. Li Y-F, Ren L-N, Guo G, Cannella LA, Chernaya V, Samuel S, et al. Endothelial progenitor cells in ischemic stroke: an exploration from hypothesis to therapy. J Hematol Oncol 2015; 8: 33-50.
34. Efimenko AY, Kochegura TN, Akopyan ZA, Parfyonova YV. Autologous stem cell therapy: How aging and chronic diseases affect stem and progenitor cells. Biores Open Access 2015; 4: 26-38.
35. Yao YL, Zhu W, Cheng M, Zhang J, Phillips MI, Qin G. Transplantation of hypoxia-preconditioned cardiac stem cells improves infarcted heart function via Cxcr4 upregulation and improved survival of implanted cells. Circ 2008; 118: 280.
36. Kicic A, Chua AC, Baker E. Effect of iron chelators on proliferation and iron uptake in hepatoma cells. Cancer 2001; 92: 3093-3110.
37. Gao W, Qiao X, Ma S, Cui L. Adipose‐derived stem cells accelerate neovascularization in ischaemic diabetic skin flap via expression of hypoxia‐inducible factor‐1α. J Cell Mol Med 2011; 15: 2575-2585.
38. Ikeda Y, Tajima S, Yoshida S, Yamano N, Kihira Y, Ishizawa K, et al. Deferoxamine promotes angiogenesis via the activation of vascular endothelial cell function. Atheroscler 2011; 215: 339-347.
39. Karuppagounder SS, Ratan RR. Hypoxia-inducible factor prolyl hydroxylase inhibition: Robust new target or another big bust for stroke therapeutics? J Cereb Blood Flow Metab 2012; 32: 1347-1361.
40. Pugh CW, Ratcliffe PJ. Regulation of angiogenesis by hypoxia: Role of the HIF system. Nat Med 2003; 9: 677-684.
41. Willam C, Koehne P, Jürgensen JS, Gräfe M, Wagner KD, Bachmann S, et al. Tie2 receptor expression is stimulated by hypoxia and proinflammatory cytokines in human endothelial cells. Circ Res 2000; 87: 370-377.
42. Choi CW, Lee J, Lee HJ, Park H-S, Chun Y-S, Kim BI. Deferoxamine improves alveolar and pulmonary vascular development by upregulating hypoxia-inducible factor-1α in a rat model of bronchopulmonary dysplasia. J Korean Med Sci 2015; 30: 1295-1301.
43. Duan L-J, Zhang-Benoit Y, Fong G-H. Endothelium-intrinsic requirement for Hif-2α during vascular development. Circ 2005; 111: 2227-2232.
44. Lee YN, Wang HH, Su CH, Lee H. Deferoxamine accelerates endothelial progenitor cell senescence and compromises angiogenesis. Aging 2021; 13: 21364- 21380
45. Holden P, Nair LR. Deferoxamine: An angiogenic and antioxidant molecule for tissue regeneration. Tissue Eng Part B Rev 2019; 25: 461-470.
46. Tepper OM, Capla JM, Galiano RD, Ceradini DJ, Callaghan MJ, Kleinman ME, et al. Adult vasculogenesis occurs through in situ recruitment, proliferation, and tubulization of circulating bone marrow–derived cells. Blood 2005; 105: 1068-1077.
47. Siavashi V, Sariri R, Nassiri SM, Esmaeilivand M,  Asadian S, Cheraghi H. Angiogenic activity of endothelial progenitor cells through Angiopoietin-1 and Angiopoietin-2. Anim Cells Syst 2016; 20: 118-129.
48. Findley CM, Cudmore MJ, Ahmed A, Kontos CD. VEGF induces Tie2 shedding via a phosphoinositide 3-Kinase/Akt–dependent pathway to modulate Tie2 signaling. Arterioscler Thromb Vasc Biol 2007; 27: 2619-2626.
49. Oses C, Olivares B, Ezquer M, Acosta C, Bosch P, Donoso M, et al. Preconditioning of adipose tissue-derived mesenchymal stem cells with deferoxamine increases the production of pro-angiogenic, neuroprotective and anti-inflammatory factors: Potential application in the treatment of diabetic neuropathy. PLoS One 2017; 12: e0178011-0178032.
Iranian Journal of Basic Medical Sciences
Volume 28, Issue 11
November 2025
Pages 1589-1597

Files

Share

How to cite

Statistics